Académique Documents
Professionnel Documents
Culture Documents
Edited by
Leland J. Cseke Ara Kirakosyan
Peter B. Kaufman Margaret V. Westfall
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v
vi Contents
Chapter 33 Methods of Stem Cell Lineage Analysis In Situ and in Cell Culture....................... 579
Andrei B. Borisov
Leland J. Cseke
Ara Kirakosyan
Peter B. Kaufman
Margaret V. Westfall
ix
Editors
Leland J. Cseke, PhD, received his doctorate in plant cellular and molecular biology through the
Department of Molecular, Cellular, and Developmental Biology at the University of Michigan, Ann
Arbor, Michigan. His dissertation research included the molecular biology, evolution, and biotechno-
logical applications of terpenoid scent compound production in Clarkia and Oenothera species in the
laboratory of Dr. Eran Pichersky. Currently, Dr. Cseke is an assistant professor in the Department of
Biological Sciences at the University of Alabama, Huntsville, where he works in conjunction with the
Department of Energy and Argonne National Laboratory to determine the molecular mechanisms of
keystone species in forest ecosystem responses to environmental changes. In addition, Dr. Cseke is work-
ing with the United States Department of Agriculture to develop cost-effective molecular methods for
plant species identification and has helped to develop the national guidelines for plant molecular identifi-
cation. Other projects include the metabolic engineering of Jatropha curcas plants for the improvement
of biodiesel with emphasis on transcriptional regulators that help to control oil biosynthesis.
Dr. Cseke spent several years as an assistant research professor at Michigan Technological
University working to discover the function of aspen (Populus tremuloides) MADS-box genes in
wood and floral development. He has also been a postdoctoral fellow in the Department of Plant
Sciences at the University of Arizona in the laboratory of Dr. Richard Jorgensen. There, he worked
to elucidate the factors involved in functional sense and antisense suppression of genes involved in
anthocyanin biosynthesis. Dr. Cseke’s interests include the biosynthesis of plant chemical products,
environmental genomics, and metabolic engineering of plant metabolism. This led to his coauthor-
ing another book, Natural Products from Plants, Second Edition (CRC Press, 2006). In addition,
he has done some work in the study of methods for improving the separation of the cancer-fighting
diterpene, taxol, in Taxus species in the laboratory of Dr. Peter B. Kaufman, and his knowledge
of such subjects has been directed toward teaching classes that emphasize biotechnology and the
chemical principles of biology.
Ara Kirakosyan, PhD, DSc, is an associate professor of biology at Yerevan State University,
Armenia, and is currently a research scientist at the University of Michigan. He received his PhD
in molecular biology in 1993 and his DSc in biochemistry and biotechnology in 2007, both from
Yerevan State University, Armenia. Dr. Kirakosyan’s primary research areas include the impact of
phytopharmaceuticals on prevention of heart failure, challenges and pitfalls in antioxidant research,
and mechanisms of synergistic action of bioactive medicinal compounds at target sites. His research
on phytochemicals and antioxidants in foodstuffs has direct implications for the health industry,
specifically in the area of cardiovascular disease. He is also interested in plant biotechnology
research to produce enhanced levels of medicinally important, value-added secondary metabolites.
He carried out postdoctoral research at Gifu Pharmaceutical University, Gifu, Japan, under the
supervision of Professor Kenichiro Inoue. The primary research topic dealt with molecular biology
of dianthrones and triterpene glucoside biosynthesis. He also took part in visiting research investi-
gator positions in Germany. First, he was visiting scientist at Heinrich Heine University, Düsseldorf
(host scientist Professor Dr. W.A. Alfermann). The research there concerned a lignan anticancer
project, i.e., the production of cytotoxic lignans from Linum (flax). The second position involved
a carbohydrate-engineering project as a DAAD fellow in the Institute of Plant Genetics and
Crop Plant Research (IPK) Gatersleben, under the supervision of Professor Dr. Uwe Sonnewald.
Dr. Kirakosyan’s collaboration with U.S. scientists started with the USDA-founded project on plant
cell biotechnology for the production of dianthrones and phloroglucinol derivatives in Hypericum
perforatum. This research was carried out with Dr. Donna Gibson at USDA Agricultural Research
xi
xii Editors
Service, Ithaca, New York. In 2002, he was a Fulbright visiting research fellow at the University
of Michigan, Department of Molecular, Cellular, and Developmental Biology in the laboratory of
Professor Peter B. Kaufman. He is the coauthor of two other books, namely, Recent Advances in
Plant Biotechnology (Springer, 2009) and Natural Products from Plants, Second Edition (CRC
Press/Taylor & Francis, 2006). Dr. Kirakosyan is principal author of over 50 peer-reviewed research
papers in professional journals and several chapters in books dealing with plant biotechnology, bio-
chemistry, pharmacology, and molecular biology.
Peter B. Kaufman, PhD, received his BSc in plant science from Cornell University, Ithaca, New York,
and his doctorate in plant biology from the University of California, Davis, California. He is cur-
rently professor emeritus of biology at the University of Michigan, Ann Arbor, and research sci-
entist with the University of Michigan Integrative Medicine (UMIM) program. He is the author of
nine books and over 230 research publications. His most recent books include Recent Advances in
Plant Biotechnology (A. Kirakosyan and P.B. Kaufman, editors) (Springer Publishers, New York,
2009); Natural Products from Plants, Second Edition (CRC Press, 2006), coauthored with Leland J.
Cseke, Ara Kirakosyan, Sara Warber, James Duke, and Harry Brielmann; and Creating a Sustainable
Future: Living in Harmony with the Earth (Researchco Book Centre, New Delhi, India, 2002), coau-
thored with James Hoyt, Christopher Coon, Barbara Madsen, Sara Warber, J.N. Govil, and Casey R.
Lu. Dr. Kaufman is a fellow of the American Association for the Advancement of Science (AAAS).
He is also past president of the Michigan Botanical Club and past chairman of the Michigan Natural
Areas Council. He served as secretary-treasurer of the American Society of Gravitational and Space
Biology (ASGSB) and was the recipient of ASGSB’s Orr Reynolds Distinguished Service Award.
He is currently doing research on natural products of medicinal value in plants with support from
UMIM, the National Institutes of Health, Xylomed Research, Denali BioTechnologies, LLC, and
the Cherry Marketing Institute (CMI) of Michigan. He has performed research with colleagues at
Lund University, Lund, Sweden; University of Calgary, Alberta, Canada; Nagoya and Kyushu uni-
versities, Nagoya and Fukuoka, Japan; International Rice Research Institute, Los Baños, Philippines;
Michigan State University, East Lansing; University of Colorado, Boulder; North Carolina State
University, Raleigh; Purdue University, West Lafayette, Indiana; USDA Plant Hormone Lab,
Beltsville, Maryland; Hawaiian Sugarcane Planters’ Association, Aiea Heights, Honolulu; the Volcani
Agricultural Research Center, Bet Degan, Israel; Denali BioTechnologies, Homer, AK; and Yerevan
State University, Yerevan, Armenia. He recently (2007) established the Hazel S. Kaufman Integrative
Medicine Retreat Center at Manchester, Michigan (hskimrc.org), and established at this same site, sus-
tainable energy projects, including a geothermal heating system, a solar photovoltaic system (14 kW),
and a wind turbine (5 kW).
Margaret V. Westfall, PhD, is an associate professor of surgery and molecular and integrative physi-
ology and an assistant professor of biomedical engineering at the University of Michigan, Ann Arbor,
Michigan. She received her BA in biology from Colorado College, Colorado Springs, Colorado; her
MSc in microbiology from the University of Montana, Missoula, Montana; and her doctorate in phys-
iology from Loyola University, Chicago, Illinois. She held postdoctoral positions at the University of
Illinois at Chicago and at the University of Michigan, Ann Arbor, Michigan. Dr. Westfall has taught
skeletal muscle physiology, and molecular and integrative cardiac physiology for engineers. Her
research is focused on heart failure and includes studies on protein kinase C modulation of contractile
function; the role of the thin filament molecular switch protein, troponin I, in modulating contractile
function; and analysis of proteomic expression in end stage heart failure. Dr. Westfall’s research
has been funded by the National Institutes of Health and the American Heart Association. She cur-
rently serves on the editorial board for the American Journal of Physiology—Heart and Circulatory
Physiology and the Journal of Molecular and Cellular Cardiology. Dr. Westfall has published over
50 peer-reviewed research publications and is the author of several book chapters. She is a fellow of
the Council on Basic Cardiovascular Sciences for the American Heart Association.
Contributors
Joy M. Agee, MSc, is a scientist in training in the PhD program for biotechnology, science, and engi-
neering at the University of Alabama in Huntsville (UAH). She is currently conducting her doctoral
research in the laboratory of Dr. Richard M. Myers at HudsonAlpha Institute for Biotechnology. Her
research is aimed at understanding the molecular biology of triple negative breast cancer (TNBC), a
form of breast cancer that lacks receptors for estrogen, progesterone, and human epidermal growth
factor 2. Agee received her MSc in molecular biology from UAH. Her research focuses on iden-
tifying the expression of RNAi genes in the ectomycorrhizal fungus, Laccaria bicolor. Agee is a
National Science Foundation Graduate research fellow.
Chuanfu An, PhD, is a postdoctoral research scientist at the University of Florida. He received his PhD
in plant genetics and genomics from Mississippi State University, Mississippi State, Mississippi in 2008,
and was a postdoctoral researcher in the School of Forestry and Natural Resources at the University of
Georgia for two years. His research interests include plant secondary metabolism and defense.
Andrei B. Borisov, PhD, is an assistant professor at Wayne State University in Detroit. He received his
doctorate from the Institute of Cell Biology of the Russian Academy of Sciences, St. Petersburg, and
later worked at the University of Michigan, Ann Arbor, before accepting his current position at Wayne
State. Dr. Borisov’s research interests are focused on the interrelations of proliferation and differentia-
tion of precursor cells during development, regeneration, and malignant growth. Another field pertain-
ing to his studies is cell and molecular biology of aging and the mechanisms underlying the impairment
of the compensatory and regenerative capability of skeletal and cardiac muscle during senescence.
Lynn Boyd, PhD, is an associate professor at the University of Alabama in Huntsville. Dr. Boyd
received his BA in Latin from Wake Forest University in 1983, and later received his PhD from the
Department of Human Genetics at the University of Utah in 1992. Research in the Boyd laboratory
focuses on the role of the ubiquitin pathway in Caenorhabditis elegans development.
Nidhi P. Chanana, PhD, fellow, The Energy and Resources Institute (TERI), New Delhi, has more
than 10 years of experience in plant cell and tissue culture. She received her doctorate from the
University of Delhi under the guidance of Professor S.S. Bhojwani. She also worked with Dr. J.B.M.
Custers at Plant Research International, Wageningen, the Netherlands (1999–2000). Her present
interests include development of haploids as well as genetic transformation system in J. curcas
and sesame. She spent three months in the laboratory of Dr. Gopi K. Podila at the University of
Alabama, Huntsville, United States, where she worked on optimizing the protocol for genetic trans-
formation of J. curcas. As an adjunct faculty member at TERI University, she teaches and super-
vises PhD students in the field of plant biotechnology.
xiii
xiv Contributors
Feng Chen, PhD, is an associate professor of plant functional genomics in the Department of Plant
Sciences, University of Tennessee, Knoxville, Tennessee. He received his BSc in molecular biology
from Nankai University, Tianjin, China; his MSc in genetics from the Institute of Genetics, Chinese
Academy of Sciences, Beijing, China; and his PhD in plant biology from the University of California,
Davis, California. Dr. Chen’s research program focuses on use of an integrated genomics approach that
combines bioinformatic analysis, metabolomics, transcriptomics, and large-scale in vitro biochemical
assays to characterize the biosynthesis, biological function, and evolution of plant secondary metabolism.
Luis Rogelio Cruz-Vera, PhD, received his BSc in chemistry at Autonomous University of
Puebla, Mexico, in 1995; his MSc in genetics and molecular biology at the Center for Research
and Advanced Studies (CINVESTAV-IPN), Mexico, in 1996; and his PhD in genetics and molec-
ular biology at the Center for Research and Advanced Studies (CINVESTAV-IPN), Mexico, in
2000. His previous appointments include postdoctoral researcher, Department of Genetics and
Molecular Biology, Center for Research and Advanced Studies, Mexico City, Mexico (2000–
2003) and postdoctoral researcher, Department of Biological Sciences, Stanford University
(2003–2007). He currently serves as assistant professor, Department of Biological Sciences,
University of Alabama in Huntsville (2007–present). His research interests include regulation of
gene expression, especially on those mechanisms involving the regulation of ribosome function.
Sarah Beth Cseke, MSc, received her BSc in biology and MSc in plant molecular biology with
an emphasis on tissue culture from the University of Alabama in Huntsville (UAH). Since 2006,
she serves as the lead research assistant in the Center for Molecular Biology at Alabama A&M
University (AAMU). She has seven years of combined experience as a research assistant. During
this period, she has worked with diverse genes in transgenic poplar trees (P. tremuloides), cot-
ton species (Gossypium spp.), and reniform nematodes (Rotylenchulus reniformis). She has also
undertaken plant transformation for over-expression and RNAi in Arabidopsis, tobacco, Populus,
and Glycine species. She is familiar with the basic molecular biology research tools, including
PCR, RNA extractions, cDNA library preparation, and plant transformation. She has also taught
graduate, undergraduate, and high school students in formal and individualized sessions.
Arun Kumar Das, PhD, is currently affiliated with the Michigan Metabolomics and Obesity Center
(MMOC), Internal Medicine Department, University of Michigan, where he is acting head of the
Lipidomics Section. Dr. Das worked for many years at MHRI (Mental Health Research Institute, now
known as The Molecular and Behavioral Neuroscience Institute), University of Michigan, in the area
of membrane lipid research. He received his PhD in India at the University of Calcutta, where his
graduate training focused on organic chemistry specialization in lipid chemistry and analysis.
Dr. Das acquired an extensive knowledge and background on various aspects of chemistry, biochemis-
try/enzymology and molecular biology of membrane lipid and its metabolic enzymes. Over the years,
he made a number of discoveries in the area of chemical synthesis of phospholipids, enzymology of
lipids, effects of hypolipidemic drugs on lipid metabolism, cloning cDNA of lipid metabolic enzyme
and so on. Dr. Das has published 30 peer-reviewed research publications and is a coauthor of several
book chapters on lipid research. He became a member of the American Oil Chemist Society in 1992.
Contributors xv
His current interest at the MMOC is on studies of structural and functional aberration of membrane
lipids, lipid metabolites, and the enzymes involved in diabetes, obesity, and related physiological
dysfunctions.
Maria R. Davis, PhD, was an associate professor of molecular biology in the Department of
Biological Sciences at the University of Alabama in Huntsville. Unfortunately, we lost Maria in
the tragic shooting that occurred on the campus of UA-Huntsville on February 12, 2010. Dr. Davis
received her bachelor of engineering degree from the University of Michigan, Ann Arbor, in chemi-
cal engineering with a bio-option minor in 1981. She worked at E.I. DuPont DeNemours in the
Chemical, Dyes and Pigments Department in East Chicago, Indiana, as a process engineer until
1983. She completed her master of engineering degree in chemical engineering with an interdisci-
plinary minor at North Carolina State University, Raleigh, in 1985. She then pursued her PhD in
biochemistry to graduate in 1992 with a minor in plant pathology. Dr. Davis did her postdoctoral
research at Monsanto Chemical Corporation in St. Louis, Missouri, where her interest in plant
sciences and bioengineering of crop plants blossomed. She worked for six years at RESGEN (an
Invitrogen Company) as a senior scientist overseeing both production and research staff, developing
large insert libraries, testing RNA/DNA isolation kits, assembling vector constructs, and engineer-
ing Escherichia coli strains. Her most recent research interests were devoted to identification of
fungal pathogenicity factors to better understand the progression of fungal diseases in animals and
plants. She chose to use a variety of proteomic, genomic, metabolic methods together with molecu-
lar biology tools to identify new factors, then address how these factors may work to enhance the
attack of the fungus on the host organism. Additional research interests were in the area of cellular
transport in order to understand mechanisms used to localize proteins extracellularly. She received
research grants from the National Science Foundation and Binational Agricultural Research and
Development (BARD) Program. Dr. Davis had more than 15 publications in peer-reviewed journals,
including reports in conference proceedings. She taught an introductory course in biology and a
course in cell and developmental biology at the University of Alabama in Huntsville.
Eric Devaney, BA, MD, is an associate professor of surgery at the University of Michigan, where
he practices as a pediatric cardiac surgeon. Dr. Devaney graduated summa cum laude from the
University of Virginia and received his MD from the University of California, Los Angeles. His
general surgery training was undertaken at the University of California, San Francisco. He received
specialty training in cardiothoracic surgery and pediatric cardiac surgery at the University of
Michigan. Dr. Devaney is board certified in general and thoracic surgery. His clinical interests are
in the surgical repair of complex neonatal cardiac defects and in the use of ventricular assist devices
in the treatment of pediatric heart failure. His primary basic science research interests focus on the
sarcomeric basis of contractile dysfunction in heart failure and cardiomyopathy, and the genetic
modification of cardiac myocytes using adenoviral gene delivery.
Scott A. Harding, PhD, is a senior research scientist with the Warnell School of Forestry and Natural
Resources at the University of Georgia. Dr. Harding received his PhD in agronomy from Kansas State
University, Manhattan, in 1990. His research interests include plant development and metabolism.
Susan Holmes, PhD, was a tenured researcher specializing in biological applications at INRIA in
Montpellier, France until 1993. She came to Stanford University for a year to teach computational
statistics, was at MIT and Harvard for two years and at Cornell for two years, and then returned
to Stanford where she is currently a professor of statistics. All of her work focuses on multivariate
statistics applied to biology. She is currently working on the phylogeny and statistical analyses of
HIV drug resistance, the interaction between the immune system and cancer, codon usage bias and
the phylogeny of Dehalococcoides, and image segmentation for cell localizations in lymph nodes.
xvi Contributors
Sarah E. Kampert, BA, graduated from Boston University in 2008 with a degree in biochemistry
and molecular biology. She is currently a graduate student in the Program in Cellular and Molecular
Biology at the University of Michigan. Kampert is pursuing research to evaluate cellular structure
and function in response to adenoviral-mediated gene transfer of sarcomeric proteins.
Ramesh V. Kantety, PhD, is a professor of plant biology and genomics in the Department of Natural
Resources and Environmental Sciences at Alabama A&M University. He received his MSc in plant
breeding from Auburn University, his PhD in plant genetics and breeding from Purdue University,
and received his postdoctoral training on plant–pathogen interactions from the Boyce Thompson
Institute for Plant Biology at Cornell University. Later, he worked as a research associate in the
Department of Plant Breeding at Cornell University, where he was instrumental in establishing
plant genomics and bioinformatics programs in several key crops important to U.S. and world food
security. His research program’s focus is to improve the productivity of food, fiber, and energy crops
to meet current and emerging global needs utilizing integrated genetic and genomic approaches.
His graduate and undergraduate level teaching responsibilities include plant genetics, genomics,
and bioinformatics.
Akira Katoh, PhD, is a senior researcher at the Core Laboratory, Nara Prefectural Small and
Medium-sized Enterprises Support Corporation in Nara, Japan. He received his PhD from the
School of Agricultural Science, Nagoya University, Aichi, and later worked at the National Institute
for Basic Biology in Aichi, at the University of Tsukuba in Ibaraki, as well as at the Nara Institute
of Science and Technology in Nara before accepting his current position at the Core Laboratory.
Dr. Katoh’s research interests focus on metabolomics, pharmacological analysis based on transcrip-
tomics, herbal medicine, and plant physiology based on molecular biology.
Evelyn H. Kim, PhD, is a postdoctoral fellow in the Department of Surgery at the University
of Michigan, Ann Arbor, Michigan. She received her BSc and MSc in chemistry from Dankook
University, South Korea (1996, 1999), and her PhD in chemistry from the University of Michigan
(2009). Dr. Kim’s research work is in the areas of proteomic analysis of human disease, including
ovarian cancer and pancreatic cancer. She has published six peer-reviewed manuscripts and pre-
sented over a dozen talks at scientific conferences.
Lance Larka, PhD, is chief operating officer of iXpressGenes, Inc. at the HudsonAlpha
Institute for Biotechnology. He received his BSc in genetics in 1995 from the University of
California, Davis. Larka has worked in the field of molecular biology for public research institu-
tions, educational institutions, private for-profit companies, and international for-profit compa-
nies, focusing on laboratory automation, liquid handling processes, oligonucleotide synthesis,
process optimization, DNA (Sanger) sequencing, and laboratory design and construction. His
company focuses on protein production and purification, DNA sequencing for service, and pro-
ducing synthetic biology bio-parts. He is a judge for the new innovative product category at
LabAutomation.
Casey R. Lu, PhD, is a professor of biology in the Department of Biological Sciences, Humboldt
State University (HSU), Arcata, California. He received his BSc with honors in biology in 1980,
his MSc in biology in 1987, and his PhD in education and biology in 1993, all from the University
of Michigan, Ann Arbor. Dr. Lu has worked to enhance the electron microscopy facilities at HSU
since 1999. He has also received numerous grants from the University of California Office of the
President for establishing and directing the Redwood Science Project at HSU. At HSU, he teaches
introductory cellular and molecular biology, plant physiology, secondary science methods, plant
tissue culture, and electron microscopy. He has presented papers/posters at annual meetings of the
American Society of Plant Biologists, California Science Teachers Association, and the National
Contributors xvii
Science Teachers Association. His research interests include ultrastructural changes in plants chal-
lenged with heavy metals and physiology of the gravitropic response in plants.
Michael Mashore is the lead technician at the Veteran’s Administration San Diego Core for Micro
Imaging. He began studying microscopy with a focused interest in electron microscopy while com-
pleting his BSc in biology at Humboldt State University, Arcata, California.
Maureen McKenzie, PhD, is the chief executive officer of Denali BioTechnologies, Inc. (DBI),
the first and only company in Alaska dedicated to pharmaceutical and nutraceutical discovery and
development from boreal territories. Dr. McKenzie founded DBI to focus on drug discovery from
unique plants, microbes, and marine organisms that thrive in harsh, psychrophilic (cold-loving)
habitats. In 2005, Dr. McKenzie received USDA funding, in conjunction with the University of
Alaska–Fairbanks, to demonstrate the feasibility of a nutraceutical industry in frontier regions
of Alaska. Her work on this initiative culminated in invited sessions before the Natural Health
Products Research Society of Canada, International Convention on Biodiversity, and the United
Nations in 2007. Dr. McKenzie has received many awards for innovation in research and business
and is the author or coauthor of numerous patents on drug discovery technology and nutraceuti-
cal products, peer-reviewed articles, and book chapters. She has served as adjunct faculty in the
Department of Pharmaceutics at the College of Pharmacy of the University of Florida, Gainesville,
since 2004. Prior to that, she was an affiliate assistant professor in the Department of Physiology
and Pharmacology at the College of Veterinary Medicine of Iowa State University, Ames, and was
an assistant professor of chemical biology and pharmacognosy and member of the Laboratory for
Cancer Research in the College of Pharmacy at Rutgers, The State University of New Jersey, New
Brunswick. She received her BSc in nutrition/food technology from Iowa State University in 1978,
her MSc in food science from Rutgers in 1982, and her PhD in biochemistry through a joint pro-
gram of Rutgers, the University Medicine and Dentistry of New Jersey and Robert Wood Johnson
Medical School, and Princeton University in 1987.
David E. Misek, PhD, is a research assistant professor in the Department of Surgery at the
University of Michigan, Ann Arbor, Michigan. Dr. Misek received his BSc in microbiology from
Colorado State University (1979) and his PhD in pathology from SUNY Downstate Medical Center
(1986). He is a member of the University of Michigan Cancer Center, the University of Michigan
Gastrointestinal Peptide Research Center, and an associate member of the Early Detection Research
Network (EDRN) of the NIH/NCI. Dr. Misek’s research focuses on proteomic and genomic analy-
sis of human disease, including breast cancer and pancreatic cancer. His research has been sup-
ported by the National Cancer Institute within NIH and the Department of Defense. Dr. Misek has
published over 95 papers in peer-reviewed journals and books. He also serves as an ad hoc reviewer
for many journals.
Kathleen R. Noon, PhD, is director of the Innovation Center Mass Spectrometry Facility at the
Medical College of Wisconsin, Milwaukee. She received her BSc in medical technology from
Alverno College, Milwaukee, Wisconsin; her MSc in chemistry from the University of Wisconsin–
Madison; and her PhD in chemistry from Michigan State University, East Lansing. She has held
postdoctoral positions at the University of Utah, Salt Lake City, and the University of Michigan,
Ann Arbor. Prior to her current position, she was the manager of the Biomedical Mass Spectrometry
Facility in the Pharmacology Department at the University of Michigan. Dr. Noon’s research focuses
on separation science and biomedical applications of mass spectrometry, including small molecule
quantitative analysis and proteomics. She has trained over 50 graduate students in the theory and
operation of mass spectrometers. Dr. Noon has published more than 14 papers in peer-reviewed
journals and 1 book chapter. She is a member of the American Society of Mass Spectrometry and
the American Chemical Society.
xviii Contributors
Taketo Okada, PhD, is an assistant professor of molecular biology and biochemistry of medicinal
plants and oriental medicine in Tokushima Bunri University at Kagawa, Japan, since 2005. He
received his BSc in 1999 from Hoshi University, Tokyo, Japan; and his MSc in 2002 and his PhD
in 2005 from Chiba University, Chiba, Japan, in pharmacy. He is also a pharmacist licensed by the
Ministry of Health, Labour and Welfare of Japan in 1999. His research interests focus on the molec-
ular characterization of secondary metabolite biosynthesis in medicinal plants and on metabolome
analysis of medicinal plants and herbal medicines. He is also working at an herbal garden for culti-
vation research of medicinal plants.
Ajay Kumar Pandey, PhD, is a postdoctoral research associate at Foreign Diseases Weed
Science Research Unit (ARS-USDA), Fort Detrick, Maryland. Dr. Pandey received his BSc from
H.N.B. Garhwal University, India, with a major in botany and chemistry in 1996. He received his MSc
in biotechnology from Himachal Pradesh University, India, in 1998, and then pursued his PhD
in biotechnology and graduated in 2003 from Thapar Institute of Engineering and Technology,
Patiala, India. Dr. Pandey started postdoctoral research work at Academia Sinica, Taiwan, and
continued in this capacity at the University of Alabama in Huntsville with research interest in
molecular aspects of plant–microbe interactions. Currently, he is working on Asian soybean rust
(ASR) and utilizing virus-induced gene silencing (VIGS) assays to investigate the role of candi-
date defense genes in soybean.
Gopi K. Podila, PhD, was professor and chair of the Department of Biological Sciences at the
University of Alabama in Huntsville, Alabama. However, we lost Gopi during the tragic shooting
that occurred on the campus of UA-Huntsville on February 12, 2010. Until May 2002, Dr. Podila had
served as professor of biochemistry and molecular biology in the Department of Biological Sciences
at Michigan Tech University, Houghton, Michigan. He received his BSc in biological sciences from
Nagarjuna University in India, his MSc in plant pathology from Louisiana State University (1983),
and his PhD in molecular biology from Indiana State University (1987). Dr. Podila’s research dealt
with plant–fungus interactions, molecular biology of plant development, plant biotechnology, and
functional genomics. His research was supported by USDA, USFS, NSF, DOE, CPBR, and industry.
He published over 80 papers in peer-reviewed journals and books, and a book, Current Advances in
Mycorrhizae Research. Dr. Podila served on the editorial boards of Symbiosis, New Phytologist, and
Physiology and Molecular Biology of Plants, was an ad hoc reviewer of USDA, NSF, USDA-BARD,
and Italian Ministry of Education and Research grant proposals. He had visiting professor appoint-
ments at INRIA-Nancy, France; the University of Torino and the University of Urbino, Italy; and the
University of Helsinki, Finland. Dr. Podila organized and chaired several international symposia on
plant–fungus interactions and served as councilor-at-large for the International Symbiosis Society.
E. Mitchell Seymour, MSc, PhD, is a research associate. Dr. Seymour received his BSc in biol-
ogy from the University of Notre Dame and his PhD in biochemical and molecular nutrition from
Michigan State University. He has conducted extensive research in both eukaryotic and prokaryotic
cell and molecular biology. His research has included projects in molecular virology, environmental
microbiology, and cancer biology. His work has utilized several animal models and cell culture
lines. His research has been supported by the NIH, the California Table Grape Commission, the
Cherry Marketing Institute, the U.S. Highbush Blueberry Council, and the U.S. Apple Association.
His current research explores the impact of phytochemical-enriched diets on heart failure pathogen-
esis, metabolic syndrome, and atherosclerosis. He is currently the manager of the Cardioprotection
Research Laboratory at the University of Michigan Medical School, Section of Cardiac Surgery.
Avinash Sreedasyam, MSc, is a PhD student in the Department of Biological Sciences, University
of Alabama in Huntsville. He received his BSc in microbiology from Kakatiya University, India,
Contributors xix
and his MSc in biotechnology from the University of Wollongong, New South Wales, Australia. His
current research focuses on understanding defense response regulation during plant–fungal symbi-
otic interactions using high throughput transcriptomic approaches.
Tamara K. Stevenson, BA, graduated from the University of Michigan in 2008. She is a research
associate in the Department of Surgery at the University of Michigan. She is currently pursuing
research on the functional response to pressure overload–induced hypertrophy and remodeling of
the adult heart.
Ramesh C. Thakur, PhD, received his PhD in forestry from the University of Horticulture and
Forestry, Solan, India, in 1989. Since then, he has worked as a member of the forestry faculty in
the Department of Tree Improvement and Genetic Resources at the University of Horticulture and
Forestry, Solan, India; as a postdoctoral fellow at the University of Freiburg, Germany; Forestry
and Forest Products Research Institute, Tsukuba, Japan; and currently holds the position of assistant
research scientist at Michigan Technological University, United States. Dr. Thakur has published
extensively in various journals, books, and conference proceedings. He has taught courses on tree
improvement and plant biotechnology. His research interests include tissue culture, genetic manipu-
lation, genetics and plant breeding, environmental pollution, and plant nutrition.
Geetika Trivedi, MSc, is a PhD student in the Department of Biological Sciences, University of
Alabama in Huntsville, Alabama. She received her BSc in biology in 2003 from C.S.J.M. University,
India, and her MSc in biotechnology in 2005 from Allahabad Agricultural Institute–Deemed
University, India. Her current research involves understanding the molecular basis of signaling and
metabolic re-programming during plant–fungal symbiosis.
Chung-Jui Tsai, PhD, is Winfred N. Haynes Professor and Georgia Research Alliance Eminent
Scholar of the Warnell School of Forestry and Natural Resources and the Department of Genetics
at the University of Georgia, Athens, Georgia. Dr. Tsai received her BSc and MSc from National
Taiwan University in Taiwan (1989 and 1991, respectively), and her PhD in forest science from
Michigan Technological University (1995). Her research is concerned with phenylpropanoid metab-
olism, wood formation, one-carbon metabolism, and growth-and-defense trade-offs in Populus.
Dr. Tsai’s research is supported through DOE, NSF, USDA, CPBR, and industry. He has published
over 50 peer-reviewed papers in journals and books.
Keerthi P. Venkataramanan, BTech, is a fourth year graduate student pursuing his PhD in the
Biotechnology Science and Engineering Program at the University of Huntsville in Alabama. His
research focuses on the anaerobic fermentation of biodiesel-derived crude glycerol using Clostridium
pasteurianum. His research interests include functional genomics and metabolomics. He received
his BTech in biotechnology from S.R.M. University, India, in 2007. He is expected to receive his
MSc in chemical engineering in the summer of 2011.
xx Contributors
Yuh-Shuh Wang, PhD, is currently a senior scientist in the Institute of Technology, University of
Tartu, Estonia. Dr. Wang received her PhD degree in forest molecular genetics and biotechnology
from Michigan Technological University, Houghton, in 2002; and her M.Sc. degree in botany at the
National Taiwan University, Taipei, Taiwan. Her research has focused on molecular mechanisms
of plant development. Using cDNA-AFLP and differential display, she isolated and characterized
suites of cDNAs that are differentially regulated during tuberization of sweet potato and vascular
development of quaking aspen as her graduate work.
Marilyn D. Yoder, PhD, is an associate professor of cell biology and biophysics in the School of
Biological Sciences, University of Missouri–Kansas City. She received her BSc in nutrition from the
University of Kentucky and her PhD in biochemistry from the University of California, Riverside.
Dr. Yoder’s research involves structural, functional, and evolutionary relationships of proteins using
x-ray crystallography and other biophysical techniques. Specific proteins of interest include plant
and bacterial pectate-degrading enzymes, eukaryotic phosphatidylinositol transfer proteins, human
semenogelin proteins, and bacterial cytolethal distending proteins.
Part I
DNA-Based Technology
GENETICS, GENOMICS
AND BREEDING OF
CUCURBITS
Series Editor
Chittaranjan Kole
Department of Genetics and Biochemistry
Clemson University
Clemson, SC
USA
Editors
Yi-Hong Wang
Department of Biology
University of Louisiana at Lafayette
Lafayette
USA
Tusar Kanti Behera
Division of Vegetable Science
Indian Agricultural Research Institute
New Delhi
India
Chittaranjan Kole
Department of Genetics and Biochemistry
Clemson University
Clemson, SC
USA
Science Publishers
Jersey, British Isles
Enfield, New Hampshire
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Chittaranjan Kole
The Family Cucurbitaceae (cucurbits hereafter) has 118 genera and 825
species distributed primarily in the tropics and subtropics. It contains
some of the most nutritious, delicious and versatile food items in the
human diet. For example, watermelon contains 40% more lycopene than
tomatoes and lycopene is a powerful antioxidant that may lower the risk
of certain cancers and heart diseases. Melon is the main source of dietary
β-carotene together with carrots and broccoli. But only seven species of
four genera are economically important: Citrullus lanatus (watermelon),
Cucumis sativus (cucumber), Cucumis melo (melon), Cucurbita pepo (squash,
gourd, and pumpkin), Cucurbita maxima (squash), Cucurbita moschata
(squash and pumpkin), and Lagenaria vulgaris (L. siceraria, bottle gourd).
Less prominent are Luffa (Luffa acutangula and L. cylindrica), bitter melon
(Momordica charantia), and waxy gourd (Benincasa hispida).
The last two decades have proved to be the most exciting period in
cucurbit research although breeding effort and genetic/genomic studies
mostly focused on the economically more important cucurbits, i.e.,
cucumber, melon and watermelon. Cucumber becomes the first cucurbit
to be sequenced, after other field crops such as rice, sorghum, soybean
and maize. Its 26,682 predicted genes will facilitate genetic studies and
marker development in other closely related cucurbits such as melon and
watermelon. High-density genetic maps are now available for cucumber,
melon, watermelon and squash. More efficient and abundant marker
systems such as SNP and SSR are being developed. Genomic resources
such as sequenced ESTs, large-insert genomic libraries, high-throughput
sequencing have been or are being developed for cucurbits. Molecular
breeding using markers linked to agronomically important traits has become
an efficient tool in speeding up the new variety release.
This book provides an indepth review of the current state-of-the-art
of genetic and genomic research conducted in cucurbits. Each chapter is
authored by specialists in their field to report the latest trends and findings.
The chapters are well documented and illustrated. The hard work of all
contributors is greatly appreciated.
The book begins with an exhaustive description of cucurbits in terms of
classification, geographical distribution, production and their importance
in our diet (Chapters 1 and 2). These are followed by a discussion on how
traditional cucurbit breeding has produced a vast number of new varieties
that meet the needs of modern consumers (Chapter 3). Chapter 4 extends
the discussion to breeding of novelty cucurbits, i.e., squash for decoration
and pumpkins for Halloween, a popular tradition for children in the United
States. Chapter 5 describes the applications of genetic markers to diversity
analysis in cucurbits. Genetic mapping and map-based cloning of cucurbit
genes are described in Chapter 6. This is followed by a discussion in
mapping of monogenic traits and molecular breeding in Chapter 7. Chapter
8 expands the discussion to mapping of quantitative traits, which include
majority of agronomically important traits in cucurbits. The following three
chapters, Chapters 9, 10, and 11, describe the progress in research using
-omics in melon, watermelon and cucumber, respectively. Chapter 12 is
devoted into an important topic of cucurbits: sex expression as both genetic
and environmental factors can change sex expression of cucurbit flowers.
And finally, Chapter 13 provides perspectives on cucurbit research areas
that may become increasingly important.
This book is a testimony to the substantial progress made in the field of
cucurbit genetics, genomics and breeding, and the definite value of cucurbits
as a model system to study niche area such as sex expression. It is true that
the tools and concepts that are presented in the book will continue to evolve
rapidly and we hope this volume will provide a solid foundation for further
development in cucurbit genetics, genomics and breeding.
Lafayette, Louisiana, USA Yi-Hong Wang
New Delhi, India Tusar Kanti Behera
Clemson, South Carolina, USA Chittaranjan Kole
T.K. Behera
Division of Vegetable Science Indian Agricultural Research Institute,
New Delhi, India.
Email: tusar@rediffmail.com
Hugo E. Cuevas
Plant Genome Mapping Laboratory, Center for Applied Genetic,
Technologies, 111 Riverbend Road, Athens, GA 30602, USA.
Email: hcuevas@uga.edu
Angela R. Davis
Wes Watkins Agricultural Research Laboratory, USDA-ARS, PO Box 159,
Hwy.3 West, Lane, OK 74555, USA.
Email: angela.davis@lane-ag.org
C. Esteras
Instituto de Conservación y Mejora de la Agrodiversidada Valenciana
(COMAV), Universidad Politécnica de Valencia, Camino de Vera 14,
Valencia 46022, Spain.
Email: criesgo@upvnet.upv.es
Tel. +34 96 387 94 15
Hiroshi Ezura
Gene Research Center, Graduate School of Life and Environmental
Sciences, University of Tsukuba, Ten-nodai 1-1-1, Tsukuba 305-8572,
Japan.
Email: ezura@gene.tsukuba.ac.jp
Zhangjun Fei
Boyce Thompson Institute for Plant Research, Cornell University, Ithaca,
NY 14853, USA.
Email: zf25@cornell.edu
Rebecca Grumet
Department of Horticulture and Graduate Program in Genetics,
Michigan State University, East Lansing, MI 48824, USA.
Email: grumet@msu.edu
Shaogui Guo
National Engineering Research Center for Vegetables Banjing, Beijing
100097, PO Box 2443, PR China.
Email: guoshaogui@nercu.org
Jun He
Institute of Vegetables and Flowers, Chinese Academy of Agricultural
Sciences, Beijing, 100081, China.
Email: hejun@caas.net.cn
Sanwen Huang
Institute of Vegetables and Flowers, Chinese Academy of Agricultural
Sciences, Beijing, 100081, China.
Email: huangsanwen@caas.net.cn
Sabina Islam
Indian Agricultural Research Institute, New Delhi, India.
Email: tokumkum@yahoo.com
Stephen R. King
Vegetable and Fruit Improvement Center, Department of Horticultural
Sciences, Texas, A & M University, College Station, TX 77843-2119, USA.
Email: srking@tamu.edu
Amnon Levi
USDA-ARS, U.S. Vegetable Laboratory, 2700 Savannah Highway,
Charleston, SC 29414, USA.
Email: Amnon-Levi@usda.ars.gov
Kai-Shu Ling
USDA-ARS, U.S. Vegetable Laboratory, 2700 Savannah Highway,
Charleston, SC 29414, USA.
Email: kai.ling@ars.usda.gov
Yang Liu
Boyce Thompson Institute for Plant Research Cornell University, Ithaca,
NY 14853, USA.
Current address: College of Medicine, Texas A&M Health Science Center,
Temple, TX 76504, USA.
Email: lyang@medicine.tamhsc.edu
J. Brent Loy
Department of Biological Sciences, University of New Hampshire,
Durham, NH 03824,USA.
Email: jbloy@unh.edu
A.D. Munshi
Division of Vegetable Science Indian Agricultural Research Institute,
New Delhi, India.
Email: anilabhm@yahoo.co.in
F. Nuez
Instituto de Conservación y Meora de la Agrodiversidada Valenciana
(COMAV), Universidad Politécnica de Valencia, Camino de Vera 14,
Valencia 46022, Spain.
Email: fnuez@btc.upv.es
B. Picó
Instituto de Conservación y Mejora de la Agrodiversidada Valenciana
(COMAV), Universidad Politécnica de Valencia, Camino de Vera 14,
Valencia 46022, Spain.
Email: mpicosi@btc.upv.es
Umesh K. Reddy
Department of Biology, West Virginia State University, Institute, WV
25112, USA.
Email: ureddy@wvstateu.edu
A.S. Sidhu
Indian Institute of Horticulture Research, Bengaluru, India.
Email: amrik_sidhu1@rediffmail.com
Jack E. Staub
USDA-ARS, Forage and Range Research Laboratory, Utah State
University, Logan, UT 84322-6300, USA.
Email: jack.staub@ars.usda.gov
Zhanyong Sun
East-West Seed International Ltd., No.92-1 Minzu Avenue, Nanning,
Guangxi, 530022, P. R. of China.
Email: zhangyong.sun@eastwestseed.com
A.K. Sureja
Indian Agricultural Research Institute, New Delhi, India.
Email: aksureja@rediffmail.com
Jessica Taft
Department of Horticulture and Graduate Program in Genetics,
Michigan State University, East Lansing, MI 48824, USA.
Email: taftjess@msu.edu
Judy. A. Thies
USDA-ARS, U.S. Vegetable Laboratory, 2700 Savannah Highway,
Charleston, SC 29414, USA.
Email: judy.thies@ars.usda.gov
Yi-Hong Wang
Department of Renewable Resources, University of Louisiana at
Lafayette, Lafayette, LA 70504, USA.
Email: yxw9887@louisiana.edu
W. Patrick Wechter
USDA-ARS, U.S. Vegetable Laboratory, 2700 Savannah Highway,
Charleston, SC, 29414, USA.
Email: pat.wechter@ars.usda.gov
Todd C. Wehner
Department of Horticultural Science, Box 7609, North Carolina State,
University, Raleigh, NC 27695-7609, USA.
Email: todd_wehner@ncsu.edu
Yiqun Weng
USDA-ARS Vegetable Crops Research Unit, Horticulture Department,
University of Wisconsin, Madison, WI 53706, USA.
Email: weng4@wisc.edu
Yong Xu
National Engineering Research Center for Vegetables Banjing, Beijing
100097, PO Box 2443, P. R. China.
Email: xuyong@nercv.org
Juan E. Zalapa
USDA-ARS, Madison WI; Dept. Horticulture, 1575 Linden Drive,
Madison, WI 53706, USA.
Email: jezlapa@wisc.edu
Xingping Zhang
Syngenta Seeds, 21435 Road 98, Woodland, CA 95695, USA.
Email: xingping.zhang@syngenta.com
Zhonghua Zhang
Institute of Vegetables and Flowers, Chinese Academy of Agricultural
Sciences, Beijing, 100081, China.
Email: zhangzhonghua@caas.net.cn
A Andromonoecious
ACC 1-amino-1-cyclopropane carboxylate
ACO ACC-oxidase
ACS 1-Amino- cyclopropane-1-carboxylate synthase
AFLP Amplified fragment length polymorphisms
AG AGAMOUS
AP APETELA
ARO Agriculture Research Organization (Israel)
AVG Aminoethoxyvinyl glycine
AVRDC Asian Vegetable Research & Development Center
(presently The World Vegetable Center)
AWF Average weight fruit
BAC Bacterial artificial chromosome
BC Backcross
BLAST Basic Local Alignment Search Tool
BR Brassinosteroids
BSA Bulked segregant analysis
CAAS Chinese Academy of Agricultural Sciences
CAPS Cleaved amplified polymorphic sequence
CATIE Centro Agronomico Tropical de Investigacion y
Ensenanza
CC Coiled domain
cDNA Complementary DNA
CMV Cucumber mosaic virus
CNPH Embrapa Hortalicas
CENARGEN Embrapa Recursos Geneticos e Biotecnologia
cM Centi-Morgan
CP Coat protein (of virus)
cpDNA chloroplast DNA
CRC CRABSCLAW
CuGI Cucumber genome initiative (China)
CuLCrV Cucurbit leaf crumple virus
CVYV Cucumber vein yellowing virus
DTF Days to flower
ABSTRACT
Cucurbit is a general term to denote all species within the Cucurbitaceae
family, which includes approximately 800 species in 118 genera.
Major cucurbit crops include cucumber, melon, watermelon, and
squash/pumpkin, which are all important vegetable crops that play
significant roles in human diet as well as rural economy. Nevertheless,
our understanding in phylogeny, genetics, biology, genomics and
many other fields of cucurbit crops, as compared with field crops and
model species, are lagging far behind. However, in recent years, with
technology innovations and instrumentation development, rapid
progress is being made in advancing our knowledge in major cucurbit
crops. In this chapter, we will give a brief review of research progress in
the past several years in four major cucurbits with focus on the economic
and biological importance of major cucurbits, the significant revision of
classical taxonomy of the genus Cucumis, new findings in domestication
and chromosome evolution of major cucurbits. We will also review the
current status of cucurbit germplasm conservation and utilization.
Keywords: Cucurbit, Cucumis, cucumber, melon, phylogenetics,
taxonomy, germplasm
1.1 Introduction
“Cucurbit” is a general term to denote all species within the Cucurbitaceae
family, which includes approximately 800 species in 118 genera. Cucurbits
are mostly annual, herbaceous, tendril-bearing and frost sensitive vines and
1
USDA-ARS Vegetable Crops Research Unit, Horticulture Department, University of Wisconsin,
Madison, WI 53706, USA; e-mail: weng4@wisc.edu
2
East-West Seed International Ltd., No.92-1 Minzu Avenue, Nanning, Guangxi, 530022, P. R.
of China.
*Corresponding author
C. mixta and C. ficifolia are used for food in Mexico and in Central and South
American countries. Squash and pumpkin are usually grown for their fruit,
harvested immature for summer squash or mature for winter squash and
pumpkin. The world total area harvested for pumpkin was approximately
1.5M ha in 2008 (Table 1-1).
Table 1-1 Five leading countries in production of four major cucurbits in 2008*.
Crops Country Area harvested (ha)
Cucumber China 1,702,777
Cameroon 120,000
Iran 82,000
Russia 73,000
Egypt 67,810
World total 2,635,058
Watermelon China 2,162,456
Iran 135,000
Turkey 139,000
Russia 133,000
Brazil 93,600
World total 3,752,568
Melon China 570,874
Turkey 103,000
Iran 80,000
Egypt 74,417
Spain 38,600
World total 1,346,962
Pumpkin India 360,000
China 330,212
Cameroon 110,000
Cuba 73,038
Russia 53,000
World total 1,529,935
*Data source: FAO Statistics 2010 (http://faostat.fao.org/).
carbohydrates, fats, and protein. Varieties with very large seeds have been
developed especially for use as food in China, where more than 200,000
tons are produced annually on 140,000 ha land (Zhang 1996).
physiology because of both the ease of sampling phloem sap and the facile
visualization of their large phloem sieve elements (Eschrich et al. 1971;
Clark et al. 1997; Zhang et al. 2010). Cucurbit phloem transport is unique
in that sucrose diffuses symplastically into intermediary cells (a form of
companion cell), where it is converted into raffinose-family oligosaccharide
sugars (Turgeon 1996).
Most cucurbits are consumed as either immature (for example
cucumber, summer squash) or mature (for example, melon, watermelon or
winter squash) fruits. A number of fruit development and ripening studies in
melon and cucumber have been conducted. Considering its morphological,
physiological, and biochemical diversity in flavor development and textural
changes during fruit ripening, melon was proposed to be a model plant
for the elucidation of key traits in fruit development (Ezura and Owino
2008).
Most species in the Cucurbitaceae family have basic chromosome
numbers of 7, 11, 12, 13, or 20, and relatively small genome sizes. The
chromosome numbers and genome size of the major cucurbit crops are listed
in Table 1-2. This also provides us a good opportunity to study chromosome
evolution in cucurbits.
Table 1-2 Chromosome number and genome size of major cucurbit crops.
Name Scientific name Chromosome # Genome size
(Mbp)*
Cucumber Cucumis sativus L. 2n = 2 x = 14 367
Melon Cucumis melo L. 2n = 2x = 24 450
Watermelon Citrullus lanatus Mats. & Nakai 2n = 2x = 22 430
Squash/pumpkin Cucurbita maxima Duch. 2n = 2x = 40 440
Cucurbita moschata Duch. 2n = 2x = 40 417
Cucurbita pepo L. 2n = 2x = 40 460–520
*Based on Arumuganathan and Earle (1991), and Tatum et al. (2006).
Figure 1-1 Major and minor cucurbit phylogenetic tree. Chromosome numbers and common
names follow each species name (when available). Molecular clock in million years ago,
if available, was shown on branching points. The tribe to which the species belongs was
shown to the right of vertical bars. Geographical occurrence of species: Green—America;
Black—mainland African; Red—Asia; Blue—Australia. The tree was redrawn after Schaefer
et al. (2009).
Color image of this figure appears in the color plate section at the end of the book.
1.4.2 Watermelon
The genus Citrullus is taxonomically complex and its composition is not
unanimously accepted by all taxonomists yet. Currently, Citrullus consists of
four diploid (2n = 2x = 22) species. C. lanatus var. lanatus is the domesticated
watermelon. Wild watermelon also known as citron is C. lanatus var. citroides
(L.H. Bailey) Mansf (but citron is known to be cultivated; Laghetti and
Hammer 2007). Three other wild species are Citrullus colocynthis (L.) Schrad.,
C. eccirrhosus Cogn and C. rehmii De Winter. The perennial C. colocynthis
grows in northern Africa, southwestern Asia and the Mediterranean,
whereas the perennial C. eccirrhosus and annual C. rehmii are endemic to the
Namib Desert (Levi et al. 2005; Dane and Liu 2006). Watermelon is thought
to have originated in southern Africa because it is found growing wild
throughout the area, and reaches maximum diversity there. Watermelon
may have been cultivated in Africa for over 4,000 years. C. colocynthis is
considered to be a wild ancestor of watermelon with small fruits and seeds,
and bitter flesh. Interspecific crosses of C. lanatus with C. colocynthis can
produce viable F1 hybrids. Although Citrullus species grow wild in southern
and Central Africa, C. colocynthis also grows wild in India. Thus, India and
China may be considered secondary centers of diversity for the genus. More
details regarding watermelon are discussed in Chapter 10.
1.4.3 Squash/pumpkin
The genus Cucurbita (squashes and pumpkins) is composed of 12–14 species
including five cultivated ones: C. pepo, C. maxima, C. moschata, C. ficifolia
and C. mixta. The latter two have less economic importance and a narrower
distribution (Robinson and Decker-Walters 1997).
While Cucurbita has an American origin in general, current genetic,
bio-geographic, and archaeological data suggest that the five cultivated
species were domesticated in different places, ranging from North America
to southern South America (Sanjur et al. 2002; Piperno and Stothert 2003).
Each species probably represents an independent domestication event from
different ancestor populations. Based on analysis of mitochondrial genes,
Sanjur et al. (2002) found that at least six independent domestication events
from distinct wild ancestors. C. mixta likely was domesticated from a wild
Mexican gourd, C. sororia. The wild ancestor of C. moschata is still unknown,
but will probably be found in lowland northern South America. C. andreana
may be the wild progenitor of C. maxima, but humid lowland regions of
Bolivia in addition to warmer temperate zones in South America from
where C. andreana was originally described should possibly be considered
as an area of origin for C. maxima.
Archaeological evidence of domestication of C. pepo in southern
Mexico dates back 10,000 years. Sanjur et al. (2002) suggested two separate
domestications in the C. pepo complex. The potential zone of domestication
for one of the domesticated subspecies, C. pepo subsp. ovifera, includes
eastern North America and should be extended to northeastern Mexico.
The wild ancestor of the other domesticated subspecies, C. pepo subsp. pepo,
is undiscovered but is closely related to C. pepo subsp. fraterna and possibly
will be found in southern Mexico.
with the domesticated ones thus being used for germplasm improvement
of cultivated species, e.g., powdery mildew and virus resistance, through
interspecific crossings. The zucchini yellow mosaic virus (ZYMV) resistance
gene(s) of C. moschata and the powdery mildew resistance gene of
C. okeechobeensis have been introgressed into C. pepo (Robinson and Decker-
Walters 1997; Whitaker and Robinson 1986). Multiple virus (ZYMV,
watermelon mosaic virus, and papaya ringspot virus) resistance genes,
derived from C. ecuadorensis, have been bred into C. maxima (Robinson and
Decker-Walters 1997; Herrington et al. 2001). The powdery mildew gene
of C. okeechobeensis spp. martinezii has been incorporated into C. moschata
(Robinson and Decker-Walters 1997; Cho et al. 2003).
References
Alverson AJ, Wei XX, Rice DW, Stern DB, Barry K, Palmer JD (2010) Insights into the evolution
of mitochondrial genome size from complete sequences of Citrullus lanatus and Cucurbita
pepo (Cucurbitaceae). Mol Biol Evol 27: 1436–1448.
Arumuganathan K, Earle ED (1991) Nuclear DNA content of some important plant species.
Plant Mol Biol Rep 9: 208–218.
Benjak A, Gonzalez V, Mir G, Arús P, Aranda M, Álvarez-Tejado M, Droege M, Du L,
Puigdomènech L, Garcia-Mas J (2010) A draft sequence of the melon genome revealed by
high throughput shotgun sequencing. Abstracts of Plant & Animal Genome Conference
XVIII (http://www.intl-pag.org/)
Boualem A, Fergany M, Fernandez R, Troadec C, Martin A, Morin H, Sari MA, Collin F,
Flowers JM, Pitrat M, Purugganan MD, Dogimont C, Bendahmane A (2008) A conserved
mutation in an ethylene biosynthesis enzyme leads to andromonoecy in melons. Science
321: 836–838.
Boualem A, Troadec C, Kovalski L, Sari M-A, Perl-Treves R, Bendahmane A (2009) A conserved
ethylene biosynthesis enzyme leads to andromonoecy in two Cucumis species. PLoS
One 4: e6144.
Cavagnaro PF, Senalik DA, Ynag LM, Simon PW, Harkins TT, Kodira CD, Huang S, Weng Y
(2010) Genome-wide characterization of simple sequence repeats in cucumber (Cucumis
sativus L.). BMC Genomics 11: 569.
Chen JF, Kirkbride JH (2000) A new synthetic species Cucumis (Cucurbitaceae) from interspecific
hybridization and chromosome doubling. Brittonia 52: 315–319.
Chen JF, Staub JE, Tashiro Y, Issiki S, Myiazaki S (1997) Successful interspecific hybridization
between Cucumis sativus L. and C. hystrix Chakr. Euphytica 96: 413–419.
Chen JF, Ling MS, Qian ChT (2001) Identification of Meloidogyne incognita (Kofoid & White)
Chitwood resistance in Cucumis hystrix Chakr. and the progenies of its interspecific
hybrid with cucumber (C. sativus L.) J Nanjing Agricult Univ 24: 21–24 (In Chinese with
English abstract).
Chen JF, Luo XD, Qian ChT, Molly MJ, Staub JE, Zhuang FY, Lou QF, Ren G (2004) Cucumis
monosomic alien addition lines: morphological, cytological, and genotypic analyses.
Theor Appl Genet 108: 1343–1348.
Cho MC, Om YH, Huh YC, Mok IG, Park HG (2003) Two oriental squash varieties resistant
to powdery mildew bred through interspecific crosses. Cucurbit Genet Coop Rep 26:
40–41.
Clark AM, Jacobsen KR, Bostwick DE, Dannenhoffer JM, Skaggs MI, Thompson GA (1997)
Molecular characterization of a phloem-specific gene encoding the filament protein,
phloem protein 1 (PP1), from Cucurbita maxima. Plant J 12: 49–61.
Dane F, Liu L (2006) Diversity and origin of cultivated andcitron type watermelon (Citrullus
lanatus). Genet Resour Genet Resour Crop Crop Evol 54: 1255–1265.
Davis AR, Penelope P-V, Sakata Y, López-Galarza S, Maroto JV, Lee S, Huh Y, Sun ZY, Miguel
A., King S. R, Cohen, R. and Lee J-M (2008) ‘Cucurbit Grafting’, Critical Reviews in Plant
Sciences 27: 50-74.
Díez MJ, Nuez F, Maggioni L, van Dooijeweert W (2007) The ECP/GR Cucurbitaceae Working
Group Acta Hort 731: 25–30.
Dijkhuizen A, Kennard WC, Havey MJ, Staub JE (1996) RFLP variability and genetic
relationships in cultivated cucumber. Euphytica 90: 79–89.
Engels JMM, Rao VR, Brown AHD, Jackson MT (2002) Managing Plant Genetic Diversity.
CABI Publishing, Oxford, UK.
Eschrich W, Evert RF, Heyser W (1971) Proteins of the sieve tube exudate of Cucurbita maxima.
Planta 100: 208–221.
Ezura H, Owino WO (2008) Melon, an alternative model plant for elucidating fruit ripening.
Critical Rev Biotech 28: 13–55.
Ezura H, Fukino N (2009) Research tools for functional genomics in melon (Cucumis melo L.):
Current status and prospects. Plant Biotechnology 26: 359–368.
FAO (2009) Draft second report on the world’s plant genetic resources for food and agriculture
(CGRFA-12/09/Inf.7Rev.1). ftp://ftp.fao.org/docrep/fao/meeting/017/ak528e.pdf.
Accessed on December 13, 2010.
FAO (2010) http://faostat.fao.org/site/567/DesktopDefault.aspx?PageID=567#ancor.
Accessed on December 10, 2010.
Feher T (1993) Watermelon. In: G Kalloo, BO Bergh (eds) Genetic Improvement of Vegetable
Crops. Pergamon Press, Oxford, England, pp 295–314.
Ferriol M, Pico B (2008) Pumpkin and winter squash. In: J Prohens, F Nuez (eds) Handbook
of Plant Breeding—Vegetables 1. Springer, Berlin, Germany, pp 317–350.
Ghebretinsae AG, Thulin M, Barber JC (2007a) Relationships of cucumbers and melons
unraveled: molecular phylogenetics of Cucumis and related genera (Benincaseae,
Cucurbitaceae). Am J Bot 94: 1256–1266.
Ghebretinsae AG, Thulin M, Barber JC (2007b) Nomenclatural Changes in Cucumis
(Cucurbitaceae). NOVON 17: 176–178.
Green, SK, Engle LM, Liu Ca, Kuo CG (2007) AVRDC’s cucurbit genetic resources and prospects
for their use as sources of disease resistance. In: Symposium of International Workshop on
the Cucurbit Diseases and Resistance Breeding. Wufeng, Taichung: The Plant Protection
Society of the Republic of China. pp 119–136.
Han YH, Zhang ZH, Liu CX, Liu JH, Huang SW, Jiang JM, Jin WW (2009) Centromere
repositioning in cucurbit species: Implication of the genomic impact from centromere
activation and inactivation. Proc Natl Acad Sci USA 106: 14937–14941.
Havey MJ (1997) Predominant paternal transmission of the cucumber mitochondrial genome.
J Hered 88: 232–235.
Havey MJ, McCreight JD, Rhodes B, Taurick G (1998) Differential transmission of the Cucumis
organellar genomes. Theor Appl Genet 97: 122–128.
Herrington ME, Prytz S, Wright RM, Walker IO, Persley DM, Greber RS (2001) ‘Dulong
QHI’ and ‘Redlands Trailblazer’, PRSV-W-, ZYMV-, and WMV- resistant winter squash
cultivars. HortScience 36(4): 811–812.
Horst EK, Lower RL (1978) Cucumis hardwickii, a source of germplasm for the cucumber
breeder. Cucurbit Genet Coop Rpt 1: 5.
Huang S, Li RQ, Zhang ZH, Li L, Gu XF, Fan W, Lucas WJ, Wang XJ et al. (2009) The genome
of the cucumber, Cucumis sativus L. Nat Genet 41: 1275–1281.
Jacks TJ, Hensarling TP, Yatsu LY (1972) Cucurbit seeds: I. Characterizations and uses of oils
and proteins, a review. Econ Bot 26: 135–141.
Jeffrey C (1980) A review of the Cucurbitaceae. Bot J Linn Soc 81: 233–247.
Sanjur OI, Piperno DR, Andres TC, Wessel-Beaver L (2002) Phylogenetic relationships among
domesticated and wild species of Cucurbita (Cucurbitaceae) inferred from a mitochondrial
gene: Implications for crop plant evolution and areas of origin. Proc Natl Acad Sci USA
99: 535–540.
Schaefer H (2008) Cucumis (Cucurbitaceae) must include Cucumella, Dicoelospermum, Mukia,
Myrmecosicyos, and Oreosyce: a recircumscription based on nuclear and plastid DNA
data. BLUMEA 52: 165–177.
Schaefer H, Heibl C, Renner SS (2009) Gourds afloat: a dated phylogeny reveals an Asian
origin of the gourd family (Cucurbitaceae) and numerous oversea dispersal events. Proc
R Soc B 276: 843–851.
Sebastian P, Schaefer H, Telford IR, Renner SS (2010) Phylogenetic relationships among
domesticated and wild species of Cucumis (Cucurbitaceae): The sister species of melon
is from Australia. Proc Natl Acad Sci USA 107: 14269–73.
Simon PW, Navazio JP (1997) Early orange mass 400, early orange mass 402, and late orange
mass 404: high-carotene cucumber germplasm. HortScience 32: 144–145.
Sisko M, Ivancic A, Bohanec B (2003) Genome size analysis in the genus Cucurbita and its use
for determination of interspecific hybrids obtained using the embryo-rescue technique.
Plant Sci 165: 663–669.
Smith BD (1997) The initial domestication of Cucurbita pepo in the Americas 10,000 years ago.
Science 276: 932–934.
Staub JE, Knerr LD, Holder DJ, May B (1992) Phylogenetic relationships among several African
Cucumis species. Can J Bot 70: 509–517.
Staub JE, Robbins MD, Wehner TD (2008) Cucumber. In: J Prohens, F Nuez (eds) Handbook
of Plant Breeding—Vegetables 1. Springer, Berlin, Germany, pp 241–282.
Tatlioglu G (1993) Cucumber. In: G Kalloo, BO Bergh (eds) Genetic Improvement of Vegetable
Crops, Pergamon Press, Oxford, England, pp 197–234.
Tatum TC, Nunez L, Kushad MM, Rayburn AL (2006) Genome size variation in pumpkin
(Cucurbita sp.). Ann Appl Biol 149: 145–151.
Trebitsh T, Staub JE, O’Neill SD (1997) Identification of a 1-aminocyclo- propane-1-carboxylic
acid synthase gene linked to the female (F) locus that enhances female sex expression in
cucumber. Plant Physiol 113: 987–995.
Turgeon R (1996) Phloem loading and plasmodesmata. Trends Plant Sci 1: 418–423.
Ward BL, Anderson RS, Bendich AJ (1981) The mitochondrial genome is large and variable in
a family of plants (Cucurbitaceae). Cell 25: 793–803.
Wehner TC (2008) Watermelon. In: J Prohens, F Nuez (eds) Handbook of Plant Breeding—
Vegetables 1, Springer, Berlin, Germany, pp 381–418.
Whitaker TW (1993) Cytological and phylogenetical studies in the Cucurbitaceae. Botanical
Gazette (Crawfordsville) 94: 780–790.
Whitaker TW, Davis GN (1962) Cucurbits—Botany, Cultivation, Utilization. Interscience Publ,
New York, p 266.
Whitaker TW, Robinson RW (1986) Squash breeding. In: MJ Basset (ed) Vegetable Breeding.
AVI Publishing Company, Westport, Connecticut, pp 209–242.
Xu Y, Guo SG, Zhang HY, Gong GY, Huang SW, Ye HP, Wu MZ, Zheng Y, Fei ZJ (2009) Latest
advances in watermelon genomics. Abstract of 4th Intl. Cucurbitaceae Symp. Sept 20–24,
Changsha, China, p 38.
Zhang BC, Tolstikov V, Turnbull C, Hicks LM, Fiehn O (2010) Divergent metabolome and
proteome suggest functional independence of dual phloem transport systems in cucurbits.
Proc Natl Acad Sci USA. doi/10.1073/pnas.0910558107.
Zhang J (1996) Breeding and production of watermelon for edible seed in China. Cucurbit
Genetics Cooperative Report 19: 66–67.
Zhou XH, Wan HJ, Qian CT, Chen JF (2008) Development and characterization of Cucumis
sativus-hystrix introgression lines exhibiting resistance to downy mildew. Cucurbitaceae
2008, Proceedings of the IXth EUCARPIA meeting on genetics and breeding of
Cucurbitaceae, M Pitrat (ed), INRA, Avignon, France, May 21–5, pp 353–358.
ABSTRACT
Minor cururbitaceous vegetables are major sources of calories, minerals
and vitamins. Among these vegetables, pumpkin is a rich source of
carotene, bitter gourd is rich in vitamin C and iron and Luffa in minerals.
Pumpkin cultivars grow fruits with the strongest flavor, highest soluble
solids, and deepest flesh color that are preferred for canned and frozen
products. Besides being used as vegetables, bitter gourd possesses
antioxidant, antimicrobial, antiviral, antihepatotoxic, antiulcerogenic
properties, while also having the ability to lower blood sugar. The young
tender fruits of Luffa acutangula (ridge gourd) and Luffa cylindica (sponge
gourd) are edible and may be eaten sliced like cucumbers, or in soups
such as okra, or cooked like squash. Nutritional value like antioxidants,
carotenoids, tocopherol, minerals and ascorbic acid are also to be
considered while breeding these crops. In pumpkin, total carotenoid
content has a positive association with the fruit-flesh color intensity.
Dark orange fleshed varieties have high carotenoid content whereas
varieties with a bright yellow color flesh have high lutein content and
low carotene content. Use of wild species for transferring economically
important traits continues to be major objective for breeders especially
in cases where resistance genes for several pathogens and pests have
not been found within the cultivated species.
Keywords: Cucurbita moschata, Momordica charantia, Luffa spp, minor
cucurbits, germplasm enhancement, genetics and breeding.
1
Indian Agricultural Research Institute, New Delhi, India.
2
Indian Institute of Horticulture Research, Bengaluru, India.
*Corresponding author: tusar@rediffmail.com, tusar@iari.res.in
2.1 Introduction
2.1.1 Distribution
Cucurbits are of tremendous economic importance and are cultivated
throughout the world from tropical to sub-temperate zones. China, Turkey,
India and Iran are important cucurbit growing countries. Pumpkin may
have been domesticated in Mexico and northern South America. Then it
migrated to the Caribbean islands and from there reached Florida, the
native Americans developed a distinct landrace called Seminole pumpkin.
More diversification on pumpkin has taken place in Asia and Africa. Bitter
gourd is a very important crop of India, the Phillipines, Malaysia, China,
Australia, Africa, the Middle East, Latin America and the Caribbean.
Momordica charantia was domesticated in eastern India or southern China.
Luffa cylindrica (smooth gourd) contains wild populations distributed from
southern Central Asia to north-eastern Australia and the South Pacific.
The domestic variety is cultivated in Asia, Africa and tropical America.
Luffa acutangula is mostly grown in South-eastern Asia and other tropical
countries (Table 2-1). In India, this diversity is concentrated in the Indo-
Gangetic plains, north-eastern regions, north-western Himalayas, the
Western and Eastern Ghats and sporadically in the tribal dominant belts
of Central India. More diversity occurs in Cucurbita spp. in the north-east
as also for ash gourd, bottle gourd while for Luffa, it is more concentrated
in the eastern peninsular tract. In case of Cucumis melo and round gourd,
it is more confined to north-western and Indo-Gangetic plains. In pointed
gourd, diversity is concentrated more in eastern part of the Indo-Gangetic
plains particularly in West Bengal and adjoining Bihar along eastern Uttar
Pradesh. Coccinia cordifolia (growing wild throughout India, raw fruits
used as vegetables), Cucumis sativus (distributed throughout India, in the
Himalayas as well), Lagenaria siceraria (African origin but domesticated
throughout India, tender fruits used as vegetables), Luffa cylindrica
(indigenous to India whereas acutangula found in western, Central and
southern India and is regarded as a wild form of cultivated species—tender
fruits used as vegetables), L. acutangula var. amara (occurs in peninsular
India and is a wild relative of cultivated spongegourd), L. echinata (western
Himalayas, Central India Gangetic plains) and L. graveolans (considered
as wild progenitor of L. hermaphrodita in Bihar and Sikkim) are potential
species. In addition, Momordica, M. balsamina occurs in semi-dry north-
western plains, northern parts of Eastern and Western Ghats, M. dioica
and M. cochinchinensis occur as wild/semi-wild in the Gangetic plains.
Trichosanthes has 21 species occurring in India and the major zones of species
concentration are (a) along the Malabar coast in the Western Ghats, (b) low
and medium elevation zones in the Eastern Ghats and the north-eastern
Minor Cucurbits 19
L. cylindrica Sponge gourd Chikni India 26 Throughout tropics, India Young fruits as vegetable,
tori mature for fiber
Table 2-1 contd....
20
Botanical name Common name(s) Origin Chromosome Geographical distribution Uses
Momordica charantia Bitter gourd Indo-Burma 22 Throughout tropics, India, S Young fruit, leaf as
Karela Region Tropical & SE Asia vegetable, seed as
Africa condiment
M. cochinchinensis Cochinchin gourd, India 28 South Asia Immature fruit as
sweet gourd vegetable, young leaf,
flower and seeds are
edible
M. dioica Kaksa, kakrol Southern tropical 28 Tropical Asia and Africa Fruit, shoot and leaf as
Asia vegetable
Praecitrullus fistulosuos Round melon, Indian India 24 North-West India Fruit as vegetable
squash, tinda
Sechium edule Chayote, chow-chow, Mexico, 24 Throughout tropics Young fruit and root
mirliton Guatemala especially Latin America (tuberous) as vegetable
Sicana odorifera Casabanana Brazil, Peru 40 Central America, northern As fruit
South America
Telfairia occidentalis Fluted pumpkin West Africa 22 Tropical Africa Leaf and shoot as
vegetable, seed is edible
T. pedata Oyster nut Africa 22 Tropical Africa Potential substitute for
almonds or Brazil nuts
Trichosanthes anguina syn. Snake gourd, India 22 South and South-East Asia Immature fruit, shoot and
T. cucumerina chichinda leaf as vegetable
2.2.1 Pumpkin
The common terms “pumpkin”, “squash”, “gourd”, “cushaw”, “ayote”,
“zapallo”, “calabaza”, etc. are often indiscriminately applied to different
cultivated species of the genus Cucurbita L.: C. pepo L., C. maxima Duchesne,
C. moschata Duchesne, C. argyrosperma C. Huber and C. ficifolia Bouche
(Ferriol and Pico 2008). “Pumpkin” is mostly used to refer to cultivars
with round fruits, which are used when mature for baking or for feeding
livestock. C. pepo is the most economically important species of Cucurbita
grown worldwide. C. maxima and C. moschata mainly includes cultivars
grown as “winter squashes” in developing countries under low-input
agricultural systems. In southern Latin America, C. maxima is largely grown
for immature fruit consumption (zapallito varieties) and some C. moschata
cultivars are also valued as “summer squashes”. C. argyrosperma and
C. ficifolia have less economic importance and a narrower distribution. In
both these species, the mature fruits are more valued, but some varieties
are eaten as a vegetable (Ferriol and Pico 2008).
Among cucurbitaceous vegetables, pumpkin (Cucurbita spp.) has been
appreciated for high yields, long storage life and high nutritive value.
C. moschata are eaten mature and can be baked, boiled, or microwaved. They
are low in saturated fat, cholesterol, and sodium, and are a good source
of dietary fiber, vitamins, and minerals (Nutrition Data 2006). Fruits with
yellow or orange flesh generally have high concentrations of carotenes,
some of which are the precursors of vitamin A (e.g., β-carotene) and play a
significant role in human nutrition, especially in tropical countries where
their consumption is high. C. moschata cultivars producing fruits with the
strongest flavor, highest soluble solids, and deepest flesh color are preferred
for canned and frozen winter squash. A variety of value added products such
as jam, jelly, marmalade, candy, puree, sauce, chutney, pickle and halwa are
prepared from pumpkin. Deep orange, carotene-rich fruits of C. moschata
and C. maxima are processed for use in the baby food industry. Pumpkin
flour can be used to supplement cereal flours in bakery products, soups,
instant noodles and natural coloring agent in pasta and flour mixes. Its
male flowers, young stems and leaves and young and ripe fruits are eaten
as a vegetable. The latter are also commonly used to prepare sweets and as
fodder. Its seeds are also consumed in many parts of Mexico and Central
America. The seeds are eaten whole, roasted or toasted and are ground
into different stews. They have high oil and protein contents. Plants of C.
moschata are also used as rootstocks for its resistance to diseases and abiotic
stresses, for the winter cultivation of watermelon, melon and cucumber
(Traka-Mavrona et al. 2000). Pumpkin seeds are a rich source of oil and
nutrients and can be consumed as food. The seed flour is used as a protein
supplement in bread and cookies. Pumpkin seeds have many health benefits
valued as medicine in China. Juice from the above ground stem of sponge
gourd is used against respiratory problems in Japan. It is also used as an
ingredient for cosmetics (Lee and Yoo 2006). They are used for bathing,
removing toxins and regenerating the skin. They help varicose veins and
cellulite by stimulating circulation. The tender sponge gourd is considered
a diuretic and lactagog. It is also considered good against diabetes (Bal et
al. 2004a). Oral administrations (2 g/kg) of ethanolic extract of L. aegyptiaca
seeds decreased blood glucose level in streptozotocin diabetic rats (Fiky et
al. 1996). The ripe fruit after burning and pulverizing is used as carminative
and anthelmintic. The fruit juice is considered a purgative. Mature seeds
are bitter, emetic and cathartic. The seed oil is said to be useful for skin
affections and in Brazil, they are suggested as possible substitutes for olive
oil (Porterfield Jr. 1955).
Research in modern medicine system involves treating disease at the
gene level, aims at blocking expression of proteins, which are responsible
for disease. A wide variety of compounds (inhibitors) have been
isolated from plants and the most extensively studied are the ribosome
inactivating proteins (RIP). The RIP luffins have been isolated from Luffa
and characterized (Kishida et al. 1983). Besides, polypeptides of about 5
kDa molecular mass, such as thionins, have been reported to inhibit cell-
free protein synthesis, probably by a nonenzymatic mechanism (Garcia-
Olmedo et al. 1983). Ramakrishnan et al. (1989) purified a protein with
molecular weight of 30 kDa from L. aegyptiaca seeds, which inhibited cell free
translation at picomolar concentrations. Chemical linkage of the protein to
a monoclonal antibody reactive to transferring receptor resulted in a highly
cytotoxic conjugate. ChangYun and ZuChuan (1998) isolated and purified a
group of novel small ribosome inactivating proteins, LuffinS1, LuffinS2 and
Luffin S3 from L. cylindrica seeds. Parkash et al. (2002) reported presence of
luffacyclin, a ribosome inactivating peptide with antifungal activity against
Mycospharella arachidicola and Fusarium oxysporum from L. cylindrica seeds.
Luffaculin 1, a ribosome inactivating protein purified from L. acutangula is
reported to possess anti-HIV1 activities (Jing et al. 2008).
2.3.3 Luffa
Luffa is a member of the subfamily Cucurbitoideae, tribe Benincaseae,
subtribe Luffinae (Jeffrey 1962, 1980a). It is the only member of the subtribe
and has species in both the Old and New Worlds. In some respects, it
resembles members of the Cyclantherinae, an entirely New World subtribe
of the Siceyeae. It thus may be the connecting link of these two tribes. In the
most comprehensive taxonomic treatment of the genus presently available,
Cogniaux and Harms (1924) accepted eight species. One of these species
Luffa variegata Cogn., has since been transferred to Lemurosieyos (Keraudren
1965) and two others, L. forskalii Schwein. Ex Harms and L. umbellata (Klein)
M.J. Roem., have been reduced to synonymy or varietal status under L.
acutangula (L.) Roxb. (Jeffrey 1980a; Heiser and Schilling 1988). Five species,
four from the Old World and one in the New World, have generally been
accepted in recent years (Jeffrey 1980b; Heiser and Schilling 1988 ). Heiser
et al. (1988), however, proposed that three species are present in the New
World.
All Luffa species are vines and bear solitary pistillate flowers and
racemes of male flowers. Analyses of the flavonoids, breeding system,
leaves, flowers and fruit resulted in the grouping of L. acutangula and
L. aegyptiaca Mill. into a single clade, apart from the other five species (Heiser
and Schilling 1990). This result is not entirely supported by chloroplast DNA
makers (Chung et al. 2003).
species and may include other species that are fully cross-compatible. The
secondary gene pool is represented by all other populations that can be
crossed with the crop; the gene flow is possible but is connected with a
reduction of fertility within hybrid generations. Species from the tertiary
gene pool cannot be crossed with the crop species except through special
biotechnological approaches because the resulting hybrids often express
abnormalities and are usually lethal or completely sterile.
Generally, the five domesticated Cucurbita species are reproductively
isolated from one another. The primary gene pools of each species are
represented by their landraces and commercial cultivars as well as by their
infraspecific taxa (Table 2-4). Experimental crosses can be made among them
but with difficulty, and interspecific progenies are usually either sterile or
sparingly fertile (Merrick 1995). Spontaneous crosses between the cultivated
Cucurbita species are uncommon, but natural interspecific hybrids can be
occasionally detected in landraces mostly from Mexico (Decker-Walters et
al. 1990; Merrick 1990). Kristkova (1991) reported a spontaneous hybrid of
C. maxima × C. pepo under Central European field conditions.
Table 2-4 Gene pools of cultivated Cucurbita species.
and wild species can occur sympatrically, and genetic interchange between
them takes place, providing a natural source of variation within populations
(Wilson 1990; Merrick 1991).
Hybridization experiments and field observations involving
C. argyrosperma and other wild and cultivated Cucurbita taxa have revealed
(Lira-Saade 1996) that, among the cultivated species, C. moschata has the
highest degree of compatibility with C. argyrosperma, placing it into its
secondary gene pool. The next level of cross-compatibility involves the wild
and cultivated taxa of C. pepo, some cultivars of C. maxima, and the wild
perennial species C. foetidissima, which collectively represent the tertiary
gene pool. The wild species that have shown some degree of compatibility
with C. argyrosperma possess genes of resistance to some viral diseases that
have a high incidence in the cultivated species (Lira-Saade 1996).
Greatest diversity in C. moschata occurs among the innumerable
landraces of the American tropics (Andres 2004a). Isozyme variation studies
in Cucurbita by Andres (1990) and Decker-Walters et al. (1990) revealed
moderate amounts of genetic differentiation within C. moschata. In the New
World, C. moschata is cultivated in a wide range of elevations, which suggests
that this species has evolved diverse adaptations to various environmental
conditions (Andres 2004b). The species is highly polymorphic (Andres
2004a) with considerable morphological diversity of its seeds and fruit
(color, shape, thickness, and durability of the fruit’s skin). The landraces
from northern Peru also exhibit high diversity. This includes the special
type called “Loche”, which typically has warty fruits and deep, orange
flesh valued highly by the locals as a flavoring for stews (Andres 2004a).
The existence of varieties with life cycles of differing phenology and the
breadth of its numerous cultivars developed in other parts of the world
(Gwanama et al. 2000) and of local varieties with excellent horticultural
characteristics strongly suggest that its collective genetic variation is very
extensive (Lira-Saade 1996).
Variation of C. moschata populations is also observed outside its center of
origin. For example, a landrace native to Nigeria, represents the only source
of resistance to certain viral diseases (Provvidenti 1993). The gene pool of
C. moschata is represented by the numerous commercial cultivars that have
been mainly developed in the United States and, to a lesser extent, in Brazil.
Some of these commercial cultivars have also different levels of resistance
and/or susceptibility to certain diseases, indicating a wide genetic variation
of this species (Lira-Saade 1996). Some of the Cuban landraces are a source of
genetic material for tolerance to marginal growth conditions (Andres 2004a).
Significant diversity is found in landraces from warmer regions of Asia or
Africa (the warty and wrinkled Japanese fruits; the Indian landraces with
large, soft-skinned fruits; the abundance of barbell-shaped fruits in Asia
Minor, the Nigerian landraces resistant to diseases, etc.) (Lira-Saade 1996;
More recently molecular markers [i.e., RAPD (Dey et al. 2006), ISSR (Singh
et al. 2007), and AFLP (Gaikwad et al. 2008)] have been used to assess the
genetic diversity of Indian bitter gourd genotypes including two promising
gynoecious lines (DBGy-201 and DBGy-202). A wide range in genetic
diversity was detected, indicating that a standard accession reference
array for future analyses might include “Pusa Do Mausami-green”, “Pusa
Do Mausami-white”, DBTG-2, Mohanpur Sel-215, and Jaynagar Sel-1.
Regardless of marker analyses type, however, Mohanpur Sel-125, DBTG-
101, and Jaynagar Sel-1 from West Bengal (an eastern state of India) are
genetically distinct (genetic similarity, GS) from other common landrace
accessions from North Indian states (GS = 0.57 to 0.72). Genetic differences
between M. charantia var. charantia (both exotic and indigenous) and M.
charantia var. muricata (indigenous) accessions are indicative of their use
as potential parents for the establishment of narrow (indigenous) and
wide-based (both exotic and indigenous) mapping populations. The exotic
(obtained from AVRDC-World Vegetable Centre) populations may be
informative for the characterization of qualitative and quantitative traits
in other cucurbit species (Serquen et al. 1997; Zalapa et al. 2007).
In bitter gourd, gynoecy is particularly interesting for hybrid
development (e.g., gynoecious x monoecious lines) and their commercial
production. The commercial deployment of gynoecy in hybrid technology
avoids the tedious step of pinching of male flowers during the production of
monoecious x monoecious hybrids. The utilization of such gynoecious lines
allows for the production of gynoecious or predominantly gynoecious lines
that provide early, uniform, high yield potential (Dey et al. 2010; Ram et al.
2002a). The hybrid, “Cuilli No.1” (China), for instance, was developed by
utilizing a gynoecious line as a maternal parent (Zhou et al. 1998). Likewise,
gynoecious lines originating in India were identified by Behera et al. (2006a;
lines DBGy-201 and DBGy-202) and Ram et al. (2002b; line Gy263B) for use
in hybrid development programs.
Commercial bitter gourd varieties and a few accessions/lines with
potentially important horticultural traits have been deposited and registered
in national germplasm collections. However, the National Bureau of Plant
Genetic Resources (NBPGR), New Delhi, India possess some unique
accession such as IC 256185, IC 248256, IC 213311, IC 248282, IC 256110 and
IC 248281 (Dhillon et al. 2005; resistant to fruit fly), NIC-12285 and VRBT-39
(resistant to downy mildew), IC 202195 (high yield and long fruited type),
TCR 404 (high yield and white fruited type), EC 399808 (high yield and
larger number of fruits), and INGR 03037 (gynoecious sex with high yield),
which can be used directly by plant breeders. Wild bitter gourd ecotypes and
botanical varieties (e.g., M. charantia var. muricata) are also important sources
of economically important traits (e.g., resistance against Dacus cucurbitae;
2.4.3 Luffa
Cytogenetic investigations have been conducted among the Old World
species and chromosome counts in all the species were found to be the
same (2n = 26) with basic chromosome number of 13 (x = 13). The New
World species L. astorii also have 26 chromosome counts, but the presence
of numerous globular inclusions in the pollen mother cells of L. operculata
and L. quinquefida made it impossible to make exact counts in these species.
From the analysis of some of the hybrids, it is apparent that the chromosome
number of both the species is n = 13. Comparative morphology of the Old
World wild and cultivated species and chromosome pairing in interspecific
hybrid suggests that L. graveolens is the prime species, which has given rise
to the two cultivated monoecious species L. acutangula and L. cylindrica (Dutt
and Roy 1971). Pathak and Singh (1949) have given a detailed account on
the cytological behavior of interspecific hybrids between L. acutangula ×
L. cylindrica. These hybrids exhibited reduced fertility. Fertility is restored
when F1 hybrid is crossed with either of the parents. Cytological studies
reveal that each chromosome of haploid complements of the two species
is sufficiently homologous, but non-homologous segments are present in
normally pairing chromosomes of F1 hybrid between the two species. The
interspecific combinations between L. graveolens, L. aegyptiaca/cylindrica,
L. acutangula and Luffa echinata resulted in sterile hybrids (Dutt and Roy
1969, 1971). In a study on interspecific hybrids between L. acutangula and
L. graveolens and their amphidiploid (2n = 4x = 52), Dutt and Roy (1976)
reported frequent occurrence of univalents and bivalents at diakinesis and
metaphase-I. The univalents ranged from 0.0–18.0, the mean being 5.83 per
pollen mother cell (PMC). The rod bivalents varied from 0.0–14.0, whereas
the ring bivalents were 8.0–20.0 per cell. The mean number of chiasmata
per PMC was 37.43 as against 17.12 in the F1. Chromosome associations,
higher than bivalents were frequently scored, trivalents in 33.3% of the cells
and quadrivalents in 30.0% of the PMCs were observed. Low frequency
(<1%) of higher quadrivalents were also noticed. At anaphase-I, regular
disjunction of chromosomes along with few irregularities like unequal
separation and lagging chromosomes were found, which resulted in empty
pollen formation and subsequent pollen sterility of the amphidiploids.
Autotetraploidy was induced in L. acutangula (4x = 52), however, such
(Hi, Hard rind inhibitor and Bi, Bitter fruit in C. maxima). In C. maxima and
C. moschata, the Butternut and Buttercup varieties are popular because they
lack a hard shell and can be easily cut with a kitchen knife. Fruit shape is
polygenic, but is controlled by a major gene in C. moschata (Bn, Butternut).
Many genes control rind color (some affecting flesh color). However, other
allelic and non-allellic genes have also been reported in C. maxima and
C. moschata. Some of these genes are multiple-allelic and interact with each
other (Paris and Brown 2005). In C. maxima Bmax (Bicolour), derived from the
subsp. andreana, is responsible for precocious yellow fruit pigmentation,
and Rd (Red skin) for the red color of “Turk’s Caps”, dominant to other
colors. Knowledge of genetics of these traits facilitates their use in breeding
programs for improvement of Cucurbita.
Local Cucurbita germplasm has been subjected to horticultural and
agronomic evaluation worldwide with the objective of identifying valuable
accessions for breeding, viz. Bulgaria (Stefanova et al. 1994), Brazil (Choer
1999; Romao et al. 1999; Ramos et al. 2000), Cuba (Rios Labrada et al. 1998),
China (Chen 1993; Zhou et al. 1995) and India (Babu et al. 1996; Kale et al.
2002; Kumar et al. 2006). Transfer of economically important characteristics
such as host-plant resistance to diseases and pests or stress tolerance from
wild Cucurbita to their cultivated counterparts is challenging for breeders.
An interspecific cross is often used to introgress such characters. In the
genus Cucurbita, several attempts have been made to produce interspecific
hybrids among five cultivated species (C. pepo, C. maxima, C. moschata,
C. argyrosperma, C. ficifolia) and a number of non-cultivated ones. Using such
an approach, one could expect novel traits to be introgressed or genotypes
with recombined characters of involved species to be created.
Generally wild Cucurbita species have viny growth habit. The Bu
allele (Bush habit), which is responsible for bush vine in C. pepo, is also
responsible for the bushy habit in C. maxima, but the conversion from
compact to a more spreading growth occurs much faster in bush-vine
heterozygotes of C. maxima than those of C. pepo. A different gene confers
determinant plant growth in C. moschata (de, determinate) (Kwack 1995).
Bushy or semibushy habit has been introduced in many varieties of C.
pepo, C. maxima and C. moschata. The bushy habit has been introduced
into some cultivars of C. maxima by developing productive short- × long-
vined hybrids. The introduction of the bushy habit in C. moschata was
first transferred to temperate types by interspecific hybridization between
C. pepo and C. moschata followed by backcrosses to C. moschata. Some
compact hybrids have been derived from these bushy C. moschata cultivars
and long-vined types with tropical fruit characteristics (adapted to tropical
conditions and resistant to diseases) have been developed (Maynard et
al. 2002). These hybrids showed higher yields, but smaller fruits than the
long-vine landraces (Chesney et al. 2004). Bush plants are characterized
1986). Ovule culture has been reported for use when making compatible
crosses in Cucurbita species (Hong and Hyo-Guen 1994). The Sakata Seed
Company of Japan offers the interspecific hybrids of C. moschata × C. maxima
(known as Kabocha) under different cultivar names (e.g., Alguri, Kikusui,
Tetsakabuto). Fruits are reported to combine favorable characters from
both species, and there is considerable heterosis for degree of female sex
expression and for yield (Whitaker and Robinson 1986).
Production of interspecific Cucurbita hybrids have been attempted using
embryo rescue, following techniques reviewed by Rakoczy-Trojanowska
and Malepszy (1989a) and Rakoczy-Trojanowska et al. (1992). Hybrid
genotypes with novel traits or entirely new characteristics may result. In
general, success of hybridization within this genus varies from combination
to combination. Successful rescue of Cucurbita hybrid embryos has
been reported for many interspecific combinations, including C. pepo ×
C. moschata (Wall 1954; de Oliveira et al. 2003; Sisko et al. 2003), C. maxima
× C. pepo (Rakoczy-Trojanowska and Malepszy 1986, 1989b; Sisko et al.
2003), C. moschata × C. maxima (Kwack and Fujieda 1987). C. ficifolia ×
C. maxima (El-Mahdy et al. 1991; Sisko et al. 2003), C. maxima × C. foetidissima
(Rakoczy-Trojanowska et al. 1992), and C. argyrosperma × C. moschata
(Sisko et al. 2003). Limited success in obtaining fully developed seeds that
germinated and produced viable plants has also been reported for a few
interspecific combinations, including C. andreana × C. ficifolia (Whitaker
1954), C. lundelliana × C. moschata (Whitaker 1959), and C. maxima ×
C. ecuadorensis (Paran et al. 1989).
The hybrid nature of plantlets obtained from interspecific crosses need
to be tested, since partenocarpic development or accidental pollination with
parental species cannot be fully excluded. Several methods are available to
confirm the hybrid nature of regenerants, such as differentiation in plant
morphological characters, chromosome counting, or the use of various
DNA-based molecular marker techniques. The genome sizes can be
compared and the hybrid nature of the rescued plants can be determined
early in the rescue process by using flow cytometry. Sisko et al. (2003)
showed that differences in genome size within the genus Cucurbita are big
enough to be used efficiently in determination of interspecific hybrids.
The main utility of genetic maps in plant improvement is their deployment
in marker-assisted selection (MAS) and breeding. The predictive value of
genetic markers used in MAS depends upon their inherent repeatability,
map position, and linkage with economically important quantitative or
qualitative traits (Staub et al. 1996). Until recently, the genetic mapping in
the genus Cucurbita was hampered by the lack of available sequence-specific
markers. Based on anonymous RAPD and AFLP markers, two maps using
interspecific crosses between C. pepo and C. moschata (Lee et al. 1995; Brown
and Myers 2002) were developed that did not allow comparative analysis
soluble solid content (> 3.10 0Brix; “MC-84”, “Preethi”, “RHRBG-5” and
“PBIG-1”), and vitamin C (> 950 mg·kg–1; “Konkan Tara” and “Hirkani”)
and fruit protein (>1.5%; ‘DVBTG-1’, “Preethi”, “Hirkani” and “Konkan
Tara”) content (Kore et al. 2003).
Polyploids can be produced by treating the seedlings at the cotyledon
stage with an emulsion of 0.2% colchicine. The seed treatment with 0.2, 0.4%
colchicine or 0.003% amiprophos-methyl was effective for chromosome
doubling, among which the treatment with 0.4% colchicine was most
effective. Amiprophos-methyl treatment also produced octoploid plants
with a high rate of seed germination. Multiple shoot treatments with
0.05% colchicine for 12 and 24 hours, and 0.1% colchicine for 24 hours also
produced octoploid plants. Leaf and guard cell size were bigger, and leaf
shape index (leaf length/leaf width) was lower in the octoploid as compared
to tetraploid plants. Leaves of the octoploid plants were uneven on the
surface with clear serrations.
Triploid plants of M. charantia were obtained by crossing the tetraploid
(colchicine-induced) and diploid plants. In seedlings of bittergourd,
colchicine at 0.2% for 18 hours to the shoot tip produced tetraploids (Kadir
and Zahoor 1965). However, polyploids were inferior to diploids in terms
of economic characters.
In M. charantia, progeny (M1) derived from radiation mutagenesis can
possess economically important unique characters, which are controlled by
single recessive genes. One such bitter gourd variety, MDU 1, developed
as a result of gamma raditation (seeds treatment) of the landrace MC
103 was found to possess improvement for yield (Rajasasekharan and
Shanmugavelu 1984). Likewise, the white bitter gourd mutant “Pusa
Do Mausami” (white-fruited type) was developed through spontaneous
mutation from the natural population “Pusa Do Mausmi” (green-fruited
type) at the Indian Agriculture Research Institute.
2.5.3 Luffa
Protein variation has been widely used for genetic studies in cucurbits
and seed protein is used mainly for species or variety differentiation as
they do not change in dry mature seed (Ladizinsky and Hymowitz 1979).
Tolentino et al. 1997 employed SDS-PAGE of seed proteins to determine
genetic diversity and taxonomic relationship of Luffa species. The similarity
indices of L. cylindrica and L. acutangula of accessions were 80.0 and 71.1%,
respectively. Domestication of the smooth gourd may have contributed to
its lesser diversity. A similarity index of 68.2% between the two species
suggests that the genome of smooth and ridge gourds do not differ
very much from each other. Contrary to this, LiLin et al. (2007) based
on morphological characters and RAPD analysis suggested that genetic
sequence (Cot value > 10–1–102) of total nucleotides in DNA and 48% unique
sequence (Cot value > 102), whereas L. cylindrica exhibited 30, 20 and 50%
highly repeated, moderately repeated and unique sequences, respectively.
The ratio of repeated to unique sequence was 1.08 for L. acutangula and 1.0
for L. cylindrica (Pasha and Sen 1995).
Hyashi et al. (1999) cloned cDNA encoding cycloartenol synthase
from L. cylindrica and deposited the sequence in the gene bank. Nucleotide
sequence of a cDNA putative oxidosqualence cyclase from L. cylindrica is
also present in the gene bank (Hyashi et al. 2000).
A novel circulating loop bioreactor with cell immobilized in L. cylindrica
sponge has been used for the bioconversion of raw cassava starch to ethanol
(Roble et al. 2002). Loofa sponge has been used as a medium for the culture
of human hepatocyte cell line (Chen et al. 2003). L. cylindrica showed a
very good performance as a solid substrate for the development of biofilm
aggregating microorganisms capable of metabolizing both organic and
inorganic compounds adsorbed in it, particularly those responsible for
nitrification (Tavares et al. 2008).
Cell immobilization technique using biological materials are eco-
friendly and have many advantages over suspended cell systems. Many
studies have shown that the adsorption method for immobilization of
microorganisms has many advantages over the entrapment method using
gel beads (Ogbonna et al. 1994; Fujii et al. 2001). L. cylindrica sponge is an
excellent carrier for immobilization of microorganisms, plants and animal
cells (Liu et al. 1999; Ogbonna et al. 2001; Roble et al. 2002; Chen et al. 2003).
In order to use a loofa sponge as an immobilization carrier in systems
containing/producing cellulose enzymes, a method of protecting loofa
from cellulose by acetylation has been developed (Akhiro et al. 2007). Loofa
sponge-immobilized fungal biosorbent have been used extensively for the
biosorption of heavy metals from olive oil mill wastewater (Ahmadi et al.
2006a) and other wastewater. Loofa sponge is a suitable natural matrix for
immobilization of P. chrysosporium. Immobilized P. chrysosporium has been
successfully used as biosorbing agent for removal of cadmium and lead
(Iqbal and Edyvean 2004, 2005; Ahmadi et al. 2006b).
Recently production of renewable energy from biomass materials
have been emphasized as a means of solving current environmental
problems such as global warming. In this regard, production of ethanol
from lignocellulosic material has great potential. The simultaneous
saccharification and fermentation process seemed to be one of the most
promising options (Ye and Jiayang 2002; Itoh et al. 2003).
2.6 Conclusion
Studies to elaborate taxonomic and phylogenetic relationships among
most wild and cultivated types are still required, and information about
biogeography and ecobiology of wild relatives is rather limited.
In spite of the progress, in many germplasm collections, basic
characterization data of these species are incomplete, limiting the utility
of these collections. Broadening the genetic diversity is essential for
development of commercial cultivars with novel traits in these species.
The germplasm collections have received considerable attention for
evaluation for pathogen, pest, and abiotic stress resistance. A complex of
viruses continues to pose a serious threat to the crop of these species in the
tropics and subtropics, necessitating the identification and pyramiding of
genes for resistance to these viruses in elite cultivars. Extensive germplasm
evaluations have to be conducted for tolerance to high temperatures, salinity,
and drought.
Transfer of economically important traits from wild species to their
cultivated counterparts continues to be challenging for breeders especially in
cases where resistance genes for several pathogens and pests have not been
found within the primary gene pools. Careful biosystematic and gene-pool
studies should continue to help in selection of genotypes for increasing the
probability of successful interspecific hybridization.
Genetic diversity analysis, genetic mapping and map construction is still
in its infancy in all of these species. The development of high-throughput
DNA markers in recent years has created a fertile ground for genetic
mapping and genomics in these crops. Current advancements in sequencing
technologies, functional genomics, and metabolomics will play important
roles in research and development in these crops in the coming decades.
References
Adebooye OC (2009) The properties of seed oil and protein of three underutilized edible
Cucurbitaceae of Southwest Nigeria. Acta Hort 806(1): 347–354.
Adolfo AC, Michael H (2005) Mexican plants with hypoglycaemic effect used in the treatment
of diabetes. J Ethnopharmacol 99: 325–348.
Ahmadi M, Vahabzadeh F, Bonakdarpour B, Mehranian M (2006a) Empirical modeling of
olive oil mill wastewater treatment using Luffa- immobilized Phanerochaete chrysosporium.
Process Biochem 41(2): 1148–1154.
Ahmadi M, Vahabzadeh F, Bonakdarpour B, Mehranian M, Mofarrah E (2006b) Phenolic
removal in olive oil mill wastewater using loofah-immobilized Phanerochaete chrysosporium.
World J Microbiol Biotechnol 22: 119–127.
Akihiro H, Ogbonna JC, Hideki A, Hideo T (2007) Acetylation of loofa (Luffa cylindrica)
sponge as immobilization carrier for bioprocesses involving cellulose. J Biosci Bioeng
103(4): 311–317.
Andres TC (1990) Biosystematics, theories on the origin and breeding potential of Cucurbita
ficifolia. In: DM Bates, RW Robinson, C Jeffrey (eds) Biology and Utilization of the
Cucurbitaceae. Cornell Univ Press, Ithaca, New York, USA, pp 102–119.
Buchbauer G, Boucek B, Nikiforov A (1998) On the aroma of Austrian pumpkin seed oil:
correlation of analytical data with olfactoric characteristics. Ernahrung/Nutrition 22(6):
246–249.
Caili F, Huan S, Quanhong L (2006) A review on pharmacological activities and utilization
technologies of pumpkin. Plant Foods Hum Nutr 61: 73–80.
Chakravarty HL (1990) Cucurbits of India and their role in the development of vegetable crops.
In: DM Bates, RW Robinson, C Jeffrey (eds) Biology and Utilization of Cucurbitaceae.
Cornell Univ Press, Ithaca, New York, USA, pp 325–334.
Chang YM, Liou PC, Hsiao CH, Shii CT (1999) Observation of fruit anatomy and development
of bitter gourd. I. Fruit anatomy and fertilization of bitter gourd. J Agri Res China 48:
23–31.
ChangYun X, ZuChuan Z (1998) Isolation, purification and characterization of a group of novel
small molecular ribosome inactivating protein—LuffinS from seeds of Luffa cylindrica.
Acta Biochim Biophys Sin 30(2): 142–146.
Chaudhari SM, Kale PN (1991) Studies on heterosis in bitter gourd (Momordica charantia L.).
Maharashtra J Hort 5: 45–51.
Chen JG (2005) Effects of sugar-removed pumpkin zymptic powders in preventing and
treating the increase of blood glucose in alloxan-induced diabetic mice. Chin J Clin
Rehabil 9: 94–95.
Chen JP, Yu SC, Hsu BR, Fu SH, Liu HS (2003) Loofa. Sponge as a scafford for the culture of
human hepatocyte cell line. Biotechnol Prog 19: 522–527.
Chen YM (1993) Preliminary study on local germplasm of pumpkins of inner Mongolia (in
Chinese). Crop Genet Resour 2: 13–14.
Chesney P, Wessel-Beaver L, Maynard DN (2004) Both traditional and semi-bush tropical
pumpkin can be intercropped with beans or cowpeas. HortScience 39: 525–528.
Cho MC, Om YH, Huh YC, Mok IG, Park HG (2003) Two oriental squash varieties resistant to
powdery mildew bred through interspecific Crosses. Cucurbit Genet Coop 26: 40–41.
Choer E (1999) Avaliacao morfologica de acessos de Cucurbita spp. Agropecuaria Clima
Temperado 2: 151–158.
Chung SM, Decker Walters DS, Staub JE (2003) Genetic relationships within the Cucurbitaceae
as assessed by consensus chloroplast simple sequence repeats (ccSSR) marker and
sequence analyses. Can J Bot 81: 814–832.
Cogniaux A, Harms H (1924) Cucurbitaecae-Cucurbiteae Cucumerinae. In: A Engler,
pflanzereich 2: 1–246 Engelmann, Leipzig.
Cohen R, Hanan A, Paris HS (2003) Single gene resistance to powdery mildew in zucchini
squash of C. pepo. Euphytica 130: 433–441.
Cutler HC, Whitaker TW (1969) A new species of Cucurbita from Ecuador. Ann MO Bot Gard
55: 392–396.
Daniel AL, Brecht JK, Sims CA, Maynard DN (2005) Sensory analysis of bush and vining types
of tropical pumpkin. Proc Fla State Hort Soc 108: 312–316.
de Wilde WJJO, Duyfjes BEE (2002) Synopsis of Momordica (cucurbitaecae) in SE-Asia and
Malesia. Bot 287: 132–148.
de Oliveira ACB, Maluf WR, Pinto JEBP, Azevedo SM (2003) Resistance to papaya ringspot
virus in summer squash Cucurbita pepo L. introgressed from an interspecific C. pepo ×
C. moschata cross. Euphytica 132: 211–215.
Decker-Walters DS, Walters TW (2000) Squash. In: KF Kiple, KC Ornelas (eds) The Cambridge
World History of Food. Cambridge Univ Press, Cambridge, United Kingdom,
pp 335–351.
Decker-Walters DS, Walters TW, Posluszny U, Kevan PG (1990) Genealogy and gene flow
among annual domesticated species of Cucurbita. Can J Bot 68: 782–789.
Desai UT, Musmade AM (1998) Pumpkins, squashes and gourds. In: DK Salunkhe, SS Kadam
(eds) Handbook of Vegetable Science and Technology: Production, Composition, Storage
and Processing. Marcel Dekker, New York, USA, pp 273–298.
Devadas VS, Ramadas S (1994) Genetic analysis of bitter principals in Momordica charantia
Linn. Cucurbit Genet Coop Rep 17: 129–131.
Dey SS, Behera TK, Munshi AD, Pal Anand (2010) Gynoecious inbred with better combining
ability improves yield and earliness in bitter gourd (Momordica charantia L.). Euphytica
173: 37–47.
Dey SS, Singh AK, Chandel D, Behera TK (2006) Genetic diversity of bitter gourd (Momordica
charantia L.) genotypes revealed by RAPD markers and agronomic traits. Sci Hort 109:
21–28.
Dhillon MK, Singh R, Naresh JS, Sharma NK (2005) Influence of physico-chemical traits of
bitter gourd, Momordica charantia L. on larval density and resistance to melon fruit fly,
Bactrocera cucurbitae (Coquillett). Journal of Appl Entomol 129: 393–399.
Dhiman AK, Sharma KD, Attri S (2009) Functional constituents and processing of pumpkin:
a review. J Food Sci Tech Mysore 46(5): 411–417.
Dutt B, Roy RP (1969) Cytogenetical studies in the interspecific hybrid of Luffa cylindrica L.
and L. graveolens Roxb. Genetica 40: 7–18.
Dutt B, Roy RP (1971) Cytogenetic investigation in Cucurbitaceae. I. Interspecific hybridization
in Luffa. Genetica 42: 139–156.
Dutt B, Roy RP (1976) Cytogenetic studies in an experimental amphidiploid in Luffa. Caryologia
29 (1): 15–22.
El-Mahdy I, Metwally EI, EL-Fadly GA (1991) In vitro differentiation of the immature
interspecific embryo derived from the crosses between Cucurbita pepo and Cucurbila
ficifolia. In: Proc 4th Natl Conf of Pest & Diseases of Vegetables & Fruits in Egypt, vol 2,
29–31 October 1991, Ismailia, Egypt, pp 598–606.
Esteras C, Diez MJ, Picó B, Sifres A, Valcarcel JV, Nuez F (2008) Diversity of Spanish landraces
of Cucumis sativus and Cucurbita ssp. In Cucurbitaceae 2008, Proceedings of the IXth
EUCARPIA meeting on genetics and breeding of Cucurbitaceae, M Pitrat (ed) INRA,
Avignon, France, May 21–24th, 2008, pp 67–76.
FAO (2010) FAOSTAT Agricultural Database. Food and Agriculture Organization of the United
Nations, Rome, Italy: http: //apps.fao.org
Ferriol M, Pico B (2008) Pumpkin and Winter Squash. In: J Prohens, F Nuez (eds) Handbook
of Plant Breeding, vol. 1: Vegetables I Springer, Heidelberg, Germany, pp 317–349.
Ferriol M, Pico B, Nuez F (2003) Genetic diversity of some accessions of Cucurbita maxima from
Spain using RAPD and SBAP markers. Genet Resour Crop Evol 50: 227–238.
Ferriol M, Pico B, de Cordova PF, Nuez F (2004a) Molecular diversity of a germplasm collection
of squash (Cucurbita moschata) determined by SRAP and AFLP markers. Crop Sci 44:
653–664.
Ferriol M, Pico B, Nuez F (2004b) Morphological and molecular diversity of a collection of
Cucurbita maxima land races. J Am Soc Hort Sci 129: 60–69.
Fiky EFK, Karam AMA, Afify EA (1996) Effect of Luffa aegyptiaca (seeds) and Carissa edulis
(leaves) extracts on blood glucose level of normal and streptozotocin diabetic rats. J
Ethnopharmacol 50(1): 43–47.
Fujii N, Oki T, Sakurai A, Suya S, Sakakibara M (2001) Ethanol production from starch by
immobilized Aspergillus awamori ans Sac-charomyces pastorianus using cellulose
carriers. J Microbiol Biotechnol 27: 52–57.
Gaikwad AB, Behera TK, Singh AK, Chandel D, Karihaloo JL, Staub JE (2008). AFLP analysis
provides strategies for improvement of bitter gourd (Momordica charantia L.). HortScience
43: 127–133.
Garcia-Olmedo F, Carbonero P, Hernandez-Lucas C, Paz-Ares J, Ponz F, Vicente O, Sierra
JM (1983) Inhibition of eukaryotic cell-free protein synthesis by thionins from wheat
endosperm. Biochim Biophys Acta 52: 740.
Gong L, Stift G, Kofler R, Pachner M, Lelley T (2008a) Microsatellites for the genus Cucurbita and
an SSR-based genetic linkage map of Cucurbita pepo L. Theor Appl Genet 117: 37–48.
Gong L, Pachner M, Kalai K, Lelley T (2008b) SSR-based genetic linkage map of Cucurbita
moschata and its synteny with Cucurbita pepo. Genome 51: 878–887.
Kurtar ES, Balkaya A (2010) Production of in vitro haploid plants from in situ induced haploid
embryos in winter squash (Cucurbita maxima Duchesne ex Lam.) via irradiated pollen.
Plant Cell Tiss Organ Cult 102: 267–277.
Kurtar ES, Balkaya A, Ozbakir M, Ofluoglu T (2009) Induction of haploid embryo and plant
regeneration via irradiated pollen technique in pumpkin (Cucurbita moschata Duchesne
ex. Poir). Afr J Biotechnol 8(21): 5944–5951.
Kwack SN (1995) Inheritance of determinate growth habit in Cucurbita moschata Poir. J Kor
Soc Hort Sci 36(6): 780–784.
Kwack SN, Fujieda K (1985) Breeding high female lines through interspecific hybridization
of Cucurbita. Cucurbit Genet Coop Rep 8: 78–79.
Kwack SN, Fujieda K (1987) Seed abortion and techniques for obtaining hybrids in interspecific
crosses of Cucurbita. J Jap Soc Hort Sci 55 (4): 455–460.
Ladizinsky G, Hymowitz T (1979) Seed protein electrophoresis in taxonomic and evolutionary
studies, Theor App Genet 54: 145–151.
Lebeda A, Kristkova E (1996) Resistance in Cucurbita pepo and Cucurbita maxima germplasms
to cucumber mosaic virus. Genet Resour Crop Evol 43: 461–469.
Lebeda A, Widrlechner MP (2004) Response of wild and weedy Cucurbita L. to pathotypes
of Pseudoperonospora cubensis (Berk. & Curt.) Rostov. (cucurbit downy mildew). In: PTN
Spencer-Phillips, M Jeger (eds) Advances in Downy Mildew Research, vol 2. Kluwer
Academic Publ, Dordrecht, The Netherlands, pp 203–210.
Lee S, Yoo JG (2006) (WO/2006/019205) method for preparing transformed Luffa cylindrica
Roem (World Intellectual property organi-zation): http: //www.wipo.int/pctdb/en/
wo.jsp
Lee YH, Jeon HJ, Hong KH, Kim BD (1995) Use of random amplified polymorphic DNA for
linkage group analysis in an interspecific cross hybrid F2 generation of Cucurbita. J Kor
Soc Hort Sci 36: 323–330.
LiLin H, ManWai L, ChiHsiung H, TzongShyan L, WenJu Y (2007) Genetic diversity among
the lines and cultivars of Luffa species. J Tai Soc Hort Sci 53(1): 99–110.
Liou TD, Chen KS, Lee SP, Lin JN, Tsao SJ, Yang YW (2002) ‘Fengshan 036’, a white bitter
gourd cultivar. HortScience 37: 1142–1143.
Lira-Saade R (1996) Estudios Taxonómicos y Ecogeográphicos de las Cucurbitaceae
Latinoamericanas de Importancia Económica. Systematic and Ecogeographic Studies
on Crop Genepools. No 9. IPGRI, Rome, Italy, pp 57.
Liu YK, Seki M, Furusaki S (1999) Plant cell immobilization in loofa sponge using two-way
bubble circular system. J Ferment Bioeng 32: 8–14.
Liu ZG, Long MH, Qin RY, Wang XY (2005) Studies on genetic variation, correlation and path
analysis in bitter gourd (Momordica charactinca L.). Guangxi Bot 25: 426–430.
Lopes JF, Brune S, Henz GP (1999) Metodologia de avaliacao da resistencia de germoplasma
de aboboras e morangas a Phytophthora capsici. Hort Brasil 17(Suppl): 30–32.
Loy JB (2004) Morpho-physiological aspects of productivity and quality in squash and
pumpkins (Cucurbita spp.). Crit Rev Plant Sci 23(4): 337–363.
Marr KL, Bhattarai NK, Yong MX (2005) Allozymic, morphological and phonological diversity
in cultivated Luffa acutangula (Cucurbitaceae) from China, Laos and Nepal and allozyme
divergence between L. acutangula and L. aegyptiaca. Econ Bot 59(2): 154–165.
Matsui T, Guth H, Grosch W (1998) A comparative study of potent odorants in peanut, hazelnut,
and pumpkin seed oils on the basis of aroma extract dilution analysis (AEDA) and gas
chromatography olfactometry of headspace samples (GCOH). Lipid Lett 100(2): 51–56.
Matsumoto S, Ito M, Konishi H, Takebe I (1987) Isolation and culture of protoplasts from Luffa
cylindrica suspension cultures. Plant Tiss Cult Lett 4(1): 35–37.
Maynard DN, Elmstrom GW, Talcott ST, Carle RB (2002) “EI Dorado” and “La Estrella”:
compact plant tropical pumpkin hybrids. HortScience 37: 831–833.
Mazali IO, Alves OL (2005) Morphosynthesis: high fidelity inorganic replica of the fibrous
network of loofa sponge (Luffa cylindrica). Ann Acad Bras Ciên 77(1): 25–31.
Porterfield WM Jr (1951) The principal Chinese vegetable foods and food plants of Chinatown
markets. Econ Bot 5: 3–37.
Porterfield WM Jr (1955) Loofah—the sponge gourd. Econ Bot 9: 211–223.
Provvidenti R (1982) Sources of resistance and tolerance to viruses in accessions of Cucurbita
maxima. Cucurbit Genet Coop Rep 5: 46–47.
Provvidenti R (1986) Viral diseases of cucurbits and sources of resistance. ASPAC Food &
Fertilizer Technology Center Tech Bull (Taipei), No 93.
Provvidenti R (1990) Viral diseases and genetic sources of resistance in Cucurbita species. In:
DM Bates, WR Robinson, C Jeffrey (eds) Biology and Utilization of the Cucurbitaceae.
Cornell Univ Press, Ithaca, New York, USA, pp 427–435.
Provvidenti R (1993) Resistance to viral diseases of cucurbits. In: MM Kyle (ed) Resistance to
Viral Diseases of Vegetables: Genetics and Breeding. Timber Press, Portland, OR, USA,
pp 8–43.
Provvidenti R (1997) Resistance to viral diseases of cucurbits conferred by biotechnological
and natural resistance genes (In Chinese). China Veg 4: 55–57.
Provvidenti R, Robinson RW, Munger HM (1978) Resistance in feral species to six viruses
infecting Cucurbita. Plant Dis Rep 62: 326–329.
Puchalski JT, Robinson RW (1978) Comparative electrophoretic analysis of isozymes in
Cucurbita species. Cucurbit Genet Coop Rep 1: 28.
Puchalski JT, Robinson RW (1990) Electrophoretic analysis of isozymes in Cucurbita and
Cucumis and its application for phylogenetic studies. In: DM Bates, ,WR Robinson, C
Jeffrey (eds) Biology and Utilization of the Cucurbitaceae. Cornell Univ Press, Ithaca,
New York, USA, pp 60–76.
Rabbani MG (2007) USDA-Bangladesh collaborative research-CVFB-Annual Research Report.
Dep of Horticulture, Bangladesh Agricultural University, Mymensing, Bangladesh,
p 35.
Rajasekharan KR, Shanmugavelu KG (1984) MDU-1 bitter gourd. S Indian Hort 32: 47.
Rakoczy-Trojanowska M, Malepszy S (1986) Obtaining of hybrids within family Cucurbitaceae
by in vitro culture of immature embryos. I. Characteristics of hybrids from Cucurbita
maxima X Cucurbita pepo. Genet Pol 27: 259–271.
Rakoczy-Trojanowska M, Malepszy S (1989a) A method for increased plant regeneration from
immature F1 and BC1 embryos of Cucurbita maxima Duch. x C. pepo L. hybrids. Plant Cell
Tiss Org Cult 18: 191–194.
Rakoczy-Trojanowska M, Malepszy S (1989b) Obtaining of hybrids within the family
Cucurbitaceae by in vitro culture of immature embryos, II. Characteristics of BC1 and F2
hybrids between Cucurbita maxima x C. pepo. Genet Pol 30 (1–2): 67–73.
Rakoczy-Trojanowska M, Pleder W, Malepszy S (1992) Embryo rescue hybrids between various
Cucurbitaceae. In: RW Doruchowski (ed) Proc Fifth EUCARPIA Cucurbitaceae Symp,
Research Institute for Vegetable Crops, Skierniewice, Poland, pp 91–94.
Ram D, Kumar S, Banerjee MK, Kalloo G (2002a) Occurence, identification and preliminary
characterisation of gynoecism in bitter gourd (Momordica charantia L.). Indian J Agri Sci
72: 348–349.
Ram D, Kumar S, Banerjee MK, Singh B, Singh S (2002b) Developing bitter gourd (Momordica
charantia L.) populations with very high proportion of pistillate flowers. Cucurbit Genetic
Coop Rep 25: 65–66.
Ramakrishnan S, Enghlid JJ, Bryant HL Jr, Xu FJ (1989) Characterization of a translation
inhibitory protein from Luffa aegyptiaca. Biochem Biophys Res Commun 160(2):
509–516.
Raman A, Lau C (1996) Anti-diabetic properties and phytochemistry of Momordica charantia
L. (Cucurbitaceae). Phytomedicine 2: 349–362.
Ramos SRR, de Queiroz MA, Casali VWD, Cruz CD (2000) Divergencia genetica em
germoplasma de abobora procedente de diferentes areas do nordeste. Hort Bras 18:
195–199.
Sisko M, Ivancic A, Bohanec B (2003) Genome size analysis in the genus Cucurbita and its use
for determination of interspecific hybrids obtained using the embryo rescue technique.
Plant Sci 165: 663–669.
Sommerburg O, Keunen JEE, Bird AC, van Kuijk FJGM (1998) Fruits and vegetables that
are major sources for lutein and zeaxanthin: the macular pigment in human eyes. J
Ophthalmol 82: 907–910.
Srivastava VK, Nath P (1972) Inheritance of some qualitative characters in Momordica charantia
L. Indian J Hort 29: 319–321.
Staub JE, Serquen F, Gupta M (1996) Genetic markers, map construction and their application
in plant breeding. HortScience 31: 729–741.
Stefanova L, Neykov S, Todorova T (1994) Genetic diversity in the cucurbit family. Plant Genet
Resour Newslet 99: 3–4.
Subratty AH, Gurib-Fakim A, Mahomoodally F (2005) Bitter melon: an exotic vegetable with
medicinal values. Nutr Food Sci 35: 143–147.
Suribabu B, Reddy NE, Ramarao M (1986) Inheritance of certain quantitative and qualitative
characters in bitter gourd (Momordica charantia L.). S Indian Hort 34: 380–382.
Tavares J, Israel NH, Rui O, Wilton SL, Valderi DL (2008) Nitrification in a submerged
attached growth bioreactor using Luffa cylindrica as solid substrate, Afr J Biotechnol
7(15): 2702–2706.
Tindall HD (1983) Vegetables in the Tropics. Macmillan Press, London, UK, p 553.
Tolentino MIS, Laude RP, Vina AC dela (1997) Genetic diversity analysis of Luffa species based
on seed protein profile using SDS-PAGE. Phil J Crop Sci 22(3): 141–146.
Traka-Mavrona E, Koutsika-Sotiriou M, Pritsa T (2000) Response of squash (Cucurbita spp.)
as a rootstock for melon (Cucumis melo L.). Sci Hort 83: 353–362.
Trivedi RN, Roy RP (1972) Cytological studies in some species of Momordica. Genetica 43:
282–291.
Vahab MA (1989) Homeostatic analysis of components of genetic variance and inheritance
of fruit colour, fruit shape, and bitterness in bitter gourd (Momordica charantia L.). PhD
Thesis, Kerala Agricultural Univ, India, p 96.
Vahab MA, Peter KV (1993) Crossability between Momordica charantia and Momordica dioica.
Cucurbit Genetics Cooperative Report No. 16: http: //cuke.hort.ncsu.edu/cgc/cgc16/
cgc16–32.html
Vinter V, Kristkova A, Lebeda A, Kristkova E (2004) Descriptor lists for genetic resources of
the genus Cucumis and cultivated species of the genus Cucurbita. In: A Lebeda, HS Paris
(eds) Progress in Cucurbit Genetics and Breeding Research, Proc Cucurbitaceae 2004, the
8th EUCARPIA Meeting on Cucurbit Genetics and Breeding. Olomouc, Czech Republic:
Palacky University in Olomouc, Czech Republic, pp 95–99.
Wall JR (1954) Interspecific hybrids of Cucurbita obtained by embryo culture. Proc Am Soc
Hort Sci 63: 427–430.
Wang H, Ng TB (2002) Luffangulin, a novel ribosome inactivating peptide from ridge gourd
(Luffa acutangula) seeds. Life sci 70(8): 899–906.
Weeden F (1984) Isozyme studies indicate that the genus Cucurbita is an ancient tetraploid.
Cucurbit Genet Coop Rep 7: 84–85.
Weeden NF, Robinson RW (1990) Isozyme studies in Cucurbita. In: DM Bates, RW Robinson,
C Jeffrey (eds) Biology and Utilization of the Cucurbitaceae. Cornell Univ Press, Ithaca,
New York, USA, pp 51–59.
Welihinda J, Karunanayake EH, Sheriff MH, Jayasinghe KS (1986) Effect of Momordica charantia
on the glucose tolerance in maturity onset diabetes. J. Ethnopharmacol 17: 277–282.
Whitaker TW (1954) A cross between an annual species and a perennial species of Cucurbita.
Madrono 12(7): 213–217.
Whitaker TW (1959) An interspecific cross in Cucurbita (C. lundelliana Bailey × C. moschata
Duch.). Madrono 15(1): 4–13.
Whitaker TW, Davis GN (1962) Cucurbits: Botany, Cultivation, and Utilization. World Crops
Books, London, New York, p 249.
Whitaker TW, Bemis WP (1976) Cucurbits. In: NW Simmonds (ed) Evolution of Crop Plants.
Longman, London, UK, pp 64–69.
Whitaker TW, Robinson RW (1986) Squash breeding. In: MJ Bassett (ed) Breeding Vegetable
Crops. AVI Publ, Westport, CT, USA, pp 209–242.
Wilson EH (1913) A naturalist in Western China, vol 1–2. Cadogan Books, London, UK
Wilson HD (1989) Discordant patterns of allozyme and morphological variation in Mexican
Cucurbita. Syst Bot 14: 612–623.
Wilson HD (1990) Gene flow in squash species. BioScience 40: 449–455.
Wilson HD, Doebley J, Duvall M (1992) Chloroplast DNA diversity among wild and cultivated
members of Cucurbita (Cucurbitaceae). Theor Appl Genet 84: 859–865.
Xiang CP, Wu CY, Wang LP (2000) Analysis and utilization of nutrient composition in bitter
gourd (Momordica charantia). J Huazhong Agricultural Univ 19: 388–390.
Xiong XM (2000) Study on extraction and separation of effective composition of pumpkin
polysaccharide and its glucatonic effect. Chin Tradit Patent Med 22(8): 563–565.
Yadav M, Chaudhary R, Yadav HS (2003) Screening of varieties of bitter gourd against fruit
fly (Dacus cucurbitae). JNKVV Res J 37: 100–101.
Yang SL, Walters TW (1992) Ethnobotany and the economic role of the Cucurbitaceae of China.
Econ Bot 46: 349–367.
Yawalkar KS (1985) Vegetable Crops of India. 3rd edn. Agric. Horticultural Publishing House,
Nagpur, Maharashtra, India, pp 166–170.
Ye S, Jiayang C (2002) Hydrolysis of lignocellulosic materials for ethanol production: a review.
Bioresour Technol 83: 1–11.
Yeung HW, Li WW, Ng TB (1991) Isolation of a ribosome-inactivating and abortifacient protein
from seeds of Luffa acutangula. Int J Pept Prot Res 38 (1): 15–19.
Yoshinari O, Sato H, Igarashi K (2009) Anti-diabetic effects of pumpkin and its components,
trigonelline and nicotinic acid, on Goto-Kakizaki rats. Biosci Biotechnol Biochem 73(5):
1033–1041.
Youn SJ, Chung HD (1998) Genetic relationship among the local varieties of the Korean native
squashes (Cucurbita moschata) using RAPD technique. (In Korean with English abstract).
J Kor Soc Hort Sci 39: 517–521.
Yuwai KE, Rao KS, Kaluwin JC, Jones GP, Rivetts DE (1991) Chemical Composition of
Momordica charantia L. Fruits J. Agric. Food Chem. 39: 1782–1763.
Zalapa JE, Staub JE, Chung SM, Cuevas H, McCreight JD (2007) Mapping and QTL analysis
of plant architecture and fruit yield in melon. Theor Appl Genet 114: 1185–1201.
Zhang C, Luo S, Guo J, Zheng X, Luo H, Xiao J (2006) Study on the genetic effects of fruit
length of bitter gourd. Guangdong Agri Sci 1: 34–35.
Zhang H (2003) Determination of γ -amino-butyric acid and amino acids in pumpkin. Food
Res Dev 24(3): 108–109.
Zhang ZJ (1998) Effects of superfine pumpkin powder on alloxaninduced Diabetes mellitus
rabbits. J Chin Cereals Oils Assoc 13(3): 52–56.
Zhang XP, Bai XM (2004) Effect of compound pumpkin powder on diabetic mice. Chin J Mod
Appl Pharmacol 21(4): 278–280.
Zhang YJ, Yao HY (2002a) Revealing the effective ingredient in pumpkin for reducing blood
sugar. J Chin Cereals Oils Assoc 17(4): 59–62.
Zhang YJ, Yao HY (2002b) Composition analysis of pumpkin polysaccharide and its glucatonic
effect. J Wuxi Univ Light Ind 21(2): 173–175.
Zhou SK, Qiu ZH, Li GX, Li Z, Li XR (1995) Study on the germplasm resources of Cucurbita
for seed and its utilization (in Chinese). Crop Genet Resour 2: 13–15.
Zhou WB, Lou S, Luo JN (1998) An early maturing and high yielding bitter gourd hybrid
Cuilli No.1. Plant Breed Abstr 68: 1002.
ABSTRACT
Much advancement has been made in traditional breeding and classical
genetics of cucurbit crops, although most significant advancement have
been related to qualitative traits. Significant improvement in many
quantitative traits have been much harder to achieve, and typically
result from several incremental advances that occur over a long period
of time. Molecular techniques have the potential to overcome many
of the obstacles presented by traditional breeding techniques, but it is
imperative that the development and utilization of these new molecular
technologies work in conjunction with traditional breeders who have
the skill set necessary to evaluate the germplasm resulting from these
new technologies.
3.1 Introduction
Cucurbit crops simultaneously bestow upon the breeder several advantages
and disadvantages. As pointed out by Whitaker and Bohn (1950), cucurbit
crops are easily grown, indeterminant plant types, which typically offer
plenty of large flowers to work with over a fairly long period of time.
Probably the greatest disadvantage is that cucurbit crops tend to be large
plants that require abundant field space for proper examination of most
agronomically important traits. Adding to the disadvantages, cucurbit
1
Vegetable and Fruit Improvement Center, Department of Horticultural Sciences, Texas A &
M University, College Station, TX 77843-2119, USA; e-mail: srking@tamu.edu
2
Wes Watkins Agricultural Research Laboratory, USDA-ARS, PO Box 159, Hwy.3 West Lane,
OK 74555, USA; e-mail: angela.davis@lane-ag.org
3
Department of Horticultural Science, Box 7609, North Carolina State University, Raleigh, NC
27695-7609, USA; e-mail: todd_wehner@ncsu.edu
*Corresponding author
3.2.1.1 Watermelon
Watermelon is a useful crop species for genetic research because of its small
genome size, and the many available gene mutants. The genome size of
watermelon is 424 million base pairs (Arumuganathan and Earle 1991).
Like some of the other cultivated cucurbits, watermelon has much genetic
variability in seed and fruit traits. Genetic investigations have been made
for some of those, including seed color, seed size, fruit shape, rind color,
rind pattern, and flesh color.
Many watermelon fruit quality genes have been identified. Fruit shape
is controlled by an incompletely dominant gene, resulting in elongate (OO),
oval (Oo), or spherical (oo) fruit (Weetman 1937; Poole and Grimball 1945).
A recessive gene (f) controls furrowed fruit surface (Poole 1944). Explosive
rind (e) causes the fruit rind to burst or split when cut (Porter 1937); these
fruit are easily crushed and are sometimes used as pollenizer cultivars
not intended for harvest. Tough rind (E) improves shipping ability. The
single recessive gene su (Chambliss et al. 1968) eliminates bitterness in C.
lanatus, and is allelic to the dominant gene (Su) for bitter flavor in Citrullus
colocynthis.
Watermelon flesh color is controlled by several genes to produce scarlet
red, coral red, orange, salmon yellow, canary yellow, pale yellow, green,
or white. Genes conditioning flesh colors are B (Shimotsuma 1963), C
(Poole 1944), Wf (Shimotsuma 1963), y (Porter 1937), y-o (Henderson 1989;
Henderson et al. 1998), and py (Bang et al. 2010). There is some confusion
in the literature regarding flesh color inheritance, possibly due to the
potential presence of two different “white flesh” phenotypes (Bang et al.
2010). Wf is reported to control white flesh in watermelon and is reported
to be epistatic to B, where genotypes Wf--- are white, wfwfB- are yellow
and wfwfbb are red (Shimotsuma 1963). Henderson et al. (1998) reported
two genes separate red flesh and canary yellow flesh, C and i-C, where a
dominant C gives canary yellow flesh except in the presence of a dominant
i-C, which would give red flesh. Bang et al. (2007) demonstrated a single
gene distinguishing red and canary yellow flesh. They also showed a pale
yellow phenotype can result between crosses of canary yellow and red
(Bang et al. 2010). The recessive py gene seems to require the presence of
a dominant C gene. This py mutant may have been confused with white
flesh, which is caused by a dominant Wf. Interactions of Wf, B, C, and now
py need further study for clarification.
The coral red gene (Y) is dominant to salmon yellow (y), and orange
flesh (y-o) is a member of a multiple allelic system at that locus, where Y
(Coral red flesh) is dominant to both y-o (orange flesh) and y (salmon yellow),
and y-o (orange flesh) is dominant to y (salmon yellow). It is reported that a
dominant Scr produces scarlet red flesh instead of the recessive coral red
flesh (summarized in Wehner 2007). Scr is now believed to be part of the
multiple allelic system at the Y locus where Scr is another allele at the Y
locus (T Wehner, unpubl. data). More study is needed that includes classical
genetics combined with biochemical and molecular data to fully understand
the genes and inheritance of flesh color in this crop. This understanding is
critical since flesh color is indicative of carotenoid content, which impacts
the nutritional benefits of watermelon.
Several genes have been identified that affect the rind of watermelon.
The gene Sp produces spotted fruit (Poole 1944). Golden yellow (go) is a
single recessive gene that causes fruit to turn yellow at maturity (Barham
1956). Intermittent stripes (ins) produces narrow dark stripes at the peduncle
end of the fruit that become irregular in the middle and nearly absent at
the blossom end of the fruit (Gusmini and Wehner 2006). Yellow belly, or
ground spot, on “Black Diamond Yellow Belly” is controlled by a single
dominant gene (Yb). Weetman (1937) proposed that three alleles at a single
locus determined rind pattern. The allelic series was renamed to G, gs,
and g by Poole (1944). The g-s gene produces a striped rind, but the stripe
width (narrow, medium, and wide stripe patterns) has not been explained
as yet. Porter (1937) found that dark green was completely dominant to
light green (yellowish white, in his description). The watermelon gene p
produces pencilled rind pattern (Robinson et al. 1976) and the m gene for
mottled rind was first described by Weetman (1937).
3.2.1.2 Cucumber
Sex expression in cucumbers has played an important role in seed
production as well as development of new fruit types. This trait is affected
by several single-gene mutants. The F locus governs gynoecy, but is
modified by other genes and the environment, and interacts with a and m
(androecious and andromonoecious, respectively) (Rosa 1928; Tkachenko 1935;
Galun 1961; Shifriss 1961; Wall 1967; Kubicki 1969a). The F gene may also
be modified by an intensifier gene (In-F) which increases the femaleness
(Kubicki 1969a). Other genes that affect sex expression are gy for gynoecious,
m-2 for andromonoecious (Kubicki 1974) and Tr for trimonoecious expression
(Kubicki 1969b).
The discovery of parthenocarpy in cucumbers (Wellington and
Hawthorn 1928) has led to the development of seedless fruit when
combined with the gynoecious trait. There is dispute over the inheritance
of parthenocarpy. Pike and Peterson (1969) suggested an incompletely
dominant gene, Pc, affected by numerous modifiers. In contrast, de Ponti
and Garretsen (1976) explained the inheritance by three major isomeric
genes with additive action.
Bitterness in cucumbers can affect fruit quality, health potential, and
insect resistance. Eliminating bitterness in this crop can be accomplished
by selecting for the bitterfree allele (bi), which inhibits biosynthesis of
cucurbitacin (Andeweg and De Bruyn 1959). Cucurbitacins can be toxic
at high levels and they may act as an attractant for cucumber beetles, but
have been shown to repel spider mites and aphids.
Disease resistance is an important trait in cucumber as diseases can
reduce yield and quality. Currently there are 15 genes known to control
disease resistance in C. sativus. Three of these genes condition virus
resistance. Wasuwat and Walker (1961) found a single dominant gene
3.2.1.3 Melon
Most melon cultivars are andromonoecious, but other expression patterns
are possible. Genes a (andromonoecious) and g (gynomonoecious) interact to
influence sex expression: a_g_ = monoecious; a_ gg = gynomonoecious; aa g_
= andromonoecious; and aa gg = hermaphrodite (Pitrat 2006). A third gene
was identified that creates a much more stable gynomonoecious phenotype
(gy), so that a+_ gg gygy = stable gynomonoecious.
Sterility is common in melon, as there are five different male-sterile
genes and two total plant sterility genes (Pitrat 2006). However, since there
are no seedling markers for these recessive male sterile genes, it is impossible
to tell which plants are sterile without growing the plants to flowering. If
the purpose is to use the male-sterile trait for hybrid seed production, by
the time you identify which plants are sterile, the fertile plants will have
contaminated your hybrid seed.
Fruit quality in melon is polygenic, but there are several genes that
have major effects. Some negative alleles for fruit quality include Bif for
Bitter fruit (Parthasarathy and Sambandam 1981), Me for Mealy flesh texture
(Ganesan 1988) and So for Sour taste (Kubicki 1962). Flesh color is dictated
by single genes but the intensity of color is quantitative. Fruit shape is also
influenced by genes for oval fruit shape (O), sutures (s) and spherical fruit shape
(sp), but sex expression may also have an influence as perfect flowers tend
to give round fruit while female flowers tend to give more elongated fruit
(Lumsden 1914; Bains and Kang 1963; Wall 1967).
There are many loci in melon for disease resistance. Fusarium wilt
resistance is provided by Fom-1 and Fom-3, which are alleles for independent
genes that provide resistance to races 0 and 2, and Fom-2 gives resistance to
races 0 and 1 (Risser 1973; Zink and Gubler 1985). Resistance to Alternaria
is provided by Ac (Thomas et al. 1990), and there are up to six genes that
provide a high to moderate level of resistance to gummy stem blight (Prasad
and Norton 1967; Zuniga et al. 1999; Frantz and Jahn 2004). There are up to
17 different genes that govern resistance to powdery mildew, depending
on the race/species involved (Jagger et al. 1938; Bohn and Whitaker 1964;
Harwood and Markarian 1968a, b; Kenigsbuch and Cohen 1989; Epinat
et al. 1993; Anagnostou et al. 2000; McCreight 2003). There are four genes
reported for downy mildew resistance (Cohen et al. 1985; Thomas et al.
1988; Epinat and Pitrat 1989; Kenigsbunch and Cohen 1992), and a fifth
gene is reported to act in combination with at least one other modifier gene
(Angelov and Krasteva 2000). There are also resistance genes for papaya
ringspot virus, zucchini yellow mosaic virus, and necrotic spot virus (See
Pitrat 2006 for gene names.).
Melon also has resistance genes for several insects. Gene Af provides
resistance to red pumpkin beetle (Vashistha and Choudhury 1974). Tolerance
to melon aphid is provided by Ag (Bohn et al. 1973), and Vat provides
resistance to viruses transmitted by aphids (Pitrat and Lecoq 1980). Melon
fruit fly resistance is provided by two genes, dc-1 and dc-2 (Sambandam
and Chelliah 1972). As with cucumber and squash, cucurbitacin content
also influences insect resistance/susceptibility, which in the case of melon
is governed by two genes, Bi and cb (Lee and Janick 1978; Nugent et al.
1984).
yet been transferred to cultivated squash. There are four male sterile genes
reported, two in each species, for C. pepo and C. maxima (Scott and Riner
1946; Eisa and Munger 1968; Superak 1987; Korzeniewska 1992), but as with
most of the other cucurbit crops there are no morphological markers, which
would allow them to be useful for seed production. There are two total plant
sterility genes, one in C. maxima and one in C. pepo (Carle 1997).
The reduced internode length that gives a bush habit in C. pepo and
C. maxima is governed by a single gene (Bu) (Shifriss 1947; Grebenšcikov
1958; Decker-Walters and Munger 1963; Wu et al. 2007). This trait is greatly
appreciated by gardeners with limited space. A unique gene found in the
Cucurbita spp. is the naked seeded trait where seed lack a lignified seed
coat and is typically used for the roasted pumpkin seed market (Schöniger
1952; Grebenšcikov 1954; Xianglin 1987; Zraidi et al. 2003, 2007).
There are far fewer identified disease resistance genes in squash than the
other major cultivated cucurbit crops. Resistance to powdery mildew (PM)
in C. okeechobeensis and C. lundelliana is controlled by a single dominant allele
and modifiers (Contin 1978; Paris and Cohen 2000; Cohen et al. 2003) and
two PM-resistant genes to race 1 and race 2 were identified in C. moschata
(Adeniji and Coyne 1983). A single gene has also been reported to provide
resistance to zucchini yellow mosaic virus. There are three complementary
dominant genes for resistance to Phytophthora capsici (Crown rot) (Padley et
al. 2009). Known virus resistance consists of one recessive gene for cucumber
mosaic virus (Brown et al. 2003), one recessive resistance gene to papaya
ringspot virus (Brown et al. 2003), two watermelon mosaic virus resistance
genes, one from C. moschata (Fulks et al. 1979; Brown et al. 2003), which may
be linked with or identical to Zym-1 and one from C. ecuadorensis (Shifriss
1989), and a total of six resistance genes and one modifier gene have been
reported for zucchini yellow mosaic virus in C. moschata, C. pepo, and/or C.
ecuadorensis (Mains 1950; Fulks et al. 1979; Paris et al. 1988; Robinson et al.
1988; Paris and Cohen 2000; Brown et al. 2003; Pachner and Lelley 2004).
There is one resistance gene reported for insects (Fr, for melon fruit fly
resistance) (Nath et al. 1976), although the gene cu (Sharma and Hall 1971),
which reduces foliar cucurbitacin content, will have a similar effect as for
the other cucurbit crops by reducing cucumber beetle preference while
making the plant more attractive to aphids and spider mites.
Fruit quality, shape and color are extremely diverse in the squash and
pumpkin species and a thorough review of the genes involved was compiled
by Paris and Kabelka (2009). A number of fruit color genes have been
identified. The B gene that was found in an ornamental gourd can be used
to give a yellow color and high vitamin A content. This same gene also has
pleiotropic effects on fruit and foliage and is affected by several modifier
genes (Ep-1, Ep-2 and Ses-B). The B gene is also complementary to L-2 to give
intense orange flesh instead of light yellow flesh color which also enhances
the carotenoid content. There are up to five different fruit bitterness genes,
but allelism tests have not been reported for all five genes.
when trying to select for quantitative traits that are heavily influenced
by the environment, such as yield. Because of environmental influences
on quantitative traits, large populations are needed to account for this
variability, which adds to time and space requirements, and cost for proper
evaluation. Breeders and geneticists have attempted to identify markers that
are associated with agronomically important traits, with the hope that the
marker can be: 1) identified at the seedling stage or shortly thereafter, and
2) have a small or no environmental influence. Traditional approaches to
identify traits early include morphological and isozyme markers, both of
which have been used to some degree in various cucurbit crops. It is likely
that both these markers have been used to maintain cultivar identities, but
there is little published information with regard to the extent that these
markers are used. Most major cucurbit seed companies have routinely used
isozyme markers to check hybrid purity (S. King, pers. comm.).
3.2.2.1 Watermelon
In watermelon 60 genes and 111 isozyme markers have been identified
(Wehner 2007), but there is only one linkage map. It includes two genes
(fruit bitterness and red flesh) and 22 isozymes that comprise seven linkage
groups covering 354 cM (Navot et al. 1990). While it is valuable to have
important fruit traits such as flesh bitterness and red flesh color linked to
markers, the utility has been limited since most breeding work is conducted
within germplasm that already lacks the bitter trait and is often conducted
within red fleshed cultivars.
Much work was performed to identify morphological markers
associated with genetic male-sterility in watermelon. This trait would
be extremely useful for the production of hybrid cultivars. It is essential,
during hybrid seed production, to identify which progeny contain the
male-sterile trait, since segregating populations are the only means to
maintain the genetic male-sterility trait. There are currently five genes for
genic male-sterility reported in this crop. One gene (gms) is associated with
glabrous foliage (Watts 1962, 1967; Ray and Sherman 1988), which makes
for an excellent morphological marker since it is simple to identify the trait
in young plants. A second male-sterility gene (ms-dw) is associated with
reduced internode length, another easy morphological marker (Huang et
al. 1998). However, these genes, as well as two other male-sterility genes,
are also associated with reduced female-fertility, limiting their suitability
for hybrid seed production. The fifth genetic male-sterility gene (ms-2) is
not associated with a reduction of female fertility (Dyutin and Sokolov
1990), but there are currently no morphological markers associated with
this gene, limiting its utility for hybrid seed production.
3.2.2.2 Cucumber
Since cucumber has just seven chromosome pairs and over 100 known
genes, it would seem that linkage maps would be fairly complete by now.
Unfortunately, we know of a few references reporting linkages of more than
two gene loci. Some of the difficulties linking genes in cucumber are due
to a portion of the nomenclature being unclear about possible duplication
of gene names originating from studies using different parental lines.
Additionally, some of the linkage relationships analyzed in previous studies
did not involve specific single genes but involved multigenic traits, or if a
single gene was involved, it was not specifically identified. Even with these
limitations, Wehner (2005) was able to summarize the literature for linkages
and describe the different linkage groups. This summary contained six
linkage groups and an assortment of linked genes that could not be placed
in any of the linkage groups. His work on cucumber linkages that include
traits is included below with modifications. The order in which the genes
were expressed in each group does not necessarily represent the order in
which they may be found on the chromosome and a question mark “?” will
follow each gene which has a questionable origin.
Lane and Munger (1985) and Munger and Lane (1987) determined that
a gene for resistance to powdery mildew (pm?) was also linked with Cca
for susceptibility to target leaf spot but that linkage, though fairly tight,
was breakable.
The last four genes in this group are Tu, D, te and u (Strong 1931). Until
recently it was believed that each in the recessive form was pleiotropic and
consistent with European type cucumbers and each in the dominant form
was pleiotropic and consistent with American type cucumbers.
Fanourakis (1984) and Fanourakis and Simon (1987) reported that
crossing-over (R = 23.7) occurred between te and the other three genes,
which still appeared to be associated. However, using triple backcrosses
they demonstrated that there is a definite order for Tu, D and u within
their chromosome segment and that the Tu end is associated with the ns
and ss end.
3.2.2.3 Melon
There are approximately 186 identified genes and isozyme markers in
melon, making it the most saturated cucurbit crop in terms of identified
genes (Pitrat 2006). Linkages have been identified between several
agronomic and morphological traits. The linkages have been assigned to
eight linkage groups (Pitrat 1991).
and Wilson 1987). There are also reports of genetic linkage between genes,
where dark stem (D) was found to be linked to mature orange fruit (mo-2)
in C. pepo (Paris 1997), mottled leaves (M) were linked to warty fruit (Wt) in
C. pepo (Paris et al. 2004), resistance to watermelon mosaic virus 2 (WMV-2)
with a plastid-specific aldolase (Aldo-p) in Cucurbita ecuadorensis, and bitter
fruit (Bi) was found to be linked to lobed leaves (Lo-2) in a C. ecuadorensis x C.
maxima interspecific cross (Herrington and Brown 1988; Paris et al. 2004).
While it is obvious how some of these linkages could be valuable in a
breeding program (e.g., select against lobed leaves to eliminate bitter fruit),
in practicality, three of these linkages have limited usage because there are
multiple genes for the trait as well as other modifier effects which affect
the phenotype which are not accounted for in the linkage.
3.3.1.1 Watermelon
Watermelon is unique among the cucurbit crops in that a significant portion
of current cultivars are seedless triploids, especially in the American market.
Triploid hybrids are produced by crossing tetraploid female with diploid
male inbred lines. Improving seedless watermelon involves selecting the
best diploid and tetraploid line, then testing them in hybrid combinations.
This method creates a new level of complexity for breeders since both
diploid and tetraploid lines must be managed. Initial breeding efforts on
tetraploids simply involved selecting the best diploids and using these to
create tetraploid parent lines. While this method has provided many current
triploid cultivars in the market, it has limitations since most diploids do not
make good tetraploid parents for triploid seed production. Fertility in the
tetraploid is an extremely important trait that has limited the use of many
tetraploid parents. Additionally, breeding within tetraploids is often more
complex than breeding diploids.
Watermelon cultivars are often monoecious, with older cultivars and
many wild accessions being andromonoecious. Watermelon is naturally
cross-pollinated like maize. However, there is little inbreeding depression
and heterosis in watermelon. It has been suggested that the lack of inbreeding
depression is due to the small population sizes used by farmers during the
domestication of the species, which forced out deleterious recessive alleles.
Therefore, even with monoecious sex expression and insect-pollinated
flowers, there would have been considerable inbreeding among the few
plants representing the population. Since there is little inbreeding depression
in watermelon, inbred lines are developed using self-pollination with little
loss of vigor from the parental population.
In studies of heterosis in watermelon, some estimates have shown a 10%
advantage of the hybrid over the high parent, but only for some parental
combinations (Wehner 1999). The small amount of heterosis observed in
watermelon makes it possible for growers to compete in the seeded market
using less expensive open-pollinated lines. However, hybrid varieties are
useful for combining multiple dominant traits from different parents.
Examples of such traits include red or canary yellow flesh, resistance to
Fusarium wilt and anthracnose, and resistance to powdery mildew. Hybrids
also protect proprietary breeding lines from unauthorized use. One of the
most important uses of hybrids is the production of seedless varieties.
Watermelon breeders today are less interested in studying heterosis or
measuring general (GCA) and specific (SCA) combining ability, because
hybrids have an advantage for protection of the proprietary breeding lines.
Furthermore, seedless cultivars are in high demand and can be produced only
as triploid hybrids. However, in the future it might be possible to develop
transgenic diploid seedless watermelons. In that case, the advantage of using
heterotic hybrids vs. inbred cultivars will again be questioned.
Environmental factors such as water availability may be important
in contrasting inbred cultivar vs. hybrid yields. A Florida study observed
that watermelon hybrids out-yielded inbred cultivars only in irrigated
fields, but quality was higher among the inbred cultivars in dry conditions
(Rhodes 1985).
Disease resistance has been, and continues to be a high priority of most
watermelon breeding programs. Resistance to Fusarium wilt has been
studied since the early 1900s (Orton 1911), and most modern cultivars have
resistance to most Fusarium races today (Henderson et al. 1970), although
the fungus continues to evolve in response to host plant resistance (Zhou
et al. 2010), necessitating continued breeding efforts. Resistance genes are
also used to provide protection from anthracnose (Layton 1937; Winstead
et al. 1959), papaya ringspot virus (Guner et al. 2008), and zucchini yellow
mosaic virus (Provvidenti 1991; Xu et al. 2004). Gummy stem blight
remains a high priority for watermelon research (King and Davis 2007),
but despite potential resistance in germplasm, protection does not hold
up in all locations.
Quality traits have been selected in watermelon for thousands of years.
These traits include size, shape, shelf-life, color, sugar content, and total
nutrient content. Watermelon is a dynamic plant with great potential for
alteration of quality traits. However, what is perceived as quality depends on
the country, demographics, and personal preference. Some people prefer the
mini melons, around 5 pounds, whereas others want giant watermelons over
100 pounds. There are unsweet, firm, white watermelons used for pickling
and preserves, and dark crimson watermelon with brix up to 14, even 15%.
More recently, breeders have been interested in phytonutrient content,
and breeding programs have focused on increasing total carotenoids,
lycopene, and improving citrulline contents (personal communication with
watermelon breeders).
3.3.1.2 Cucumber
Early studies on cucumber report considerable heterosis and/or inbreeding
depression within this crop, so long as the parents are not closely related
(Hayes and Jones 1916; Hutchins 1938; Ghaderi and Lower 1979a, b).
However, Rubino and Wehner (1986) indicated that inbreeding depression
was not important in cucumber and that midparent heterosis was noted
for most traits in many hybrids obtained from crossing S6 lines with a
gynoecious inbred line. In a similar but larger study, Cramer and Wehner
(1999) demonstrated in one out of four crosses, heterosis for fruit yield was
associated with a decreased correlation between percentage of fruit set
and fruit weight, an increased negative correlation between percentage of
fruit set and both the number of branches per plant and the percentage of
pistillate nodes, and an increased negative correlation between the number
of nodes per branch and total fruit weight. Inbreeding depression was
associated with a weakening of the strong negative correlations between
percentage of fruit set and the number of branches per plant, and between
the number of nodes per branch and total fruit weight. Those correlations
were associated with high-parent heterosis and inbreeding depression
only for the one cross, and would not necessarily apply to future crosses
in which heterosis may be observed for yield. More recently, Munshi et al.
(2006) suggest from their results that heterosis breeding is important for
effective utilization of non-additive gene actions; and Godoy et al. (2008)
showed both positive heterosis of hybrids over parent lines.
Cucumber breeders have combined several of the sex expression and
fruit quality genes to create improved varieties. It was discovered that
gynoecious sex expression created an earlier maturity, since there are no
male flowers. Many pickling cucumber hybrids were created that combined
gynoecious varieties with a monoecious variety to create a blended hybrid
composed of two distinct varieties. The monoecious variety provides pollen
to create fruit set in the gynoecious variety.
Gynoecious sex expression has also been combined with parthenocarpy
as a way to set fruit without pollen. It has been discovered that parthenocarpic
fruit has an improved shelf life and quality, and when combined with the
bitterfree allele the resultant fruit are of high quality. The combination of
gynoecious sex expression along with parthenocarpy is responsible for the
development of the extensive greenhouse cucumber industry, since this
eliminates the need for pollination and produces a superior quality fruit.
Gynoecious sex expression can also be used for hybrid seed production since
the female plants do not produce male flowers. Since it has been found that
sex expression can easily be changed with growth regulators, maintaining
the female line is relatively straight forward.
Cucumber breeders have been able to create varieties with multiple
disease resistances. Resistance to angular leaf spot, Anthracnose, downy
mildew, powdery mildew scab and target leaf spot are common in the
market. There are also varieties with multiple virus resistance available,
including cucumber mosaic virus, papaya ringspot virus, watermelon
mosaic virus and zucchini yellows mosaic virus. These disease resistant
varieties were created by screening thousands of segregating plants in
multiple generations using tedious inoculations.
3.3.1.3 Melon
Early reports differed in their analysis of whether melon showed heterosis
in yield and quality traits. The cumulative data suggests that well chosen
parental lines can produce hybrids with improved quality and yield (See
Robinson 1999 for a complete review of this subject in melons). More
recent findings demonstrated that in snake melon (C. melo L. var. flexuosus)
heterosis increased ascorbic acid and carotenoid content (Pandey et al.
2010), and heterosis could alter fruit shape in C. melo (Fernández-Silva et
al. 2009). An elegant study by Luan et al. (2010) demonstrated dramatic
performance differences between parents from diverse geographic origins
and among F1 hybrid progeny, a strong relationship between genetic
distance (determined using molecular techniques) and heterotic effects was
not consistently detected.
Breeders have created a diversity of types of melon, with well over a
dozen distinct forms currently on the market in various regions around
the world. The different types included a variety of shape, skin and flesh
color and texture. Breeders have also successfully combined a number of
quantitative traits such as sweetness and level of aromatic compounds,
despite the lack of efficient markers for these quantitative traits.
The discovery of gynoecious melons created a lot of excitement in the
seed industry, since the ability to produce gynoecious inbreds should be an
advantage for seed production in the same way it is for cucumber; however,
our experience has been that the gynoecious trait is influenced by genetic
background so that occasional perfect flowers may sometimes develop.
When a strong gynomonoecious genotype is identified, converting its sex
expression with hormones is much more difficult than with cucumber
(S King, unpubl. data).
Breeders have also created multiple disease resistant varieties of melon.
Resistance to Fusarium wilt and powdery mildew are common, but the
pathogens for these diseases continue to evolve making continued breeding
efforts necessary.
3.4 Conclusion
Traditional genetics and plant breeding have made great strides in our
understanding and improvement of cucurbit crops. We have used classical
References
Abul-Hayja Z, Williams PH, Peterson CE (1978) Inheritance of resistance to anthracnose and
target leaf spot in cucumbers. Plant Dis Rep 62: 43–45.
Adeniji AA, Coyne DP (1983) Genetics and nature of resistance to powdery mildew in crosses
of butternut with calabaza squash and ‘Seminole Pumpkin’. J Am Soc Hort Sci 108:
360–368.
Ahmed EA, Ibn Oaf HS, El Jack AE (2003) Combining ability and heterosis in Line x tester
crosses of summer squash (Cucurbita pepo L.). Cucurbit Genet Coop Rpt 26: 54–56.
Akhilesh T, Ram HH (2006) Qualitative inheritance of segmented leaf shape in bottle gourd
(Lagenaria siceraria (Molina) Standl.). Veg Sci 33: 117–121.
Anagnostou K, Jahn M, Perl-Treves R (2000) Inheritance and linkage analysis of resistance
to zucchini yellow mosaic virus, watermelon mosaic virus, papaya ringspot virus and
powdery mildew in melon. Euphytica 116: 265–270.
den Nijs APM, Boukema IW (1983) Results of Linkage Studies and the Need for a Cooperative
Effort to Map the Cucumber Genome. Cucurbit Genet Coop Rep 6: 22–23.
den Nijs APM, Boukema IW (1985) Short petiole, A useful seedling marker for genetic studies
in cucumber. Cucurbit Genet Coop Rpt 8: 7–8.
de Ponti OM, Garretsen F (1976) Inheritance of parthenocarpy in pickling cucumbers (Cucumis
sativus L.) and linkage with other characters. Euphytica 25: 633– 642.
Dessert JM, Baker LR, Fobes JF (1982) Inheritance of reaction to Pseudomonas lachrymans in
pickling cucumber. Euphytica 31: 847–856.
Dossey BF, Bemis WP, Scheerens JC (1981) Genetic control of gynoecy in the buffalo gourd.
J Hered 72: 355–356.
Dyutin KE, Sokolov SD (1990) Spontaneous mutant of watermelon with male sterility. Cytol
Genet (Tsitologiya i Genetika) 24: 56–57.
Eisa HM, Munger HM (1968) Male sterility in Cucurbita pepo. Proc Am Soc Hort Sci 92:
473–479.
Epinat C, Pitrat M (1989) Inheritance of resistance of three lines of muskmelon (Cucumis
melo) to downymildew (Pseudoperonospora cubensis). In: CE Thomas (ed) Proceedings of
Cucurbitaceae 89: Evaluation and Enhancement of Cucurbit Germplasm. US Dep Agric
Agric Res Serv Charleston, SC, USA, pp 133–135.
Epinat C, Pitrat M, Bertrand F (1993) Genetic analysis of resistance of five melon lines to
powdery mildews. Euphytica 65: 135–144.
Fanourakis NE (1984) Inheritance and linkage studies of the fruit epidermis structure and
investigation of linkage relations of several traits and of meiosis in cucumber. PhD Diss,
Univ of Wisconsin, Madison, USA.
Fanourakis NE, Simon PW (1987) Analysis of genetic linkage in the cucumber. J Hered 78:
238–242.
Fernández-Silva I, Moreno E, Eduardo I, Arús P, Álvarez JM, Monforte AJ (2009) On the genetic
control of heterosis for fruit shape in melon (Cucumis melo L.). J Hered 100: 229–235.
Frantz JD, Jahn MM (2004) Five independent loci each control monogenic resistance to gummy
stem blight in melon (Cucumis melo L.). Theor Appl Genet 108: 1033–1038.
Firpo IT, Lopez Anido F, Garcia SM, Cointry E (1998) Heterosis in summer squash (Cucurbita
pepo L.). Cucurbit Genet Coop Rep 21: 43–45.
Fulks BK, Scheerens JC, Bemis WP (1979) Sex expression in Cucurbita foetidissima HBK. Cucurbit
Genet Coop Rep 2: 36.
Galun E (1961) Study of the inheritance of sex expression in the cucumber. The interaction of
major genes with modifying genetic and non-genetic factors. Genetica 32: 134–163.
Ganesan J, Sambandam CN (1985) Inheritance of leaf shape in muskmelon (Cucumis melo L.)
I. A qualitative approach. Annamalai Univ Agric Res Annals 12: 53-–58.
Ganesan T (1988) Genetic studies on certain characters of economic importance in musk melon
(Cucumis melo L.). PhD Thesis, Dept Agri Botany, Annamalai Univ, Tamil Nadu, India.
Ghaderi A, Lower RL (1979a) Analysis of generation means for yield in six crosses of cucumber.
J Am Soc Hort Sci 104: 567–572.
Ghaderi A, Lower RL (1979b) Heterosis and inbreeding depression for yield in populations
derived from six crosses of cucumber. J Am Soc Hort Sci 104: 564–567.
Godoy AR, Higuti ARO, Cardosa AII (2008) Yield and heterosis in cucumber inbred lines
crosses. Bragantia 67: 839–844.
Grebenšcikov I (1954) Zur Vererbung der Dünnschaligkeit bei Cucurbita pepo L. Züchter 24:
162–166.
Grebenšcikov I (1958) Notulae cucurbitologicae III. Kulturpflanze 6: 38–60.
Guner N, PesicVan-Esbroeck Z, Wehner TC (2008) Inheritance of resistance to Papaya ringspot
virus-watermelon strain in watermelon. J Hered 95: 498–502.
Gusmini G, Wehner TC (2006) Qualitative inheritance of rind pattern and flesh color in
watermelon. J Hered 97: 177–185.
Gusmini G, Song R, Wehner TC (2005) New sources of resistance to gummy stem blight in
watermelon. Crop Sci 45: 582–588.
Gwanama C, Botha AM, Labuschagne MT (2001) Genetic effects and heterosis of flowering
and fruit characteristics of tropical pumpkin. Plant Breed 120: 271–272.
Hall MR, Sumner DR (1999) Managing pests with resistant crops. In: Proc 1999 Georgia
Vegetable Conf 30: 34–36.
Harwood RR, Markarian D (1968a) The inheritance of resistance to powdery mildew in the
cantaloupe variety Seminole. J Hered 59: 126–130.
Harwood RR, Markarian D (1968b) A genetic survey of resistance to powdery mildew in
muskmelon. J Hered 59: 213–217.
Hayes HK, Jones DF (1916) First generation crosses in cucumbers. Connecticut Agri Exp Sta
Annu Rep, pp 319–322.
Henderson WR (1989) Inheritance of orange flesh color in watermelon. Cucurbit Genet Coop
Rep 12: 59–63.
Henderson WR, Jenkins SF Jr, Rawlings JO (1970) The inheritance of Fusarium wilt resistance
in watermelon, Citrullus lanatus (Thunb.) Mansf. J Am Soc Hort Sci 95: 50–53.
Henderson WR, Scott GH, Wehner TC (1998) Interaction of flesh color genes in watermelon.
J Hered 89: 50–53.
Herrington ME, Brown JP (1988) Inheritance of leaf and fruit characteristics in Cucurbita
maxima Duch. cv. Queensland Blue x C. ecuadorensis Cutler and Whitaker. Queensl J
Agri Anim Sci 45: 45–48.
Huang H, Zhang X, Wei Z, Li Q, Li X (1998) Inheritance of male-sterility and dwarfism in
watermelon [Citrullus lanatus (Thunb.) Matsum. and Nakai]. Sci Hort 74: 175–181.
Hujieda K, Akiya R (1962) Inheritance of powdery mildew resistance and spine color of fruit
in cucumber. J Hort Assoc Jpn 31: 30–32.
Hutchins AE (1938) Some examples of heterosis in cucumber, Cucumis sativus L. Proc Am Soc
Hort Sci 36: 660–664.
Hutchins AE (1940) Inheritance in the cucumber. J Agri Res 60: 117–128.
Iezzoni AF, Peterson CE (1979) Linkage of bacterial wilt resistance and sex expression genes
in cucumber. Cucurbit Genet Coop Rep 2: 8.
Iezzoni AF, Peterson CE (1980) Linkage of bacterial wilt resistance and sex expression in
cucumber. HortScience 15: 257–258.
Iezzoni AF, Peterson CE, Tolla GE (1982) Genetic analysis of two perfect flowered mutants in
cucumber. J Am Soc Hort Sci 107: 678–681.
Ingart F, Weeden NF (1984) Allozyme variation in cultivars of Cucurbita pepo L. Euphytica
33: 779–785.
Jagger IC, Whitaker TW, Porter DR (1938) Inheritance in Cucumis melo of resistance to powdery
mildew (Erysiphe cichoracearum). Phytopathology 28: 761.
Jenkins JM Jr (1946) Studies on the inheritance of downy mildew resistance. J Hered 37:
267–276.
Kabelka E, Ullah Z, Grumet R (1997) Multiple alleles for zucchini yellow mosaic virus resistance
at the zym locus in cucumber Theor Appl Genet 95: 997–1004.
Kauffman CS, Lower RL (1976) Inheritance of an extreme dwarf plant type in the cucumber.
J Am Soc Hort Sci 101: 150–151.
Kenigsbuch D, Cohen Y (1989) Independent inheritance of resistance to race 1 and race 2 of
Sphaerotheca fuliginea in muskmelon. Plant Dis 73: 206–208.
Kenigsbuch D, Cohen Y (1992) Inheritance of resistance to downy mildew in Cucumis melo PI
124112 and commonality of resistance genes with PI124111F. Plant Dis 76: 615–617.
Keinath AP, Duthie JA (1998) Yield and quality reductions in watermelon due to anthracnose,
gummy stem blight, and black rot. In: Recent Research and Development in Plant
Pathology Research, vol 2. Signpost, Trivandrum, India, pp 77–90.
King SR, Davis AR (2007) Pilot survey results to prioritize research needs in the watermelon
industry. Cucurbit Genet Coop Rep 28–29: 43–46.
Kirkpatrick DJ, Decker DS, Wilson HD (1985) Allozyme differentiation in the Cucurbita pepo
complex: C. pepo var. medullosa vs. C. texana. Econ Bot 39: 289–299.
Koch PS, Costa CP (1991) Inheritance of plant and fruit characters in gherkin. Hort Bras 9(2):
73–77.
Kooistra E (1968) Powdery mildew resistance in cucumber. Euphytica 17: 236–244.
Kooistra E (1971) Inheritance of flesh and skin colors in powdery mildew resistant cucumbers
(Cucumis sativus L.). Euphytica 20: 521–523.
Korzeniewska A (1992) New genes in Cucurbita maxima Duch. In: RW Doruchowski, E Kozik,
K Niemirowicz-Szczytt (eds) Proc Cucurbitaceae 1992: the 5th EUCARPIA Meeting on
Cucurbit Genetics & Breeding, pp 75–78.
Kubicki B (1962) Inheritance of some characters in muskmelons (Cucumis melo). Genet Polon
3: 265–274.
Kubicki B (1969a) Investigations of sex determination in cucumber (Cucumis sativus L.). V.
Genes controlling intensity of femaleness. Genet Polon 10: 69–86.
Kubicki B (1969b) Investigations of sex determination in cucumber (Cucumis sativus L.). VII.
Trimonoecism. Genet Polon 10: 123–143.
Kubicki B (1974) New sex types in cucumber and their uses in breeding work. Proc XIX Intl
Hort Congr 3: 475–485.
Lane KP, Munger HM (1985) Linkage between Corynespora leafspot resistance and powdery
mildew susceptibility in cucumber (Cucumissativus). HortScience 20(3): 593.
Layton DV (1937) The parasitism of Colletotrichum lagenarium (Pass.) Ells & Halst. Iowa Agri
Exp Sta Annu Bul, p 223.
Lee CW, Janick J (1978) Inheritance of seedling bitterness in Cucumis melo. HortScience 13:
193–194.
López Anido F, Cravero V, Asprelli P, Firpo T, García SM, Cointry E (2004) Heterotic patterns
in hybrids involving cultivar-groups of summer squash, Cucurbita pepo L. Euphytica
135: 355–360.
Loy JB (1972) A comparison of stem peroxidases of squash (Cucurbita maxima Duch. and C.
pepo L.). J Exp Bot 23: 450–457.
Luan F, Sheng Y, Wang Y, Staub JE (2010) Performance of melon hybrids derived from parents
of diverse geographic origins. Euphytica 173: 1–16.
Lumsden D (1914) Mendelism in melons. New Hampshire Agri Exp Sta Bull 172: 1–58.
Mains EB (1950) Inheritance in Cucurbita pepo. Papers Mich Acad Sci Arts Lett 36: 27–30.
McCreight JD (1983) Linkage of red stem and male sterile-1 in muskmelon. Cucurbit Genet
Coop Rep 6: 48.
McCreight JD (2003) Genes for resistance to powdery mildew races 1 and 2U.S. in melon PI
313970. HortScience 38: 591–594.
Miller GA, George Jr WL (1979) Inheritance of dwarf determinate growth habits in cucumber.
J Am Soc Hort Sci 104: 114–117.
Munger HM, Lane DP (1987) Source of combined resistance to powdery mildew and orynespora
leafspot in cucumber. Cucurbit Genet Coop Rep 10: 1.
Munger HM, Wilkinson RE (1975) Scab resistance in relation to fruit length in slicing cucumbers.
Veg Imp Nwsl 17: 2.
Munshi AD, Kumar R, Panda B (2006) Combining ability in cucumber (Cucumis sativus).
Indian J Agri Sci 76(12): 750–752.
Nath P, Dutta OP, Velayudhan S, Swamy KRM (1976) Inheritance of resistance to fruit fly in
pumpkin. SABRAO J 8: 117–119.
Navot N, Sarfatti M, Zamir D (1990) Linkage relationships of genes affecting bitterness and
flesh color in watermelon. J Hered 81: 162–165.
Netzer D, Niegro S, Galun F (1977) A dominant gene conferring resistance to Fusarium wilt
in cucumber. Phytopathology 67: 525–527.
Norton JD, Cosper RD, Smith DA, Rymal KS (1986) ‘AU-Jubilant’ and ‘AU-Producer’
watermelons. HortScience 21: 1460–1461.
Nugent PE, Cuthbert FPJ, Hoffman JC (1984) Two genes for cucumber beetle resistance in
muskmelon. J Am Soc Hort Sci 109: 756–759.
Nuttall WW, Jasmin JJ (1958) The inheritance of resistance to bacterial wilt (Erwinia tracheiphila
[E. F. Sm.] Holland) in cucumber. Can J Plant Sci 38: 401–404.
Odland ML, Groff DW (1963b) Linkage of vine type and geotropic response with sex forms
in cucumber Cucumis sativus L. Proc Am Soc Hort Sci 82: 358–369.
Orton WA (1911) The development of disease resistant varieties of plants. In: Vilmorin (ed) IV
Conf Int de Genetique, Paris, Comptes Rendus et Rapports, Masson et Cie, 1913, Paris,
France, pp 247–265.
Owens KW, Peterson CE (1982) Linkage of sex type, growth habit and fruit length in 2 cucumber
inbred backcross populations. Cucurbit Genet Coop Rep 5: 12.
Pachner M, Lelley T (2004) Different genes for resistance to zucchini yellow mosaic virus
(ZYMV) in Cucurbita moschata. In: A Lebeda, HS Paris (eds) Proc Cucurbitaceae 2004,
Palacký Univ, Olomouc, Czech Republic, pp 237–243.
Padley LD Jr, Kabelka EA, Roberts PD (2009) Inheritance of resistance to crown rot caused by
Phytophthora capsici in Cucurbita. HortScience 44: 211–213.
Pandey S, Dhillon NPS, Sureja AK, Singh D, Malik AA (2010) Hybridization for increased
yield and nutritional content of snake melon (Cucumis melo L. var. flexuosus). Plant Genet
Resour Characteris Utilis 8: 127–131.
Paris HS (1994) Genetic analysis and breeding of pumpkins and squash for high carotene
content. In: HF Linskens, JF Jackson (eds) Modern Methods of Plant Analysis, vol 16:
Vegetables and Vegetable Products. Springer, Berlin, Germany, pp 93–115.
Paris HS (1997) Genes for developmental fruit coloration of acorn squash. J Hered 88:
52–56.
Paris HS, Cohen S (2000) Oligogenic inheritance for resistance to zucchini yellow mosaic virus
in Cucurbita pepo. Ann Appl Biol 136: 209–214.
Paris HS, Kabelka E (2009) Gene list for Cucurbita species, 2009. Cucurbit Genet Coop Rep
31–32: 44–69.
Paris HS, Cohen S, Burger Y, Yoseph R (1988) Single gene resistance to zucchini yellow mosaic
virus in Cucurbita moschata. Euphytica 37: 27–29.
Paris HS, Hanan A, Baumkoler F (2004) Assortment of five gene loci in Cucurbita pepo. In: A
Lebeda, HS Paris (eds) Proc Cucurbitaceae 2004, Palacký Univ, Olomouc, Czech Republic,
pp 389–394.
Parthasarathy VA, Sambandam CN (1981) Inheritance in Indian melons. Indian J Genet 41:
114–117.
Pike LM, Peterson CE (1969) Inheritance of parthenocarpy in the cucumber (Cucumis sativus
L.). Euphytica 18: 101–105.
Pitrat M (1991) Linkage groups in Cucumis melo L. J Herdity 82: 406–411.
Pitrat M (1994) Linkage groups in Cucumis melo L. Cucurbit Genet Coop Rep 17: 148–149.
Pitrat M (2006) Gene list for melon. Cucurbit Genet Coop Rep 28–29: 142–163.
Pitrat M, Lecoq H (1980) Inheritance of resistance to cucumber mosaic virus transmission by
Aphis gossypii in Cucumis melo. Phytopathology 70: 958–961.
Pitrat M, Lecoq H (1982) Relations genetiques entre les resistance par non acceptation et par
antibiose du melon a Aphis gosypii. Recherche de liaisons avec d autres genes Agronome
2: 503–508.
Pitrat M, Lecoq H (1984) Inheritance of Zucchini Yellow Mosaic Virus resistance in Cucumis
melo L. Euphytica 33: 57–61.
Pitrat M, Risser G, Ferrière C, Olivier C, Ricard M (1991) Two virescent mutants in melon
(Cucumis melo). Cucurbit Genet Coop Rep 14: 45.
Poole CF (1944) Genetics of cultivated cucurbits. J Hered 35: 122–128.
Poole CF, Grimball PC (1945) Interaction of sex, shape, and weight genes in watermelon. J
Agr Res 71: 533–552.
Porter DR (1937) Inheritance of certain fruit and seed characters in watermelons. Hilgardia
10: 489–509.
Prasad K, Norton JD (1967) Inheritance of resistance to Mycosphaerella citrullina in muskmelon.
Proc Am Soc Hort Sci 91: 396–400.
Sharma GC, Hall CV (1971) Cucurbitacin B and total sugar inheritance in Cucurbita pepo related
to spotted cucumber beetle feeding. J Am Soc Hort Sci 96: 750–754.
Shifriss O (1947) Developmental reversal of dominance in Cucurbita pepo. Proc Am Soc Hort
Sci 50: 330–346.
Shifriss O (1961) Sex control in cucumbers. J Hered 52: 5–12.
Shifriss O (1989) Relationship between the B genes of two Cucurbita species, II. Cucurbit Genet
Coop Rep 12: 75–78.
Shifriss O, Myers CH, Chupp C (1942) Resistance to mosaic virus in cucumber. Phytopathology
32: 773–784.
Shimizu S, Kanazawa K, Kato A (1963) Studies on the breeding of cucumber for resistance to
downy mildew, part 2. Difference of resistance to downy mildew among the cucumber
varieties and the utility of the cucumber variety resistance to downy mildew (in Japanese).
Bul Hort Res Sta Jpn. Ser. A, 2: 80–81.
Shimotsuma M (1963) Cytogenetical studies in the genus Citrullus. VII. Inheritance of several
characters in watermelons. Jpn J Breed 13: 235–240.
Sing AK (1972) Inheritance of some seed characters in Melothria medraspatana (L.) Conz. Balwant
Vidyapeeth J Agri Sci Res 14: 56–57.
Sowell GB Jr (1975) An additional source of resistance to gummy stem blight in watermelon.
Plant Dis Rep 59: 413–415.
Sowell G Jr, Pointer GR (1962) Gummy stem blight resistance of introduced watermelons.
Plant Dis Rep 46: 883–885.
Srivastava VK, Nath P (1972) Inheritance of some qualitative characters in Momordica charantia
L. Indian J Hort 29: 319–321.
Stephenson AG, Hayes CN, Jóhannsson MH, Winsor JA (2001) The performance of
microgametophytes is affected by inbreeding depression and hybrid vigor in the
sporophytic generation. Sexual Plant Reprod 14: 77–83.
Staub JE, Meglic V, McCreight JD (1998) Inheritance and linkage relationships of melon
(Cucumis melo L.) isozymes. J Am Soc Hort Sci 123: 264–272.
Strong WJ (1931) Breeding experiments with the cucumber (Cucumis sativus L.). Sci Agri 11:
333–346.
Superak TH (1987) A green corolla mutant in Cucurbita pepo. Cucurbit Genet Coop Rep 10: 103.
Taja M, Wehner TC (2008) Gene list for other genera of Cucurbitaceae, 2008. Cucurbit Genetics
Cooperative Report 2007. 31–32: 41–43.
Thomas CE, Cohen Y, McCreight JD, Jourdain EL, Cohen S (1988) Inheritance of resistance to
downy mildew in Cucumis melo. Plant Dis 72: 33–35.
Thomas CE, McCreight JD, Jourdain EL (1990) Inheritance of resistance to Alternaria cucumerina
in Cucumis melo line MR-1. Plant Dis 74: 868–870.
Tkachenko NN (1935) Preliminary results of a genetic investigation of the cucumber, Cucumis
sativus L. Bull Appl Plant Breed 9: 311–356.
Tyagi ID (1976) Inheritance of some qualitative characters in bottle gourd (Lagenaria siceraria
Standal). Indian J Hort 33: 78–82.
van Vliet GJA, Meysing WD (1974) Inheritance of resistance to Pseudoperonospora cubensis Rost.
in cucumber (Cucumis sativus L.). Euphytica 23: 251–255.
van Vliet GJA, Meysing WD (1977) Relation in the inheritance of resistance to Pseudoperonospora
cubensis Rost. and Sphaerotheca fuliginea Poll. in cucumber (Cucumis sativus L.). Euphytica
26: 793–796.
Vashishta RN, Choudhury B (1972) Inheritance of resistance to red pumpkin beetle in muskmelon,
bottle gourd and watermelon. In: KL Chadha (ed) Proc 3rd Int Symp Sub-tropical and
Tropical Hort, vol 1. Horticultural Society of India, Bangalore, India, pp 75–81.
Vashistha RN, Choudhury B (1974) Inheritance of resistance to red pumpkin beetle in
muskmelon. SABRAO J 6: 95–97.
Wai T, Grumet R (1995) Inheritance of resistance to the watermelon strain of papaya ringspot
virus in the cucumber line TMG-1. HortScience 30: 338–340.
Wai T, Staub JE, Kabelka E, Grumet R (1997) Linkage analysis of potyvirus resistance alleles
in cucumber J Heredity 88: 454–458.
Wall JR (1967) Correlated inheritance of sex expression and fruit shape in Cucumis. Euphytica
23: 251–255.
Wang YJ, Provvidenti R, Robinson RW (1984) Inheritance of resistance in cucumber to
watermelon mosaic virus. Phytopathology 51: 423–428.
Wang YJ, Provvidenti R, Robinson RW (1987) Inheritance of resistance to watermelon mosaic
virus 1 in cucumber. HortScience 19: 582–588.
Wasuwat SL, Walker JC (1961) Inheritance of resistance in cucumber to cucumber mosaic
virus. Phytopathology 51: 423–428.
Watts VM (1962) A marked male-sterile mutant in watermelon. Proc Am Soc Hort Sci 81:
498–505.
Watts VM (1967) Development of disease resistance and seed production in watermelon stocks
carrying the msg gene. Proc Am Soc Hort Sci 91: 579–583.
Weetman LM (1937) Inheritance and correlation of shape, size, and color in the watermelon,
Citrullus vulgaris Schrad. Iowa Agri Exp Sta Res Bull 228: 222–256.
Wehner TC (1999) Heterosis in vegetable crops. In: JG Coors, S Pandey (eds) Genetics and
Exploitation of Heterosis in Crops. Am Soc Agron, Madison, WI, USA, pp 387–397.
Wehner TC (2005) Gene list for cucumber. Cucurbit Genet Coop Rep 28–29: 105–141.
Wehner TC (2007) Gene list for watermelon, 2007. Cucurbit Genet Coop Rep 30: 96–120.
Wellington R, Hawthorn LR (1928) A parthenocarpic hybrid derived from a cross between and English
forcing cucumber and the Arlington White Spine. Proc Am Soc Hort Sci 25: 97–100.
Whelan EDP (1973) Inheritance and linkage relationship of two radiationinduced seedling
mutants of cucumber. Can J Genet Cytol 15: 597–603.
Whelan EDP (1974) Linkage of male sterility, glabrate and determinate plant habit in cucumber.
HortScience 9: 576–577.
Whitaker TW, Bohn GW (1950) The taxonomy, genetics, production and uses of the cultivated
species of Cucurbita. Econ Bot 4: 52–81.
Whitaker TW, Davis GN (1962) Cucurbits. Interscience Publ, New York, USA.
Wilson JM (1968) The relationship between scab resistance and fruit length in cucumber,
Cucumis sativus L. MS Thesis, Cornell Univ, Ithaca, NY, USA.
Winstead NN, Goode MJ, Barham WS (1959) Resistance in watermelon to Colletotrichum
lagenarium races 1, 2, and 3. Plant Dis Rep 43: 570–577.
Wu T, Zhou J, Zhang Y, Cao J (2007) Characterization and inheritance of a bush-type in tropical
pumpkin (Cucurbita moschata Duchesne). Sci Hort 114: 1–4.
Xianglin Z (1987) A study on the breeding of naked kernel pumpkin and its genetic behavior.
Acta Hort Sin 14: 115–118 (in Chinese with English summary).
Xu Y, Kang D, Shi Z, Shen H, Wehner TC (2004) Inheritance of resistance to zucchini yellow
mosaic virus and watermelon mosaic virus in watermelon. J Hered 96: 498–502.
Youngner VB (1952) A study of the inheritance of several characters in the cucumber. PhD
Diss, Univ of Minnesota, St. Paul, MN, USA.
Zhou XG, Everts KL, Bruton BD (2010) Race 3, a new and highly virulent race of Fusarium
oxysporum f.sp. niveum causing fusarium wilt in watermelon. Plant Dis 94: 92–98.
Zijlstra S, den Nijs APM (1986) Further results of linkage studies in Cucumis sativus L. Cucurbit
Genet Coop Rep 9: 55.
Zink FW, Gubler WD (1985) Inheritance of resistance in muskmelon to Fusarium wilt. J Am
Soc Hort Sci 110: 600–604.
Zraidi A, Pachner M, Lelley T, Obermayer R (2003) On the genetics and histology of the hull-less
character of Styrian oil-pumpkin (Cucurbita pepo L.). Cucurbit Genet Coop Rep 26: 57–61.
Zraidi A, Stift G, Pachner M, Shojaeiyan A, Gong L, Lelley T (2007) A consensus map for
Cucurbita pepo. Mol Breed 20: 375–388.
Zuniga TL, Jantz JP, Zitter TA, Jahn MK (1999) Monogenic dominant resistance to gummy
stem blight in two melon (Cucumis melo) accessions. Plant Dis 83: 1105–1107.
ABSTRACT
Within the genus Cucurbita (Cucurbitaceae) there are five domesticated
species, three of which, C. pepo, C. maxima, and C. moschata, are important
crop species grown world-wide. Although cultigens of all three species are
found in temperate and sub-tropical climates, C. maxima and C. pepo are
considered best adapted to temperate climates; whereas, many cultigens
of C. moschata are restricted to tropical and subtropical climates. Cucurbita
species have a vining habit of growth, and are monoecious with large
showy flowers that are bee pollinated. Horticulturally, domesticated
members of this genus are conveniently classified into three broad
groupings: (1) summer squash cultigens, the fruit of which is consumed
immature, about three to five days after fruit set; (2) winter squash, the
fruit of which is harvested when mature, about 50 to 60 days after fruit set,
but which may require additional storage to reach optimum sugar levels
for desirable eating quality; and (3) gourd and pumpkin cultigens that
are used mainly for ornamental purposes. Although Cucurbita cultigens
are considered highly outcrossed under natural conditions, they can be
highly inbred without readily apparent inbreeding depression. They are
often classified in breeding books along with other self-pollinating crops.
For this reason, the pedigree system of breeding has been widely and
successfully adopted by most cucurbit breeders. For wide, interspecific
crosses, the backcross system, together with selection, has also been
utilized together with the pedigree system. Most breeding efforts have
been for qualitative traits such as fruit appearance (size, color, shape),
bush or vine habit of growth, and resistance to a few major diseases.
Important quantitative traits are fruit size, seed size, and % dry matter
of flesh, the latter an important parameter of eating quality. Prior to 1980,
most varieties of squash and pumpkins were open-pollinated. However,
in North America there has been a plethora of new F1 hybrids introduced
into the seed trade during the past 30 years, with the majority of cultivars
having a bush or semi-bush habit of growth.
4.1 Introduction
The genus Cucurbita within the family Cucurbitaceae comprises five
domesticated species, three of which, C. pepo L., C. maxima Duchesne, and
C. moschata Duchesne, represent economically important species cultivated
worldwide (Whitaker and Davis 1962; Robinson and Decker-Walters 1996).
The other two domesticated species, C. argyrosperma C. Huber and C. ficifolia
Bouché have more limited cultivation and use and will not be discussed
here. Squash, pumpkins, and to a lesser extent gourds, are economically
important crops, with world production estimated to be in excess of 20
million mtons grown on over 1.5 million hectares (FAOSTAT 2008). Decker-
Walters and Walters (2000) provide a comprehensive description of fruit of
both wild and domesticated forms of Cucurbita, along with an overview
of phytogeographic origins of the different species and general food and
ornamental uses. In North America in 2008, 859 thousand mtons of squash
and pumpkins were produced on 39.4 thousand hectares, and the farm value
was approximately 373 million U.S dollars (Statistics Canada 2008; USDA
2008). The agricultural statistical reporting does not separate ornamental
pumpkin production from that used for canning and does not distinguish
among winter and summer squash production.
There have been different interpretations on usage of the terms squash,
pumpkin, and gourd beginning with the time these crops were introduced
into Europe in the early part of the 16th century. In the first English printing
of Vilmorin-Andreieux’s “The Garden Vegetables” (Robinson 1885), all
members of the Cucurbita genus are described under the heading “Gourds,”
equivalent to the French name “courge,” and presumably, categories of the
different species followed the classification scheme proposed by Charles
Naudin. Although “marrows” clearly referred to fruit, mainly within
C. pepo, eaten immature, there was no clear distinction between pumpkins
and squash, except that pumpkins were generally mentioned as bearing
large fruit. In current North American usage “gourds” are defined as
various hard-rind forms of fruit within the Cucurbitaceae family, and in
the Cucurbita genus, most common within the species pepo (Bailey 1937).
“Pumpkin” typically refers to cultigens with round or oval fruit grown for
ornamental purposes, for pie processing, and less commonly for livestock
feed; whereas, “squash” is applied to any members of the three species
having culinary use. Summer squash includes cultivars in which the fruit
is consumed while immature and typically harvested within 3 to 5 days
after pollination, while fruit of winter squash is consumed when mature. A
squash can be defined as mature when its reproductive cycle is completed
in terms of embryo development (seed fill), which occurs about 55 days
after fruit set in temperate climates (Loy 2004). The three most important
classes of winter squash consumed in North America are acorn (C. pepo),
Figure 4-1 Illustration of the three major squash types grown in North America: acorn (C.
pepo, A), kabocha/buttercup (C. maxima, B) and butternut (C. moschata, C).
Color image of this figure appears in the color plate section at the end of the book.
A wild progenitor of C. pepo subsp. pepo, has not been found. The wild
species C. pepo subsp. ovifera var. texana is endemic to Texas, and another
closely related wild species, C. pepo subsp. fraterna (L.H. Bailey) Andres,
is native to northeastern Mexico. Domesticated forms of subspecies pepo
include ornamental pumpkins, zucchini, caserta, and vegetable marrow
summer squash, and a few ornamental gourds such as “Orange” and
“Miniature Ball”. Subspecies ovifera includes acorn, “Delicata”, and “Sweet
Dumpling” types of winter squash, yellow crookneck and straightneck
summer squash, and various gourds, including egg, pear and spoon gourds.
A third subspecies, C. pepo subsp. gumala Teppner, possibly endemic to
Guatemala, has been described by Teppner (2004). Fruit of this species are
characterized by a thick, hard, warty rind, and 10 main ribs with mostly
double interstitial ribbing. Teppner is of the opinion that cultivars of subsp.
gumala may be closely related to the original wild progenitors of subsp.
gumula and subsp. pepo.
Species diversity in C. moschata appears to be greatest in northern South
America (Nee 1990; Wessel-Beaver 2000; Andres 2004). Moreover, Dillehay
et al. (2007) have provided compelling evidence from radiocarbon dating
of macro plant remains of squash from a tropical dry forest region in the
Nanchoc Valley in Peru that domesticated C. moschata squash was being
used as far back as 10,000 years ago. This also agrees with the age (10,000
to 7,000 years BP) of phytoliths of C. moschata recovered from Ecuador and
Columbia, and believed, based on size, to represent domesticated species
(Piperno and Stothert 2003).
Cucurbita maxima is clearly a South American domesticate. There are few
archaeological sites documenting early use of C. maxima, but according to
Sauer (1993), the earliest archaeological remains of C. maxima are from the
coastal Viru Valley in Peru and are dated about 3800 BP. C. maxima subsp.
andreana (Naud.) Filov, native to Argentina and Uruguay, is considered to be
the wild progenitor of the domesticated subsp. maxima. The two subspecies
can readily hybridize and produce fertile offspring, and recent molecular
data support the view that subsp. andreana is the progenitor of domesticated
C. maxima (Sanjur et al. 2002).
Figure 4-2 Pistillate (left) and staminate (right) flowers of monoecious C. maxima.
Color image of this figure appears in the color plate section at the end of the book.
from the perimeter of a nectary cup at the base of the perianth tube and
terminates in a three-lobed stigma. Staminate flowers are borne on long,
slender pedicels and have three stamens borne on single filaments that are
united by two tetrasporangiate and one bisporangiate anther (Hayward
1938; Kirkwood 1905; Fig. 4-2). Flower morphology differs among the
three species (Fig. 4-3). C. maxima is characterized by a long floral tube, and
edges of moderately lobed corollas are serrated. Flowers of C. pepo have
deep and sharply angled corolla lobes; whereas, in C. moschata, the floral
tube is shallow to moderate in length and the lobes tend to be rounded.
The nectaries in Cucurbita maxima are distinctly more fragrant than in the
other species. The fruit of Cucurbita is a multi-seeded berry. The peduncle
becomes highly lignified during the initial 40 days of fruit development
Figure 4-3 Flower morphology of C. maxima (left), C. pepo (middle) and C. moschata (right),
represented by pistillate flowers.
Color image of this figure appears in the color plate section at the end of the book.
Figure 4-4 Illustration of peduncle types in C. maxima (bottom laft), C. moschata (bottom right),
C. pepo subsp. ovifera (top laft), C. pepo subsp. pepo (top right). See text for explanation.
Color image of this figure appears in the color plate section at the end of the book.
Figure 4-5 Pistillate flower (left) tied off with a Twist-ems tie one day prior to anthesis;
pistillate flower retied and tagged after pollination.
Color image of this figure appears in the color plate section at the end of the book.
ups, vinyl colored flags on PVC stakes work well, and can be reused. One
color flag can be used for set-ups for self-pollinations, another color can
be used for female flowers set up for crosses, and an additional colored
flag can be placed next to fruit that have been pollinated. It is useful to
periodically walk through breeding plots to check for fruit set and remove
flags from unsuccessful pollinations. Pistillate flowers that have been self
or cross pollinated are conveniently labeled with ¾ x 9 inch polyethylene
bands (Sato Labeling Solutions, America, Inc.) placed around the pedicel
or the internode preceding the flower node. Biodegradable labeling tags
are available.
Squash pollen is not long lived, and typical recommendations are to
complete hand-pollinations by noon (Whitaker and Robinson 1986). Hayase
(1956b) reported that fruit set in C. maxima is greatest when pollinations
occur shortly after anthesis and progressively declines until mid-day. It has
been repeatedly observed that pollination success declines rapidly when
flowers begin to wilt or lose turgidity (JB Loy, pers. observation). In a study
reported by Nepi and Pacini (1993), wilting and flower closure in C. pepo
occurred by 11:30 to 12:30 hours, about six hours after anthesis, and this
was followed by rapid loss of pollen viability, presumably due to pollen
dehydration. In agreement with their results, it has been observed that
under overcast conditions with high humidity, flowers stay turgid much
longer, and it is possible to make successful pollinations much later in the
day (JB Loy, pers. observation).
There are also species and genotypic differences in duration of pollen
viability. Flowers of C. pepo will often begin to lose turgidity by 9:00 hours,
and flowers of certain groups of C. pepo, such as the small ornamental
gourds (C. pepo subsp. ovifera), generally wilt prior to those of ornamental
pumpkins (C. pepo subsp. pepo). In summer squash homozygous for the
glabrous gene (gl-2), flowers remain noticeably turgid longer than those on
non-glabrous genotypes, and successful pollinations have been obtained
on these genotypes beyond 12:30 hours on overcast days (JB Loy, unpubl.
results). Flowers of C. moschata stay turgid longer than those of C. pepo, and
closure of C. maxima flowers is usually last among the three species (JB Loy,
unpubl. observations). With judicious planning of planting schedules with
the different species of Cucurbita, it is possible to spread out pollinations
among the three species for more efficient use of labor. In addition, more
pollinations can be accomplished per day by taking advantage of differences
in length of pollen viability and flower closure among and within the three
species.
A further technical problem in pollinating squash is that of dichogamy,
in which flowers of one sex type reach anthesis prior to the other. For
example, early summer squash lines tend to exhibit pistillate flowering
as much as a week to 10 days before staminate flowering. In butternut
temperate climates (Vining and Loy 1998). Fruits harvested when immature
may require up to two to three months of storage to alleviate potential
seed dormancy problems (Young 1949). Seed development continues in
immature stored fruit, but seed fill may not be as complete as with intact fruit
(Vining and Loy 1998; Loy 2000, 2006). Well-filled seed has been obtained
from squash harvested as early as 18 days after pollination and stored for
6 or 7 weeks (J.B. Loy, unpubl. observations). Occasionally seed extracted
from mature fruits show dormancy (Odland 1937), which can usually be
broken by storing fruits for an additional period at room temperature prior
to extraction.
Seeds are easily removed from fruit of C. pepo and C. moschata, and
can be washed in a strainer. The moisture content of the embryos removed
from mature fruit is about 40 to 50% (Cui and Loy 2002). Seed should be
immediately dried on a screen in a forced air dryer at 30 to 35oC for 48 to 72
hours to about 12% moisture (Hawthorne and Pollard 1954). Seed extraction
from fruit of C. maxima is often difficult, especially when the seed cavity
is tightly filled with seed, and placental tissue often remains attached to
the micropylar end of the seed. Dried seed can be further cleaned, and
misshapen and unfilled seed can be removed. An air column separator is
useful for a final cleaning step, and aids in removing the clear, membrane-
like endocarp layer which adheres to seed.
1.5 kg fruit with more or less globular shape, shallow to deep ribs and a
distinct striping pattern, characterized by narrow green and wide white
stripes early in development changing to wide tan and narrow orange
stripes after storage. Another member of this group, more popular in the
past, is the “Delicata” type, characterized by oblong fruit about 8 to 10
inches long and 3 to 4 inches in diameter with the same striping pattern as
“Sweet Dumpling” squash.
Seed Pumpkins: In terms of fruit type and plant habit, seed pumpkins fall
into the same category as ornamental pumpkins (subspecies pepo). However,
seed pumpkins carry the hull-less seed trait which first appeared in oil seed
pumpkins grown in the Styrian region of Austria toward the turn of the
19th century (Teppner 2000). In these types, the seed coat is reduced to a
thin membranous covering of the seed resulting from reduced amounts of
lignin and cellulose in certain cell layers within the seed coat (Stuart and
Loy 1983), rendering the seed more efficient for oil extraction. In addition
to pumpkin seed oil, hull-less pumpkins seed are consumed as a snack,
and used in trail mixes, in crackers, in the confectionary trade, and also by
the pharmaceutical industry.
Spaghetti squash: Spaghetti squash is considered to be a member of the
marrow group in terms of origin and fruit type (Maynard et al. 2001). It
is a unique cucurbit in that the flesh separates into strands when cooked,
and has been marketed as a low calorie substitute for semolina products.
The flesh character has been ascribed to a recessive gene, sp (Mazurek and
Niemirowicz-Szczytt 1992). Bush cultivars and cultivars with orange flesh
(Paris et al. 1985) have been developed. Productivity and nutritional value
have also been evaluated (Beany et al. 2002).
Calabaza: Calabaza are ovoid squash, often with tan and green mottling,
and low to moderate dry matter content (Daniel et al. 1995; Maynard et
al. 2002), and are adapted to tropical regions. They are popular in parts of
Florida, the Caribbean basin, and Central and South America.
sources (Larry Hollar, Hollar Seeds; Rob Johnston, Johnny’s Selected Seeds;
pers. comm.), presumably due to mutational events.
Fertile F1 hybrids have been obtained from the cross of C. moschata x
C. martinezii (Contin and Munger 1977; Munger and Washek 1983; Cho et al.
2003), allowing for introgression of PMR and virus resistance into C. moschata
through subsequent backcrossing and selfing. Compatibility between C. pepo
and C. moschata is limited, and for the most part, researchers have relied on
the hope that two chosen parents will “nick.” Wall and York (1960) obtained
successful crosses between C. pepo (pistillate parent) and C. moschata using
Yankee Hybrid, an F1 hybrid between two yellow summer squash lines, as
the C. pepo parent. Cucurbita pepo is largely incompatible with C. martinezii
(Vaulx and Pitrat 1979), but by crossing C. martinezii with C. moschata, the
F1 hybrid has been used as a genetic bridge to transfer PMR and resistance
to cucumber mosaic virus (CMV) into C. pepo (Munger and Washek 1983;
Whitaker and Robinson 1986). Using embryo culture, Robinson (1997) was
able to recover F1 progeny from crosses of C. pepo to C. ecuadorensis. Through
a laborious series of backcrossing and screening, Robinson was eventually
successful in transferring zucchini yellow mosaic virus (ZYMV), papaya
ringspot virus (PRSV), and CMV resistance into C. pepo, with the subsequent
release of the variety “Whitaker”.
Crosses between C. maxima and C. pepo have been largely unsuccessful.
For example, Erwin and Haber (1929) performed nearly 2,000 reciprocal
crosses between different cultivars of C. pepo and C. maxima, resulting in
177 fruits and 23 fertile seeds. Twenty-one of the fertile seeds resulted from
crosses using Hubbard as either the male (10 seeds) or female (11 seeds)
parent. Using bud-pollination and embryo culture, Hayase (1961b) obtained
F1 hybrid plants with intermediate characters of C. pepo and C. maxima
parents. The F1 plants were gynoecious, but could be backcrossed to C. pepo.
Cucurbita maxima is, however, fully fertile with C. ecuadorensis, to which it is
closely related taxonomically (Sanjur et al. 2002). Genetic resistance to PRSV,
ZYMV, and watermelon mosaic virus (WMV) in C. ecuadorensis has been
transferred by backcrossing into C. maxima with the release of the varieties
“Redlands Trailblazer” and “Dulong QHI” (Herrington et al. 1991, 2001).
Gene transfer between C. maxima and C. moschata has been difficult.
Crosses between some cultigens of the two species yield neither seed nor
fruit. In other crosses, however, both fruit and ample, viable F1 seeds are
obtained (Erwin and Haber 1929; Hayase 1956; Robinson et al. 1978; Loy
and Uretsky 2010). The F1 hybrid progeny resulting from such crosses are
vigorous and fruit production is prolific, but the F1 hybrids are largely male-
sterile (Erwin and Haber 1929; Hayase 1956; Robinson et al. 1978). Japanese
breeders have produced interspecific F1 hybrids of C. maxima x C. moschata
on a commercial basis, most notably “Tetsukabuto”, a cross between
“Delicious” (C. maxima) x “Kurokawa No. 2” (C. moschata) (Robinson
disadvantage depending upon whether they are linked to the trait being
transferred. There are numerous genes contributing to differences in fruit
coloration and patterns of fruit coloration (Paris and Brown 2005), and
variation in these traits may differ substantially in subspecies crosses,
depending upon the particular cultigens being crossed.
1962), and progeny show no readily perceptible loss in vigor and fertility
with inbreeding. Given the focus on qualitative traits and considering the
reproductive characteristics in Cucurbita species, the pedigree system of
breeding has been widely adopted. When transferring only a few genes
into a desirable genotype from a wide or interspecific cross, the backcross
system is often used. The number of backcross generations needed prior
to selfing depends upon the genetic distance between parents used for
the initial cross, the size of the segregating backcross populations and the
degree of selection intensity for the phenotype desired.
Bush plants are considerably easier to manipulate than vine plants
because staminate and pistillate flowers are readily identified on individual
plants and higher plant populations can be grown per unit area. Of all the
cultivar groups, bush summer squash are easiest to breed because of their
exceedingly compact but open growth habit, together with the convenience
of being able to select for numerous immature fruit traits in segregating
progeny prior to making controlled pollinations.
Hull-less oil seed pumpkins (C. pepo subsp. pepo) are an important crop
in eastern Europe and China, but are not widely grown in North America.
Pumpkin seed oil will not likely become an important commodity in North
America, but the market for snackseeds, either alone or in trail mixes, and
in confectionary items, could expand. In breeding seed pumpkins, seed
yield is a major breeding focus, along with obtaining sufficient seed size
and uniformity (170 mg or greater), and good tip fill. Obtaining consistent
seed fill in high yielding cultigens is challenging because seed fill continues
until at least 55 days after pollination (DAP) in temperate climates, three
to four weeks after peak mesocarp dry matter is attained (Vining and Loy
1998), and 40 to 50 days after plants reach maximum leaf area (Loy 2004; Cui
2005). A major factor in achieving high yields has been developing inbred
lines and hybrids with relatively small fruit (1.0–1.5 kg) that partition a
large proportion (45 to 55% when expressed as kJ glucose energy per kg
fruit fresh weight) of photosynthate into seeds rather than mesocarp tissue
(Cui and Loy 2002; Loy 2004). A thorough review of the genetics, breeding,
and seed composition of oil seed pumpkins has recently been published
(Lelly et al. 2010).
There has been meager effort in breeding quantitative traits in squash
and pumpkin, and in the case of traits such as dry matter content of flesh,
seed size and fruit size which may exhibit quantitative inheritance (Singh
1949; Carle and Loy 1994), genetic manipulations of these traits can be done
in breeding cycles, such that no more than two or possibly three genes for
a particular trait are segregating in a breeding population.
Techniques for pollination, tagging fruit, and marking plants to be
pollinated were discussed above. Row and within row spacing varies
with the particular growth habit of the types of squash being bred, size
and number of fruit needed for evaluation, and the space available to the
breeder. Rows at least 3.0 m or more apart are preferable for vine genotypes.
To aid in pollinations, it is desirable to train vines in one direction. Related
genotypes are normally grown adjacent to one another, both for evaluation
in the same soil type and topography and for ease of making any necessary
crosses. Control lines of known performance for traits being selected should
be interspersed within breeding plots of segregating material.
Because of extensive space requirements, evaluation of elite germplasm
for possible commercial release can consume both time and resources
needed for breeding, especially in a mature breeding program. It is
frequently not possible to replicate evaluation plots involving dozens of
hybrid combinations, and in many instances, judgments concerning the
rating of pumpkin cultigens are based more on plant vigor and growth
habit and fruit appearance than on replicated yield records. Plot size for
comparing vining and semi-bush cultigens is often in the range of 10 to 15
plants spaced 1.0 m apart in rows 2.1 to 2.4 m apart. For summer squash, a
within-row spacing of 0.6 m and row spacing of 2.0 m is sufficient, and guard
rows are not needed because inter-row competition is minimal. Guard rows
for each cultigen are rarely used when evaluating winter squash or pumpkin
either because of space or seed limitations. It is therefore important to place
all cultigens with a similar growth habit (bush, semi-bush, or vine) in the
same block to minimize growth competition between adjacent cultigens.
Cultigens which look promising in initial non-replicated evaluations
can then be evaluated in replicated plots and in different locations in the
following years.
with low dry matter and reduced integrity of peduncles (Berg 2004).
Branching habit appears to be under simple genetic control in Cucurbita,
but with some modifying genes (J.B. Loy, unpubl. results). In a small
population of acorn plants we observed a 3:1 segregation of plants with
a strong main stem and few laterals as compared to plants with multiple
branching habit. In numerous hybrid F1 plants resulting from crosses of
breeding lines with a dominant main stem to lines with multiple branching,
single stem is dominant (J.B. Loy, unpubl. observations). In summer squash,
there are clearly additional modifying genes that affect the intensity of a
strong single stem. In winter squash and pumpkin, unlike summer squash,
multiple branching habit is generally considered a desirable trait because
multiple growing points contribute to more rapid and uniform leaf canopy
development. Determinate growth habit, only recently reported in squash
(Kwack 1995), has not been utilized in breeding.
the strong association of high soluble solids and starch levels to perceived
quality and the cost and time of group culinary evaluation, taste panels may
be an overly cumbersome and unnecessary addition to a squash breeding
program. On the other hand, once a breeder has identified a potentially
good cultigen, squash samples should be made available for evaluation
by other persons.
The stage of maturity of squash strongly influences evaluation of eating
quality and nutrition. Based on rind color alone, many squash cultivars
appear mature relatively early in development. Acorn squash, for example,
turn dark green and attain nearly full size within about two weeks after fruit
set. Tan rind color in butternut squash occurs by 35 to 40 days after fruit set.
Although peak dry matter is attained in all three species of squash by 30 to 35
DAP, acceptable soluble solids levels are usually not attained in acorn squash
until 50–55 DAP (Loy 2006), and in C. moschata, may not be attained until
several weeks of storage following harvest at 55 to 60 DAP (Noseworthy
and Loy 2008). Solids contents greater than 26% occur commonly in some
kabocha squash. Cooked squash with an over-abundance of starch and
low sugar has a dry flaky texture, and may score low in cooking tests. With
additional storage, accompanied by elevated sugar levels and some loss in
dry matter, texture and sweetness will become acceptable.
Carotenoids are an abundant dietary component of squash. Many
squash cultigens are not only a good source of β-carotene, important in
development and eye function, but also an excellent source of lutein,
which has an important photoprotective function in the macular region
of the retina (Azevedo-Meleiro and Rodriguez-Amaya 2007). Cultigens
with high carotenoid levels have been bred mostly using a visual scale
(Noseworthy and Loy 2008). However, accurate analysis of carotenoids
requires spectrophotometric determination of total carotenoids combined
with high pressure liquid chromatography (HPLC) to examine carotenoids
profiles. Increases in total carotenoid levels during fruit maturation and
subsequent storage correspond well with changes in soluble solids, so it
is appropriate to evaluate carotenoids when squash are near peak eating
quality (Noseworthy and Loy 2008).
There is considerable variability in percent dry matter of pericarp
tissue among cultigens, and even among fruits from the same plant. In
the classical paper of Culpepper and Moon (1945) in which 35 cultivars of
C. pepo, C. maxima and C. moschata were compared, percent solids varied
from as high as 15.7% for Table Queen to as low as 6.4% for King of the
Mammoths, a show pumpkin. Plant breeders have increased dry matter
content, and therefore eating quality, in some of the better cultivars. There
are now numerous cultivars of kabocha and buttercup squash with dry
matter consistently above 20%, and a few of the better C. pepo winter
squash cultigens average nearly 20%. The most popular C. moschata cultivar,
Waltham Butternut, often exhibits dry matter in the 17 to 21% range in New
England (J. Noseworthy and J.B. Loy, unpubl. results). The wide range in
dry matter in winter squash is undoubtedly under multigenic control; this,
together with seasonal, within plant, and plant to plant variability within
a cultigen, render inheritance studies of dry matter content a complex
genetic area to broach. Singh (1949) conducted a thorough genetic study
of solids content in C. maxima; however, the mean solids content (% DM)
of the two parental cultigens was exceedingly low (2.74 and 6.73%). His
results fit a two gene model and additive gene action. Observations have
been that for F1 hybrids to have high dry matter, both parents must have
high dry matter, and in F2 selections exhibiting high dry matter, progeny
in later generations retain the high dry matter trait. This suggests that high
dry matter is under control of either recessive or additive alleles (J.B. Loy,
unpubl. observation).
scheme. Seven out of 15 hybrids showed high parent heterosis for mean
fruit weight, 13 out of 15 were heterotic for seed weight per fruit, and all
hybrids showed heterosis for seed weight per plant. Cui and Loy (2002)
evaluated high parent heterosis for different components of seed yield in
two hybrids and four parental strains. Total fruit dry biomass per plot was
significantly higher in both hybrids than in parental strains. Seed yield per
plot was significantly higher in one of the two hybrids in comparison to
parental inbreds, and seed number per fruit was 24 and 36% higher in the
two hybrids as compared to the two highest parental lines.
Open-pollinated strains still dominate commercial acreage of C.
moschata cultigens, and there have been few detailed studies of heterosis
in this species (Robinson 1999).
characteristic of black rot. Gummy stem blight can overwinter on crop debris
and can be seed transmitted (Zitter et al. 1996). Specific cultivars resistant
to gummy stem blight (GSB) have not been released; however, Zhang et
al. (1995) screened 308 accessions for GSB resistance and reported seven
accessions of C. martinezii, two of C. moschata and three of C. Pepo, that
showed high resistance to the disease. Angular leaf spot is a widespread
bacterial disease of squash and pumpkin, often causing severe foliar
destruction but less fruit rot than bacterial leaf spot. In favorable weather
following a disease outbreak, new plant growth will often appear free of
the disease. Considerable variability in susceptibility of Cucurbita cultigens
to this disease on both foliage and fruit has been observed (J.B. Loy, pers.
observation). However, the reliance of natural field infestation for evaluation
of this disease has prevented a reliable ranking of cultivars for degree of
susceptibility (J.B. Loy, unpubl. results). Bacterial leaf spot often occurs
during weather episodes that favor angular leaf spot. This disease is very
subtle to detect because leaf symptoms, appearing as small necrotic spots
with a yellow halo, are often fairly obscure, and if lesions coalesce, may
look similar to angular leaf spot (Zitter et al. 1996). Initial symptoms on
fruit appear as tiny, corky tan dots, but many lesions expand into slightly
sunken tan spots, surrounded by a dark ring. Many of the lesions become
encapsulated with callose tissue at the fruit surface, but some eventually
penetrate into the flesh and seed cavity, causing severe fruit rot. Fruits of
zucchini and ornamental pumpkin (C. pepo subsp. pepo) are more susceptible
to both angular leaf spot and bacterial leaf spot than those of subspecies
ovifera. Oil seed pumpkins appear to be particularly susceptible. Tolerance
of fruit to this disease has been observed among ornamental pumpkins (J.B
Loy, unpubl. observations), and it appears that plants showing medium
resistance or tolerance to angular leaf spot are more tolerant to bacterial
leaf spot.
three viruses (Fuchs and Gonsalves 2007). Once incorporated into breeding
lines, the CP gene constructs can be transferred to other breeding lines
and recombined with other CP genes to achieve multiple virus resistance.
Yield trials comparing cultivars with transgenic virus resistance to those
without resistance have generally shown favorable results under conditions
of natural virus pressure or with inoculated plants (Webb and Tyson 1997;
Fuchs et al. 1998; Schultheis and Walters 1998; Rowell et al. 1999). In 2005,
transgenic squash accounted for 12% of the summer squash acreage in the
US (Fuchs and Gonsalves 2007).
The development of in vitro techniques for producing haploid plantlets
via androgenesis and gynogenesis has been a significant development in
commercial breeding for rapid generation of homozygous lines. Among
cucurbits, this technology has been refined and utilized for cucumber and
melon breeding, but is still in its infancy in squash and pumpkin. Most
in vitro studies have focused on obtaining haploid plants from C. pepo,
using both ovule (Metwally et al. 1998a; Shalaby 2007) and anther culture
(Metwally et al. 1998b). More recently, pollen irradiation has been used to
induce haploid plants in C. pepo (Kurtar et al. 2002), C. maxima (Kurtar and
Balkaya 2010) and C. moschata (Kurtar et al. 2009). Further improvement is
needed in cultural techniques to improve the frequency of obtaining haploid
plants for wide scale adoption of androgenic and gynogenic techniques for
breeding purposes.
Acknowledgement
I wish to express my sincere thanks to Jacob B.Uretsky for his critical review
of this manuscript.
References
Aboul-Nasr MH, Damarany AM, Abdalla MMA (2002) Yield and its components of some
summer squash (Cucurbita pepo L.) genotypes. In: DN Maynard (ed) Cucurbitaceae 2002.
ASHS Press, Alexandria, VA, USA, pp 88–94.
Adneniji AA and Coyne DP (1983) Genetics and nature of resistance to powdery mildew in
crosses of butternut with calabaza squash and ‘Seminole Pumpkin.’ J Am Soc Hort Sci
108: 360–368.
Ahmed EA, Ibn Oaf HS, El Jack AE (2003) Combining ability and heterosis in line x tester
crosses of summer squash (Cucurbita pepo L.). Cucurbit Genet Coop Rep 26: 54–56.
Amaya AT, Ortega SG (1996) High yields of summer squash lines and hybrid combinations.
Cucurbit Genet Coop Rep 19: 78–80.
Andres TC (2004) Diversity in tropical pumpkin (Cucurbita moschata): A review of infraspecific
classifications. In: A Lebeda, HS Paris (eds) Progress in Cucurbit Genetics and Breeding
Research. Proc Cucurbitaceae 2004, 8th EUCARPIA Meeting on cucurbit genetics and
breeding, Olomouc, Czech Republic, pp 107–112.
Fuchs M, Gonsalves D (2007) Safety of virus-resistant transgenic plants two decades after their
introduction: Lessons from realistic field risk assessment studies. Annu Rev Phytopathol
45: 173–202.
Fuchs M, Tricoli DM, Carney KJ, Schesser M, McFerson JR, Gonsalves D (1998) Comparative
virus resistance and fruit yield of transgenic squash with single and multiple coat protein
genes. Plant Dis 82: 1350–1356.
Gaba V, Zelcer A, Gal-On A (2004) Cucurbit biotechnology—the importance of virus resistance.
In Vitro Cell Dev Biol-Plant 40: 346–358.
Gong L, Pachner M, Kalai K, Lelley T (2008a) SSR-based genetic linkage map of Cucurbita
moschata and its synteny with Cucurbita pepo. Genome 51: 878–887.
Gong L, Stift G, Kofler R, Pachner M, Lelley T (2008b) Microsatellites for the genus Cucurbita and
an SSR-based genetic linkage map of Cucurbita pepo L. Theor Appl Genet 117: 37–48.
Gonsalves C, Xue B, Gonsalves D (1995) Somatic embryogenesis and regeneration from
cotyledon explants of six squash cultivars. HortScience 30: 1295–1297.
Grebenščikov I (1958) Notulae cucurbitalogicae III. Kulturpflanze (Berlin) 6: 38–60.
Grebenščikov I (1975) Notulae cucurbitologicae VIII. Zur Frage der Reziprokenunterschiede
bei den quantitiativen Ertragsmerkmalen vom Kürbis. Kulturpflanze 23: 139–155.
Haber ES (1928) Inbreeding in Table Queen (DesMoines) squash. Proc Am Soc Hort Sci 25:
111–114.
Harvey WJ, Grant DG, Lammerink JP (1997) Physical and sensory changes during development
and storage of buttercup squash. NZ J Crop Hort Sci 25: 341–351.
Hawthorn LR, Pollard LH (1954) Vegetable and flower seed production. Blakiston Co, New
York, USA.
Hayase H (1956a) Cucurbita crosses. VII. The commencing time of pollen germination on
stigmata and anther dehiscence. Jpn J Breed 5: 261–267.
Hayase H (1956b) Cucurbita crosses. VIII. On the reciprocal interspecies hybrids between
C. maxima and C. moschata. Res. Bull. Hokkaido Natl Agri Exp Sta 70: 15–29.
Hayase H (1961) Cucurbita-crosses. XIII, Utilization of bud pollination in obtaining interspecific
hybrids of C. pepo x C. maxima. Jpn J Breed 11: 25–32.
Hayase H, Ueda T (1956) Cucurbita crosses. IX. Hybrid vigor of reciprocal F1 crosses in Cucurbita
maxima. Hokkaido Natl Agri Sta Res Bull 71: 125–128.
Hayes CN, Winsor JA, Stephenson AG (2005) Environmental variation influences the
magnitude of inbreeding depression in Cucurbita pepo ssp. texana (Cucurbitaceae). J Evol
Biol 18: 147–155.
Hayward HE (1938) The Structure of Economic Plants. MacMillan Co, New York, USA.
Herrington ME, Brown JP (1988) Inheritance of leaf and fruit characteristics in Cucurbita
maxima Duch. cv. Queensland Blue x C. ecuadorensis Cutler and Whitaker, Queenl J Agri
Anim Sci 45: 45–48.
Herrington ME, Prytz S, Brown P, Persley DM, Greber R (1991) Resistance to papaya ringspot
virus-W, zucchini yellow mosaic virus, and watermelon mosaic virus-2 in C. maxima.
Cucurbit Genet Coop Rep 14: 123–124.
Herrington ME, Prytz S, Wright RM, Walker IO, Brown P, Persley DM, Greber R (2001) ‘Dulong
QUI’ and ‘Redlands Trailblazer’, PRSV-W-, ZYMV- and WMV-resistant winter squash
cultivars. HortScience 36: 811–812.
Hurst PL, Corrigan VK, Hannan PJ, Lill RE (1995) Storage rots, compositional analysis, and
sensory quality of three cultivars of buttercup squash. NZ J Crop Hort Sci 23: 89–95.
Hutchins AA, Croston FE (1941) Productivity of F1 hybrids in squash, C. maxima. Proc Am
Soc Hort Sci 39: 332–336.
Hutchins AE (1936) The interaction of blue and green color factors in hubbard squash. Proc
Am Soc Hort Sci 33: 514.
Inamdar JA, Gangadhara M, Shenoy KN (1990) Structure, ontogeny, organographic distribution,
and taxonomic significance of trichomes and stomata in the Cucurbitaceae. In: DM Bates,
RW Robinson, C Jeffrey (eds) Biology and utilization of the Cucurbitaceae. Cornell Univ
Press, Ithaca, NY, USA, pp 290–224.
Isakeit T (2007) Phytophthora blight caused by Phytophthora capsici on pumpkin and winter
squash in Texas. Plant Dis 91: 633. Abs.
Islam SZ, Babadoost M, Lambert KN, Ndeme A (2005) Characterization of Phytophthora capsici
isolates from processing pumpkin in Illinois. Plant Dis 89: 191–197.
Jahn M, Munger HM, McCreight JD (2002) Breeding cucurbit crops for powdery mildew
resistance. In: RR Belanger, WR Bushnell, AJ Dik, TLW Carver (eds) The powdery
mildews: A comprehensive treatise. Am Phytopathol Soc, St. Paul, MN, USA.
Jalaska S (1972) Embryoid formation by fragments of cotyledons and hypocotyls in Cucurbita
pepo. Planta 103: 278–280.
Jalaska S (1986) Cucurbits. In: YPS Bajaj (ed) Biotechnology in agriculture and forestry, vol 2.
Crops. Springer, New York, USA, pp 371–386.
Johannsson M, Gates J, Stephenson AG (2002) Inbreeding depression affects pollen performance
in Cucurbita texana. J Evol Biol 11: 579–588.
Julier HE, Roulston TH (2009) Wild bee abundance and pollinations service in cultivated
pumpkins: Farm management, nesting behavior, and landscape effects. J Econ Entomol
102: 563–573.
Kintzios S, Sereti E, Bluchos P, Drossopoulos JB, Kitsake CK, Liopa-Tsakalidis A (2002) Growth
regulator pretreatment improves somatic embryogenesis in from leaves of squash
(Cucurbita pepo L.) and melon (Cucumis melo L.). Plant Cell Rep 21: 1–8.
Kirkwood JC (1905) The comparative embryology of the Cucurbitaceae. Bull NY Bot Gard
3: 13–402.
Korzeniewska A, Niemirowicz-Szczyt K (1993) Combining ability and heterosis effect in winter
squash (Cucurbita maxima Duch.). Genet Polon 34: 259–272.
Krístková E, Lebeda A (1997) Variation in field resistance of Cucurbita spp. accessions to the
powdery mildew of cucurbits. In: F Kobza, M Pidra, R Pokluda (eds) Biological and
technological development in horticulture. Proc Int. Hort Sci Conf, Lednice na Morav,
Czech Republic, p 331.
Krístková E, Lebeda A (2000) Resistance in Cucurbita pepo and Cucurbita maxima germplasm
to watermelon mosaic potyvirus-2. Plant Genet Resour Newsl 121: 47–52.
Křístková E, Lebeda A, Sedláková B (2007) Temporal and spatial dynamics of powdery mildew
species on cucurbits in the Czech Republic. Acta Hort 731: 337–343.
Kurtar ES, Balkaya A (2010) Production of in vitro haploid plants from in situ induced haploid
embryo in winter squash (Cucurbita maxima Duchesne ex Lam.). via irradiated pollen.
Plant Cell Tiss Org Cult 102: 267–277.
Kurtar ES, Sari N, Abak K (2002) Obtention of haploid embryhos and plants through irradiated
pollen technique in squash (Cucurbita pepo L.). Euphytica 127: 335–344.
Kurtar ES, Balkaya A, Ozbakir M, Ofluoglu T (2009) Induction of haploid embryo and plant
regeneration via irradiated pollen technique in pumpkin (Cucurbita moschata Duchesne
ex. Poir). Afr J Biotechnol 8: 5944–5951.
Kwack SN (1995) Inheritance and morphology of determinate growth habit in Cucurbita
moschata Poir. J Kor Soc Hort Sci 36: 1096–1101.
Kwack SN, Fujieda K (1987) Seed abortion and techniques for obtaining hybrids in interspecific
crosses of Cucurbita. J Jpn Soc Hort Sci 55: 455–460.
Lebeda A, Krístková E (1996a) Resistance in Cucurbita pepo and Cucurbita maxima germplasms
to cucumber mosaic virus. Genet Resour Crop Evol 43: 461–469.
Lebeda A, Krístková E (1996b) Genotypic variation in field resistance of Cucurbita pepo cultivars
to powdery mildew (Erysiphe cichoracearum). Genet Resour Crop Evol 43: 79–84.
Lee YH, Jeon JJ, Hong KH, Kim BD (1995) Use of random amplified polymorphic DNAs for
linkage group analysis in interspecific hybrid F2 generation of Cucurbita. J Kor Soc Hort
Sci 36: 323–330.
Lee YK, Chung WI, Ezura H (2003) Efficient plant regeneration via organogenesis in winter
squash (Cucurbita maxima Duch.). Plant Sci 164: 413–418.
Lelley T, Loy B, Murkovic M (2010) Breeding oil seed pumpkins. In: J Vollman, I Rajcan (eds)
Handbook on Plant Breeding, vol 4: Oil Crops. Springer, New York, USA, pp 469–492.
Noseworthy J, Loy B (2008) Improving eating quality and carotenoid content of squash. In: M
Pitrat (ed) Cucurbitaceae 2008, Proceedings of the IXth EUCARPIA Meeting on Genetics
and Breeding of Cucurbitaceae, INRA, Avignon, France, pp 521–528.
Odlund MJ (1937) Observations on dormancy in vegetable seed. Proc Am Soc Hort Sci 35:
562–566.
Oliveira ACB, de, Maluf WR, Pinto JEBP, Azevedo SM (2003) Resistance to papaya ringspot
virus in summer squash Cucurbita pepo L. introgressed from an interspecific C. pepo x
C. moschata cross. Euphytica 132: 211–215.
Pachner M, Lelley T (2004) Different genes for resistance to zucchini yellow mosaic virus
(ZYMV) in Cucurbita moschata. In: A Lebeda, HS Paris (eds) Progress in genetics and
breeding research, Proc Cucurbitaceae 2004. 8th EUCARPIA Meeting on cucurbit genetics
and breeding, Olomouc, Czech Republic, pp 237–243.
Padley LD, Kabelka EA (2009) Inheritance of resistance to crown rot caused by Phytophthora
capsici in Cucurbita. HortScience 44: 211–213.
Paris HS (1986) A proposed subspecific classification for Cucurbita pepo. Phytologia 61:
133–138.
Paris HS (1989) Historical records, origins, and development of the edible cultivar groups of
Cucurbita pepo (Cucurbitaceae). Econ Bot 43: 423–443.
Paris HS (1997) Genes for developmental fruit coloration of acorn squash. J Hered 88:
52–56.
Paris HS (2008) Summer squash,. In: J Prohens, F Nuez (eds) Vegetables I, Handbook of Plant
Breeding, Springer, New York, pp 352–379.
Paris HS, Nerson H (1986) Genes for intense pigmentation of squash. J Hered 77: 403–409.
Paris HS, Cohen R (2000) Oligogenic inheritance for resistance to zucchini yellow mosaic virus
in Cucurbita pepo. Ann Appl Biol 136: 209–214.
Paris HS, Brown RN (2005) The genes of pumpkin and squash. HortScience 40: 1620–1630.
Paris HS, Edelstein M, Nerson H, Burger Y, Karchi Z, Lozner D (1985) ‘Orangetti’ and ‘Go-
ghetti’, two new spaghetti squash hybrids. Hassadeh 66: 254–256.
Paris HS, Nerson H, Burger Y (1987) Leaf silvering of Cucurbita. Can J Plant Sci 67: 593–598.
Paris HS, Yonash N, Portnoy V, Mozes-Daube N, Tzuri G, Katzir N (2003) Assessment of
genetic relationships in Cucurbita pepo (Cucurbitaceae) using DNA markets. Theor Appl
Genet 106: 971–978.
Paris HS, Hanan A, Baumkoler F (2004) Assortment of five gene loci in Cucurbita pepo In: A
Lebeda, HS Paris (eds) Proc Cucurbitaceae 2004. 8th EUCARPIA Meeting on cucurbit
genetics and breeding, Olomouc, Czech Republic, pp 389–392.
Pearson OH (1968) Unstable gene systems in vegetable crops and implications for selection.
HortScience 3: 271–274.
Pearson OH (1983) Heterosis in vegetable crops. In: R Frankel (ed) Heterosis. Springer, Berlin,
Germany, pp 138–188.
Phillips TG (1946) Changes is the composition of squash during storage. Plant Physiol 21:
533–541.
Piperno DR, Stothert KE (2003) Phytolith evidence for early Holocene Cucurbita domestication
in Southwest Ecuador. Science 299: 1054–1057.
Pitrat M, Dogimont C, Bardin M (1998) Resistance to fungal diseases of foliage in melon. In: JD
McCreight (ed) Cucurbitaceae 98: Evaluation and enhancement of cucurbit germplasm.
ASHS Press, Alexandria, VA, USA, pp 167–173.
Provvidenti R (1982) Sources of resistance or tolerance to viruses in accessions of Cucurbita
maxima. Cucurbit Genet Coop Rep 5: 46–47.
Provvidenti R (1990) Viral diseases and genetic sources of resistance in Cucurbita species. In:
DM Bates, RW Robinson, C Jeffrey (eds) Biology and utilization of the Cucurbitaceae.
Cornell Univ Press, Ithaca, NY, USA, pp 427–435.
Provvidenti R (1997) New American summer squash cultivars possessing a high level of
resistance to a strain of zucchini yellow mosaic virus from China. Cucurbit Genet Coop
Rep 20: 57–58.
Rhodes AM (1959) Species hybridization and interspecific gene transfer in the genus Cucurbita.
Proc Am Soc Hort Sci 74: 546–551.
Rhodes AM (1964) Inheritance of powdery mildew resistance in the genus Cucurbita. Plant
Dis Rep 48: 54–56.
Robinson RW (1987) Inheritance of fruit skin color in Cucurbita moschata. Cucurbit Genet
Coop Rep 10: 84.
Robinson RW (1997) Barriers to gene transfer in an interspecific Cucurbita cross. HortScience
32: 495.
Robinson RW (1999) Rationale and methods for producing hybrid cucurbit seed. In: AS Basra
(ed) Hybrid seed production in vegetables: rationale and methods in selected crops. Food
Products Press, New York, USA, pp 1–47.
Robinson RW, Decker-Walters DS (1997) Cucurbits. CAB International, New York, USA.
Robinson RW, Whitaker TH, Bohn GW (1970) Promotion of pistillate flowering in Cucurbita
by 2-chloroethylphosphonic acid. Euphytica 19: 180–183.
Robinson RW, Boettger MA, Shail JW (1978) Gynoecious sex expression in Cucurbita resulting
from an interspecific cross. Cucurbit Genet Coop. Rep 1: 31–32.
Robinson W (1885) The Vegetable Garden. English translation of book written by Vilmorin-
Andrieux, J Murray, London, UK.
Rowell B, Nesmith W, Snyder JC (1999) Yields and disease resistance of fall-harvested transgenic
and conventional summer squash in Kentucky. HortTechnology 9: 282–288.
Rudich J, Halevy AH, Kedar N (1969) Increase in femaleness of three cucurbits by treatment
with ethrel, an ethylene releasing compound. Planta 86: 69–76.
Rudich J, Kedar N, Halevy AH (1970) Changed sex expression and possibilities for F1-hybrid
seed production in some cucurbits by application of ethrel and alar (B995). Euphytica
19: 47–53.
Salehi-Mohammadi R, Kashi A, Lee SG, Huh YC, Lee JM, Babalar M, Delshad M (2009)
Assessing the survival and growth performance of Iranian melon to grafting onto
Cucurbita rootstocks. Kor J Hort Sci Technol 27: 1–6.
Sanjur OI, Piperno DR, Andres TC, Wessel-Beaver L (2002) Phylogenetic relationships among
domesticated and wild species of Cucurbita (Cucurbitaceae) inferred from a mitochondrial
gene: Implications for crop plant evolution and areas of origin. Proc Natl Acad Sci 99:
535–540.
Sauer JD (1993) Historical geography of crop plants. CRC Press, Boca Raton, FL, USA.
Schaffer AA, Boyer CD, Paris HS (1986) Inheritance of rind lignification and warts in Cucurbita
pepo L. and a role for phenylalanine ammonia lyase in their control. Z Pflanzenzuchtg
96: 147–153.
Schultheis J, Walters SA (1998) Yield and virus resistance of summer squash cultivars and
breeding lines in North Carolina. HortTechnology 8: 31–39.
Schultheis J, Hassell R, Kelley T, Kumar R, Olson S, Wehner TC (2008) Grafted watermelon:
evaluation of planting density for high yield. National Watermelon Association: http:
//www.nationalwatermelonassociation.com/scientific_research.php.
Scott GW (1933) Sex ratios and fruit production studies in bush pumpkins. Proc Am Soc Hort
Sci 30: 520–525.
Scott GW (1934) Observations on some inbred lines of bush types of C. pepo. Proc Am Soc
Hort Sci 32: 480.
Shalaby TA (2007) Factors affecting haploid induction through in vitro gynogenesis in summer
squash (Cucurbita pepo L.). Sci Hort 115: 1–6.
Sharma GC, Hall CV (1971) Influence of cucurbitacins, sugars and fatty acids on cucurbit
susceptibility to spotted cucumber beetle. J Am Soc Hort Sci 96: 675–680.
Shriffriss O (1947) Developmental reversal of dominance in Cucurbita pepo. Proc Am Soc Hort
Sci 50: 330–346.
Shiffriss O (1981) Origin, expression, and significance of gene B in Cucurbita pepo L. J Am Soc
Hort Sci 106: 220–232.
Shifriss O (1989) Relationship between the B genes of two Cucurbita species, II. Cucurbit Genet
Coop Rep 12: 75–78.
Shiffriss O (1990) Relationship between the B genes of two Cucurbita species, III. Cucurbit
Genet Coop Rep 13: 50–54.
Shiffriss O (1991) The two B genes of Cucurbita are unlinked. Cucurbit Genet Coop Rep 14:
116–122.
Shiffriss O, Paris HS (1981) Identification of modifier genes affecting the extent of precocious
fruit pigmentation in Cucurbita pepo L. J Am Soc Hort Sci 106: 653–660.
Shridhar, Singh D (1990) Palynology of the Indian Cucurbitaceae. In: DM Bates, RW Robinson,
C Jeffrey (eds) Biology and utilization of the Cucurbitaceae. Cornell Univ Press, Ithaca,
NY, USA, pp 200–208.
Shuler RE, Roulston TH, Farris GE (2005) Farming practices influence wild pollinator
populations on squash and pumpkin. J Econ Entomol 98: 790–795.
Singh D (1949) Inheritance of certain economic characters in the squash, Cucurbita maxima
Duch. MN. Agri Exp Sta Tech Bull 186.
Sinnott EW, Durham GB (1922) Inheritance in the summer squash. J Hered 13: 177–186.
Smith BD (1997) The initial domestication of Cucurbita pepo in the Americas 10,000 years ago.
Science 276: 932–934.
Statistics Canada (2008) http: //www.statcan.gc.ca.
Stephenson AG, Devlin B, Horton JB (1988) The effects of seed number and prior fruit
dominance on the pattern of fruit production in Cucurbita pepo (Zucchini: squash). Ann
Bot 62: 653–661.
Stevenson DG, Yoo S, Hurst PL, Jane J (2005) Structural and physicochemical characteristics
of winter squash (Cucurbita maxima D.) fruit starches at harvest. Carbohydr Polymers
59: 153–163.
Stuart SG, Loy JB (1983) Comparison of testa development in normal and hull-less seeded
strains of Cucurbita pepo L. Bot Gaz 144: 491–500.
Tapley WT, Enzie WD, Van Eseltine GP (1937) The vegetables of New York—The Cucurbits,
vol. I, part 4. J.N. Lyon Co., Albany, New York.
Teppner H (2000) Cucurbita pepo (Cucurbitaceae)—History, seed coat types, thin coated seeds
and their genetics. Phyton 40: 1–42.
Teppner H (2004) Notes on Lagenaria and Cucurbita (Cucurbitaceae)—Review and new
contributions. Phyton 44: 245–308.
USDA (2008) http: //www.usda.gov/QuickStats/index2.jsp
Valdez-Melara M, Garcia A, Delgado M, Gatica-Arias AM, Ramirez-Fonseca P (2009) In vitro
plant regeneration system for tropical butternut squash genotypes (Cucurbita moschata).
Rev Biol Trop 57(suppli. 1): 119–127.
Van Eseltine GP (1936) Cucurbita hybrids. Proc Am Soc Hort Sci 34: 577–581.
Vaulx RD, Pitrat M (1979) Interspecific cross between Cucurbita pepo and C. marinezii. Cucurbit
Genet. Coop Rpt 2: 35.
Vining KJ, Loy JB (1998) Seed development and seed fill in hull-less seeded cultigens of
pumpkin (Cucurbita pepo L.). In: JM McCreight (ed) Cucurbitaceae 98: Evaluation and
enhancement of cucurbit germplasm, ASHS Press, Alexandria, VA USA, pp 64–69.
Wall JR (1954) Interspecific hybrids of Cucurbita obtained by embryo culture. Proc Am Soc
Hort Sci 63: 427–430.
Wall JR (1961) Recombination in the genus Cucurbita. Genetics. 46: 1677–1685.
Wall JR, York TL (1960) Gametic diversity as an aid to interspecific hybridization in Phaseolus
and in Cucurbita. Proc Am Soc Hort Sci 75: 419–428.
Weaver JE, Bruner WE (1927) Root development of vegetable crops. McGraw-Hill, New York,
USA.
Webb SE, Tyson RV (1997) Evaluation of virus-resistant squash varieties. Proc Fla State Hort
Soc 110: 299–302.
Weeden NF, Robinson RW (1986) Allozyme segregation ratios in the interspecies cross Cucurbita
maxima x C. ecuadorensis suggest that hybrid breakdown is not caused by minor alteration
in chromosome structure. Genetics 114: 593–609.
Wessel-Beaver L (1993) Powdery and downy mildew resistance in Cucurbita moschata accessions.
Cucurbit Genet Coop Rep 16: 73–74.
Wessel-Beaver L (1998) Sources of whitefly-induced silvering resistance in Cucurbita. In: JM
McCreight (ed) Cucurbitaceae 98: Evaluation and enhancement of cucurbit germplasm,
ASHS Press, Alexandria, VA, USA, pp 118–124.
Wessel-Beaver L (2000) Evidence for the center of diversity of Cucurbita moschata in Columbia.
Cucurbit Genet Coop Rep 23: 54–55.
Whitaker TW, Davis GN (1962) Cucurbits. Interscience Publ., New York.
Whitaker TW, Robinson RW (1986) Squash breeding. In: MJ Bassett (ed) Breeding vegetable
crops, AVI Publ Boca Raton, FL, USA, pp 209–242.
Whitcomb WD, Garland WJ (1948) Susceptibility of Cucurbitaceae to squash borer. Proc Am
Soc Hort Sci 48: 417–422.
Wisemann BR, Hall CV, Painter RH (1961) Interactions among the cucurbit varieties and feeding
responses of striped and spotted cucumber beetles. Proc Am Soc Hort Sci 78: 379–384.
Wien HC (1997) Correlative growth in vegetables. In: HC Wien (ed) The physiology of vegetable
crops. CABI Publ., Wallingford, England, pp 181–206.
Wien H C, Stapleton SC, Maynard DN, McClurg C, Nyankanga R, Riggs D (2002) Regulation
of female flower development in pumpkin (Cucurbita spp.) by temperature and light. In:
DN Maynard (ed) Cucurbitaceae 2002, ASHS Press, Alexandria, VA, USA, pp 307–315.
Wu T, Zhou J, Zhang Y, Cao J (2007) Characterization and inheritance of a bush-type in tropical
pumpkin (Cucurbita moschata Duchesne). Sci Hort 114: 1–4.
Xiao QB, Loy JB (2007) Inheritance and characterization of a glabrous trait in summer squash.
J Am Soc Hort Sci 132: 327–333.
Young RE (1949) The effect of maturity and storage on germination of butternut squash. Proc
Am Soc Hort Sci 53: 345–346.
Zack CD, Loy JB (1980) The effect of light quality and photoperiod on vegetative growth of
Cucurbita maxima. J Am Soc Hort Sci 105: 939–943.
Zack CD, Loy JB (1981) Effect of fruit development on vegetative growth of squash. Can J
Plant Sci 61: 673–676.
Zhang Y, Anagnostou K, Kyle MM (1995) Seedling screens for resistance to gummy stem blight
in squash. Cucurbit Genet Coop Rep 18: 59–61.
Zhou XL (1987) A study on the breeding of naked kernel pumpkin and its genetic behavior.
Acta Hort Sin 14: 114–118.
Zitter TA, Hopkins DL, Thomas CE (1996) Compendium of cucurbit diseases. APS Press, St.
Paul, MN, USA.
Zraidi A, Stift G, Pachner M, Shojaeiyan A, Gong L, Lelley T (2007) A consensus map for
Cucurbita pepo. Mol Breed 20: 375–388.
ABSTRACT
Cucurbitaceae are among the largest and most diverse plant families.
These include several economically important cucurbits such as
watermelon (Citrullus lanatus), cucumber (Cucumis sativus), melon
(Cucumis melo) and squashes (Cucurbita spp.), but also minor crops
and wild species distributed worldwide. It is assumed to be of Asian
origin, but cucurbits have diversified around the world. In this chapter,
we describe studies that aim to know the extant genetic diversity
among cucurbits. Main tribes, comprising the most important genera
are first described, Benincaseae (Cucumis, Citrullus, Benincasa and
Lagenaria) and Cucurbiteae (Cucurbita). Most of the studies deal with
variability within the main crops of each genus, melon and cucumber
(Cucumis), watermelon (Citrullus), and pumpkins, C. pepo, C. moschata
and C. maxima (Cucurbita), but also studies dealing with minor crops
are included, African horned cucumber (Cucumis metuliferus), pickling
cucumber (Cucumis anguria), white gourd (Benincasa hispida), bottle
gourd (Lagenaria siceraria) and malabar melon (Cucurbita ficifolia). In
addition to the main tribes, we also review Joliffieae, Luffeae and
Sicyeae, including minor crops such as bitter melon (Momordica
charantia), chayote (Sechium edule) and sponge gourd (Luffa cylindrica),
respectively.
For each genus, a general view is provided on the taxonomy, distribution
and intra-genus relationships. For each species, we summarize the
information on taxonomy, morphological studies and molecular assays
using different marker systems. Reported works describe the variability
1
Instituto de Conservación y Mejora de la Agrodiversidada Valenciana (COMAV), Universitat
Politécnica de Valéncia, Camino de Vera 14, Valencia 46022, Spain.
a
e-mail: fnuez@btc.upv.es
b
e-mail: mpicosi@btc.upv.es
*Corresponding author: criesgo@upvnet.upv.es
5.1 Introduction
The family Cucurbitaceae comprises some of the most economically
important crops. Cucurbits have been used by humans as food, containers,
musical instruments and as a source of medicine, for more than 12,000
years (Whitaker and Davis 1962; Brothwell and Brothwell 1969; Lira-Saade
1995).
Jeffrey (1990) and Robinson and Decker-Walters (1997) described 118
genera and about 825 species within the family. However, according to
the most recent classifications (Jeffrey 2005; Jeffrey and De Wilde 2006)
Cucurbitaceae comprises 130 genera, including about 800 species. The
family is divided into two subfamilies: Nhandiroboideae, also called
Zanonioideae (Jeffrey 1990), with 19 genera and about 60 species without
economic value; and Cucurbitoideae, with 111 genera and 740 species.
The subfamily Cucurbitoideae comprises some of the most important
tribes and genera within cucurbits: Benincaseae (Benincasa, Citrullus,
Coccinia, Lagenaria, Cucumeropsis, Cucumis), Luffeae (Luffa), Cucurbiteae
(Cucurbita), Sicyeae (Cyclanthera, Sechium), Joliffieae (Momordica, Telfairia),
Schizopeponeae (Schizopepon), Trichosantheae (Hodgsonia, Trichosanthes),
Bryonieae (Ecballium), Herpetospermae and Coniandreae (Jeffrey 1990;
Rubatzky and Yamaguchi 1997; Jeffrey 2005). There are some recent
phylogenetic studies of the family Cucurbitaceae using chloroplast DNA
sequences (cpDNA) (Kocyan et al. 2007). Molecular data weakly support the
traditional subfamilies Cucurbitoideae and Nhandiroboideae, and recover
most of the 11 tribes, but almost none of the subtribes. Within the subfamily
Cucurbitoideae, a clade comprising all the tribes but Joliffieae (polyphyletic)
was reported as a “fused stamen” clade due to their fused filaments or
connectives. In this clade several tribes clustered together in four groups:
Herpetospermae-Schizopeponeae group, Bryonieae group, Coniandreae-
Benincaseae-Cucurbiteae group and Luffeae-Sicyeae-Trichosantheae group
(Kocyan et al. 2007). The most important diagnostic characters for the
genera and tribes of Cucurbitaceae come from androecium and gynoecium
morphology (number of free styles, fusion of filaments and/or anthers),
tendril type, pollen structure and seed coat. Subfamily Nhandiroboideae
is characterized by free styles, small pollen grains with a striate exine and
branched tendrils with a sensitive basal part. Subfamily Cucurbitoideae
is characterized by having styles united in a single column, tectate to
semitectate pollen with a reticulate or echinate exine and simple, bifid or
multifid tendrils with non-spiralling basal part. These traits correlate quite
well with the phylogeny obtained with molecular data.
The genera Cucumis, Cucurbita and Citrullus include species (cucumber,
melon, watermelon and squash) that are among the most widely cultivated
crops worldwide. Apart from them, there are other notable cucurbits of local
or regional economic importance, such as Lagenaria, Momordica, Benincasa,
Luffa and Sechium (Lira Saade and Montes-Hernández 1994; Bates et al.
1995; Lira-Saade 1995).
Cucurbits have diversified around the world. A large amount of genetic
resources adapted to many different environmental and growing conditions
can be found in different areas. Knowing the extant genetic diversity among
cucurbits is important in order to optimize collection and conservation
programs and to facilitate the ongoing efforts by plant breeders worldwide
to improve melon, cucumber, watermelon and squash with traits from wild
relatives.
Figure 5-1 Distribution map of Cucumis spp. Principal diversity centers based on Kirkbride
(1993).
C. sativus L. and C. sativus L. var. hardwickii (Royle) Alef.
C. hystrix Chakravarty
C. myriocarpus Naudin ▲
C. africanus L. ▲
C. anguria L.
C. dipsaceus Ehrenberg ex Spach
C. zeyheri Sonder
C. ficifolius A. Richard
C. metuliferus E. Meyer ex Naudin
♦
C. melo L. ssp. Melo and C. melo L. ssp. agrestis (Naudin) Pangalo ♦
C. sagittatus Peyritsch
Color image of this figure appears in the color plate section at the end of the book.
2004) and chloroplast SSRs and sequence analysis (Chung et al. 2003, 2006).
One of the most recent studies, conducted by Renner et al. (2007), tested
the genetic relationships among Cucumis, combining cpDNA sequences
and nuclear ITS. This study extended the analysis performed in previous
studies considerably, using representatives of other potentially related
genera (Cucumella, Dicaelospermum, Mukia, Muellerargia, Myrmecosicyos,
and Oreosyce) included in the most recent morphology-based classification
of Cucurbitaceae (Jeffrey 2005). The Cucumis species relationships
found by these authors differ from those found in earlier studies. The
deepest divergence lies between the common ancestor of C. hirsutus and
C. humifructus and the stem lineage of the remainder of the genus. The
authors also concluded that the closest relative of Cucumis is Muellerargia,
and showed that the genera Cucumella, Dicaelospermum, Mukia, Myrmecosicyos
and Oreosyce are grouped among species of Cucumis. Similar results were
obtained by Ghebretinsae et al. (2007) and Schaefer (2007), who proposed
the expansion of the genus Cucumis to include these related genera. Renner
et al. (2007) extend the range of the genus throughout the Malesian region
and into Australia. According to this study, Cucumis comprises an old
Australian/Asian component. Cucumis sativus would have evolved within
this Australian/Asian clade. Cucumis melo is sister to this Australian/Asian
clade, rather than being close to African species as previously thought. In
fact, a more recent research, carried ourt by Sebastian et al. (2010), reports a
species from Australia (Cucumis picrocarpus) as the melon’s closest relative.
According to these studies (Renner et al. 2007; Sebastian et al. 2010), C. melo
might have originated in Asia and then arrived in Africa.
Africa from there (Renner et al. 2007). In fact, wild melons are very frequent
in East and West Africa, but also from Central Asia to India (Whitaker and
Bemis 1976; Staub et al. 1987; McCreight et al. 1993; McCreight and Staub
1993; Rubatzky and Yamaguchi 1997). It is also possible that in Africa and
Asia different domestication events took place (Bates and Robinson 1995),
the most extensive of which may have occurred in Asia (more edible
types). Independent domestication has also been proposed for the two
subspecies.
Melon has suffered an intense process of diversification, and today
shows great variation in morphological and physiological characters.
Primary and secondary centers of genetic diversity are located from eastern
Asia to the Mediterranean Sea (Afghanistan, Iran, Iraq, Saudi Arabia, Turkey,
China, Russia and India) (Robinson and Decker-Walters 1997; Akashi et al.
2002). Seeds of wild C. melo ssp. agrestis and possibly selected forms more
closely resembling currently cultivated C. melo could have been introduced
from Africa into the Middle East (Turkey, Iraq and Iran) and Asia (India,
China, and Japan) through land and sea commerce routes (Kajale 1979;
Walters 1989; Fujishita 1992). It seems that melon was cultivated in Iran
and China 3000 BC, in India 2000 BC, in Egypt 1500 BC and western Japan
as early as 100 BC (Walters 1989; Fujishita 1992; Decker-Walters 1999;
Stepansky et al. 1999; Karchi 2000; Luan et al. 2008). Melon spread through
Asia and subsequently to Europe (Roman-Greek periods) (Szabó et al.
2005). Three independent introductions to Europe (from the east: Russia,
Bulgaria, Hungary; from the south-east: Greece, Albania, Romania; from
the south: Italy) were hypothesized by Pitrat et al. (1999). Later, Columbus
introduced this crop to America, where it became popular and dispersed
quickly, producing a wide range of new cultivars.
C. melo is considered to be the most variable species in the genus
Cucumis (Kirkbride 1993; Bates and Robinson 1995). Great diversity in fruit
shape, size, color and taste exists among melons. This variability was first
classified as different species (C. melo, C. flexuosus, C. dudaim, C. callosus,
C. chate, C. conomon and C. momordica) that now are considered varieties
or morphotypes within the species. Naudin (1859) defined 9 “tribes” of
cultivated melons and one wild form. Many scientists have since added or
merged types (reviewed in Pitrat et al. 2000). Munger and Robinson (1991)
proposed a simplified version of Naudin´s taxonomy, which is still used
in many studies, with seven cultivar groups: agrestis Naud. (wild melon),
cantalupensis Naud. (cantaloupe or muskmelon; Middle East), inodorus
Naud (winter melons, honeydew, Cassaba; Middle East, southern Europe),
conomon Mak. (pickling melon, Chinese white cucumber; Asia), chito-dudaim
Naud (mango melon-queen’s pocket melon; Asia), flexuosus Naud. (snake
melon; Middle East), and momordica (Phoot or snap melon; Asia) (Robinson
and Decker-Walters 1997). More recently, Pitrat et al. (2000) described 16
botanical varieties: var. conomon, var. makuwa, var. chinensis, var. momordica,
var. acidulus (included in ssp. agrestis) and var. cantalupensis, var. reticulatus,
var. adana, var. chandalak, var. ameri, var. inodorus, var. flexuosus, var. chate,
var. tibish, var. dudaim, var. chito (included in ssp. melo). In later revisions,
Pitrat (2008) merged some groups. The description of the main botanical
groups is included in Table 5-1. Many of these botanical groups include
different cultivar-groups that are highly popular in different parts of the
world. Cantalupensis and inodorus are of commercial interest in the United
States, as well as in many European, Mediterranean and Asian countries
(McCreight et al. 1993), and include cultivars belonging to different market
classes. For example, while the popular market classes Charentais, Shipper,
Ogen and Galia are in the cantalupensis group, the inodorus group houses
an array of Cassaba market class types (e.g., Rochet, Piel de Sapo, Tendral,
Crensahw, Honeydew, Kirkagac and Yellow Canari).
Variety Distribution Sex type Fruit (shape/ Flesh color Rind Sweetness Aroma Climateric
color)
CONOMON Eastern Asia Andromonoecious Elongated White Smooth, thin No No No
148
Variety Distribution Sex type Fruit (shape/ Flesh color Rind Sweetness Aroma Climateric
RETICULATUS Europe, Asia, Andromonoecious Round-oval Orange Netted Yes Yes Yes
North and (sometimes
South America ribbed)
AMERI Western and Andromonoecious Elongated-oval White-light Slightly Yes Little Yes
Central Asia Yellow-light orange netted
green
Melon has become a model within Cucurbits for genetic and genomic
studies (see Chapter 9). Different initiatives are generating sequence
collections (González et al. 2010, González-Ibeas et al. 2007, http://www.
melogen.upv.es; www.icugi.org) which are facilitating single nucleotide
polymorphism (SNP) detection. Morales et al. (2004) first detected SNPs
in melon using a small set of available ESTs. These authors indicated an
average frequency of 1 SNPs per 441 bp between two inodorus genotypes.
In a more complete study, screening 30,000 ESTs sequences from four
genotypes, 356 high-quality SNPs were found (González-Ibeas et al. 2007).
Some of them proved to study diversity in a set of melon accessions, giving
genetic relationships similar to that found with SSRs (Deleu et al. 2009).
In addition to genetic diversity studies, these markers have been used to
construct several melon maps (Baudracco-Arnas and Pitrat 1996; Wang et
al. 1997; Liou et al. 1998; Brotman et al. 2000; Oliver et al. 2001; Danin-Poleg
et al. 2002; Périn et al. 2002; Gonzalo et al. 2005; Deleu et al. 2009) that are
being merged within the International Cucurbit Genomics Initiative (ICuGI)
(http://www.icugi.org) (see “Molecular maps” in Chapter 6 and Chapter 9
on melon genetic maps) . The availability of increasing amounts of melon
sequences is allowing the application of new methods to study variability
in candidate genes of breeding interest (Nieto et al. 2007). In a recent study
reported by Esteras et al. (2009b) EcoTILLING techniques are being applied
to study polymorphisms in genes involved in quality and ripening processes
using a highly variable core collection of melons.
Molecular markers have been used to characterize elite melon
germplasm (commercial cultivars, hybrids and breeding lines, mainly
from the USA and European markets) mostly belonging to cantalupensis
(Charentais, Shipper, European, western and eastern USA types, Galia and
Ogen) and inodorus (Honey dew type and Cassaba Rochet, Piel de Sapo
and Yellow Canari types) types. Despite the fact that molecular analysis
discriminates between these market cultivars, the groupings were somewhat
ambiguous, most likely due to intogressions during plant breeding. They
also found a limited genetic diversity in some groups (Garcia et al. 1998;
Staub et al. 2000).
Much higher genetic diversity is reported when exotic germplasm (wild,
feral, landraces) from these and other botanic groups is also included. Most
of the molecular studies support quite well the division into two major
groups (ssp. melo and ssp. agrestis) (Staub et al. 1997; Silberstein et al. 1999;
Danin-Poleg et al. 2001; Monforte et al. 2003; Nakata et al. 2005; Deleu
et al. 2009; Esteras et al. 2009a). In general, higher molecular variability
(number of alleles and polymorphic loci) is reported in Central Africa and
India than in the extremes of melon distribution (the Mediterranean area
and China Sea). One of the most complete studies covering representatives
of most of the botanical groups was conducted by Stepansky et al. (1999).
They combined both phenotypic and molecular data (RAPDs and ISSRs),
analyzing accessions from 23 countries, including wild, feral and cultivated
forms representing the primary and secondary centers of diversity (Africa,
southern and western Asia and the Far East). Phenotyping was based on
a set of traits usually employed to define different botanical groups (seed
size, stem thickness, pubescence, sex type, ovary shape, ovary pubescence,
fruit shape and size, skin color, texture and design, splitting, abscission,
external aroma, flesh color, taste, sucrose, glucose and fructose and pH).
According to these traits, the subdivision into most of the varietal groups
persisted, indicating that the traditional classification is mainly based on
consistent and highly informative characters. Some traits with taxonomic
value show high variability. For example, wild types defined by small fruit
size, thought to be agrestis, can show ovaries with long or short hairs. Also
within the flexuosus types there exist both kinds of ovaries. The molecular
results did not substantially contradict the phenotype-based dendrogram.
The sweet-fruited cantalupensis and inodorus clustered together, in spite of
their ripening differences, and the nonsweet varieties agrestis, conomon and
momordica grouped together. There exist discrepancies in the classification
of some botanical groups. For example, dudaim and chito cultivars often are
grouped with agrestis types, although they have been reported to belong to
ssp. melo. The flexuosus types clustered closer to the non-sweet genotypes in
the phenotypic tree, and dispersed with the “dessert” ones in the molecular
tree. The non existence of reproductive barriers within the species makes
the crosses among the different cultivar groups possible, giving rise to a
continuous distribution-pattern of variation. Esteras et al. (2009a) found
similar results in a recent analysis with AFLPs of a melon core collection
of 212 accessions, representing all the genetic diversity of the species, with
dudaim and chito types being intermediate between ssp. melo and agrestis and
momordica and flexuosus types which, being highly variable, interspersed
among both groups.
length to diameter ratio) (Horejsi and Staub 1999). RAPDs have also been
successful in discriminating elite accessions, but have detected limited
genetic diversity (Bernet et al. 2003; Duca et al. 2008; Onto et al. 2008).
Similar to what occurred in melons, several authors have made a big
effort to develop cucumber SSRs. Katzir et al. (1996) reported seven highly
polymorphic genomic SSRs in cucumber and melon. Later, Danin-Poleg et
al. (2001), Fazio et al. (2002) and Kong et al. (2006) increased the number of
microsatellites. SSR-enriched genomic libraries have also been developed
to increase SSR availability (Fukino et al. 2008; Watcharawongpaiboon and
Chunwongse 2008). Transferability to melon, bitter gourd, watermelon and
pumpkin has been assessed. These markers have been tested in diversity
studies using a different set of cucumber cultivars and have also been used
for constructing genetic maps (Park et al. 2000; Bradeen et al. 2001; Fazio
et al. 2003) (see “Molecular maps” in Chapter 6). EST-derived SSRs have
also been used (Kong et al. 2006). The draft genome sequence of Cucumis
sativus var. sativus was recently published. The availability of this sequence
will facilitate the high-throughput discovery of new markers, such as SNPs
(Huang et al. 2009).
In general, molecular studies report a low degree of genetic diversity
within C. sativus var. sativus compared to other cross-fertilized species of
the genus, such as melons (Dane 1976, 1983; Esquinas-Alcazar 1977; Knerr
et al. 1989; Dijkhuizen et al. 1996; Horejsi et al. 1999), but they also describe
higher levels of polymorphisms in var. hardwickii and consistently separate
both varieties (Dijkhuizen et al. 1996; Meglic et al. 1996; Horejsi et al. 1999).
Indeed, Staub et al. (2005) tried to identify a useful reference marker array
(from a set of 155 markers, SSRs and Sequence Characterized Amplified
Region (SCARs)) in order to distinguish very closely related varieties and
elite breeding lines, which is essential for variety protection. They found
difficulties in discriminating this genetic material suggesting that other
markers, such as SNPs are needed to better define these cultivars.
5.2.1.3.2 C. anguria
C. anguria L., commonly called “pickling cucumber”, has been considered
to have originated in America, but now it is supposed to come from tropical
Africa, having been introduced into the New World by African slaves
(reviewed by Baird and Thieret 1988). This species has been screened for
resistances to several diseases, like powdery and downy mildew (Nikolova
et al. 2002) and increased yield (Oliveira et al. 2009). Crosses between
C. anguria and C. anguria var. longaculeatus have been attempted to obtain
elite lines (Modolo and Costa 2003). Some phylogenetic studies on
Cucurbitaceae have also included this species, as well as C. myriocarpus
Naudin, to better represent genus Cucumis, but few accessions have been
assayed (Renner et al. 2007; Singh and Matta 2008). C. myriocarpus, which
has toxic fruits, has also been investigated for resistances, having been
reported as resistant to CVYV (Marco et al. 2003), and adequate mineral
ratios (Flyman and Afolayan 2007).
Figure 5-2 Distribution map of Citrullus spp. Principal diversity centers based on Jeffrey
(2001).
C. colocynthis (L.) Schrad.
C. ecirrhosus Cogn.
C. lanatus ((Thunb.) Matsum & Nakai) var. lanatus ▲ and var. citroides ▲
C. rehmii De Winter ♦
Color image of this figure appears in the color plate section at the end of the book.
5.2.2.1.Citrullus lanatus
5.2.2.1.1 Origin and Taxonomy
C. lanatus ((Thunb.) Matsum & Nakai) includes wild, cultivated and feral
forms. Fursa (1972) described three subspecies: ssp. vulgaris (divided into
var. vulgaris and var. cordophanus, including red sweet fruited cultivated
forms), ssp. lanatus (including tsamma types from the Kalahari Desert (var.
caffer), and the citron) and ssp. mucosospermus (including the egusi types
from West Africa). Recently, the species has been reclassified and divided
into two botanical varieties: var. lanatus (the cultivated forms, including
egusi types), distributed in tropical and subtropical regions worldwide, and
var. citroides (Bailey) Mansf. (including citron and tsamma types), which
grows in southern Africa.
Despite the numerous studies, the origin, distribution and domestication
of this species still remains unclear. Jeffrey (1967, 2001) and Zeven and
Zhukovsky (1975) proposed var. caffer as the ancestor of the species while
Navot and Zamir (1987) proposed the var. citroides. Other theories propose
C. colocynthis or, as previously noted, C. ecirrhosus (Dane and Liu 2007).
Primitive watermelons are supposed to have had nonsweet, bitter, white-
fleshed fruits, similar to those of citron or colocynth. Today it is generally
accepted that watermelon originated in Africa where it reaches maximum
diversity (DeCandolle 1883). The two putative ancestors, var. citroides and
C. colocynthis, can be found growing wild in Africa.
Different regions in Africa have been postulated as centers of origin
of this species: southern Africa, principally around the Kalahari Desert
(Meeuse 1962; Esquinas-Alcázar and Gulick 1983), Central Africa (Mallick
and Masui 1986) and northern Africa (Keay and Hepper 1985). In fact,
watermelon seeds (about 5,000 years old) recently discovered at an
archaeological site in southwest Libya (Wasylikowa and van der Veen 2004)
support northern Africa as the most probable domestication center. There
is evidence of watermelon cultivation in the Nile Valley by 2000 BC, when
southwest Africans did not yet practice farming (Zohary and Hopf 2000).
Colocynth seeds have been found at early sites in Egypt, Libya and the
Near East, indicating that they could have been used first. Some authors
assume that cultivation of watermelon began in ancient Egypt and India,
from where it spread to the Mediterranean area, Near East and Asia. The
Romans introduced this crop to Europe, and later the Muslims increased the
number of varieties on the continent. However, watermelon did not become
as popular there as it did in China where it arrived in the 10th–12th centuries.
Subsequently, watermelon reached America (17th century) (Rubatzky 2001;
Wehner 2008). Today, southern Africa and, to some extent, western Africa,
are considered primary centers of diversity. China constitutes a secondary
center of diversity, whereas a great variety of landraces and wild accessions
can also be found in other regions; India, where C. colocynthis grows wild,
the Middle East and countries in the Mediterrranean basin.
whose fruits present bitterness and firm white flesh. Its seeds are coated
by an adherent layer of tissues. This type is used for its nutritional seeds
and for cattle consumption.
Despite this morphological variability, molecular variation is limited in
commercial cultivars (Jarret et al. 1997; Maggs-Kolling et al. 2000; Levi et al.
2004; Levi and Thomas 2005). This poor variability can be increased using
germplasm from the diversification areas. Germplasm of Citrullus from
China and South Africa is represented in genebank collections, but India,
south, southwest and tropical Africa and the southern areas of the former
USSR and Iran are still priorities for collection (Wehner 2008).
Molecular markers have been used to estimate genetic relatedness of
watermelon cultivars, and can be used to evaluate inbred lines for purity.
Studies with isozymes (Zamir et al. 1984; Navot and Zamir 1987; Biles et al.
1989; Walters et al. 1991) reflect a low degree of genetic diversity, and only a
few informative isozyme markers are available. Diversity within C. lanatus
(20 cultivated, 70 citron watermelon and reference types) has been recently
assessed by Dane and Liu (2007) by using PCR-RFLPs and sequencing of
cpDNA of several non-coding regions. RAPD markers were more efficient
at detecting genetic variation (Hashizume et al. 1993; Zhang et al. 1994). Lee
et al. (1996) used RAPDs with a representative collection of watermelon
cultivars, obtaining four groups. Molecular clusters correlated with fruit
quality traits such as sugar content. Moreover, Levi et al. (2001b) used this
marker system to characterize a collection of C. lanatus and C. colocynthis
exhibiting several disease resistances. They found three clusters, one with
watermelon cultivars, one with the C. lanatus var. citroides accessions, and
the third with C. colocynthis accessions. Levi and Thomas (2005) performed
their study using citoplasmatic markers, with which they also obtained
a clear differentiation of the accessions belonging to var. lanatus (the five
cultivars assayed), var. citroides and C. colocynthis as previously reported.
They found a closer relationship between citroides and colocynth vs lanatus
and colocynth. Levi et al. (2001a) found higher levels of variability in
C. colocynthis and var. citroides than in cultivated watermelon and differentiated
some watermelon accessions with introgressions from var. citroides. Similar
studies have been performed with ISSRs and AFLPs (Levi et al. 2004). This
study showed that AFLPs and ISSRs are highly informative and much more
efficient at differentiating between American heirloom cultivars with a
narrow genetic base. AFLPs were also successfully used to detect variability
among watermelon cultivars (Ke-peng et al. 2003). The polymorphism rate
detected with this kind of markers proved higher than with other previously
tested kinds, such as isozymes, and three groups were obtained among the
30 genotypes surveyed. Although low genetic diversity was found, classical
American ecotypes, breeding and selected lines and cultivars originating
from Japanese and Chinese pedigrees could be differentiated.
Figure 5-3 Distribution map of Cucurbita spp. Principal diversity centers based on Lira-Saade
(1995) and Sanjur et al. (2002).
C. argyrosperma C. Huber
C. ficifolia Bouché
C. maxima Duchesne ▲
C. moschata Duchesne ▲
C. pepo L.
C. ecuadorensis
C. okeechobeensis (J.K. Small) L.H. Bailey
C. lundelliana L.H. Bailey ♦
C. digitata A. Gray, C. cylindrata L.H. Bailey and C. palmata S. Watson ♦
C. foetidissima Kunth ♦
Color image of this figure appears in the color plate section at the end of the book.
sequences. They found extensive allele sharing among these species, which
led to an inconclusive phylogenetic analysis, suggesting a high frequency of
introgression during domestication or polyploidization events in the genus.
Analysis using ISSRs and microsatellite DNAs has also been reported (King
et al. 1995; Katzir et al. 2000a, b).One of the most relevant studies has been
that by Sanjur et al. (2002), using mtDNA, which suggests six independent
domestication events from distinct wild ancestors.
Cocozelle Europe (Italy), Far East, Long, bulbous fruits Smooth Immature
Turkey, Yugoslavia Striped, light green non-striped, Female
ribbed or non ribbed flowers
172
Morphotype Distribution Fruit (shape/color) Rind Consumption
Color image of this table appears in the color plate section at the end of the book.
report a high variability within the species. When different species are
compared, allelic diversity is greatest in C. pepo and C. moschata. Katzir
et al. (2000a), using SSRs and ISSRs grouped the cultigens of C. pepo ssp.
ovifera more-or-less according to fruit shape, whereas in C. pepo ssp. pepo,
subclustering differentiated the cocozelle group from the zucchini group.
Some of the SSR loci used for Cucurbita analysis were transferred from
Cucumis, as only a few microsatellites were available for Cucurbita. Paris
et al. (2003) attempted to apply cucumber and melon SSRs described
by Katzir et al. (1996) and Danin-Poleg et al. (2001) attempted to assess
diversity in C. pepo. A set of 102 Cucumis-SSR primers developed by Fazio
et al. (2002) were also proved in this species for mapping, but none turned
out to be polymorphic. Sixty Cucumis SSRs (gSSRs and EST-SSRs) were
tested in Cucurbita spp. accessions, but over 63% did not amplify in any
of them (Picó et al. 2005-2006). Transferability from Cucumis EST-SSRs
has also been assayed (Fernández-Silva et al. 2008), but the fact that these
markers are located in expressed regions of the genome did not increase the
transferability rate and only 5.4% were polymorphic. Therefore, SSRs are
not easily transferable between genera within Cucurbitaceae, and because
of this, a wide collection of SSRs has been developed recently using SSR-
enriched partial genomic libraries from C. pepo ssp. pepo and C. moschata.
These markers show high inter-species transferability and have been used
to construct the first published C. pepo map (Zraidi et al. 2007; Gong et al.
2008a, b). Stift et al. (2004) had already reported a better transferability rate
from C. pepo-SSRs to C. moschata, C. maxima and C. ecuadorensis and Gong
et al. (2008a) found with their new set of 500 SSRs (from SSR-enriched
partial genomic libraries) a higher percentage of C. pepo markers transferred
to C. ecuadorensis in comparison to C. moschata, which implies a closer
relatedness between C. pepo and C. ecuadorensis than between C. moschata
and C. ecuadorensis.
Two of the most complete studies performed to date in C. pepo are those
by Paris et al. (2003) and Ferriol et al. (2003a). Paris et al. (2003) assayed a set
of C. pepo accessions belonging to the three subspecies with three different
marker systems: AFLPs, ISSRs and SSRs, finding a high correlation between
them. Whereas previous studies describe high levels of variation in wild
genotypes (Decker-Walters et al. 2002b), Paris et al. (2003) found higher
variation among domesticated than among wild populations. In general,
their results were coherent with botanical and horticultural classification
and with other studies with allozymes (Ignart and Weeden 1984) and DNA
markers (Torres Ruiz and Hemleben 1991; Katzir et al. 2000a). Clustering
agreed with the division into ssp. pepo, ssp. ovifera (syn. texana) and ssp.
fraterna. Subspecies fraterna and ovifera appeared more closely related to each
other than to ssp. pepo. Results show that the cultivar-groups are genetically
quite distinct. In fact subclusters within ssp. ovifera were in accordance
recently few SSR markers have been developed for C. moschata. In this
genus, most molecular tools had been generated mainly for C. pepo or
transferred from more important species in the family such as Cucumis
melo. The inter-genus transferability was quite low, both for genomic and
EST-SSRs (Picó et al. 2005-2006; Watcharawongpaiboon and Chunwongse
2007; Fernández-Silva et al. 2008). Recently, Gong et al. (2008a) developed
a set of 500 SSRs from SSR-enriched partial genomic libraries of C. pepo
and C. moschata, reporting a high interspecific transferability between both
species. These newly developed SSRs have been used to construct the first
SSR-based map for C. moschata (Gong et al. 2008b). A high level of macrosynteny
was found comparing both C. pepo and C. moschata maps, revealing that
the transferability of markers may be more easily accomplished (see
Chapter 8).
from America. Spain acted as a bridge between America and Europe after
the discovery of the continent, but other types first arrived in Australia,
Africa and Asia where they diversified (secondary centers) and were later
exported to Europe. However, it seems that commercial types were selected
in the USA from materials collected in South America.
5.3.1.4.2 C. argyrosperma
C. argyrosperma C. Huber (syn. C. mixta Pang.) was first considered a
different species from C. moschata by Pangalo (1930). It is believed that its
domestication took place in southwestern and Central Mexico, since the
oldest archeological remains found in different caves of the region dated
back between 3085 and 115 BC (Merrick 1990; Smith 2005). The wild ancestor
of cultivated forms (ssp. argyrosperma) is supposed to be C. argyrosperma
ssp. sororia (L.H. Bailey) Merrick and Bates (Sanjur et al. 2002), which is
distributed from Mexico to Central America.
Decker-Walters and Walters (2000) described a low variability within
the species with few commercial cultivars existing. Due to the poor quality
of the flesh, many cultivars are only grown for their seeds and for cattle
consumption. Within ssp. argyrosperma (cultivated forms) three varieties
may be distinguished: var. argyrosperma (the more primitive one, commonly
striped with bright color), var. callicarpa (the most recent and variable one,
comprising most of the commercial cultivars and landraces) and stenosperma
(striped fruits, mainly used for its edible seeds).
(wild, free-living types). A more recent work (Kole et al. 2010) with AFLPs,
fruit traits and three healthy compounds (cucurbitacin-C, charantin and
plant insulin), reported a high genetic diversity among 22 genotypes from
six countries. The knowledge of the phytomedicinal compounds variation
in bitter gourd germplasm and the pathways and genes involved, makes
this species a model for the new phytomedomics era.
With regard to within-genus studies, the most recent one is a DNA
sequence phylogenetic research that includes 58 Momordica species (Schaefer
and Renner 2010). They conclude this genus is monophyletic and consists
of 11 well-supported clades.
Acknowledgements
The authors acknowledges support from the INIA projects RTA2008-00035-
C02-02 and RF2008-00003-C02-02.
References
Abdelnour A, Rocha OJ (2008) Genetic characterization of a collection of chayote, Sechium
edule (Jacq.) Swartz, in Costa Rica by using isozyme markers. Genet Resour Crop Evol
55: 163–170.
Akashi Y, Fukunda N, Wako T, Masuda M, Kato K (2002) Genetic variation and phylogenetic
relationships in East and South Asian melons, Cucumis melo L., based on the analysis of
five isozymes. Euphytica 125: 385–396.
Akashi Y, Tanaka K, Nishida H, Kato K, Khaning MT, Yi SS, Chou TT (2006) Genetic diversity
and phylogenetic relationship among melon accessions from Africa and Asia revealed
by RAPD analysis. In: GJ Holmes (ed) Proc of Cucurbitaceae 2006. Asheville, North
Carolina, USA, pp 317–325.
Ashurmetov OA (1995) On morphology and taxonomy of the genera Cucumis L. and Melo
Mill. FEDDES Rep 106: 155–159.
Baird JR, Thieret JW (1988) The Bur Gherkin (Cucumis anguria var. anguria, Cucurbitaceae).
Econ Bot 42(3): 447–451.
Baranek M, Stift G, Vollmann J, Lelley T (2000) Genetic diversity within and between the
species Cucurbia pepo, C. moschata and C. maxima as revealed by RAPD markers. Cucurbit
Genet Coop Rep 23: 73–77.
Bates DM, Robinson RW (1995) Cucumbers, melons and water-melons. In: J Smartt, NW
Simmonds (eds) Evolution of Crop Plants. 2nd edn. Longman Scientific, Harlow, Essex,
UK, pp 89–96.
Bates DM, Merrick LC, Robinson RW (1995) Minor cucurbits—Benincasa, Lagenaria, Luffa,
Sechium, and other genera (Cucurbitaceae). In: J Smartt, NW Simmonds (eds) Evolution
of Crop plants. 2nd edn. Longman Scientific, Harlow, Essex, UK, pp 105–111.
Baudracco-Arnas S, Pitrat M (1996) A genetic map of melon (Cucumis melo L.) with RFLP, RAPD,
isozyme, disease resistance and morphological markers. Theor Appl Genet 93: 57–64.
Beharav A, Cohen Y (1995) Attempts to overcome the barrier of interspecific hybridization
between Cucumis melo and C. metuliferus. Isr J Plant Sci 43(2): 113–123.
Behera TK, Dey SS, Sirohi PS (2006) ‘DBGy-201’ and ‘DBGy-202’: two gynoecious lines in
bitter gourd (Momordica charantia L.) isolated from indigenous source. Indian J Genet
66: 61–62.
Behera TK, Singh AK, Staub JE (2008) Comparative analysis of genetic diversity in Indian
bitter gourd (Momordica charantia L.) using RAPD and ISSR markers for developing crop
improvement strategies. Sci Hort 115: 209–217.
Bemis WP, Rhodes AM, Whitaker TW, Carmer SG (1970) Numerical taxonomy applied to
Cucurbita relationships. Am J Bot 57(4): 404–412.
Bernet GP, Bramardi S, Calvache D, Carbonell EA, Asins MJ (2003) Applicability of molecular
markers in the context of protection of new varieties of cucumber. Plant Breed 122:
146–152.
Biles CL, Martyn RD, Wilson HD (1989) Isozymes and general proteins from various
watermelon cultivars and tissue types. HortScience 24: 810–812.
Bisht IS, Bhat KV, Tanwar SPS, Bhandari DC, Joshi K, Sharma AK (2004) Distribution and
genetic diversity of Cucumis sativus var. Hardwickii (Royle) Alef in India. J Hort Sci
Biotechnol 79(5): 783–791.
Bradeen JM, Staub JE, Wye C, Antonise R, Peleman J (2001) Towards an expanded and
integrated linkage map of cucumber (Cucumis sativus L.). Genome 44: 111–119.
Brothwell D, Brothwell P (1969) Food in Antiquity: A Survey of the Diet of Early Peoples.
Thames and Hudson (eds) Fredrick A. Praeger Publ, New York, USA.
Brotman Y, Silberstein L, Kovalski J, K1ingler J, Thompson G, Katzir N, Perl-Treves R (2000)
Linkage groups of Cucumis melo, including resistance gene homologues and known genes.
In: N Katzir, HS Paris (eds) Proc of Cucurbitaceae 2000, Ma’aleh Hahamisha, Israel, 19–23
March 2000, Acta Hort 510: 441–44.
Cadena-Iñiguez J, Avendaño-Arrazate CH, Soto-Hernández M, Ruiz-Posadas LM, Aguirre-
Medina JF, Arévalo-Galarza L (2008) Infraspecific variation of Sechium edule (Jacq.) Sw.
in the state of Veracruz, Mexico. Genet Resour Crop Evol 55: 835–847.
Canul J, Ramírez P, Castillo F, Chávez JL (2005) Diversidad morfológica de calabaza cultivada
en el centro-oriente de Yucatán, México. Revista Fitotecnia Mexicana. Sociedad Mexicana
de Citogenética. AC. Chapingo, México, 28(004): 339–349.
Carle RB, Maynard DN, Wessel-Beaver L (2000) Tropical pumpkin hybrid development:
landraces to hybrid cultivars. In: N Katzir, HS Paris (eds) Proc of Cucurbitaceae 2000,
Ma’aleh Hahamisha, Israel, 19–23 March 2000, Acta Hort 510: 95–100.
Castetter EF (1925) Horticultural groups of cucurbits. Proc Am Soc Hort Sci 22: 338–340.
Chen JF, Staub JE, Tashiro Y, Isshiki S, Miyazaki S (1997a) Successful interspecific hybridization
between Cucumis sativus L. and Cucumis hystrix Chakr. Euphytica 96: 413–419.
Chen JF, Isshiki S, Tashiro Y, Miyazaki S (1997b) Biochemical affinities between Cucumis hystrix
Chakr and two cultivated Cucumis species (C. sativus L. and C. melo L.) based on isozyme
analysis. Euphytica 97: 139–141.
Chiba N, Suwabe K, Numone T, Hirai M (2003) Development of microsatellite markers in
melon (C. melo L.) and their application to major cucurbit crops. Breed Sci 53: 21–27.
Chung HD, Youn SJ, Choi YJ (1998) Ecological and morphological characteristics of the Korean
native squash (Cucurbita moschata). J Kor Soc Hort Sci 39(4): 377–384.
Chung S, Decker-Walters DS, Staub JE (2003) Genetic relationships within the Cucurbitaceae as
assessed by consensus chloroplast simple sequence repeats (ccSSR) marker and sequence
analyses. Can J Bot 81: 1–19.
Chung SM, Staub JE, Chen JF (2006) Molecular phylogeny of Cucumis species as revealed
by consensus chloroplast SSR marker length and sequence variation. Genome 49:
219–229.
Costa J, Catalá MS, Cortés C, Nuez F, Abadía J, Cuartero J (1989) Evaluación de la variabilidad
en los principales tipos de melón cultivados en España. Invest Agri Prod Prot Veg 4:
43–57 (in Spanish).
Cross HB (2003) Evolution, systematics, and domestication in Sechium and genera related
(Sicyeae, Cucurbitaceae). PhD Thesis, Columbia Univ, NY, USA, 182 p.
Cross HB, Lira R, Motley TJ (2006) Origins and diversification of Chayote. In: TJ Motley, NJC
Zerega, HB Cross (eds) Darwin’s Harvest: New Approaches to the Origins, Evolution,
and Conservation of Crops. Columbia Univ Press, New York, USA, pp 171–194.
Cutler H, Whitaker T (1967) Cucurbits from the Tehuacan caves. In: DS Byers (ed) The
Prehistory of the Tehuacan Valley, Environment and Subsistence, vol 1. Univ of Texas
Press, Austin, TX, USA, pp 212–219.
Dane F (1976) Evolutionary studies in the genus Cucumis. PhD Dissert. Colorado State Univ,
Fort Collins, TX, USA.
Dane F (1983) Cucurbits. In: SD Tanksley, TJ Orten (eds) Isozymes in Plant Genetics and
Breeding, part B. Elsevier, Amsterdam, The Netherlands, pp 369–390.
Dane F (2002) Chloroplast DNA investigations in Citrullus using PCR-RFLP analysis. In: DN
Maynard (ed) Cucurbitaceae 2002. ASHS Press, Naples, FL, USA, pp 100–108.
Dane F, Lang P (2004) Sequence variation at cpDNA regions of watermelon and related
wild species: implications for the evolution of Citrullus haplotypes. Am J Bot 91(11):
1922–1929.
Dane F, Liu J (2007) Diversity and origin of cultivated and citron type watermelon (Citrullus
lanatus). Genet Resour Crop Evol 54: 1255–1265.
Dane F, Lang P, Bakhtiyarova R (2004) Comparative analysis of chloroplast DNA variability
in wild and cultivated Citrullus species. Theor Appl Genet 108: 958–966.
Danilchenko H, Paulauskiene A, Dris R, Niskanen R (2000) Biochemical composition and
processability of pumpkin cultivars. In: N Katzir, HS Paris (eds) Proc of Cucurbitaceae
2000, Ma’aleh Hahamisha, Israel, 19–23 March 2000. Acta Hort 510: 493–497.
Danin-Poleg Y, Reis N, Tzuri G, Katzir N (2001) Development and characterisation of
microsatellite markers in Cucumis. Theor Appl Genet 102: 61–72.
Danin-Poleg Y, Tadmor Y, Tzuri G, Resi N, Hirschberg J, Katzir N (2002) Construction of a
genetic map of melon with molecular markers and horticultural traits, and location of
genes associated with ZYMV resistance. Euphytica 125(3): 373–384.
Davis CC, Fritsch PW, Li J, Donoghue MJ (2002) Phylogeny and biogeography of Cercis
(Fabaceae): evidence from nuclear ribosomal ITS and chloroplast ndhF sequence data.
Syst Bot 27: 289–302.
DeCandolle A (1883) Origine des Plantes Cultivées. Baillière, Paris, France.
DeCandolle A (1959) Origin of Cultivated Plants. (Reprint of the 2nd edn, 1886). Hafner Publ,
New York, USA 478 p.
Decker DS (1985) Numerical analysis of allozyme variation in Cucurbita pepo. Econ Bot 39(3):
300–309.
Decker DS (1988) Origin(s), evolution, and systematics of Cucurbita pepo (Cucurbitaceae). Econ
Bot 42(1): 4–15.
Decker-Walters DS (1999) Cucurbits, Sanskrit, and the Indo-Aryas. Econ Bot 53: 98–112.
Decker-Walters DS, Walters TW (2000) Squash. In: KF Kipple, KC Ornelas (eds) The Cambridge
World History of Food. Cambridge Univ Press, New York, USA, pp 335–351.
Decker-Walters DS, Walters TW, Posluszny U, Kevan PG (1990) Genealogy and gene flow
among annual domesticated species of Cucurbita. Can J Bot 68: 782–789.
Decker-Walters DS, Walters TW, Cowan CW, Smith BD (1993) Isozymic characterization of
wild populations of Cucurbita pepo. J Ethnobiol 13: 55–72.
Decker-Walters D, Staub J, López-Sesé A, Nakata E (2001) Diversity in landraces and cultivars
of bottle gourd (Lagenaria siceraria; Cucurbitaceae) as assessed by random amplified
polymorphic DNA. Genet Resour Crop Evol 48: 369–380.
Decker-Walters DS, Chung SM, Staub JE, Quemada HD, López-Sessé AI (2002a) The origin
and genetic affinities of wild populations of melon (Cucumis melo, Cucurbitaceae) in
North America. Plant Syst Evol 233: 183–197.
Decker-Walters DS, Staub JE, Chung SM, Nakata E, Quemada HD (2002b) Diversity in free-
living populations of Cucurbita pepo (Cucurbitaceae) as assessed by random amplified
polymorphic DNA. Syst Bot 27(1): 19–28.
Deleu W, Esteras C, Roig C, González-To M, Fernández-Silva I, Gonzalez-Ibeas D, Blanca J,
Aranda MA, Arús P, Nuez F, Monforte AJ, Picó B, Garcia-Mas J (2009) A set of EST-SNPs
for map saturation and cultivar identification in melon. BMC Plant Biol 9: 90.
Demesure B, Sozi N, Petit RJ (1995) A set of universal primers for amplification of polymorphic
non-coding regions of mitochondrial and chloroplast DNA in plants. Mol Ecol 4:
129–131.
De Veaux JS, Schultz EB Jr (1985) Development of buffalo gourd (Cucurbita foetidissima) as a
semiaridland starch and oil crop. Econ Bot 39: 454–472.
De Winter B (1990) A new species of Citrullus (Benincaseae) from the Namib Desert, Namibia.
Bothalia 20: 209–211.
Dey SS, Singh AK, Chandel D, Behera TK (2006) Genetic diversity of bitter gourd (Momordica
charantia L.) genotypes revealed by RAPD markers and agronomic traits. Sci Hort 109:
21–28.
Dhillon NPS, Ranjana R, Singh K, Eduardo I, Monforte AJ, Pitrat M, Dhillon NK, Singh PP
(2007) Diversity among landraces of Indian snapmelon (Cucumis melo var. momordica).
Genet Resour Crop Evol 54: 1267–1283.
Dhillon NPS, Singh J, Fergany M, Monforte AJ, Sureja AK (2009) Phenotypic and molecular
diversity among landraces of snapmelon (Cucumis melo var. momordica) adapted to the
hot and humid tropics of eastern India. Plant Genetic Resources: Characterization and
Utilization 7(3): 291–300 doi: 10.1017/S1479262109990050.
Dijkhuizen A, Kennard WC, Havey MJ, Staub JE (1996) RFLP variation and genetic relationships
in cultivated cucumber. Euphytica 90: 79–87.
Duca M, Port A, Leivitchi A (2008) Characteristics of RAPD markers in breeding of Cucumis
sativus L. Roum Biotechnol Lett 13(4): 3843–3850.
Ellul P, Lelivelt C, Naval MM, Noguera FJ, Sanchez S, Atarés A, Moreno V, Corella P, Dirks R
(2007) Watermelon. In: T Nagata, H Lörz, JM Widholm (eds) Biotechnology in Agriculture
and Forestry, vol 60. Transgenic Crops V EC Pua, MR Davey (eds) Springer-Verlag Berlin,
Heidelberg, Germany, pp 129–165.
Engels JMM (1983) Variation in Sechium edule in Central America. J Am Soc Hort Sci 108:
706–707.
Erwin AT (1931) Nativity of the cucurbits. Bot Gaz 91: 105–108.
Escribano S, Lázaro A, Staub JE (2008) Genetic diversity of Spanish melons (Cucumis melo)
of the Madrid provenance. In: M Pitrat (ed) Cucurbitaceae 2008, Proc IX EUCARPIA
Meeting on Genetics and Breeding of Cucurbitaceae, INRA, Avignon, France, 21–24
May 2008, pp 301–305.
Esquinas-Alcázar JT (1977) Alloenzyme variation and relationships in the genus Cucumis. PhD
Diss. Univ of California, Davis, USA.
Esquinas-Alcázar JT (1981) Allozyme variation and relationships among Spanish landraces
of Cucumis melo L. Kulturpflanze 22: 337–352.
Esquinas-Alcázar JT, Gulick PJ (1983) Genetic Resources of Cucurbitaceae – A Global Report.
International Board for Plant Genetic Resources, Rome, Italy.
Esteras C, Díez MJ, Picó B, Sifres A, Valcarcel JV, Nuez F (2008) Diversity of Spanish landraces
of Cucumis sativus and Cucurbita ssp. In: M Pitrat (ed) Cucurbitaceae 2008, Proc IX
EUCARPIA Meeting on Genetics and Breeding of Cucurbitaceae. INRA, Avignon, France,
21–24 May 2008, pp 67–76.
Esteras C, Lunn J, Sulpice R, Blanca J, Garcia-Mas J, Pitrat M, Nuez F, Picó B (2009a) Phenotyping
a highly diverse core melon collection to be screened using Ecotilling. 8th Plant Genomics
European Meetings (Plant Gem), Lisbon, Portugal, 7–10 Oct 2009, p 214.
Esteras C, Pascual L, Saladie M, Dogimont C, Garcia-Mas J, Nuez F, Picó B (2009b) Use of
Ecotilling to identify natural allelic variants of melon candidate genes involved in fruit
ripening. 8th Plant Genomics European Meetings (Plant Gem), Lisbon, Portugal, 7–10
Oct 2009, p 213.
Esteras C, Sifres A, Nuez F, Picó B (2009c) Variabilidad de Cucurbita maxima en su zona de
origen: un recurso de interés para la mejora de esta hortaliza. In: A Pardo, ML Suso, N
Vázquez (eds) Resúmenes. Actas de Horticultura 54. VI Congreso Ibérico, XII Nacional
de Ciencias Hortícolas, Logroño, Spain, 25–29 May 2009, pp 1284–1290 (in Spanish).
Fanourakis N, Tsekoura Z, Nanou E (2000) Morphological characteristics and powdery
mildew resistance of Cucumis melo landraces in Greece. In: N Katzir, HS Paris (eds)
Proc Cucurbitaceae 2000, Ma’aleh Hahamisha, Israel, 19–23 March 2000, Acta Hort 510:
241–245.
Fantaccione S, Woodrow P, Pontecorvo G (2008) Molecular authentication of three Italian melon
accessions by ARMS-PCR and ITS1 (internal transcribed spacer 1) secondary structure
prediction. Bioinformation 2(7): 311–315.
Fazio G, Chung SM, Staub JE (2002) Development and characterization of PCR markers in
cucumber (Cucumis sativus L.). J Am Soc Hort Sci 127: 545–557.
Fazio G, Staub JE, Stevens MR (2003) Genetic mapping and QTL analysis of horticultural
traits in cucumber (Cucumis sativus L.) using recombinant inbred lines. Theor Appl
Genet 107: 864–874.
Fernández-Silva I, Eduardo I, Blanca J, Esteras C, Picó B, Nuez F, Arús P, Garcia-Mas J, Monforte
AJ (2008) Bin mapping of genomic and EST-derived SSRs in melon (Cucumis melo L.).
Theor Appl Genet 118: 139–150.
Ferriol M (2003) Análisis de la variabilidad morfológica y molecular en el género Cucurbita.
Tesis doctoral. Univ Politécnica de Valencia, Spain (in Spanish).
Ferriol M, Picó B (2008) Pumpkin and Winter squash. In: J Prohens, F Nuez (eds) Handbook
of Plant Breeding, vol I: Vegetables I. Springer, New York, USA, pp 317–349.
Ferriol M, Picó B, Nuez F (2003a) Genetic diversity of a germplasm collection of Cucurbita pepo
using SRAP and AFLP markers. Theor Appl Genet 107: 271–282.
Ferriol M, Picó B, Nuez F (2003b) Genetic diversity of some accessions of Cucurbita maxima
from Spain using RAPD and SBAP markers. Genet Resour Crop Evol 50: 227–238.
Ferriol M, Picó B, Fernández de Córdova P, Nuez F (2004a) Molecular diversity of a germplasm
collectionof squash (Cucurbita moschata) determined by SRAP and AFLP markers. Crop
Sci 44: 653–664.
Ferriol M, Picó B, Nuez F (2004b) Morphological and molecular diversity of a collection of
Cucurbita máxima landraces. J Am Soc Hort Sci 129(1): 60–69.
Ferriol M, Picó B, Nuez F (2005) Genetic diversity of Cucurbita spp. in the Canary Islands:
a bridge between America and Europe. XVII EUCARPIA Genetic Resources Section
Meeting, Castelsardo, Italy, 30 March–2 April 2005, pp 25–32.
Filov AI (1966) Ekologija I klassifikatzija tykuy. Bjulleten´ Glavnogo botaniceskogo sada 63:
33–41 (in Russian).
Flyman MV, Afolayan AJ (2007) The implication of the mineral ratios of Cucumis myriocarpus
Naud. and Pergularia daemia (Forsk.) Chiov. in human diets. J Med Food 10(3): 548–551.
Fujishita N (1992) Melons in the ancient Japan, revealed by excavated melon seeds. Archaeol
J 354: 7–13.
Fukino N, Yoshioka Y, Kubo N, Hirai M, Sugiyama M, Sakata Y, Matsumoto S (2008)
Development of 101 novel SSR markers and construction of an SSR-based genetic linkage
map in cucumber (Cucumis sativus L.). Breed Sci 58: 475–483.
Fursa TB (1972) K Sistematike roda Citrullus Schrad. Bot Z 57: 31–41.
García E, Jamilena M, Álvarez JI, Arnedo T, Oliver JL, Lozano R (1998) Genetic relationships
among melon breeding lines revealed by DNA markers and agronomic traits. Theor
Appl Genet 96: 878–885.
Garcia-Mas J, Oliver M, Gómez-Paniagua H, De Vicente MC (2000) Comparing AFLP, RAPD
and RFLP markers for measuring genetic diversity in melon. Theor Appl Genet 101:
860–864.
Garcia-Mas J, Monforte AJ, Arús P (2004) Phylogenetic relationships among Cucumis species
based on the ribosomal internal transcribed spacer sequence and microsatellite markers.
Plant Syst Evol 248: 191–203.
Ghebretinsae AG, Thulin M, Barber JC (2007) Relationships of cucumbers and melons
unraveled: molecular phylogenetics of Cucumis and related genera (Benincaseae,
Cucurbitaceae). Am J Bot 94(7): 1256–1266.
Gichimu BM, Owuor BO, Mwai GN, Dida MM (2009) Morphological characterization of
some wild and cultivated watermelon (Citrullus sp.) accessions in Kenya. ARPN J Agri
Biol Sci 4: 2.
Goldberg RB, Bemis WP, Siegel A (1972) Nucleic acid hybridization studies within the genus
Cucurbita. Genetics 72: 253–266.
Gómez-Guillamón ML, Abadía J, Cuartero J, Corté C, Nuez F (1985) Characterization of melon
cultivars. Cucurbit Genet Coop Rep 8: 39–40.
Gómez-Guillamón ML, Torés JA, Soria C, Sesé AIL (1995) Screening for resistance to Sphaerotheca
fuliginea and two yellowing diseases in Cucumis melo and related Cucumis species. In: G
Lester, J Dunlap (eds) Proc Cucurbitaceae 1994. Evaluation and Enhancement of Cucurbit
Germplasm. South Padre Island, Texas, USA, pp 205–208.
Gómez-Guillamón ML, Sánchez F, Fernández-Muñoz R (1998) Caracterización de cultivares
de melón (Spanish). Actas de las Jornadas de Selección y Mejora de Plantas Hortícolas.
Córdoba, Spain (in Spanish).
Gong L, Stift G, Kofler R, Pachner M, Lelley T (2008a) Microsatellites for the genus Cucurbita and
an SSR-based genetic linkage map of Cucurbita pepo L. Theor Appl Genet 117: 37–48.
Gong L, Pachner M, Kalai K, Lelley T (2008b) SSR-based genetic linkage map of Cucurbita
moschata and its synteny with Cucurbita pepo. Genome 51: 878–887.
González-Ibeas D, Blanca J, Roig C, González-To M, Picó B, Truniger V, Gómez P, Deleu W,
Caño-Delgado A, Arús P, Nuez F, Garcia-Mas J, Puigdomènech P, Aranda MA (2007)
MELOGEN: an EST database for melon functional genomics. BMC Genom 8: 306. doi:
10.1186/1471–2164-8-306.
González VM, Rodríguez-Moreno L, Centeno E, Benjak A, Garcia-Mas J, Puigdomènech P,
Aranda MA (2010) Genome-wide BAC-end sequencing of Cucumis melo using two BAC
libraries. BMC Genomics 11: 618.
Gonzalo MJ, Oliver M, Garcia-Mas J, Monfort A, Dolcet-Sanjuan R, Katzir N, Arús P, Monforte
AJ (2005) Simple-sequence repeat markers used in merging linkage maps of melon
(Cucumis melo L.). Theor Appl Genet 110(5): 802–811.
Grondin I, Smadja J, Armougom R (2002) The qualitative and quantitative composition of
triacylglycerols from four Cucurbitaceae seed oils of the Lagenaria and Luffa species.
OCL—Oleagineux, Corps Gras, Lipides 9(2/3): 169–173.
Guerra-Sanz JM (2002) Citrullus simple sequence repeats markers from sequence databases.
Mol Ecol Notes 2: 223–225.
Günay A (1993) Vegetable production. V.A.Ü. Ziraat Fak. Ankara. 117s. (in Turkish).
Gwanama C, Labuschagne MT, Botha AM (2000) Analysis of genetic variation of Cucurbita
moschata by random amplified polymorphic DNA (RAPD) markers. Euphytica 113:
19–24.
Gwanama C, Nichterlein K, Lungu D, Simabwachi W (2002) Variation of fruit b-carotene
content of tropical pumpkin [Cucurbita moschata (Duchesne) Poirot] landraces in Zambia.
Plant Genet Resour Newsl 129: 44–46.
Hashizume T, Sato T, Hirai M (1993) Determination of genetic purity of hybrid seed in
watermelon (Citrullus lanatus) and tomato (Lycopersicon esculentum) using random
amplified polymorphic DNA (RAPD). Jpn J Breed 43: 367–375.
Heikal AH, Abdel-Razzak HS, Hafez EE (2008) Assessment of genetic relationships among and
within Cucurbita species using RAPD and ISSR markers. J Appl Sci Res 4(5): 515–525.
Heiser CB, Schilling EE (1990) The genus Luffa: A problem in phytogeography. In: DM Bates,
RW Robinson, C Jeffrey (eds) Biology and Utilization of the Cucurbitaceae. Cornell Univ
Press, Ithaca, New York, USA, pp 120–133.
Helm MA, Hemleben V (1997) Characterization of a new prominent satellite DNA of Cucumis
metuliferus and differential distribution of satellite DNA in cultivated and wild species of
Cucumis and in related genera of Cucurbitaceae. Euphytica 94(2): 219–226.
Hoque S, Rabbani MG (2009) Assessment of genetic relationship among landraces of
Bangladeshi ridge gourd (Luffa acutangula Roxb.) using RAPD markers. J Sci Res 1(3):
615–623.
Hord M, Villalobos W, Macaya-Lizano AV, Rivera C (1997) Chayote mosaic, a new disease in
Sechium edule caused by a tymovirus. Plant Dis 81: 374–378.
Horejsi T, Staub JE (1999) Genetic variation in cucumber as assessed by random amplified
polymorphic DNAs. Genet Resour Crop Evol 46: 337–350.
Horejsi T, Box JM, Staub JE (1999) Efficiency of randomly amplified polymorphic DNA to
sequence characterized amplified region marker conversion and their comparative
polymerase chain reaction sensitivity in cucumber. J Am Soc Hort Sci 124: 128–135.
Horst EK, Lower RL (1978) Cucumis hardwickii: A source of germplasm for the cucumber
breeder. Cucurbit Genet Coop Rep 1: 5.
Hsieh L, Liu M, Hsiao C, Lin T, Yang W (2007) Genetic diversity among the lines and cultivars
of Luffa species. J Tai Soc Hort Sci 53(1): 99–110.
Huang S, Li R, Zhang Z, Li L, Gu X, Fan W, Lucas WJ, Wang X, Xie B, Ni P, Ren Y, Zhu H, Li
J, Lin K, Jin W, Fei Z, Li G, Staub J, Kilian A, van der Vossen EAG, Wu Y, Guo J, He J, Jia
Z, Ren Y, Tian G, Lu Y, Ruan J, Qian W, Wang M, Huang Q, Li B, Xuan Z, Cao J, Asan,
Wu Z, Zhang J, Cai Q, Bai Y, Zhao B, Han Y, Li Y, Li X, Wang S, Shi Q, Liu S, Cho WK,
Kim JY, Xu Y, Heller-Uszynska K, Miao H, Cheng Z, Zhang S, Wu J, Yang Y, Kang H,
Li M, Liang H, Ren X, Shi Z, Wen M, Jian M, Yang H, Zhang G, Yang Z, Chen R, Liu S,
Li J, Ma L, Liu H, Zhou Y, Zhao J, Fang X, Li G, Fang L, Li Y, Liu D, Zheng H, Zhang Y,
Qin N, Li Z, Yang G, Yang S, Bolund L, Kristiansen K, Zheng H, Li S, Zhang X, Yang H,
Wang J, Sun R, Zhang B, Jiang S, Wang J, Du Y, Li S (2009) The genome of the cucumber,
Cucumis sativus L. Nat Genet (Adv online) 41: 1275–1281.
Huh YC, Solmaz I, Sari N (2008) Morphological characterization of Korean and Turkish
watermelon germplasm. In: M Pitrat (ed) Cucurbitaceae 2008, Proc IXth EUCARPIA
Meeting on Genetics and Breeding of Cucurbitaceae. INRA, Avignon, France, 21–24 May
2008, pp 327–333.
Ignart F, Weeden NF (1984) Allozyme variation in cultivars of Cucurbita pepo L. Euphytica
33: 779–785.
Ivancic A, Sisko M, Bohanec B, Siftar S (2004) Morpho-agronomic characteristics of the
interspecific hybrid Cucurbita ficifolia x C. maxima. Agricultura (Slovenia) 3(1): 1–5.
Jarret RL, Newman M (2000) Phylogenetic relationships among species of Citrullus and the
placement of C. rehmii De Winter as determined by Internal Transcribed Spacer (ITS)
sequence heterogeneity. Genet Resour Crop Evol 47: 215–222.
Jarret RL, Merrick LC, Holms T, Evans J, Aradhya MK (1997) Simple sequence repeats in
watermelon (Citrullus lanatus (Thunb.) Matsum. & Nakai). Genome 40: 433–441.
Jeffrey C (1967) Cucurbitaceae. In: E Milne-Redhead, RM Polhill (eds) Flora of East Tropical
Africa. Crown Agents for Oversea Governments and Administrations, London, UK 17:
1–156.
Jeffrey C (1980) A review of the Cucurbitaceae. Bot J Linn Soc 81: 233–247.
Jeffrey C (1990) Appendix: An outline classification of the Cucurbitaceae. In: DM Bates, RW
Robinson, C Jeffrey (eds) Biology and Utilization of the Cucurbitaceae., Univ Press, Ithaca,
New York, USA, pp 449–463.
Jeffrey C (2001) Cucurbitaceae. In: P Hanelt (ed) Mansfeld’s Encyclopedia of agricultural and
horticultural crops. Institute of Plant Genetics and Crop Plant Research. Springer, Berlin,
Germany, 3: 1510–1557.
Jeffrey C (2005) A new system of Cucurbitaceae. Bot Zhurn 90: 332–335.
Jeffrey C, De Wilde WJJO (2006) A review of the subtribe Thladianthinae (Cucurbitaceae). Bot
Zhurn 91: 766–776.
Jeon HJ, Been CG, Hong KH, Om YH, Kim BD (1994) Identification of Cucurbitaceae cultivars
by using RAPD markers. J Kor Soc Hort Sci 35(5): 449–456.
Jobst J, King K, Hemleben V (1998) Molecular evolution of the internal transcribed spacers
(ITS1 and ITS2) and phylogenetic relationships among species of the family Cucurbitaceae.
Mol Phylogenet Evol 9(2): 204–219.
Joobeur T, Gusmini G, Zhang X, Levi A, Xu Y, Wehner TC, Oliver M, Dean RA (2006)
Construction of a watermelon BAC library and identification of SSRs anchored to melon
or Arabidopsis genomes. Theor Appl Genet 112: 1553–1562.
Joshi DC, Das SK, Mukherjee RK (1993) Physical properties of pumpkin seeds. J Agri Engg
Res 54: 219–229.
Júnior ATA (1999) Divergencia genética entre acessos de moranga do banco de germoplasma
de hortaliças de Universidade Federal de Viçosa. Hort Bras 17: 3–6 (in Portuguese).
Kajale MD (1979) On the occurrence of ancient agricultural patterns during the Chalcolithic
period (ca. 1,600–1,000 BC) at Apegaon, district Aurangabad in the central Godavari
valley, Maharashtra. In: SB Deo, MK Dhavalikar, ZD Ansari (eds) Apegaon excavation.
Pune Deccan College Postgraduate and Research Institute, Pune, India, pp 50–56.
Karchi Z (2000) Development of melon culture and breeding in Israel In: N Katzir, HS Paris
(eds) Proc Cucurbitaceae 2000, Ma’aleh Hahamisha, Israel, 19–23 March 2000, Acta Hort
510: 13–17.
Katzir N, Danin-Poleg Y, Tzuri G, Karchi Z, Lavi U, Cregan PB (1996) Length polymorphism
and homologies of microsatellites in several cucurbitacean species. Theor Appl Genet
93: 1282–1290.
Katzir N, Tadmor Y, Tzuri G, Leshzeshen E, Mozes-Daube N, Danin-Poleg Y, Paris H S (2000a)
Further ISSR and preliminary SSR analysis of relationships among accessions of Cucurbita
pepo. In: N Katzir, HS Paris (eds) Proc Cucurbitaceae 2000, Ma’aleh Hahamisha, Israel,
19–23 March 2000, Acta Hort 510: 433–439.
Katzir N, Mozes-Daube N, Danin-Poleg Y, Paris HS (2000b) Potential usefulness of SSR
markers for studying infraspecific variability in Cucurbita pepo. Cucurbit Genet Coop
Rep 23: 71–72.
Keay WJ, Hepper FN (1985) Cucurbitaceae. In: WJ Keay, FN Hepper (eds) The flora of west
tropical Africa. 2nd edn. Royal Botanic Gardens, London, UK, pp 570–574.
Keinath AP, DuBose VB (2000) Evaluation of pumpkin cultivars for powdery and downy
midew resistance, virus tolerance, and yield. HortScience 35(2): 281–285.
Ke-peng C, Liang C, Wang Y, Jin D, Wang B, Xu Y, Kang G, Zhang H (2003) Genetic assessment
of watermelon germplasm using the AFLP technique. HortScience 38: 81–84.
King K, Jobst J, Hemleben V (1995) Differential homogenization and amplification of two
satellite DNAs in the genus Cucurbita (Cucurbitaceae). J Mol Evol 41: 996–1005.
Kirkbride JH Jr (1993) Biosystematic monograph of the genus Cucumis (Cucurbitaceae).
Parkway Publ, Boone. North Carolina, USA, 159 p.
Kirkpatrick KJ, Wilson HD (1988) Interspecific gene flow in Cucurbita: C. texana vs C. pepo.
Am J Bot 75: 517–525.
Kirkpatrick KJ, Decker DS, Wilson HD (1985) Allozyme differentiation in the Cucurbita pepo
complex: C. pepo var. medullosa vs C. texana. Econ Bot 39: 289–299.
Kitamura S (1950) Notes on Cucumis of Far East. Acta Phytotaxon Geobot 14: 41–44.
Knerr LD, Staub JE, Holder DJ, May BP (1989) Genetic diversity in Cucumis sativus L. assessed
by variation at 18 allozyme coding loci. Theor Appl Genet 78: 119–128.
Kocyan A, Zhang LB, Schaeffer H, Renner SS (2007) A multi-locus chloroplast phylogeny for
the Cucurbitaceae and its implications for character evolution and classification. Mol
Phylogenet Evol 44: 553–577.
Kohpayegani JA (2004) Study of genetic diversity of some Iranian melon and effects of seed
production on genetic erosion. PhD Thesis, Univ Tehran, Iran, 124 p.
Kohpayegani JA, Behbahani M (2008) Genetic diversity of some populations of Iranian melon
using SSR markers. Biotechnology 7(1): 19–26.
Kole C, Olukolu B, Kole P, Abbott AG (2010) Towards Phytomedomics with Bitter Melon
(Momordica charantia L.) as a Model. Abstract. Plant &Animal Genomes XVIII Conference,
San Diego CA, 9–13 January 2010.
Kong Q, Xiang C, Yu Z (2006) Development of EST-SSRs in Cucumis sativus from sequence
database. Mol Ecol Notes 6: 1234–1236.
Kong Q, Xiang C, Yu Z, Zhang C, Liu F, Peng C, Peng X (2007) Mining and charactering
microsatellites in Cucumis melo expressed sequence tags from sequence database. Mol
Ecol Notes 7: 281–283.
Krauze-Baranowska M, Cisowski W (2001) Flavonoids from some species of the genus Cucumis.
Biochem Syst Ecol 29(3): 321–324.
Krawinkel MB, Keding GB (2006) Bitter gourd (Momordica charantia): a dietary approach to
hyperglycemia. Nutr Rev 64: 331–337.
Kristkova E, Lebeda A (2000) Resistance in C. pepo and C. maxima germplasm to watermelon
mosaic potyvirus-2. Plant Genet Resour Newsl 121: 47–52.
Lebeda A, Kristkova E, Dolezal K (1999) Peroxidase isozyme polymorphism in Cucurbita pepo
cultivars with various morphotypes and different level of field resistance to powdery
mildew. Sci Hort 81: 103–112.
Lebeda A, Widrlechner MP, Staub JE, Ezura H, Zalapa JE, Krístková H (2006) Cucurbits
(Cucurbitaceae; Cucumis spp., Cucurbita spp., Citrullus spp.). In: RJ Singh (ed) Genetic
Resources, Chromosome Engineering, and Crop Improvement. Taylor & Francis Group,
Boca Raton, FL, USA, pp 271–376.
Lee SJ, Shin JS, Park KW, Hong YP (1996) Detection of genetic diversity using RAPD-PCR
and sugar analysis in watermelon [Citrullus lanatus (Thunb.) Mansf.] germplasm. Theor
Appl Genet 92: 719–725.
Leppik EE (1966) Searching gene centers of the genus Cucumis through host-parasite
relationship. Euphytica 15: 323–328.
Levi A, Thomas CE (2005) Polymorphisms among chloroplast and mitochondrial genomes of
Citrullus species and subspecies. Genet Resour Crop Evol 52: 609–617.
Levi A, Thomas CE, Wehner TC, Zhang XP (2001a) Low genetic diversity indicates the need
to broaden the genetic base of cultivated watermelon. HortScience 36: 1096–1101.
Levi A, Thomas CE, Keinath AP, Wehner TC (2001b) Genetic diversity among watermelon
(Citrullus lanatus and Citrullus colocynthis) accessions. Genet Resour Crop Evol 48:
559–566.
Levi A, Thomas CE, Newman M, Reddy OUK, Zhang X, Xu Y (2004) ISSR and AFLP markers
differ among American watermelon cultivars with limited genetic diversity. J Am Soc
Hort Sci 129(4): 553–558.
Levi A, Thomas CE, Simmons AM, Thies JA (2005) Analysis based on RAPD and ISSR markers
reveals closer similarities among Citrullus and Cucumis species than with Praecitrullus
fistulosus (Stocks) Pangalo. Genet Resour Crop Evol 52: 465–472.
Levi A, Thies J, Ling K, Simmons AM, Kousik C, Hassell R (2009) Genetic diversity among
Lagenaria siceraria accessions containing resistance to root-knot nematodes, whiteflies,
ZYMV or powdery mildew. Plant Genet Resour 7: 216–226.
Liou PC, Chang YM, Hsu WS, Cheng YH, Chang HR, Hsiao CH (1998) Construction of a
linkage map in Cucumis melo L. using random amplified polymorphic DNA markers.
In: RA Drew (ed) ISHS Acta Horticulturae 461. Int Symp on Biotechnology of Tropical
and Subtropical Species, part 2. International Society for Horticultural Science, Brisbane,
Australia, pp 123–131.
Lira-Saade R (1995) Cucurbita L. Estudios Taxonómicos y Ecogeográficos de las Cucurbitaceae
Latinoamericanas de Importancia Económica. Systematics and Ecogeographic Studies on
Crop Genepools. International Plant Genetic Resources Institute, Rome, Italy 9: 281.
Lira-Saade R (1996) Chayote. Sechium edule (Jacq.) Sw. Promoting the conservation and use of
underutilized and neglected crops. Institute of Plant Genetics and Crop Plant Research,
Gatersleben, Germany/International Plant Genetic Resources Institute. Rome, Italy, 8:
58 p.
Lira-Saade R, Montes-Hernández S (1994) Cucurbits (Cucurbita spp.). In: JE Hernández Bermejo,
J León (ed) Neglected Crops: 1492 from a Different Perspective. Plant Production and
Protection Series. FAO, Rome, Italy, 26: 63–77.
López-Sesé AI, Staub J, Katzir N, Gómez-Guillamón ML (2002) Estimation of between and
within accession variation in selected Spanish melon germplasm using RAPD and SSR
markers to assess strategies for large collection evaluation. Euphytica 127: 41–51.
López-Sesé AI, Staub JE, Gómez-Guillamón ML (2003) Genetic analysis of Spanish melon
(Cucumis melo L.) germplasm using a standardized molecular-marker array and
geographically diverse reference accessions. Theor Appl Genet 108(1): 41–52.
Lotti C, Albo M, Ricciardi L, Conversa G, Elia A (2005) Genetic diversity in ‘Carosello’ and
‘Barattiere’ ecotypes (Cucumis melo L.). Colture Protette N5 (Suppl): 44–46.
Luan F, Delannay I, Staub JE (2008) Chinese melon (Cucumis melo L.) diversity analyses provide
strategies for germplasm curation, genetic improvement, and evidentiary support of
domestication patterns. Euphytica 164: 445–461.
Maggs-Kolling G, Madsen S, Christiansen JL (2000) A phenetic analysis of morphological
variation in Citrullus lanatus in Namibia. Genet Resour Crop Evol 47: 385–393.
Mallick MFR, Masui M (1986) Origin, distribution and taxonomy of melons. Sci Hort 28:
251–261.
Marco CF, Aranda MA, Montoro T, Gomez-Guillamon ML (2003) Evaluation of several
accessions and wild relatives of Cucumis melo against cucumber vein yellowing virus
(CVYV). Cucurbit Genet Coop Rep 26: 7–8.
Marr KL, Mei XY, Bhattarai NK (2004) Allozyme, morphological and nutritional analysis
bearing on the domestication of Momordica charantia L. (Cucurbitaceae). Econ Bot 58:
435–455.
Marr KL, Bhattarai NK, Xia YM (2005) Allozymic, morphological, and phonological diversity
in cultivated acutangula (Cucurbitaceae) from China, Laos, and Nepal, and allozyme
divergence between L. acutangula and L. aegyptiaca. Econ Bot 59(2): 154–165.
Marsh DB (1993) Evaluation of Cucumis metuliferus as a speciality crop for Missouri. New
crops. Proc 2nd Natl Symp on New Crops, Exploration, Research and Commercialization,
Indianapolis, IN, USA, 6–9 Oct 1991–1993, pp 558–559.
Maynard DN (2001) Variation among tropical pumpkin (Cucurbita moschata) cultivars in
susceptibility to silverleaf. Cucurbit Genet Coop Rep 24: 71–72.
McCreight JD, Staub JE (1993) Indo–US Cucumis germplasm expedition. HortScience 28:
467.
McCreight JD, Nerson H, Grumet R (1993) Melon, Cucumis melo L. In: G Kallos, BO Bergh (eds)
Genetic Improvement of Vegetable Crops. Pergamon Press, Oxford (GB), pp 267–294.
Meeuse AD (1962) The Cucurbitaceae of Southern Africa. Bothalia 8: 1–111.
Meglic V, Serquen F, Staub JE (1996) Genetic diversity in cucumber (Cucumis sativus L.): I. A
reevaluation of the US. germplasm collection. Genet Resour Crop Evol 43: 533–546.
Meng XD, Wei YY, Ma H, Zhang WH, Li JR (1996) Identification of Chinese wax gourd and
chieh-qua cultivars using RAPD markers. Acta Agri Shanghai 12: 45–49.
Merrick LC (1990) Systematics and evolution of a domesticated squash, Cucurbita argyrosperma,
and its wild and weedy relatives. In: DM Bates, RW Robinson, C Jeffrey (eds) Biology and
Utilization of the Cucurbitaceae. Cornell Univ Press, Ithaca, NY, USA, pp 77–95.
Mliki A, Staub JE, Zhangyong S, Ghorbel A (2001) Genetic diversity in melon (Cucumis melo
L.): An evaluation of African germplasm. Genet Resour Crop Evol 48: 587–597.
Mliki A, Staub JE, Zhangyong S, Ghorbel A (2003) Genetic diversity in African cucumber
(Cucumis sativus L.) provides potential for germplasm enhancement. Genet Resour Crop
Evol 50: 461–468.
Modolo VA, da Costa CP (2003) Avaliação de linhagens de maxixe paulista cultivadas em
canteiros com cobertura de polietileno. Hort Bras 21(3): 534–538 (in Portuguese).
Mohanty A, Martin JP, Aguinagalde I (2001) A population genetic analysis of chloroplast DNA
in wild populations of Prunus avium L. in Europe. Heredity 87: 421–427.
Monforte AJ, Garcia-Mas J, Arús P (2003) Genetic variability in melon based on microsatellite
variation. Plant Breed 122: 153–157.
Montes-Hernández S, Eguiarte LE (2002) Genetic structure and indirect estimates of gene
flow in three taxa of Cucurbita (Cucurbitaceae) in western Mexico. Am J Bot 89(7):
1156–1163.
Montes-Hernández S, Merrick LC, Eguiarte LE (2005) Maintenance of squash (Cucurbita
spp.) landrace diversity by farmers’ activities in Mexico. Genet Resour Crop Evol 52(6):
697–707.
Morales M, Roig E, Monforte A J, Arús P, Garcia-Mas J (2004) Single-nucleotide polymorphisms
detected in expressed sequence tags of melon (Cucumis melo L.). Genome 47: 352–360.
Morimoto Y, Maundu P, Fujimaki H, Morishima H (2005) Diversity of landraces of the
white-flowered gourd (Lagenaria siceraria) and its wild relatives in Kenya: fruit and seed
morphology. Genet Resour Crop Evol 52(6): 737–747.
Mo-Suk Y, Im-Sung H, Go-Gawn D, Ann-Chong M, Kim-Doo H, Mo-Suk Y, Im SH (1999) RAPD
analsysis of genetic diversity of melon species. Kor J Hort Sci Technol 16: 21–24.
Munger HM, Robinson RW (1991) Nomenclature of Cucumis melo L. Cucurbit Genet Coop
Rep 14: 43–44.
Mutschleer MA, Pearson OH (1987) The origin, inheritance, and instability of Butternut squash
(Cucurbita moschata Duchesne). HortScience 22(4): 535–539.
Nakata E, Staub JE, López-Sesé AI, Katzir N (2005) Genetic diversity of Japanese melon
cultivars (Cucumis melo L.) as assessed by random amplified polymorphic DNA and
simple sequence repeat markers. Genet Resour Crop Evol 52: 405–419.
Naudin C (1856) Nouvelles recherches sur les caractères spécifiques et les variétes des plantes
du genre Cucurbita. Ann Sci Nat IV 6: 5–73 (in French).
Naudin C (1859) Essais dune monographiedes espèces et des varieties du genre Cucumis. Ann
Sci Nat 11: 5–87 (in French).
Navot N, Zamir D (1987) Isozyme and seed protein phylogeny of the genus Citrullus
(Cucurbitaceae). Plant Syst Evol 156: 61–67.
Nee M (1990) The domesticationof Cucurbita (Cucurbitaceae). Econ Bot 44(3): 56–68.
Nerson H, Paris HS, Paris MP (2000) Fruit shape, size and seed yield in Cucurbita pepo. In:
N Katzir, HS Paris (eds) Proc of Cucurbitaceae 2000, Ma’aleh Hahamisha, Israel, 19–23
March 2000, Acta Hort 510: 227–230.
Neuhausen SL (1992) Evaluation of restriction fragment length polymorphisms in Cucumis
melo. Theor Appl Genet 83: 379–384.
Nieto C, Piron F, Dalmais M, Marco CF, Moriones E, Gómez-Guillamón ML, Truniger V, Gómez
P, Garcia-Mas J, Aranda MA, Bendahmane A (2007) EcoTILLING for the identification
of allelic variants of melon eIF4E, a factor that controls virus susceptibility. BMC Plant
Biol 7: 34. doi: 10.1186/1471-2229-7-34.
Nikolova V, Alexandrova M, Stoeva V (2002) Possibilities for the use of remote hybridization
in the genus Cucumis for the development of genetic diversity. Acta Hort 579: 39–43.
Nimmakayala P, Tomason YR, Jeong J, Vajja G, Levi A, Gibson P, Reddy UK (2009) Molecular
diversity in the Ukrainian melon collection as revealed by AFLPs and microsatellites.
Plant Genet Resour 7: 127–134.
Nuez F, Anastasio G, Cortés C, Cuartero J, Gómez-Guillamón ML, Costa J (1986) Germplasm
resources of Cucumis melo L from Spain. Cucurbit Genet Coop Rep 9: 60–63.
Nuez F, Ferrando C, Díez MJ, Costa J, Catalá MS, Cuartero J, Gómez-Guillamón ML (1988)
Collecting Cucumis melo L. in Spain. Cucurbit Genet Coop Rep 11: 54–56.
Oliszeweski N (2005) Archaeobotany of mound structures in Campo del Pucará, Catamarca,
Argentina (1750–1450 b.p.): ceremonial use or rubbish dumps? Veg Hist Archaeobot
14(4): 465–471.
Oliveira ANP, Oliveira AP, Leonardo FAP, Cruz IS, Silva DF (2009) Yield of gherkin in response
to doses of bovine manure. Hort Brasil 27(1): 100–102.
Oliver M, Garcia-Mas J, Cardus M, Pueyo N, López-Sesé AI, Arroyo M, Gómez-Paniagua
H, Arús P, De Vicente MC (2001) Construction of a reference linkage map for melon.
Genome 44: 836–845.
Onto S, Laosat N, Suksawat W, Popluechai S, Eungwanichayapant PD, Chukeatirote E (2008)
Phylogenetic analysis of Cucumis sativus using RAPD molecular markers. J Plant Sci
3(1): 105–110.
Pandey S, Kumar S, Mishra U, Rai A, Singh M, Rai M (2008) Genetic diversity in Indian ash
gourd (Benincasa hispida) accessions as revealed by quantitative traits and RAPD markers.
Sci Hort 118: 80–86.
Pangalo KI (1930) A new species of cultivated pumpkin. Bull Appl Bot Genet Plant Breed
23: 253–265.
Parducci L, Szmidt AE (1999) PCR-RFLP of cpDNA in the genus Abies. Theor Appl Genet
98: 802–808.
Paris HS (1986) A proposed subspecific classification for Cucurbita pepo. Phytologia 61:
133–138.
Paris HS (1989) Historical records, origins, and development of the edible cultivar groups of
Cucurbita pepo (Cucurbitaceae). Econ Bot 473(4): 423–443.
Paris HS (1998) Some observations concerning diversity in the subspecies and horticultural
groups of Cucurbita pepo. Cucurbit Genet Coop Rep 21: 51–53.
Paris HS (2000) History of the cultivar-groups of Cucurbita pepo. In: J Janick (ed) Horticulture
Review. John Wiley, New York, USA 25: 71–170.
Paris HS (2001a) Characterization of the Cucurbita pepo collection at the Newe Yaár Research
Center, Israel. Plant Genet Resour Newsl 126: 41–45.
Paris HS (2001b) History of the cultivar-groups of Cucurbita pepo. Hort Rev 25: 71–170.
Paris HS (2008) Summer squash. In: J Prohens, F Nuez (eds) Handbook of Plant Breeding, vol
I: Vegetables I. Springer, New York, USA, pp 351–379.
Paris HS, Nerson H (1998) Association of seed size and dimensions with fruit shape in Cucurbita
pepo. In: JD McCreight (ed) Cucurbitaceae 1998, Evaluation and Enhancement of Cucurbit
Germplasm. Alexandria, Va, USA, pp 230–234.
Paris HS, Yonash N, Portnoy V, Mozes-Daube N, Tzuri G, Katzir N (2003) Assessment of
genetic relationships in Cucurbita pepo (Cucurbiatceae) using DNA markers. Theor Appl
Genet 106: 971–978.
Park YH, Sensoy S, Wye C, Antonise R, Peleman J, Havey MJ (2000) A genetic map of cucumber
composed of RAPDs, RFLPs, AFLPs, amd loci coditioning resistance to papaya ringspot
and zucchini yellow mosaic viruses. Genome 43: 1003–1010.
Parkash C, Singh KP, Kalloo G (2000) Variability analysis and cause and effect relationship in
ash gourd [Benincasa hispida (Thunb.) Cogn.]. Indian J Plant Genet Res 13: 298–301.
Périn C, Hagen S, De Conto V, Katzir N, Danin-Poleg Y, Portnoy V, Baudracco-Arnas S,
Chadoeuf J, Dogimont C, Pitrat M (2002) A reference map of Cucumis melo based on two
recombinant inbred line populations. Theor Appl Genet 104: 1017–1034.
Perl-Treves R, Galun E (1985) The Cucumis plastome: physical map, intrageneric variation and
phylogenetic relationships. Theor Appl Genet 71: 417–429.
Perl-Treves R, Zamir D, Navot N, Galun E (1985) Phylogeny of Cucumis based on isozyme
variability and its comparison with plastome phylogeny. Theor Appl Genet 71:
430–436.
Petersen JB, Sidell NA (1996) Mid-Holocene evidence of Cucurbita sp. from central Maine.
Am Antiq 61: 685–698.
Picó B, Sifres A, Esteras C, Nuez F (2005–2006) Cucumis SSRs markers applied to the study of
the genetic diversity in the Cucurbita genus. Cucurbit Genet Coop Rep 28–29: 70–72.
Piperno DR, Andres TC, Stothert KE (2000) Phytolits in Cucurbita and other neotropical
Cucurbitaceae and their occurrence in early archaeological sites from the lowland
American tropics. J Archeol Sci 27: 193–208.
Pitrat M (2008) Melon (Cucumis melo L.). In: J Prohens, F Nuez (eds) Handbook of Crop
Breeding, vol I: Vegetables. Springer, New York, USA, pp 283–315.
Pitrat M, Chauvet M, Foury C (1999) Diversity, history, and production of cultivated cucurbits.
Acta Hort 492: 21–28.
Pitrat M, Hanelt P, Hammer K (2000) Some comments on infraspecific classification of cultivar
of melon In: N Katzir, HS Paris (eds) Proc of Cucurbitaceae 2000, Ma’aleh Hahamisha,
Israel, 19–23 March 2000, Acta Hort 510, 29–36.
Poe RP, Coyne DP, Swisher BA, Clegg MD (1988) Differential Cucurbita spp. Tolerance to the
herbicide trifluralin. J Am Soc Hort Sci 113(1): 35–40.
Puchalski JT, Robinson RW (1990) Electrophoretic analysis of isozymes in Cucurbita and
Cucumis and its application for phylogenetic studies. In: DM Bates, RW Robinson, C
Jeffrey (eds) Biology and Utilization of the Cucurbitaceae. Cornell Univ Press, Ithaca,
New York, USA, pp 60–76.
Ramos SRR (2007) Genetic diversity based on AFLP molecular markers and indicators for the
establishment of a core collection for pumpkin (Cucurbita moschata) for north-east Brazil.
Plant Genetic Resources Newsletter. Thesis Abstr 149(43)145: 66–67.
Renner SS, Schaefer H, Kocyan A (2007) Phylogenetics of Cucumis (Cucurbitaceae): Cucumber
(C. sativus) belongs in an Asian/Australian clade far from melon (C. melo). BMC Evol
Biol 7: 58. doi: 10.1186/1471-2148-7-58.
Ríos H, Fernández A, Batista O (1997) Cuban pumpkin genetic variability under low input
conditions. Cucurbit Genet Coop Rep 20: 48–49.
Ritschel PS, Lins TCL, Tristan RL, Buso GSC, Buso JA, Ferreira ME (2004) Development of
microsatellite markers from an enriched genomic library for genetic analysis of melon
(Cucumis melo L.). BMC Plant Biol 4: 9.
Robinson RW, Decker-Walters DS (1997) Cucurbits. Crop Production Science in Horticulture.
CAB Int, New York, USA, 6: 226 p.
Robinson RW, Decker-Walters DS (1999) Cucurbits. CABI Publ, Wallingford, Oxford, UK,
226 p.
Romao RL (2000) Northeast Brazil: A secondary center of diversity for watermelon (Citrullus
lanatus). Genet Resour Crop Evol 47: 207–213.
Romero-Rodriguez MA, Vazquez-Oderiz ML, Lopez-Hernandez J, Simal-Lozano J (1992)
Physical and analytical characteristics of the kiwano. J Food Compos Anal 5(4):
319–322.
Rubatzky VE (2001) Origin, distribution and uses. In: DN Maynard (ed) Watermelons.
Characteristics, Production and Marketing. ASHS Press, Alexandria, Va, USA, pp
21–26.
Rubatzky VE, Yamaguchi M (1997) World Vegetable Principles, Production and Nutritive
Values. 2nd edn. Chapman and Hall, International Thompson Publ, New York, USA,
853 p.
Rubatazky VE, Yamaguchi M (1999) World Vegetables. Chapman and Hall, New York,
USA.
Sanjur OI, Piperno DR, Andres TC, Wessel-Beaver L (2002) Phylogenetic relationships among
domesticated and wild species of Cucurbita (Cucurbitaceae) inferred from a mitochondrial
gene: implications for crop plant evolution and areas iof origin. Proc Nat Acad Sci 99:
535–540.
Sanwal SK, Yadav RK, Singh PK, Rai N (2008) Variability and genetic diversity studies in
indigenous chow-chow genotypes of northeast India. Indian J Hort 65(2): 167–170.
Schaefer H (2007) Cucumis (Cucurbitaceae) must include Cucumella, Dicoelospermum, Mukia,
Myrmecosicyos, and Oreosyce: a recircumscription based on nuclear and plastid DNA
data. Blumea 52: 165–177.
Schaefer H, Renner SS (2010) A three-genome phylogeny of Momordica (Cucurbitaceae) suggests
seven returns from dioecy to monoecy and recent long-distance dispersal to Asia. Mol
Phylogenet Evol 54(2): 553–560.
Sebastian P, Schaefer H, Telford IR, Renner SS (2010) Cucumber (Cucumis sativus) and melon
(C. melo) have numerous wild relatives in Asia and Australia, and the sister species of
melon is from Australia. Proc Natl Acad Sci 107(32): 14269–14273.
Sensoy S, Buyukalaca S, Abak K (2007) Evaluation of genetic diversity in Turkish melons
(Cucumis melo L.) based on phenotypic characters and RAPD markers. Genet Resour
Crop Evol 54: 1351–1365.
Silberstein L, Kovalski I, Huang R, Anagnostou K, Jahn MK, Perl-Treves R (1999) Molecular
variation in melon (Cucumis melo L.) as revealed by RFLP and RAPD markers. Sci Hort
79: 101–111.
Singh DK (2002) Genetic analysis of yield and its components in ash gourd [Benincasa hispida
(Thunb.) Cogn.]. PhD Thesis. UP College, Varanasi, India.
Singh NP, Matta NK (2008) Variation studies on seed storage proteins and phylogenetics of
the genus Cucumis. Plant Syst Evol 275: 209–218.
Smith BD (1997) The initial domestication of Cucurbita pepo in the Americas 10.000 years ago.
Science 276: 932–934.
Smith BD (2005) Reassessing Coxcatlan Cave and the early history of domesticated plants in
Mesoamerica. Proc Natl Acad Sci USA 102(27): 9438–9445.
Stachel M, Csanádi G, Vollmann J, Lellet T (1998) Genetic diversity in pumpkins (Cucurbita
pepo L.) as revealed in inbred lines using RAPD markers. Cucurbit Genet Coop Rep 21:
48–50.
Staub JE, Meglic V (1993) Molecular genetic markers and their relevance for cultivar
discrimination: A case study in cucumber. HortTechnology 3: 291–299.
Staub JE, Bacher J (1997) Cucumber as a processed vegetable. In: DS Smith, JN Cash, W Nip,
YH Hui (eds) Processing vegetables: Science and Technology IV. Technomic Publishing
Co, Inc Lancaster, PA, USA, pp 129–193.
Staub JE, Ivandic V (2000) Genetic assessment of the United States National cucumber
collection. In: N Katzir, HS Paris (eds) Proc of Cucurbitaceae 2000, Ma’aleh Hahamisha,
Israel, 19–23 March 2000, Acta Hort 510: 113–122.
Staub JE, Kupper RS, Schuman D, Wehner TC, May B (1985) Electrophoretic variation and
enzyme storage stability in cucumber. J Am Soc Hort Sci 110: 426–431.
Staub JE, Fredrick L, Marty T (1987) Electrophoretic variation in cross-compatible wild diploid
species of Cucumis. Can J Bot 65: 792–798.
Staub JE, Box J, Meglic V, Horejsi TF, McCreight JD (1997) Comparison of isozyme and random
amplified polymorphic DNA data for determining intraspecific variation in Cucumis.
Genet Resour Crop Evol 44: 257–269.
Staub JE, Serquen FC, Horejsi T, Chen J (1999) Genetic diversity in cucumber (Cucumis sativus
L.): IV. An evaluation of Chinese germplasm. Genet Resour Crop Evol 46: 297–310.
Staub JE, Danin-Poleg Y, Fazio G, Horejsi T, Reis N, Katzir N (2000) Comparative analysis of
cultivated melon groups (Cucumis melo L.) using random amplified polymorphic DNA
and simple sequence repeat markers. Euphytica 115: 225–241.
Staub JE, López-Sesé I, Fanourakis N (2004) Diversity among melón landraces (Cucumis melo
L.) from Greece and their genetic relationships with other melon germplasm of diverse
origins. Euphytica 136: 151–166.
Staub JE, Chung S-M, Fazio G (2005) Conformity and genetic relatedness estimation in crop
species having a narrow genetic base: the case of cucumber (Cucumis sativus L.). Plant
Breed 124: 44–53.
Staub JE, Robbins MD, Wehner TC (2008) Cucumber. In: J Prohens, F Nuez (eds) Handbook
of Plant Breeding, vol I: Vegetables I. Springer, New York, USA, pp 241–282.
Stepansky A, Kovalski I, Perl-Treves R (1999) Intraspecific classification of melons (Cucumis melo
L.) in view of their phenotypic and molecular variation. Plant Syst Evol 217: 313–332.
Stift G, Zraidi A, Lelley T (2004) Development and characterization of microsatellite markers
(SSR) in Cucurbita species. Cucurbit Genet Coop Rep 27: 61–65.
Sureja AK, Sirohi PS, Behera TK, Mohapatra T (2006) Molecular diversity and its relationship
with hybrid performance and heterosis in ash gourd [Benincasa hispida (Thunb.) Cogn.].
J Hort Sci Biotechnol 81(1): 33–38.
Szabó Z, Gyulai G, Humphreys M, Horvath L, Bittsansky A, Lagler R, Heszky L (2005) Genetic
variation in melon (C. melo) compared to an extinct landrace from the Middle Ages
(Hungary). I. RDNA, SSR, and SNP analysis of 47 cultivars. Euphytica 146: 87–94.
Szabó Z, Gyulai G, Tóth Z, Heszky L (2008) Morphological and molecular diversity of 47
melon (Cucumis melo) cultivars compared to an extinct landrace excavated from the 15th
century. In: M Pitrat (ed) Cucurbitaceae 2008, Proc IXth EUCARPIA Meeting on Genetics
and Breeding of Cucurbitaceae. INRA, Avignon, France, 21–24 May 2008, 313–321.
Tabei Y (1997) Study on breeding of Cucurbitaceae using biotechnology. Bull Natl Inst Agrobiol
Resour 11: 1–107.
Tanaka K, Nishitani A, Akashi Y, Sakata Y, Nishida H, Yoshino H, Kato K (2007) Molecular
characterization of South and East Asian melon, Cucumis melo L., and the origin of Group
Conomon var. makuwa and var. conomon revealed by RAPD analysis. Euphytica 153:
233–247.
Telford IR (1982) Cucurbitaceae. Flora Aust 205: 158–198.
Teppner H (2004) Notes on Lagenaria and Cucurbita (Cucurbitaceae)—review and new
contributions. Phyton 44(2): 245–308.
Thulin M, Al-Gifri AN (1994) Cucumis canoxyi (Cucurbitaceae): a new species from Yemen.
Nord J Bot 14: 315–317.
Tolentino MIS, Laude RP, dela Viña AC (1997) Genetic diversity analysis of Luffa species based
on seed protein profile using SDS-PAGE. Phil J Crop Sci 22(3): 141–146.
Torres Ruiz RA, Hemleben V (1991) Use of ribosomal DNA spacer probes to distinguish
cultivars of Cucurbita pepo L. and other Cucurbitaceae. Euphytica 53: 11–17.
Trumbull JH (1876) Vegetables cultivated by the American Indians. Bull Torrey Bot Club 6:
69–71.
Verma M, Arya L (2008) Development of EST-SSRs in watermelon (Citrullus lanatus var. lanatus)
and their transferability to Cucumis spp. J Hort Sci Biotechnol 83(6): 732–736.
Verma VK, Behera TK, Munshi AD, Parida SK, Mohapatra T (2007) Genetic diversity of ash
gourd [Benincasa hispida (Thunb.) Cogn.] inbred lines based on RAPD and ISSR markers
and their hybrid performance. Sci Hort 113: 231–237.
Walters SA, Wehner TC (2002) Incompatibility in diploid and tetraploid crosses of Cucumis
sativus and Cucumis metuliferus. Euphytica 128(3): 371–374.
Walters TW (1989) Historical overview on domesticated plants in China with special emphasis
on the Cucurbitaceae. Econ Bot 43: 297–313.
Walters TW, Decker-Walters DS, Posluszny U, Kevan PG (1991) Determination and
interpretation of comigrating allozymes among genera of the Benincaseae (Cucurbitaceae).
Syst Bot 16: 30–40.
Wang YH, Thomas CE, Dean RA (1997) A genetic map of melon (Cucumis melo L.) based
on amplified fragment length polymorphism (AFLP) markers. Theor Appl Genet 95:
791–798.
Wasylikowa K, van der Veen M (2004) An archaeobotanical contribution to the history of
watermelon, Citrullus lanatus (Thunb.) Matsum. & Nakai (syn. C. vulgaris Schrad.). Veg
Hist Archaeobot 13: 213–217.
Watcharawongpaiboon N, Chunwongse J (2007) Development of microsatellite markers from
an enriched genomic library of pumpkin (Cucurbita moschata L.). Songklanakarin J Sci
Technol 29(5): 1217–1223.
Watcharawongpaiboon N, Chunwongse J (2008) Development and characterization of
microsatellite markers from an enriched genomic library of cucumber (Cucumis sativus).
Plant Breed 127: 74–81.
Wehner TC (2008) Watermelon. In: J Prohens, F Nuez (eds) Handbook of Plant Breeding, vol
I: Vegetables I. Springer, New York, USA, pp 381–418.
Wessel-Beaver L (1993) Powdery and downy mildew resistance in Cucurbita moschata accessions.
Cucurbit Genet Coop Rep 16: 73–74.
Wessel-Beaver L (2000) Evidence for the cener of diversity of Cucurbita moschata in Colombia.
Cucurbit Genet Coop Rep 23: 54–55.
Whitaker TW (1947) American origin of the cultivated cucurbits. Ann MO Bot Gard 34:
101–111.
Whitaker TW (1971) Endemism and pre-Columbian migration of the bottle gourd, Lagenaria
siceraria (Mol.) Standl. In: JC Kelley, CW Pennington, RL Rands (eds) Man Across the
Sea. Univ of Texas Press, Austin, TX, USA, pp 320–327.
Whitaker TW, Davis GN (1962) Cucurbit, Botany, Cultivation and Utilization. Interscience
Publ, New York, USA, 249 p.
Whitaker TW, Bemis WP (1976) Cucurbits, Cucumis, Citrullus, Cucurbita, Lagenaria
(Cucurbitaceae). In: NW Simmonds (ed) Evolution of Crop Plants. Longrams, New
York, USA, pp 64–69.
Wilson HD, Doebley J, Duvall M (1992) Chloroplast DNA diversity among wild and cultivated
members of Cucurbita (Cucurbitaceae). Theor Appl Genet 84: 859–865.
Wu T, Zhou J, Zhang Y, Cao J (2007) Characterization and inheritance of a bush-type in tropical
pumpkin (Cucurbita moschata Duchesne). Sci Hort 114: 1–4.
Yanev Z, Shabelsky E, Schafferman D (1999) Colocynth: potential arid land oilseed from an
ancient cucurbit. In: J Janick (ed) Perspectives on New Crops and New Uses. ASHS Press,
Alexandria, Va, USA, pp 257–261.
Yang Y-W, Tai P-Y, Chen Y, Li W-H (2002) A study of the phylogeny of Brassica rapa, B. nigra,
Raphanus sativus and their related genera using noncoding regions of chloroplast DNA.
Mol Phylogenet Evol 23: 268–275.
ABSTRACT
Mapping of plant genomes has progressed rather rapidly in the
last two decades. The initial low density mapping with isozyme or
morphological markers has been replaced with high density mapping
using DNA markers. Such a trend is more visible within the last decade
when at least 24 maps have been created in cucurbits for melon,
cucumber, watermelon and Cucurbita ssp. Most maps were produced
for melon since 2004: 11, and six, three, and four were generated for
cucumber, watermelon and Cucurbita, respectively. In sync with this
trend, increasingly sophisticated DNA marker systems were also used:
from less abundant and radioactive RFLP to more abundant and user-
friendly SSR and SNP. The increased technical advance also facilitated
cloning of genes underlying agriculturally important traits such as
sex expression and disease resistance. The power of next generation
sequencing technology in uncovering more SSRs in the plant genome
has just been demonstrated in cucumber. This will no doubt increase
the speed of marker identification and mapping in cucurbits.
Keywords: cucurbit, genetic mapping, SSR, SNP, AFLP, RFLP, RAPD
6.1 Introduction
Genetic maps have been continuously developed for cucurbits in the last
two decades mostly using molecular markers. Maps are important for all
important agricultural crops because of two reasons. Firstly, these maps
can be used to track inheritance of traits of interest, be they single-gene
Population Parents Marker type and Number Average Length of Mapping software Reference
number of linkage marker the map
groups distance (cM)
(cM)
Melon 2n = 2x = 24
99 RILs PI414723 (C. melo subsp. 386 SSRs, 76 SNPs, 12 2.672 1222 Harel-Beja et al. 2010
agrestis) × ‘Dulce’ (C. melo six INDELs and 200
subsp. melo) AFLPs
116 F2 Q 3-2-2 (Chinese accession) 155 SSRs, 9 ESTs, 7 12 6.4 1095 MAPMAKER 3.0 Cuevas et al. 2009
202
Population Parents Marker type and Number Average Length of Mapping software Reference
Cucurbita 2n = 2x = 40
94 F2 Waltham Butternut (WB) 205 SSRs and 2 27 7 1445.4 MAPMAKER/EXP Gong et al. 2008a
(from each × Nigerian Local (NL) and phenotypic traits 3.0 and JoinMap
mapping ZHOU (hull-less) × WB
population;
merged
map)
Table 6-1 contd....
204
Population Parents Marker type and Number Average Length of Mapping software Reference
number of linkage marker the map
fragment sets that perfectly match the adapters and adjacent nucleotide(s)
(Vos et al. 1995). Although it is a portable and efficient marker system and
has been used in merging maps (Périn et al. 2002), it is included in 16 of
the 42 maps presumably because of its technical sophistication. Three maps
were constructed mostly with AFLP markers (Wang et al. 1997; Park et al.
2000; Yi et al. 2004).
Both RAPD and AFLP can be amplified by PCR, facilitating rapid large-
scale genotyping. However, these markers are usually genotype-specific
and are dominant, which limit integration of independently constructed
genetic maps. Microsatellite or simple sequence repeat (SSR) markers, on
the other hand, are hypervariable, multiallelic, often codominant, evenly
distributed through the genome, highly reproducible and they can be used
as anchor points for molecular linkage group comparisons and map merging
(Danin-Poleg et al. 2000; Gonzalo et al. 2005). For these reasons, SSRs have
been used more frequently in the recently developed maps, including
four maps almost exclusively constructed with SSR markers (Fukino et al.
2008a, b; Gong et al. 2008a; Ren et al. 2009). This may explain that 21 of the
42 maps contain SSR markers. SSR markers can be designed from cucurbit
genomic (gSSRs; Katzir et al. 1996; Jarret et al. 1997; Danin-Poleg et al. 2001;
Fazio et al. 2002; Chiba et al. 2003; Fukino et al. 2008; Gong et al. 2008b; Ren
et al. 2009) or expressed sequence tags (EST, also denominated EST-SSRs)
sequences (Katzir et al. 1996; Fernandez- Silva et al. 2008). EST-SSRs have
several advantages: (i) development costs are relatively low; (ii) they are
related to genes, being functional markers that can be used as candidate
genes to study their association with phenotypic variation, (iii) the flanking
sequences are more likely to be conserved among close or distant species
than those derived from genomic sequences, making their use as markers
for comparative mapping easier (Katzir et al. 1996; Fernandez- Silva et
al. 2008). The use of next generation sequencing significantly speed up
SSR discovery in cucurbits. Cavagnaro et al. (2010) resequenced the Gy14
cucumber genome and detected a total of 112,073 perfect repeats and from
these developed 83,000 SSRs for mapping the cucumber genome (see also
Chapter 11).
The latest marker system is called single nucleotide polymorphisms
(SNPs), which are most abundant in the plant genome (Brookes 1999;
Deleu et al. 2009). For example, in a survey of a 15 kb melon sequence
between Piel de sapo and PI 161375, there is one SNP for every 441 bp on
average (Morales et al. 2004). This would make it an ideal marker system to
saturate a map. For example, a bovine genetic map has been created with
6,769 SNPs that mapped to 3,078 unique positions with average distance
of 1.01 cM between these positions (Arias et al. 2009). In plants, Sato et al.
(2009) mapped 1,717 EST sequences as SNP markers to create an ultra-high-
density barley genetic map. But because of the expense of high-throughput
technology for SNP discovery and detection, they have not been widely
used in cucurbits (Morales et al. 2004). However, four maps in Table 6-1
did employ SNP markers including the one in melon that used 115 SNPs
(Harel-Beja et al. 2010).
It is clear from Table 6-1 that genetic map construction typically requires
linkage analyses of hundreds of Mendelian loci (molecular markers and
phenotypic traits), using a relatively large mapping population that is in
linkage disequilibrium. It can be expensive and time-consuming especially
when the objective is to obtain a high-density map or to incorporate a large
number of markers into an existing map. Vision et al. (2000) proposed a two-
step strategy (called “bin mapping”) that uses a much smaller population
size to solve the problem. First, a mapping population of standard size
(60–250; see Table 6-1) is used to construct a saturated framework map,
and second, new markers are added to this map with lower resolution
using a selected subset of highly informative plants (the bin set) from the
mapping population. The objective is to lower the cost of genotyping new
markers with a minimal loss of resolution. The optimal bin set of a given
size has the maximum possible number of breakpoints evenly spaced
throughout the genome, resulting in a high number of small bins of uniform
size (Howad et al. 2005). Genetic mapping using this strategy have been
carried out in melon (Fernandez-Silva et al. 2008; Moreno et al. 2008). A
bin-map was constructed including 80 RFLPs, 212 SSRs, 3 SNPs and the
Nsv locus, distributed in 122 bins with an average bin length of 10.2 cM
and a maximum bin length of 33 cM. Map density was 4.2 cM/marker or
5.9 cM/SSR (Fernandez-Silva et al. 2008).
In addition to markers developed from genomic sequences, EST
sequences have been increasingly used in cucurbit genetic mapping. ESTs
are partial cDNA sequences from expressed genes. Since genetic mapping of
ESTs establishes locations of known genes, a high-density EST map provides
the foundation for map-based genome analysis such as map-based cloning,
gene tagging and comparative mapping. Fernandez-Silva et al. (2008)
developed 126 SSR markers from about 30,000 melon ESTs and applied these
in bin mapping of the melon genome. Similarly, Levi et al. (2008) identified
40 SSRs from 4,700 non-redundant watermelon fruit ESTs. On the other
hand, half of the markers (187 of 370) in the latest melon map developed
by Harel-Beja et al. (2010) are derived from melon fruit EST sequences. In
addition to EST-SSR markers, EST-derived sequence-tagged site markers
(eSTS) have been extensively explored in other plants. For example, Sato
et al. (2009) designed 10,366 eSTS primer sets based on analysis of 60,000
barley ESTs. Of the 10,366 sets, 7,700 amplified useful products, 3,975 of
these detected polymorphism between the mapping parents and 2,890
eSTS markers were mapped to the barley genome. Significant cucurbit EST
resources have been generated. There are 3,5547 melon ESTs, 7,757 cucumber
ESTs, 7,891 watermelon ESTs and 879 Cucurbita ESTs already deposited in
GenBank. The International Cucurbit Genomics Initiative (www.icugi.org)
aims to produce additional 100,000 melon ESTs. And effort to map ESTs has
already begun (Harel-Beja et al. 2010). It will be a matter of time before EST
mapping in cucurbits catches up those in other crop plants.
A marker system also utilizing EST is an adapted AFLP named cDNA-
AFLP. The technique is similar to the normal AFLP but uses double-stranded
cDNA derived from mRNA as a template instead of genomic DNA. The
obtained cDNA–AFLP fragments called transcriptome derived fragments
(TDFs; Bachem et al. 1996) target coding regions of the genome, which can
be used to construct a genetic map (Brugmans et al. 2002; Li et al. 2003;
Ritter et al. 2008). For example, the potato map created by Ritter et al.
(2008) contains nearly 700 TDFs. Since TDFs are expressed genes, a map so
created can help identify genes involved in, or controlling, various biological
processes ranging from development to responses to environmental cues.
But because TDFs represent mainly highly expressed house-keeping genes
(Ritter et al. 2008), polymorphisms detected by TDFs are due to genetically
different gene expression, not due to differential expression caused by
environmental/physiological factors (i.e., plants at slightly different
physiological stages or grown in slightly different microenvironments).
Based on results from cucumber and potato, the vast majority of absence/
presence polymorphisms in transcripts are caused by genomic sequence
polymorphisms (Brugmans et al. 2002; Bae et al. 2006). The technique may
also be useful for gene tagging using bulked segregant analysis (BSA;
Michelmore et al. 1991) in addition to the whole-genome mapping, which
has not been done in cucurbits.
All the marker systems except cDNA-AFLP discussed above have been
used in cucurbit genome mapping. If genetic maps are to be populated with
markers representing coding regions, then markers developed from ESTs
and cDNAs should be considered. Since a number of maps are already
using EST-based markers, it is time to consider cDNA-AFLP to explore its
utility in cucurbits. Additional marker system such as sequence related
amplified polymorphism (SRAP; Li and Quiros 2001), inter-microsatellite
amplification or inter-SSR (IMA or ISSR; Zietkiewicz et al. 1994) and
sequence characterized amplified regions (SCARs; Paran and Michelmore
1993) are used in six, 12 and nine maps, respectively (Table 6-1) and are not
discussed further in this chapter. A recent review of these marker systems
can be found in Semagn et al. (2006).
of all recent molecular maps are listed in Table 6-1. (see Ezura and Fukino
2009 and Wang et al. 2006 for earlier maps in cucurbits). Each map was
developed with a particular mapping population (backcross, F2 or its
derivative RILs/DHLs) from two chosen parental lines, which usually
possess two different sets of phenotypes. For example, one parent may
have a number of desirable agronomic traits but lacks resistance to multiple
diseases while the other parent is completely opposite. Maps created from
such parents are useful to identify markers linked to these traits, which may
eventually lead to cloning of the underlying genes. The cloned genes can
be used in a breeding program either as a tool in marker-assisted selection
(MAS; see Chapter 7) or through genetic engineering.
Tremendous efforts by various research groups have been devoted
to mapping of the cucurbit genomes, which are small and comparable
to that of rice in size (Arumuganathan and Earle 1991). In total, 9,324
markers including 80 phenotypic traits have been mapped to the cucurbit
genomes. Melon genome is the most mapped with 3,444 markers, followed
by cucumber, Cucurbita, and watermelon (Table 6-2). RAPD, AFLP, SSR
and RFLP account for over 85% of the mapped markers. While RAPD is
the predominant marker system used in watermelon and Cucurbita, AFLP
was predominantly used in melon and SSR was predominantly used in
cucumber. On the other hand, RFLP and SNP were mostly used in melon
and SRAP was mostly used in cucumber. It is possible that some of these
markers are redundant and were used in different maps so the number of
unique markers may be lower. In addition, the actual number of mapped
phenotypic traits may be higher because in a lot of cases a particular trait was
mapped using bulked segregant analysis (Michelmore et al. 1991), which
may not be included in the map of the whole genome. This is especially
true when the purpose of mapping was to positionally clone the gene
underlying the mapped trait, not the whole genome. For example, Fom-2
was not included in the map by Wang et al. (1997). Instead, it was further
mapped with additional markers using BSA (Wang et al. 2000), which
eventually led to cloning of the gene (Joobeur et al. 2004).
There is great interest to create a reference map for each cucurbit
species. Markers from such a map may be used to facilitate mapping/
cloning of new genes. This is especially useful if a map is high-density with
transferable markers such as SSR. Ren et al. (2009) tested 995 cucumber
SSRs mapped in the cucumber genome in other cucurbits. Among them,
49, 26 and 22% of the cucumber SSRs amplified PCR products in melon,
watermelon and pumpkin, respectively with polymorphism rate at 39.6, 46.5
and 54.8%. Obviously, these markers can be used to enhance the mapping
efforts in these cucurbits. An example of Cucurbita genetic map is shown
in Fig. 6-1.
Figure 6-1 A genetic map of Cucurbita pepo. The new map contains 659 loci: 178 SSR, 244
AFLP, 230 RAPD, five SCAR markers, and two morphological traits (h and B) (From Gong
et al. 2008b).
Color image of this figure appears in the color plate section at the end of the book.
RAPD AFLP SSR RFLP SRAP ISSR SNP SCAR STS PT** Total
Melon 436 1,212 741 616 152 101 121 9 0 56 3,444
Cucumber 247 577 1,162 109 446 33 1 64 2 17 2,658
Watermelon 1,069 221 14 54 93 95 0 6 0 2 1,554
Cucurbita 821 451 386 0 0 0 0 5 0 5 1,668
Total 2,573 2,461 2,303 779 691 229 122 84 2 80 9,324
Note: *Isozyme marker is not included in the table. The numbers include those in earlier maps not listed in Table 6-3. **PT-phenotypic trait.
between newly derived markers STS296 and SSR451. The two recombination
events found between STS411 and Fom-2 were confined to 5.5 kb between
STS411 and another new marker SSR430. Thus Fom-2 was assigned to 75
kb-size interval between STS411 and STS296. Three putative genes were
found in this 75 kb interval and only two were found to be complete. One
was similar to Arabidopsis AtCPSF73-II, which was primarily expressed
in flower tissue (Xu et al. 2004). The other was highly homologous to
previously characterized NBS-LRR class of resistance genes such as I2 in
tomato and was thus designed as Fom-2 (Joobeur et al. 2004).
Fom-2 was also cloned by Pitrat and colleagues (Pech et al. 2007) using
segregating populatins from Védrantais × PI 161375. A BAC contig was
built using clones identified from the MR-1 BAC library (Luo et al. 2001)
based on linked markers. Sequencing identified three candidate genes, one
of which shares high homology to the NBS-LRR class of resistance genes.
This gene is 3 kb in length and encodes an intronless protein of 1,073 amino
acids. Sequencing analysis also revealed variation in LRR region of the
gene between resistant MR-1 and susceptible Védrantais, AY and Durango
varieties (Pech et al. 2007).
The melon Vat gene is the second cloned cucurbit gene that mediates
resistance to the melon/cotton aphid Aphis gossypii. Two segregating
populations were used to clone the Vat gene (Pauquet et al. 2004). A
population of 200 RILs from Védrantais × PI 161375 was used to first fine-
map the region and another population of 6,000 backcross progeny from
(Védrantais × PI 161375) × Védrantais was screened for recombination
events within 1.7 cM delimited by markers flanking Vat to fine-map the
region further. Markers tightly linked to the gene were used to screen a
PI 161375 BAC library. Markers were generated from end sequences of
the identified BAC clones and fine-mapping of these markers using the
backcross population further delimited a physical interval that contains
a single gene of 5.9 kb. The gene has five exons and four introns and
encodes a protein which belongs to the coiled coil (CC)—NBS—LRR family.
Transferring an 11-kb genomic fragment carrying Vat and its own promoter
into the susceptible Védrantais confers to resistance to aphids (Pauquet et
al. 2004).
The recessive nsv gene, which confers complete resistance to melon
necrotic spot virus (MNSV), is another gene cloned by the map-based
cloning approach in cucurbits, except that the cloning process also took
advantage of microsynteny between melon and Arabidopsis. Using two
mapping population of 408 F2 (PI 161375 × Piel de sapo) and 2,727 BC1
([Védrantais × PI 161375] × PI 161375), nsv was mapped in a 3.2 cM region
flanked by CAPS markers M29 and M132 (Morales et al. 2005). Additional
markers were developed from BAC clones identified by linked markers
and one of these markers (52K20sp6) cosegregated with nsv in the mapping
populations. A single BAC clone of 100 kb was thus identified that covered
a genetic distance of 0.73 cM (Morales et al. 2005). Using microsynteny
between melon and Arabidopsis (van Leeuwen et al. 2003), markers linked
to nsv (including 1R3 and 1L3) were compared to the Arabidopsis sequence
by BLAST analysis, which identified a region in Arabidopsis chromosome
4 as the most probable nsv syntenic region (Nieto et al. 2006). 1R3 and 1L3
mapped within a 182-kb Arabidopsis genomic region located between genes
At4g17770 and At4g18100. Among the genes in the region was eIF4E at
position 18040. Degenerate PCR primers designed based on eIF4E sequences
from other species amplified a product of 1.9 kb which was sequenced and
was used to design CAPS marker (M-CmeIF4E) primers from the resistant
and susceptible parents. M-CmeIF4E cosegregated with nsv among more
than 3,000 progeny of the F2 and BC1. Full length Cm-eIF4E cDNA from
the homozygous dominant (Védrantais—susceptible) and homozygous
recessive (PI 161375—resistant) parents were sequenced. The cDNAs were
1,153 bp in length with a 5’-UTR of 122 bp, a coding region of 708 bp and
a 3’-UTR of 323 bp. Sequence comparison of Cm-eIF4E proteins from the
resistant (PI) and the susceptible (Ved) cultivars revealed a single amino acid
substitution at position 228. Védrantais carries a Histidine and PI 161375
carries a Leucine. A single nucleotide change lead to this amino acid change
in the protein. A molecular marker derived from the SNP cosegregated with
nsv in the mapping population of more than 3,000 segregating plants and
differentiated seven resistant genotypes from six susceptible ones (Nieto
et al. 2006).
The first non-resistance gene cloned by map-based cloning is the
gene andromonoecious (a), which together with gynoecious (g) governs sex
determination in melon. Monoecious (A_G_) and andromonoecious (aaG_)
plants bear male flowers on the main stem and female or hermaphrodite
flowers on axillary branches while gynoecious (AAgg) and hermaphrodite
(aagg) plants only produce female and hermaphrodite flowers (Kenigsbuch
and Cohen 1990). But these patterns can be modified by hormones such
as ethylene and environmental factors (Byers et al. 1972). To fine-map a,
Boualem et al. (2008) crossed a monoecious melon cultivar PI124112 (AAGG)
and an andromonoecious cultivar Védrantais (aaGG), and backcrossed
the resultant F 1 plants with Védrantais. To identify plants carrying
recombination events linked to a locus, DNA samples were extracted from
7,000 plants from the backcross population and analyzed with the a locus
flanking markers M64 and M47. The sexual phenotype was determined for
all the 235 recombinant plants. The fine-mapping and ensued chromosome
walking delimited the a locus to a single BAC clone that had seven genes.
The two closest flanking markers (L41 and R5) were used to identify a
14 kb region containing a single gene encoding a 1-aminocyclopropane-
1-carboylic acid synthase (ACS) designated CmACS-7. ACS catalyzes a
Acknowledgement
I thank Hugo E. Cuevas for his contribution to Table 6-1.
References
Arumuganathan K, Earle ED (1991) Nuclear DNA content of some important plant species.
Plant Mol Biol Rep 9: 208–219.
Arias JA, Keehan M, Fisher P, Coppieters W, Spelman R (2009) A high density linkage map of
the bovine genome. BMC Genet 10: 18.
Bachem C, van der Hoeven RS, de Bruijn SM, Vreugdenhil D, Zabeau M, Visser RG (1996) Visualization
of differential gene expression using a novel method of RNA fingerprinting based on AFLP:
analysis of gene expression during potato tuber development. Plant J 9: 745–753.
Bae KM, Kwon YS, Cho IH, Yi SI (2006) Use of cDNA-AFLP for transcript profiling in narrow
genetic pools; for example, cucumber (Cucumis sativus L.). Plant Breed 125: 488–492.
Baudracco-Arnas S, Pitiat M (1996) A genetic map melon (Cucumic melo L.) with RFLP, RAPD,
Isozyme, disease resistance and morphological markers. Theor Appl Genet 93: 57–64.
Botstein D, White RL, Skolnick M, Davis RW (1980) Construction of a genetic map in man
using restriction fragment length polymorphisms. Am J Hum Genet 32: 314–331.
Boualem A, Fergany M, Fernandez R, Troadec C, Martin A, Morin H, Sari MA, Collin F, Flowers
JM, Pitrat M, Purugganan MD, Dogimont C, Bendahmane A (2008) A conserved mutation in
an ethylene biosynthesis enzyme leads to andromonoecy in melons. Science 321: 836–838.
Brookes AJ (1999) The essence of SNPs. Gene 234: 177–186.
Brugmans B, Fernandez del Carmen A, Bachem CW, van Os H, van Eck HJ, Visser RG (2002) A novel
method for the construction of genome wide transcriptome maps. Plant J 31: 211–222.
Byers RE, Baker LR, Sell HM, Herner RC, Dilley DR (1972) Ethylene: A Natural Regulator of
Sex Expression of Cucumis melo L. Proc Natl Acad Sci USA 69(3): 717–720.
Cavagnaro PF, Senalik DA, Yang L, Simon PW, Harkins TT, Kodira CD, Huang S, Weng Y
(2010) Genome-wide characterization of simple sequence repeats in cucumber (Cucumis
sativus L.). BMC Genomics 11: 569.
Chiba N, Suwabe K, Nunome T, Hirai M (2003) Development of microsatellite markers in melon
(Cucumis melo L.) and their application to major cucurbit crops. Breed Sci 53: 21–27.
Cuevas HE, Staub JE, Simon PW, Zalapa JE, McCreight JD (2008) Mapping of genetic loci that
regulated quantity of β-carotene in fruit of U.S. Western Shipping melon (Cucumis melo
L.). Theor Appl Genet 117: 1345–1359.
Cuevas HE, Staub JE, Simon PW, Zalapa JE (2009) A consensus linkage map identifies genomic
regions controlling fruit maturity and beta-carotene-associated flesh color in melon
(Cucumis melo L.) Theor Appl Genet 119: 741–756.
Danin-Poleg Y, Reis N, Baudracco-Arnas S, Pitrat M, Staub JE, Oliver M, Arus P, de Vincente
CM, Katzir N (2000) Simple sequence repeats in Cucumis mapping and map merging.
Genome 43: 963–974.
Danin-Poleg Y, Reis N, Tzuri G, Katzir N (2001) Development and characterization of
microsatellite markers in Cucumis. Theor Appl Genet 102: 61–72.
Deleu W, Esteras C, Roig C, González-To M, Fernández-Silva I, Gonzalez-Ibeas D, Blanca J,
Aranda MA, Arús P, Nuez F, Monforte AJ, Picó MB, Garcia-Mas J (2009) A set of EST-SNPs
for map saturation and cultivar identification in melon. BMC Plant Biol 9: 90.
Esquinas JT (1981) Alloenzyme variation and relationships among Spanish land-races of
Cucumis melo L. Kulturpflanze 29: 337–352.
Ezura H, Fukino N (2009) Research tools for functional genomics in melon (Cucumis melo
L.): Current status and prospects. Plant Biotechnol 26: 359–368.
Fanourakis NE, Simon PW (1987) Analysis of genetic linkage in cucumber. J Hered 78: 238–242.
Fazio G, Staub JE, Chung SM (2002) Development and characterization of PCR markers in
cucumber (Cucumis sativus L.). J Am Soc Hort Sci 127: 545–557.
Fernandez-Silva I, Eduardo I, Blanca J, Esteras C, Picó B, Nuez F, Arús P, Garcia-Mas J, Monforte
AJ (2008) Bin mapping of genomic and EST-derived SSRs in melon (Cucumis melo L.).
Theor Appl Genet 118: 139–150.
Fukino N, Ohara T, Monforte AJ, Sugiyama M, Sakata Y, Kunihisa M, Matsumoto S (2008a)
Identification of QTLs for resistance to powdery mildew and SSR markers diagnostic
for powdery mildew resistance genes in melon (Cucumis melo L.). Theor Appl Genet
118: 165–175.
Fukino N, Yoshioka Y, Kubo N, Hirai M, Sugiyama M, Sakata Y, Matsumoto S (2008b)
Development of 101 novel SSR markers and construction of an SSR-based genetic linkage
map in cucumber (Cucumis sativus L.). Breed Sci 58: 475–483.
Gong L, Pachner M, Kalai K, Lelley T (2008a) SSR-based genetic linkage map of Cucurbita
moschata and its synteny with Cucurbita pepo. Genome 51: 878–887.
Gong L, Stift G, Kofler R, Pachner M, Lelley T (2008b) Microsatellites for the genus Cucurbita and
an SSR-based genetic linkage map of Cucurbita pepo L. Theor Appl Genet 117: 37–48.
Gonzalo MJ, Oliver M, Garcia-Mas J, Monfort A, Dolcet-Sanjuan R, Katzir N, Arús P, Monforte
AJ (2005) Simple-sequence repeat markers used in merging linkage maps of melon
(Cucumis melo L.). Theor Appl Genet 110: 802–811.
Harel-Beja R, Tzuri G, Portnoy V, Lotan-Pompan M, Lev S, Cohen S, Dai N, Yeselson L, Meir A,
Libhaber SE, Avisar E, Melame T, van Koert P, Verbakel H, Hofstede R, Volpin H, Oliver
M, Fougedoire A, Stalh C, Fauve J, Copes B, Fei Z, Giovannoni J, Ori N, Lewinsohn E,
Sherman A, Burger J, Tadmor Y, Schaffer AA, Katzir N (2010) A genetic map of melon
highly enriched with fruit quality QTLs and EST markers, including sugar and carotenoid
metabolism genes. Theor Appl Genet 121: 511–533.
Howad W, Yamamoto T, Dirlewanger E, Testolin R, Cosson P, Cipriani G, Monforte AJ, Georgi
L, Abbott AG, Arús P (2005) Mapping with a few plants: using selective mapping for
microsatellite saturation of the Prunus reference map. Genetics 171: 1305–1309.
Huang S, Li R, Zhang Z, Li L, Gu X, Fan W, Lucas WJ, Wang X, Xie B, Ni P, Ren Y, Zhu H, Li
J, Lin K, Jin W, Fei Z, Li G, Staub J, Kilian A, van der Vossen EA, Wu Y, Guo J, He J, Jia
Z, Ren Y, Tian G, Lu Y, Ruan J, Qian W, Wang M, Huang Q, Li B, Xuan Z, Cao J, Asan,
Wu Z, Zhang J, Cai Q, Bai Y, Zhao B, Han Y, Li Y, Li X, Wang S, Shi Q, Liu S, Cho WK,
Kim JY, Xu Y, Heller-Uszynska K, Miao H, Cheng Z, Zhang S, Wu J, Yang Y, Kang H,
Li M, Liang H, Ren X, Shi Z, Wen M, Jian M, Yang H, Zhang G, Yang Z, Chen R, Liu S,
Li J, Ma L, Liu H, Zhou Y, Zhao J, Fang X, Li G, Fang L, Li Y, Liu D, Zheng H, Zhang Y,
Qin N, Li Z, Yang G, Yang S, Bolund L, Kristiansen K, Zheng H, Li S, Zhang X, Yang H,
Wang J, Sun R, Zhang B, Jiang S, Wang J, Du Y, Li S (2009) The genome of the cucumber,
Cucumis sativus L. Nat Genet 41: 1275–1281.
Jarret RL, Merrick LC, Holms T, Evans J, Aradhya MK (1997) Simple sequence repeats in
watermelon (Citrullus lanatus (Thunb.) Matsum. & Nakai). Genome 40: 433–441.
Jones CJ, Edwards KJ, Castaglione S, Winfield MO, Sala F, Wiel CVd, Bredemeijer G, Vosman
B, Matthes M, Daly A, Brettschneider R, Bettini P, Buiatti M, Maestri E, Malcevschi
A, Marmiroli N, Aert R, Volckaert G, Rueda J, Linacero R, Vazquez A, Karp A (1997)
Reproductibility testing of RAPD, AFLP and SSR markers in plants by a network of
European laboratories. Mol Breed 3: 381–390.
Joobeur T, King JJ, Nolin SJ, Thomas CE, Dean RA (2004) The Fusarium wilt resistance locus Fom-2
of melon contains a single resistance gene with complex features. Plant J 39: 283–297.
Katzir N, Danin-Poleg Y, Tzuri G, Karchi Z, Lavi U, Cregan PB (1996) Length polymorphism
and homologies of microsatellites in several Cucurbitaceae species. Theor Appl Genet
93: 1282–1290.
Kenigsbuch D, Cohen Y (1990) The inheritance of gynoecy in muskmelon. Genome 33:
317–320.
Kennard WC, Poetter K, Dijkhuizen A, Megtic V, Staub JE, Harvey MJ (1994) Linkage among
RFLP, RAPD, isozyme, disease-resistance, and morphological markers in narrow and
wide crosses of cucumber. Theor Appl Genet 89: 42–48.
Knerr LD, Staub JE (1992) Inheritance and linkage relationships of isozyme loci in cucumber
(Cucumis sativus L.). Theor Appl Genet 84: 217–224.
Lander ES, Green P, Abrahamson J, Barlow A, Daly MJ, Lincoln SE, Newburg L (1987)
MAPMAKER: an interactive computer package for constructing primary genetic linkage
maps of experimental and natural populations. Genomics 1: 174–181.
van Leeuwen H, Monfort A, Zhang HB, Puigdomenech P (2003) Identification and
characterisation of a melon genomic region containing a resistance gene cluster from a
constructed BAC library. Microcolinearity between Cucumis melo and Arabidopsis thaliana.
Plant Mol Biol 51: 703–718.
Levi A, Thomas CE, Trebitsh T, Salman A , King J, Karalium J, Newman M, Reddy OUK, Xu
Y, Zhang X (2006) An extended linkage map for watermelon based on SRAP, AFLP, SSR,
ISSR, and RAPD markers. J Am Soc Hort Sci 131: 393–402.
Levi A, Wechter PW, Davis A (2008) EST-PCR markers representing watermelon fruit genes
are polymorphic among watermelon heirloom cultivars sharing a narrow genetic base.
Plant Genet Resour 7: 16–32.
Li G, Quiros CF (2001) Sequence-related amplified polymorphism (SRAP), a new marker
system based on a simple PCR reaction: its application to mapping and gene tagging in
Brassica. Theor Appl Genet 103: 455–461.
Li G, Gao M, Yang B, Quiros CF (2003) Gene for gene alignment between the Brassica and
Arabidopsis genomes by direct transcriptome mapping. Theor Appl Genet 107: 168–180.
Li Z, Pan J, Guan Y, Tao Q, He H, Si L, Cai R (2008) Development and fine mapping of three
co-dominant SCAR markers linked to the M/m gene in the cucumber plant (Cucumis
sativus L.). Theor Appl Genet 117: 1253–1260.
Luo M, Wang Y-H, Frisch D, Joobeur T, Wing R, Dean RA (2001) Melon BAC library construction
using improved methods and identification of clones linked to the locus conferring
resistance to melon fusarium wilt (Fom-2). Genome 44: 154–162.
McCallum CM, Comai L, Greene EA, Henikoff S (2000) Targeted screening for induced
mutations. Nat Biotechnol 18(4): 455–457.
Meglic V, Staub JE (1996) Inheritance and linkage relationships of allozyme and morphological
loci in cucumber (Cucumis sativus L.). Theor Appl Genet 92: 865–872.
Michelmore RW, Paran I, Kesseli RV (1991) Identification of markers linked to disease-resistance
genes by bulked segregant analysis: a rapid method to detect markers in specific genomic
regions by using segregating populations. Proc Natl Acad Sci USA 88: 9828–9832.
Morales M, Roig E, Monforte AJ, Arús P, Garcia-Mas J (2004) Single-nucleotide polymorphisms
detected in expressed sequence tags of melon (Cucumis melo L.). Genome 47: 352–360.
Morales M, Orjeda G, Nieto C, van Leeuwen H, Monfort A, Charpentier M, Caboche M, Arús
P, Puigdomènech P, Aranda MA, Dogimont C, Bendahmane A, Garcia-Mas J (2005) A
physical map covering the nsv locus that confers resistance to Melon necrotic spot virus
in melon (Cucumis melo L.). Theor Appl Genet 111: 914–922.
Moreno E, Obando J, Dos-Santos N, Fernández-Trujillo JP, Monforte AJ, Garcia-Mas J (2008)
Candidate genes and QTLs for fruit ripening and softening in melon. Theor Appl Genet
116: 589–602.
Neuhausen SL (1992) Evaluation of restriction fragment length polymorphism in Cucumis
melo. Theor Appl Genet 83: 379–384.
Nieto C, Morales M, Orjeda G, Clepet C, Monfort A, Sturbois B, Puigdomènech P, Pitrat M,
Caboche M, Dogimont C, Garcia-Mas J, Aranda MA, Bendahmane A (2006) An eIF4E
allele confers resistance to an uncapped and non-polyadenylated RNA virus in melon.
Plant J 48(3): 452–62.
Navot N, Zamir D (1987) Isozyme and seed protein phylogeny of the genus Citrullus
(Cucurbitaceae). Plant Syst Evol 156: 61–67.
Navot N, Sarfatti M, Zamir D (1990) Linkage relationships of genes affecting bitterness and
flesh color in watermelon. J Hered 81: 162–165.
Oliver M, Garcia-Mas J, Carús M, Pueyo N, López-Sesé AL, Arroyo M, Gómez-Paniagua H, Arús P, de
Vicente MC (2001) Constriction of a reference linkage map for melon. Genome 44: 836–845.
Paran I, Michelmore RW (1993) Development of reliable PCR-based markers linked to downy
mildew resistance genes in lettuce. Theor Appl Genet 85: 985–993.
Park YH, Sensoy S, Wye C, Antonise R, Peleman J, Havey MJ (2000) A genetic map of cucumber
composed of RAPDs, RFLPs, AFLPs, and loci conditioning resistance to papaya ringspot
and zucchini yellow mosaic viruses. Genome 43: 1003–1010.
Pauquet J, Burget E, Hagen L, Chovelon V, Le Menn A, Valot N, Desloire S, Caboche M, Rousselle
P, Pitrat M, Bendahmane A, Dogimont C (2004) Map-based cloning of the Vat gene from
melon conferring resistance to both aphid colonization and aphid transmission of several
viruses. In: A Lebeda, H Paris (eds) Proc Cucurbitaceae 2004, 8th EUCARPIA Meeting on
Cucurbit Genetics and Breeding, Palaky Univ, Olomouc, Czech Republic, pp 325–329.
Pech JC, Bernadac A, Bouzayen M, Latche A, Dogimont C, Pitrat M (2007) Melon. In: EC Pua,
MR Davey (eds) Biotechnology in Agriculture and Forestry, vol 60: Transgenic Crops V.
Springer, Berlin, Heidelberg, Germany, pp 209–240.
Perchepied L, Dogimont C, Pitrat M (2005) Strain-specific and recessive QTLs involved in the
control of partial resistance to Fusarium oxysporum f. sp. melonis race 1.2 in a recombinant
inbred line population of melon. Theor Appl Genet 111: 65–74.
Périn C, Hagen S, De Conto V, Katzir N, Danin-Poleg Y, Portnoy V, Baudracco-Arnas S,
Chadoeuf J, Dogimont C, Pitrat M (2002) A reference map of Cucumis melo based on
two recombinant inbred line populations. Theor Appl Genet 104: 1017–1034.
Perl-Treves T, Zamir D, Navot N, Galun E (1985) Phylogeny of Cucumis based on isozyme
variability and its comparison with plastome phylogeny. Theor Appl Genet 71: 430–436.
Pierce LK, Wehner TC (1990) Review of genes and linkage groups in cucumber. HortScience
25: 605–615.
Pitrat M (1991) Linkage groups in Cucumis melo L. J Hered 82: 406–411.
Ren Y, Zhang Z, Liu J, Staub JE, Han Y, Cheng Z, Li X, Lu J, Miao H, Kang H, Xie B, Gu X,
Wang X, Du Y, Jin W, Huang S (2009) An integrated genetic and cytogenetic map of the
cucumber genome. PLoS One 4(6): e5795.
Ritter E, Ruiz de Galarreta JI, van Eck HJ, Sánchez I (2008) Construction of a potato transcriptome
map based on the cDNA-AFLP technique. Theor Appl Genet 116: 1003–1013.
Rounsley S, Marri PR, Yu Y, He R, Sisneros N, Goicoechea JL, Lee SL, Angelova A, Kudrna
D, Luo M, Affourtit J, Desany B, Knight J, Niazi F, Egholm M, Wing RA (2009) De novo
next generation sequencing of plant genomes. Rice 2: 35–43.
Sato K, Nankaku N, Takeda K (2009) A high-density transcript linkage map of barley derived
from a single population. Heredity 103: 110–117.
Semagn K, Bjørnstad A, Ndjiondiop MN (2006) An overview of molecular marker methods
for plants. Afri J Biotechnol 5: 2540–2568.
Shattuck-Eidens DM, Bell RN, Neuhausen SL, Helentjaris T (1990) DNA sequence variation
within maize and melon: observations from polymerase chain reaction amplificcation
and direct sequencing. Genetics 126: 207–217.
Stam P (1993) Construction of integrated genetic linkage maps by means of a new computer
package: JOINMAP. Plant J 3: 739–744.
Staub JE, Meglic V (1993) Molecular genetic markers and their legal relevance for cultivar
discrimination: a case study in cucumber. HortTechnology 3: 291–300.
Vakalounakis DJ (1992) Heart leaf; a recessive leaf shape marker in cucumber: Linkage with
disease resistance and other traits. J Hered 83: 217–221.
Vision TJ, Brown DG, Shmoys DB, Durrett RT, Tanksley SD (2000) Selective mapping: a strategy
for optimizing the construction of high-density linkage maps. Genetics 155: 407–420.
Vos P, Hogers R, Bleeker M, Reijans M, van de Lee T, et al. (1995) AFLP: a new technique for
DNA fingerprinting. Nucl Acids Res. 23: 4407–4414.
Wang G, Pan J, Li X, He H, Wu A, Cai R (2005) Construction of a cucumber genetic linkage
map with SRAP markers and location of the genes for lateral branch traits. Sci China Sr
C Life Sci 48: 213–220.
Wang J, Yao J, Li W (2008) Construction of a molecular map for melon (Cucumis melo L.) based
on SRAP. Front Agri China 2: 451–455.
Wang YH, Thomas CE, Dean RA (1997) A genetic map of melon (Cucumis melo L.) based on amplified
fragment length polymorphism (AFLP). Theoretical and Applied Genetics 95: 791–798.
Wang YH, Thomas CE, Dean RA (2000) Genetic mapping of a Fusarium wilt resistance gene
(Fom-2) in melon (Cucumis melo L.). Mol Breed 6: 379–389.
Wang YH, Dean RA, Joobeur T (2006) Genetic mapping and molecular breeding in cucurbits.
Plant Breed Rev 27: 213–244.
Watcharawongpaiboon N, Chunwongse J (2008) Development and characterization of
microsatellite markers from an enriched genomic library of cucumber (Cucumis sativus).
Plant Breed 127: 74–81.
Weeden NF, Robinson RW (1986) Allozyme segregation ratios in the interspecific cross Cucurbita
maxima × C. ecuadorensis suggest that hybrid breakdown is not caused by minor alteration
in chromosome structure. Genetics 114: 593–609.
Williams JGK, Kubelik AR, Livak KJ, Rafalski JA, Tingey SV (1990) DNA polymorphisms amplified
by arbitrary primers are useful as genetic markers. Nucl Acids Res 18: 6531–35.
Xu R, Ye X, Li QQ (2004) AtCPSF73-II gene encoding an Arabidopsis homolog of CPSF 73 kDa
subunit is critical for early embryo development. Gene 324: 35–45.
Yeboah MA, Chen X, Chen RF, Liang G, Gu M (2007) A genetic linkage map of cucumber
(Cucumis sativus L.) combining SRAP and ISSR markers. Afri J Biotechnol 6: 2784–2791.
Yi K, Xu Y, Lu XY, Xiao LT, Xu XL, Gong GY, Zhang HY (2004) Construction of AFLP molecular
genetic map for RIL population of watermelon. Acta Hort Sin 31: 53–58 (in Chinese with
English summary).
Yuan XJ, Pan JS, Cai R, Guan Y, Liu LZ, Zhang WW, Li Z, He HL, Zhang C, Si LT, Zhu LH
(2008a) Genetic mapping and QTL analysis of fruit and flower related traits in cucumber
(Cucumis sativus L.) using recombinant inbred lines. Euphytica 164: 473–491.
Yuan XJ, Li XZ, Pan JS, Wang G, Jiang S, Li XH, Deng SL, He HL, Si MX, Lai L, Wu AZ,Zhu
LH, Cai R (2008b) Genetic linkage map construction and location of QTLs for fruit-related
traits in cucumber. Plant Breed 127: 180–188.
Zalapa JE, Staub JE, McCreight JD, Chung SM, Cuevas HE (2007) Detection of QTL for yield-
related traits using recombinant inbred lines derived from exotic and elite US Western
Shipping melon germplasm. Theor Appl Genet 114: 1185–1201.
Zamir D, Navot N, Rudich J (1984) Enzyme polymorphism in Citrullus lanatus and C. colocynthis
in Israel and Sinai. Plant Syst Evol 146: 163–170.
Zhang R, Xu Y, Yi K, Zhang H, Lie L, Gong G, Levi A (2004) Genetic linkage map for watermelon
derived from recombinant inbred lines (RILs). J Am Soc Hort Sci 129: 237–243.
Zietkiewicz E, Rafalsky A, Labuda D (1994) Genome fingerprinting by simple sequence repeat
(SSR)-anchored polymerase chain reaction amplification. Genomics 20: 176–183.
Zraidi A, Stift G, Pachner M, Shojaeiyan A, Gong L, Lelley T (2007) A consensus map for
Cucurbita pepo. Mol Breed 20: 375–388.
ABSTRACT
Molecular breeding using tightly linked genetic markers has been
widely adopted. The new tool has offered the breeder unprecedented
advantages such as efficiency and time-saving. Like in other crop
plants, these markers have been extensively studied in cucurbits for
monogenic traits. During the last decade, markers linked to or genes
controlling 28 traits of agronomic importance have been identified
and are listed in this chapter. Based on germplasm characterization
results, markers more closely linked to target genes are more robust
in phenotype prediction. In an extreme case, a marker converted from
the a gene itself, which determines andromonoecy, correctly predicted
sex types of all 497 melon varieties. This further indicates that breeding
efficiency can be dramatically increased if tightly linked markers are
used to select for the underlying phenotypes.
Keywords: cucurbits, DNA markers, linkage, bulked segregant analysis,
trait
Gene Locus Parents Population Linked Marker/Distance (cM) Marker Type Reference
Cucumber
Target leaf Q5 (resistant) × P57-1 241 F2 EST-SSR/2.9 SSR Wang et al. 2010
spot (susceptible)
de Gy-7 × H-19 CSWCTT14b/1.4; SSR13251/4.2 SSR Weng et al. 2010
230
Gene Locus Parents Population Linked Marker/Distance (cM) Marker Type Reference
BSA can also be used to identify markers linked to a QTL although not
reported yet in cucurbits. Kanagaraj et al. (2010) used 11 drought-tolerant
and 12 drought-susceptible rice recombinant inbred lines (RILs) to construct
the two bulks. The parents were first screened for polymorphism using 1,206
rice simple sequence repeat (SSR) markers. Out of 134 SSR polymorphic
primers between parents, three primers showed polymorphism between
bulks. These three primers co-segregated among the individual RI lines
constituting the respective bulks. The genomic regions flanked by these
markers have been reported to be associated with several drought resistance
component traits (Kanagaraj et al. 2010). In sugarbeet, root elongation rate
controlled by QTLs was tagged with three AFLP markers using BSA on an F2
population generated from a cross between a high and a low root elongation
parent (Stevanato et al. 2010). The technique can be used in cucurbits to
identify markers linked to important quantitative traits.
varieties against Fusarium wilt (Lin et al. 2009). Another dominant SCAR
marker developed from an AFLP marker linked to a gynoecy gene at 6.7
cM was tested on 21 genotypes with known sex phenotypes. The marker
correctly identified 14 of 15 gynoecious genotypes but it misidentified
one monoecious genotype as gynoecious (Lou et al. 2007). When two
markers linked to Tu (warty fruit) locus at 2.8 (C_SC69) and 3.2 (C_SC24)
cM were tested on 28 warty fruit (TuTu) and 34 non-warty fruit (tutu)
cucumber germplasms, C_SC24 and C_SC69 could correctly predict the
fruit phenotype of 55 and 58, respectively (Zhang et al. 2010b).
It is clear from these examples that MAS is a very effective tool for
selection of plants with desired phenotype. This is the reason that MAS
has been adopted by breeding companies to release commercial varieties
sooner and at lower cost although one does not see it in the literature. For
example, Harris Moran stated in its website (http: //www.harrismoran.
com/products/biotechstatement.htm) that “Harris Moran employs many
advanced techniques to assist and improve the classic plant breeding
work that is the core of our business. One example is the use of Molecular
Markers to allow the identification of specific genes and plant traits. Our
breeders use markers to assist in selection of new plant lines with important
characteristics, our plant pathologists use markers to identify plant diseases
and our quality assurance folks use markers to help insure the seed you
buy is true to type—truly versatile and valuable technology”. Syngenta
also stated in its 2009 Annual Review that “modern technologies such as
marker-assisted breeding enable Syngenta scientists to identify genetic traits
that relate to certain plant characteristics. Using DNA testing, plants with
the desired traits can be identified at an early stage of growth, enabling
much faster development of varieties with enhanced flavor, color, nutrition
and agronomic performance” (Syngenta 2009). The seed company Pioneer
also uses MAS extensively in its commercial variety development program
(Cahill and Schmid 2004). In their experience, MAS is most efficient in early
generation single plant selections.
suitable for the Fom-1 genotype prediction. The Fom-1 genotype in var.
chinensis, conomon and makuwa seemed to be predicted accurately by CAPS2
(5.2 cM), NBS1-CAPS, C-MRGH12 (0.4 cM) and 62-CAPS (1.2 cM). CAPS2
was identical in all cultivars and lines excluding MR-1 as also reported in
their earlier study (Tezuka et al. 2009). The Fom-1 genotype in var. reticulatus
could be predicted accurately by C-TCG/GGT-400, C-MRGH12 and the
cosegregating S-TAG/GCC-470 (Tezuka et al. 2009, 2010). What is interesting
is that C-TCG/GGT-400 and CAPS2 are both CAPS markers derived from
the same AFLP marker TCG/GGT-400. Yet C-TCG/GGT-400 could predict
the resistant phenotype in var. reticulatus and var. cantalupensis but CAPS2
could only do so in var. chinensis, conomon and makuwa (Tezuka et al. 2009,
2010). Similar results were reported in two other studies (Brotman et al. 2005;
Oumouloud et al. 2008), i.e., markers linked to Fom-1 do not always predict
the resistance phenotype in all varieties, only in certain variety groups as
described above. However, one can always combine all the markers to select
the right resistant genotype in MAS.
These results may revive the issue of Fom-3, another gene conferring
resistance to F. oxysporum f.sp. melonis races 0 and 2 in the cultivar “Perlita
FR” (Zink and Gubler 1985). In “Tortuga”, a Spanish cantalupensis accession,
resistance to races 0 and 2 of F. oxysporum f.sp. melonis is found to be
conferred by two independent genes: one dominant and the other recessive.
The dominant gene was shown to be Fom-1 but the recessive gene was
suggested to be named fom-4 (Oumouloud et al. 2010). It should be noted
that both “Perlita FR” and “Tortuga” belong to var. cantalupensis. Obviously,
this is a very intriguing finding and more investigation is warranted to
resolve the issue.
Despite this shortcoming, MAS remains to be an effective breeding tool
for selection of qualitatitive traits controlled by single genes or quantitative
traits controlled by major genes. Genomics and genome-wide study in other
crop plants have shown in the last two years that more markers can result
from the next generation sequencing. By resequencing 517 rice landraces,
Huang et al. (2010) identified 3.6 million SNPs and found markers associated
with 14 agronomic traits of rice. These markers no doubt will be an asset for
MAS. In soybean, Lam et al. (2010) resequenced 17 wild and 14 cultivated
soybeans and concluded that marker-assisted breeding of soybean will
be less challenging than map-based cloning. Not surprisingly, the new
sequencing technology has already been used in cucurbits. Cavagnaro et
al. (2010) resequenced the Gy14 cucumber genome and detected a total
of 112,073 perfect repeats and from these developed 83,000 SSRs. Such a
large number of markers will speed up marker discovery for MAS in the
immediate future.
References
Al-Faifi S, Meyer JD, Garcia-Mas J, Monforte AJ, Havey MJ (2008) Exploiting synteny in
Cucumis for mapping of Psm: a unique locus controlling paternal mitochondrial sorting.
Theor Appl Genet 117(4): 523–9.
Bang H, Kim S, Leskovar D, King S (2007) Development of a codominant CAPS marker for
allelic selection between canary yellow and red watermelon based on SNP in lycopene
β-cyclase (LCYB) gene. Mol Breed 20: 63–72.
Behera TK, Staub JE, Behera S, Mason S (2010) Response to phenotypic and marker-assisted
selection for yield and quality component traits in cucumber (Cucumis sativus L.).
Euphytica 171: 417–425.
Boualem A, Fergany M, Fernandez R, Troadec C, Martin A, Morin H, Sari MA, Collin F,
Flowers JM, Pitrat M, Purugganan MD, Dogimont C, Bendahmane A (2008) A conserved
mutation in an ethylene biosynthesis enzyme leads to andromonoecy in melons. Science
321: 836–838.
Brotman Y, Kovalski I, Dogimont C, Pitrat M, Portnoy V, Katzir N, Perl-Treves R (2005)
Molecular markers linked to papaya ring spot virus resistance and Fusarium race 2
resistance in melon. Theor Appl Genet 110: 337–345.
Cahill DJ, Schmid DH (2004) Use of marker assisted selection in a product development
breeding program. New directions for a diverse planet. Proc 4th Int Crop Science
Congress, 26 Sep–1 Oct 2004, Brisbane, Australia: Published on CDROM-www.
cropscience.org.au, pp 1–9.
Cavagnaro PF, Senalik DA, Yang L, Simon PW, Harkins TT, Kodira CD, Huang S, Weng Y
(2010) Genome-wide characterization of simple sequence repeats in cucumber (Cucumis
sativus L.). BMC Genomics 11: 569.
Chi XR, Gu XF, Zhang SP, Wang XW, Wang Y (2007) Identification of molecular markers linked
to foliage non-bitterness gene (bi) in Cucumis sativus L. Acta Hort Sin 34: 1177–1182.
Danin-Poleg Y, Burger Y, Schreiber S, Katzir N, Cohen R (1999) Identification of the gene for
resistance to Fusarium wilt races 0 and 2 in Cucumis melo Dulce. Cucurbit Genet Coop
Rep 22: 19–20.
Essafi A, Díaz-Pendón JA, Moriones E, Monforte AJ, Garcia-Mas J, Martín-Hernández AM
(2009) Dissection of the oligogenic resistance to Cucumber mosaic virus in the melon
accession PI 161375. Theor Appl Genet 118: 275–284.
Fukino N, Kunihisa M, Matsumoto S (2004) Characterization of recombinant inbred lines
derived from crosses in melon (Cucumis melo L.), AR 5_ ‘Harukei No. 3’. Breed Sci 54:
141–145.
Fukino N, Ohara T, Monforte AJ, Sugiyama M, Sakata Y, Kunihisa M, Matsumoto S (2008)
Identification of QTLs for resistance to powdery mildew and SSR markers diagnostic
for powdery mildew resistance genes in melon (Cucumis melo L.). Theor Appl Genet
118: 165–175.
Galun E (1961) Study of the inheritance of sex expression in the cucumber: the interaction of
major genes with modifying genetic and non-genetic factors. Genetica 32: 134–163.
Giovannoni JJ, Wing RA, Ganal MW, Tanksley SD (1991) Isolation of molecular markers from
specific chromosomal intervals using DNA pools from existing mapping population.
Nucl Acid Res. 19: 6553–6558.
Gu XF, Zhang SQ, Zhang SP (2006) The AFLP markers linked with the bitter fruit gene (Bt) in
cucumber. Acta Hort Sin 33: 140–142 (In Chinese with English summary).
Harris KR, Ling KS, Wechter WP, Levi A (2009) Identification and utility of markers linked
to the zucchini yellow mosaic virus resistance gene in watermelon. J Am Soc Hort Sci
134: 529–534.
Huang X, Wei X, Sang T, Zhao Q, Feng Q, Zhao Y, Li C, Zhu C, Lu T, Zhang Z, Li M, Fan D,
Guo Y, Wang A, Wang L, Deng L, Li W, Lu Y, Weng Q, Liu K, Huang T, Zhou T, Jing Y,
Li W, Lin Z, Buckler ES, Qian Q, Zhang QF, Li J, Han B (2010) Genome-wide association
studies of 14 agronomic traits in rice landraces. Nat Genet 42: 961–967.
Kabelka EA, Young K (2010) Identification of molecular markers associated with resistance to
squash silverleaf disorder in summer squash (Cucurbita pepo). Euphytica 173: 49–54.
Kanagaraj P, Prince KSJ, Sheeba JA, Biji KR, Paul SB, Senthil A, Babu RC (2010) Microsatellite
markers linked to drought resistance in rice (Oryza sativa L.). Curr Sci 98: 836–839.
Kenigsbuch D, Cohen Y (1990) The inheritance of gynoecy in muskmelon. Genome 33:
317–320.
Kubicki B (1969) Investigation of sex determination in cucumber (Cucumis sativus L.). Genet
Pol 10: 5–143.
Lam HM, Xu X, Liu X, Chen W, Yang G, Wong FL, Li MW, He W, Qin N, Wang B, Li J, Jian M,
Wang J, Shao G, Wang J, Sun SS, Zhang G (2010) Resequencing of 31 wild and cultivated
soybean genomes identifies patterns of genetic diversity and selection. Nat Genet 42:
1053–1059.
Li SJ, Wang HZ, Huo ZR, Guan W (2008) Development of SCAR marker linked to cucumber
anthracnose resistance-related gene from an AFLP marker. Acta Hort Sin 35: 123–126 (In
Chinese with English summary).
Li YL, Li HZ, Cui CS, Zhang HY, Gong GY (2007) Molecular markers linked to the dwarf gene
in squash. J Agri Biotechnol (Beijing) 15: 279–282.
Li Z, Pan J, Guan Y, Tao Q, He H, Si L, Cai R (2008) Development and fine mapping of three
co-dominant SCAR markers linked to the M/m gene in the cucumber plant (Cucumis
sativus L.). Theor Appl Genet 117(8): 1253–60.
Lin HY, Chen KS, Liou TD, Huang JW, Chang PFL (2009) Development of a molecular method
for rapid differentiation of watermelon lines resistant to Fusarium oxysporum f.sp. niveum.
Bot Stud 50: 273–280.
Liu S, Xu L, Jia Z, Xu Y, Yang Q, Fei Z, Lu X, Chen H, Huang S (2008) Genetic association of
ETHYLENE-INSENSITIVE3-like sequence with the sex-determining M locus in cucumber
(Cucumis sativus L.). Theor Appl Genet 117(6): 927–933.
Luo JH, Zhang HY, Mao AJ, Zhang F, Wang YJ, Pu TL (2006) The resistance genetic analysis
and tightly linked molecular markers for ZYMV-CH in cucumber. Acta Hort Sin 33:
1001–1006.
Lou QF, Chen JF, Chen LZ, Wolukau JN, Kang BC, Jahn M (2007) Identification of an AFLP
marker linked to a locus controlling gynoecy in cucumber and its converstion into SCAR
markers useful for plant breeding. Acta Hort 763: 75–82.
Ma SQ, Xu Y, Zhang HY, Gong GY, Shen HL (2006) Identification of the molecular markers
linked to the resistance gene to Zucchini yellow mosaic virus Chinese strain in watermelon.
Acta Phytopathol Sin 36: 68–73.
Michelmore RW, Paran I, Kesseli RV (1991) Identification of markers linked to disease-resistance
genes by bulked segregant analysis: a rapid method to detect markers in specific genomic
regions by using segregating populations. Proc Natl Acad Sci USA 88: 9828–9832.
Nieto C, Morales M, Orjeda G, Clepet C, Monfort A, Sturbois B, Puigdomènech P, Pitrat M,
Caboche M, Dogimont C, Garcia-Mas J, Aranda MA, Bendahmane A (2006) An eIF4E
allele confers resistance to an uncapped and non-polyadenylated RNA virus in melon.
Plant J 48: 452–462.
Noguera FJ, Capel J, Alvarez JI, Lozano R (2005) Development and mapping of a codominant
SCAR marker linked to the andromonoecious gene of melon. Theor Appl Genet 110:
714–720.
Oumouloud A, Arnedo-Andres MS, Gonzalez-Torres R, Alvarez JM (2008) Development of
molecular markers linked to the Fom-1 locus for resistance to Fusarium race 2 in melon.
Euphytica 164: 347–356.
Oumouloud A, Arnedo-Andrés MS, González-Torres R, Álvarez JM (2010) Inheritance of
resistance to Fusarium oxysporum f. sp. melonis races 0 and 2 in melon accession Tortuga.
Euphytica DOI: 10.1007/s10681-010-0201-4.
Park SO, Crosby KM, Huang RF, Mirkov TE (2004) Identification and confirmation of RAPD
and SCAR markers linked to the ms-3 gene controlling male sterility in melon (Cucumis
melo L.). J Am Soc Hort Sci 129: 819–825.
Périn C, Hagen LS, Giovinazzo N, Besombes D, Dogimont C, Pitrat M (2002) Genetic control
of fruit shape acts prior to anthesis in melon (Cucumis melo L.). Mol Genet Genom 266:
933–941.
Poole CF, Grimball PC (1939) Inheritance of new sex forms in Cucumis melo. J. Hered. 30:
21–25.
Risser G (1987) Controversy on resistance to fusarium wilt in ‘Perlita’ (Cucumis melo L.).
Cucurbit Genet Coop Rep 10: 60–63.
Robbins MD, Staub JE (2009) Comparative analysis of marker-assisted and phenotypic selection
for yield components in cucumber. Theor Appl Genet 119: 621–634.
Robinson RW, Munger HM, Whitaker TW, Bohn GM (1976) Genes of Cucubitaceae. HortScience
11: 554–568.
Stevanato P, Trebbi D, Saccomani M (2010) Root traits and yield in sugar beet: identification
of AFLP markers associated with root elongation rate. Euphytica 173: 289–298.
Syngenta (2009) Annual Review, p 13: http: //www2.syngenta.com/en/media/publications.
html.
Teixeira APM, Camargo LEA (2006) A molecular marker linked to the Prv1 gene that confers
resistance to Papaya ringspot virus-type W in melon. Plant Breed 125: 187–190.
Teixeira APM, da Silva Barreto FA; Camargo LEA (2008) An AFLP marker linked to the Pm-1
gene that confers resistance to Podosphaera xanthii race 1 in Cucumis melo. Genet Mol
Biol 31: 547–550.
Tezuka T, Waki K, Yashiro K, Kuzuya M, Ishikawa T, Takatsu Y, Miyagi M (2009) Construction
of a linkage map and identification of DNA markers linked to Fom-1, a gene conferring
resistance to Fusarium oxysporum f.sp. melonis race 2 in melon. Euphytica 168: 177–188.
Tezuka T, Waki K, Kuzuya M, Ishikawa T, Takatsu Y, Miyagi M (2010) Development of new
DNA markers linked to the Fusarium wilt resistance locus Fom-1 in melon. Plant Breed
DOI: 10.1111/j.1439-0523.2010.01800.x
Wang HZ, Li SJ, Liu XF, Li P, Hun ZR, Guan W (2007) AFLP markers of cucumber anthracnose
resistance-related gene. Acta Hort Sin 34: 213–216 (in Chinese with English summary).
Wang H, Li S, Yang R, Wei Guan W (2010) EST-SSR marker linked to cucumber target leaf spot
resistance-related gene. Acta Hort 871: 49–56.
Wang JS, Song SH, Tang XW, Chen GL (2005) Genetics and molecular marker of the gene for
resistance to powdery mildew in Cucumis melo. Acta Agri Boreali-Sin 20: 89–92.
Wang Y-H, Dean RA, Joobeur T (2006) Genetic mapping and molecular breeding in cucurbits.
Plant Breed Rev 27: 213–244.
Weng Y, Staub JE, Johnson S, Huang S (2010) An Extended Intervarietal Microsatellite Linkage
Map of Cucumber, Cucumis sativus L. HortScience 45: 882–886.
Wolukau JN, Zhou XH, Chen JF (2009) Identification of amplified fragment length
polymorphism markers linked to gummy stem blight (Didymella bryoniae) resistance in
melon (Cucumis melo L.) PI 420145. HortScience 44: 32–34.
Yamasaki S, Fujii N, Matsuura S, Mizusawa H, Takahashi H (2001) The M locus and ethylene-
controlled sex determination in andromonoecious cucumber plants. Plant Cell Physiol
42: 608–619.
Yin T, Quinn JA (1995) Tests of a mechanistic model of one hormone regulating both sexes in
Cucumis sativus (Cucurbitaceae). Am J Bot 82: 1537–1546.
Yuste-Lisbona FJ, Capel C, Capel J, Lozano R, Gómez-Guillamón ML, López-Sesé AI (2008)
Conversion of an AFLP fragment into one dCAPS marker linked to powdery mildew
resistance in melon. In: M Pitrat (ed) Cucurbitaceae 2008, Proc IXth EUCARPIA Meeting
on Genetics and Breeding of Cucurbitaceae, INRA, Avignon, France, pp 143–148.
Zhang GH, Du SL, Wang M, Ma DH (2004) AFLP markers of cucumber powdery mildew
resistance-related gene. Acta Hort Sin 31: 189–192.
Zhang GH, Han YK, Sun XH, Li SJ, Wei AM, Du SL (2006) Molecular marker linked to the
resistant gene of cucumber scab. Sci Agri Sin 39: 2250–2254.
Zhang HY, Mao AJ, Zhang F, Xu Y, Wang YJ (2005) Mapping of three major virus resistance
genes in cucumber. J Agri Biotechnol (Beijing) 13: 709–712.
Zhang HY, Wang ZG, Mao AJ, Zhang F, Wang YJ, Xu Y (2008) SSR markers linked to the resistant
gene of cucumber powdery mildew. Acta Agri Boreali-Sin 23: 77–80.
Zhang HY, Zhang J, Mao AJ, Zhang F, Xu Y, Wang YL (2009) Sequence-characterized amplified
region (SCAR) markers linked to Watermelon mosaic virus resistant gene in cucumber. J
Agri Biotechnol (Beijing) 17: 312–316.
Zhang S, Du Y, Huang S, Wehner TC, Sun R, Wang X, Gu XF, Miao H, Xie B, Yang Y (2010a)
Genetic Mapping of the Scab Resistance Gene in Cucumber. J Am Soc Hort Sci 135:
53–58.
Zhang W, He H, Guan Y, Du H, Yuan L, Li Z, Yao D, Pan J, Cai R (2010b) Identification and
mapping of molecular markers linked to the tuberculate fruit gene in the cucumber
(Cucumis sativus L.). Theor Appl Genet 120: 645–654.
Zhao FK, Zhou H, Gao XH, Lin C (2007) Identification of RAPD molecular markers linked to
CMV resistance gene in squash. Acta Agri Boreali-Sin 22: 90–94.
Zink FW, Gubler WD (1985) Inheritance of resistance in muskmelon to Fusarium wilt. J Am
Soc Hort Sci 110: 600–604.
ABSTRACT
The cucurbits are among the most culturally and commercially
important plant species cultivated worldwide. The considerable genetic
diversity within the major cultivated species, such as melon (Cucumis
melo L.), cucumber (Cucumis sativus L.), watermelon (Citrullus lanatus)
and squash (Cucurbita moschata), provides a reservoir of genes for the
development of new cultivars in breeding programs. The development
and use of genomic tools for the genetic improvement of cucurbits
has been one of the major objectives of research programs across the
world. In fact, 34 linkage maps and more than 400 genes or quantitative
trait loci (QTLs) associated with economic important traits have been
indentified in cucurbits during the last 15 years. Subsequently, an
extensive number of publications related to genome mapping and QTL
analysis are dispersed in the literature. The objective of this chapter
is to summarize and illustrate the importance of DNA marker and
genomic analysis (e.g., mapping, QTL detection, and map-based gene
cloning) in the study of diseases, yield and fruit quality components
in cucurbit crop species. Moreover, it integrated independent research
results in order to improve the utility of genomic analysis in cucurbit
breeding programs.
1
Plant Genome Mapping Laboratory, Center for Applied Genetic Technologies, 111 Riverbend
Road, Athens, GA 30602, USA; e-mail: hcuevas@uga.edu
2
USDA-ARS, Forage and Range Research Laboratory, Utah State University, Logan, UT
84322-6300, USA.
3
USDA-ARS, Madison WI; Dept. Horticulture, 1575 Linden Drive, Madison, WI 53706,
USA.
*Corresponding author
8.1 Introduction
The globally ubiquitous, cross-pollinated family Cucurbitaceae includes
825 species representing 118 principally tropical genera of which some
have economic importance (Jeffrey 1990). For instance, species in the genera
Cucumis (cucumber; C. sativus var. sativus L; melon; Cucumis melo L.),
Citrullus [watermelon; C. lanatus (Thumb.) Matsum & Nakai], and Cucurbita
[squash (C. pepo L. and C. moschata Duchesne)] provide an array of market
types that are grown worldwide (~17.9 x 107 metric tons of marketable
product; FAO 2004). In fact, taken collectively species of the Cucurbitaceae
are among the most culturally and commercially important plant species
(Nayar and More 1998).
The origin and domestication of cucurbit crop species differ dramatically.
For example, while squash was domesticated in America more than 8,000
years ago (Smith 1997), cucumber, melon and watermelon originated in
the Near East, India, and North Africa (Kirkbride 1993, Robinson and
Decker-Walters 1997). Genetic improvement of cucurbits relies on the
incorporation of economically important alleles from genetically diverse
exotic germplasm. Consequently, the documentation (i.e., taxonomy,
nutrition, and culinary attributes) and conservation (i.e., collection and
curation) of unique accessions (e.g., Cucumis hystrix Char.; Chen and Staub
1997) is essential. Recently, the intra- and interspecific variation in Cucumis
(Staub et al. 1997, 1999, 2000, 2002, 2004; Horejsi and Staub 1999; Mliki et
al. 2001; Akashi et al. 2002; López-Sensé et al. 2002, 2003; Nakata et al. 2005;
Sensoy et al. 2007; Luan et al. 2008), Cucurbita (Merrick 1991; Paris et al.
2003), and Citrullus (Levi et al. 2001a, b) germplasm has been described and
summarized (Lebeda et al. 2007).
Contemporary/modern cucurbit domestication efforts since 19th
century primarily involved farmer-selection of landraces. However, genetic
diversity in “commercial” germplasm has more recently (after 19th century)
been augmented by the strategic introduction of novel traits (e.g., disease
resistance and plant architecture) housed in exotic germplasm (Lebeda
et al. 2007). Such initial hybridization with elite lines was followed by
intensive selection efforts primarily focused on improving the fruit yield
and quality traits. Recent increases in yield and quality in many cucurbit
crop species have, in fact, resulted from complementary improvements in
cultural practices, increased pest and disease resistances, and modifications
of plant architecture and sex expression (e.g., increased gynoecy). While
traditional breeding has been successful, it is clear that in some cucurbit
species, this process might be accelerated (i.e., increased effectiveness and
efficiency) by the application genomic tools including marker assisted
selection (MAS) (Fazio and Staub 2003b; Fan et al. 2006; Robbins and Staub
2009). Fully implemented molecular marker technologies, such as use of
tightly linked marker/trait associations (< 5 cM), can permit plant selection
without exposure to environmental (abiotic) or pathogen (biotic) challenges
(Helentjaris et al. 1986).
A wide range of DNA markers have been employed in cucurbits for
diversity analysis (Lebeda et al. 2007), genetic map construction [melon
(Baudraco-Arnas and Pitrat 1996; Wang et al. 1997; Liou et al. 1998; Oliver
et al. 2001; Danin-Poleg et al. 2002; Perin et al. 2002; Monforte et al. 2004;
Gonzalo et al. 2005; Perchepied et al. 2005a, b; Zalapa et al. 2007; Fernández-
Silva et al. 2008; Cuevas et al. 2008, 2009; Fukino et al. 2008), cucumber
(Kennard et al. 1994; Serquen et al. 1997a; Park et al. 2000; Bradeen et al.
2001; Fazio et al. 2003a; Yaun et al. 2008; Ren et al. 2009), squash (Brown and
Myers 2002; Zraide et al. 2007; Gong et al. 2008a, b), and watermelon (Levi
et al. 2001a, 2002, 2006; Hashizume et al. 2003; Zhang et al. 2004)] and MAS
(Fazio and Staub 2003b; Fan et al. 2006; Robbins and Staub 2009). Marker
analysis technologies in cucurbits have employed restriction fragment
length polymorphism (RFLP), random amplified polymorphic DNAs
(RAPD), simple sequence repeats (SSR), inter-simple sequence repeats
(ISSR), amplified fragment length polymorphism (AFLP), sequence-related
amplified polymorphisms (SRAP), sequence characterized amplified regions
(SCAR), cleaved amplified polymorphic sequence (CAPS), and single
nucleotide polymorphism (SNP). The joint analysis of marker genotyping
and trait phenotyping enables the detection and location of loci controlling
quantitative traits (QTL) (Asíns 2002). The importance of DNA marker (e.g.,
genotyping) and genomic (e.g., mapping, QTL detection, and map-based
gene cloning) analysis in major cucurbit crop species (i.e., melon, cucumber,
watermelon, and squash) are presented and discussed herein.
Fruit diameter 13 n/a ‘Piel de Sapo’ x PI 161375 NIL Eduardo et al. 2007
6 06.0–28.0 ‘USDA 841-1’ x ‘Top Mark’ RIL Paris et al. 2008
Fruit length 13 10.0–29.0 ‘Piel de Sapo’ x PI 161375 NIL Eduardo et al. 2007
Fruit area 5 n/a ‘Piel de Sapo’ x PI 161375 NIL Obando et al. 2008
Fruit shape 5 n/a ‘Védrantais’ x PI 161375 RIL Perin et al. 2002
8 08.0–33.0 ‘Piel de Sapo’ x PI 161375 DHL, F2 Monforte et al. 2004
16 11.0–52.0 ‘Piel de Sapo’ x PI 161375 NIL Eduardo et al. 2007
8 05.0–29.0 ‘USDA 841-1’ x ‘Top Mark’ RIL Paris et al. 2008
Fruit quality components Fruit netting 5 10.0–18.0 ‘USDA 841-1’ x ‘Top Mark’ RIL Paris et al. 2008
7 n/a ‘Piel de Sapo’ x PI 161375 NIL Obando et al. 2008
Fruit hardness 4 n/a ‘Piel de Sapo’ x PI 161375 NIL Obando et al. 2008
Ovary shape 6 n/a ‘Piel de Sapo’ x PI 161375 NIL Eduardo et al. 2007
Skin color and ground spot color 25 n/a ‘Piel de Sapo’ x PI 161375 NIL Obando et al. 2008
Organic acids 21 n/a ‘Piel de Sapo’ x PI 161375 NIL Obando et al. 2009
Sugar content 30 n/a ‘Piel de Sapo’ x PI 161375 NIL Obando et al. 2009
Soluble solid content 5 07.0–30.0 ‘Piel de Sapo’ x PI 161375 DHL, F2 Monforte et al. 2004
19 n/a ‘Piel de Sapo’ x PI 161375 NIL Eduardo et al. 2007
8 07.0–18.0 ‘USDA 841-1’ x ‘Top Mark’ RIL Paris et al. 2008
Diseases Resistance Resistance to Fusarium 9 05.0–36.0 ‘Védrantais’ x ‘Isabella’ RIL Perchepied et al.
oxysporum sp. melonis 2005a, Brotman et
al. 2005
Resistance to Pseudoperonospora 11 05.0–38.0 ‘Védrantais’ x PI 124112 RIL Perchepied et al.
cubensis 2005b
Resistance to Podosphaera 2 12.0–46.0 ‘AR 5’ x ‘Harukei 3’ RIL Perchepied et al.
xanthii 2005a, Fukino et al.
2008
Resistance to Melon necrotic spot 1 n/a ‘Piel de Sapo’ x PI 161375 F2 Oliver et al. 2001
virus
Resistance to Cucumber mosaic 7 12.0–79.0 ‘Védrantais’ x PI 161375 RIL Dogimont et al.
virus 2000
a
Trait name according to their published designation.
b
Percentage of phenotypic variation explain by QTL association.
c
Population type used to construct linkage maps, where F2, RIL, NIL, DHL, and F2-3 refer to F2 population, recombinant inbred lines, nearly isogenic
lines, double haploid lines, and F3 family analysis, respectively.
Figure 8-1 A bin map [framework map (FM)] of Cucumis melo obtained by genotyping 14
selected doubled haploid lines (DHL). The vertical bars represent linkage groups (LG) that
are characterized by bins defined by the genotype of the selected DHL. Markers in bold were
used for the estimation of bin size. Genetics distance (GD) is shown on the left side of LG,
indicating the postion of the last marker included in the bin according to the FM. Markers
in italic define “new bins”, and the hypothetical position of the last marker in each bin is
indicated by a dashed horizontal line within the LG bar without the GD specification. Figure
adapted with kind permission from Springer Science and Business Media: Fernández-Silva
et al. 2008, Theor Appl Genet 118: 139–150.
20%). Likewise, six QTLs controlling fruit weight were detected at the same
level of stringency, where three were considered to contribute major effects.
Subsequently, four fruit weight (fw) QTLs (i.e., fw5.1, fw5.2, fw12.1, and
fw12.2) were confirmed by Eduardo et al. (2007) using near-isogenic lines
(NILs) in the same genetic background. Twelve additional QTLs for fruit
weight and 11 genomic regions associated with maturity were subsequently
identified employing these NILs (Obando et al. 2008).
The genetics of plant architecture in melon was studied by Zalapa et al.
(2007; Group Cantalupensis) using RILs evaluated at two contrasting US
environments (Wisconsin and California). Thirty-seven QTLs associated
with primary branch number (PB = 6), percent of maturity fruit (PMF =
5), fruit weight/plot (FW = 12), average weight fruit (AWF = 5), and fruit
numbers (FN = 9) were detected at both the locations. Although, genotype x
environment interactions (G x E) were identified for those traits, 16 QTLs (PB
= 4, PMF = 1, FW = 4, AWF = 2, and FN = 5) were stable across the growing
environments examined. More recently, two major (R2 > 20%) and one minor
(R2 > 8%) QTLs associated with fruit maturity were detected using F2-3
family analysis, where progeny were derived from Chinese (early flowering,
smooth-skinned epidermis, horticultural group undetermined) and US
Western Shipping (late flowering, netted epidermis, Group Cantalupensis)
parents (Cuevas et al. 2009).
identified eight QTLs, where the QTL in LG VI (SSR CMTC123) was colinear
with a previously reported fruit color QTL (ofc12.1; Monforte et al. 2004).
Moreover, QTL analysis of F3 progeny derived from the cross between line
“Q 3-2-2” (Chinese; white flesh) and “Top Mark” (US Western Shipping;
Group Cantalupensis; orange flesh) identified QTLs colinear with gf and
wf and with the previously identified QTLs (Cuevas et al. 2008) mapped to
LG VI (Cuevas et al. 2009). The discrepancies between these studies (i.e.,
Monforte et al. 2004 and Cuevas et al. 2008, 2009) are likely due to differences
in parental constitution and the cross-specific genetic control (i.e., allelic
constitution) conditioning the flesh-colored phenotype.
The morphological diversity present in melon fruits have allowed for
QTL analysis of other important quality traits, such as seed cavity size, dry
matter content, the color of extracted mesocarp juice and pH, organic acid
components, mesocarp flesh rigidity (i.e., pressure to compress), ovary
shape, fruit hardness (pressure to compress), and netting (Table 8-1). The
lack of consistency (i.e., data repeatability) for many fruit quality traits
has contributed to the incomplete genetic dissection (i.e., gene cloning) of
QTL. For instance, the expression of each of the above traits is dramatically
affected by environment and epistasis and, thus, their inheritance is complex
[i.e., multi-genic (> 4) highly interactive QTL].
located on the distal region of LG XI. The gene conferring resistance to races
0 and 1 (Fom-1) is linked to the gene conferring resistance to papaya ring
spot virus (Brotman et al. 2002). The most closely linked markers (NBS1-
CAPS and 62-CAPS) are within 1.7 and 1.4 cM of the gene, respectively. This
genomic region is located at the terminal region of LG IX, near SSR marker
CMTC47. Nevertheless, Fusarium race 1.2 has overcome the resistance
provided by these genes (Fom-1 and Fom-2). Although, partial resistance to
race 1.2 has been identified (Risser and Rode 1973), it is under polygenetic
control (Perchepied et al. 2005a). Nine QTLs have been identified that
explain a significant portion of the phenotypic variation associated with
resistance to race 1.2 (LOD > 3.0; R2 = 41–67%), and seven of these QTLs
demonstrate digenic epistatic interactions, indicating that this resistance
has a complex genetic basis (Perchepied et al. 2005a).
During early experimentation, the inheritance of resistance to downy
mildew in melon was found to be partially dominant and under control
of many genes (Cohen et al. 1985; Kenigsbuch and Cohen 1989, 1992a;
Epinat and Pitrat 1994a, b). The identification of trait/marker associations
(i.e., via QTL analyses) using RILs derived from PI 124112 (resistance) and
“Védrantais” (susceptible) has identified one major QTL (pcXII.1; LOD > 5)
located in LG XII that explains from 12 to 38% of the resistance to P. cubensis
(Perchepied et al. 2005b). In addition, 10 minor QTL (LOD > 3.0; R2 = 5–20%)
variable express themselves under differing experimental conditions,
suggesting that this resistance is dramatically affected by environmental
conditions and epistasis.
Resistance to powdery mildew in melon has been studied (Kenigsbuch
and Cohen 1992b; Epinat et al. 1993; Pitrat et al. 1998; McCreight 2003), and
major genes and QTLs have been identified (Perin et al. 2002; Perchepied
et al. 2005b; Fukino et al. 2008). Although two genes for powdery mildew
resistance, Pm-x, and Pm-w, have been located in LG II, and V, respectively
(Perin et al. 2002), the two major QTLs controlling resistance, PmV.I and
PmXII.I, are located on LG V and XII, respectively (Perchepied et al. 2005b).
While QTL PmV.I (LOD > 12.0) explains from 33 to 89% of the observed
phenotypic variance associated with resistance to P. xanthii races 1, 2, and
3, PmXII.I (LOD > 10.0) explains from 25 to 93% of the phenotypic variance
associated with resistance to P. xanthii races 1, 2, 5, and G. cichoracearum
race 1. In addition, interactions between PmV.I and PmXII.I explain for 68
and 80% of the variance of resistance to P. xanthii races 1 and 2, respectively
(Perchepied et al. 2005b). More recently, Fukino et al. (2008) identified two
major QTLs (LOD > 5; R2 = 12–46%) on LG II (SSR interval CMBR008-
CMBR120) and LG XII (SSR interval CMN01_38 & CMBR150), which confer
resistance to P. xanthii races 1 and N1. However, due to the lack of sufficient
“anchor” markers among these genetic maps (i.e., Perchepied et al. 2005b
then used an RIL population derived from the cross of “TMG1” and “ST8”
[resistance and susceptible to Zucchini yellow mosaic virus (ZYMV) and
papaya ringspot virus (PRSV-W), respectively], to generate a 353-point
map (RAPD, RFLP, AFLP, and loci conditioning virus resistances) defining
12 LGs with a mean marker interval of 4.2 cM. Information from these
maps was subsequently used in map merging experiments to create two
consensus maps (narrow and wide-base maps) by Bradeen et al. (2001)
using common AFLP anchor markers. The narrow-based consensus map
was constituted by merging the maps of Kennard et al. (1994), Serquen et al.
(1997a) and others previously published maps constructed with isozymes
and morphological characters (Fanourakis and Simon 1987; Pierce and
Wehner 1990; Meglic and Staub 1996; Horejsi et al. 2000). This map included
255 markers distributed over 10 LGs having a total length of 538.6 cM and
an average of 2.1 cM between markers. The wide-based consensus map
(C. sativus x C. sativus var. hardwickii) was constituted by merging wide-based
map of Kennard et al. (1994) with the maps of Pierce and Wehner (1990),
Knerr and Staub (1992), and Meglic and Staub (1996). This map included
197 markers that were distributed over 15 LG having a total length of 450.1
cM with an average of 2.3 cM between markers.
The recently dramatic increase in genomic resources [e.g., expressed
sequence tag (EST) libraries] and the development of new types of molecular
makers (e.g., SSR, SNP, CAPS, SCAR, and SRAP) have contributed to the
development of moderately saturated maps. The first map possessing
seven LGs (the theoretical haploid chromosome number of cucumber)
was constructed by Fazio et al. (2003a) using RILs derived from a cross
between ‘G421” and “H19”. This map possessed 128 molecular markers
(14 SSRs, 24 SCARs, 27 AFLPs, 62 RAPDs, and one SNP), three morphological
markers, and spanned 706 cM with a mean marker interval of 5.6 cM.
Subsequently, a 173-point linkage map (116 SRAPs, 33 RAPDs, 11 SSRs, 9
SCARs, 3 ISSRs, and 1 STS) was constructed using an F2 population derived
from the indeterminate line “S94”’ (northern China open-field type) and
the gynoecious indeterminate Chinese line “S06” (European greenhouse
types) (Yuan et al. 2008). This map spanned seven LG having a total length of
1,016 cM with a mean marker interval of 5.9 cM. More recently, a saturated
SSR-based cucumber map was assembled by Ren et al. (2009) using 77 RILs
derived from the wide-based cross between “Gy14” (C. sativus) x PI 183967
(C. sativus var. hardwickii) (Kennard et al. 1994), and 130 RILs derived from
the intraspecific (C. sativus) cross between line “9930” and “9110 Gt”. Nine
hundred sixty-six (966) novel (previously unmapped) SSR markers (Ren
et al. 2009) and 29 SSR markers previously reported (Danin-Poleg et al.
2000; Fazio et al. 2002; Kong et al. 2007) were utilized to construct a map
possessing seven LGs spanning 573 cM with a mean marker interval of 0.58
cM, that defined ~680 recombinant breakpoints.
Gua’
Resistance to zucchini yellow mosaic virus 1 - ‘Straight 8’ x ‘Taichung Mou RIL Park et al. 2000
Gua’
a
Trait name according to their published designation.
b
Percentage of phenotypic variation explain by QTL association.
c
Population type used in linkage map construction, where F2-3 and RIL refer to F3 family analysis and recombinant inbred lines, respectively.
mosaic virus (CMV) and watermelon mosaic virus (WMV) are highly
quantitative, while resistance to papaya ring spot virus (PRSV-W), zucchini
mosaic virus (ZYMV), and Moroccan watermelon mosaic virus (MWMV)
are control by relatively few genes (Wai and Grumet 1995; Kabelka et al.
1997; Kabelka and Grumet 1997; Grumet et al. 2000). Moreover, only a few
of these resistances have been mapped to define associated QTL (Kennard
et al. 1994; Grumet et al. 2000; Horesji et al. 2000; Park et al. 2000; Bradeen
et al. 2001).
Resistance genes to downy mildew (dm) and scab (Ccu) were initially
mapped using F3 families derived from wide and narrow-based crosses in
greenhouse evaluations (Kennard et al. 1994). These two genes segregate
independently, and were located on different linkage groups. More recently,
Horesji et al. (2000) evaluated two narrow-based F3 populations [“WI
1983G” × “Straight 8” (Pop1), and “Zudm1” × “Straight 8” (Pop2)] in five
open-field and greenhouse environments in North America and Europe
to identify RAPD markers linked to dm. Bulked segregant analysis (BSA)
of susceptible and resistance families identified five markers (OPAS5-800,
BC526-1000, BC519-1100, OPG14-800 and OPX15-1100) linked to dm in
Pop1 and Pop 2. Subsequent map merging experiments (Bradeen et al.
2001) positioned the Ccu gene close to RFLP marker CMTC51 (~1 cM) in
LG A, and the dm gene within an 1 cM region flanked by AFLP (E11/M62-
F-170) and RAPD (BC526-1000) markers in LG C. Given their relatively
close linkage associations with resistance genes, these markers will likely
be exploitable in MAS.
Inheritance studies have demonstrated close linkage association among
potyvirus resistance genes (e.g., zymv, PRSV-W, wmv, MWMV) (Wai et al.
1997; Grumet et al. 2000; Park et al. 2000). For instance, studies by Wai et
al. (1997) demonstrated that zymv and PRSV-W co-segregate in populations
(F2 and backcross analysis) derived from inbred lines “TMG1” (resistance)
and “WI-2757” (susceptible). A subsequent linkage analysis using RILs
derived from “TMG1” and “Straight 8” (susceptible) confirmed that zymv
and PRSV-W were tightly linked (2.2 cM), and located in the distal region
of an LG (LG Q; Park et al. 2000). In addition, one AFLP marker (E15/M47-
F-197) co-segregated with zymv gene. Moreover, co-segregation among
four potyvirus resistance genes (zymv, PRSV-W, Wmv, and MWMV) was
detected during the evaluation of F2 and backcross populations derived
from three resistance sources (“TMG1”, “Dina-1” and “Surinan”) (Grumet
et al. 2000). These results suggest that multiple potyvirus resistance in
cucumber may be due to either the action of different alleles at a single
potyvirus resistance gene or multiple resistances conferred by a tightly
linked cluster of resistance genes.
spanned over 1,295 cM across 17 LGs, where the mean marker interval was
8.3 cM. A testcross population [(PI 14113 x “New Hampshire Midget”) x PI
386015] was also used by Levi et al. (2002) to construct a map spanning over
1,166 cM, where 141 RAPD, 27 ISSR and one SCAR marker were distributed
across 25 LGs with a mean marker interval of 7.9 cM. In an effort to construct
a useable map comprised largely of codominant markers, Levi et al. (2006;
Table 6-1) then added 71 AFLP, 93 SRAP and 14 SSR markers to the map of
Levi et al. (2001c) which yielded a 347- point map spanning over 1,976 cM
across 19 LGs, where the mean marker interval was 5.7 cM. More recently, an
RIL-based watermelon map (elite inbred line “97103” x PI 296341) consisting
of 104 loci (87 RAPDs, 13 ISSRs, and 4 SCARs) was constructed (Zhang et
al. 2004). This map spanned over 1,028 cM across 15 LGs, where the mean
marker interval was 9.9 cM.
Initially, it has been difficult in watermelon to obtain genetic maps with
an LG number equivalent to its haploid chromosome number (n = x = 11).
This was achieved by Hashizume et al. (2003) who employed F2 and BC1
mapping populations derived from the same parents [“H-7” elite inbred
line x “SA-1” (African wild accession)] to construct a moderately saturated
map that defined 11 LGs. While the F2 mapping population was genotyped
with 554 loci (477 RAPD, 53 RFLP, 23 ISSR, and one isozyme marker), 240
selected markers (facilitating genome coverage given F2 map placement)
were used to create a BC1-based map. These F2 and BC1 maps spanned over
2,384 cM (mean marker interval = 4.3 cM) and 1,729 cM (mean marker
interval = 7.2 cM), respectively. Given the marker saturation obtained in
such maps, additional inbreeding might be appropriate leading to the
construction of immortalized mapping populations [i.e., RIL (F2-based)
and NIL (BC1-based)].
sequences derived markers (L41 and R5) were used to delimit the region to
14 kbp containing a gene encoding for a 1-aminocyclopropane-1carboxylic
acid synthase (ACS), designated as CmACS-7. Genomic sequence analysis
of PI 124112 and “Védrantais” identified a single missense mutation in
one conserved region of the protein, which affects the enzymatic activity,
and in turn, the development of male organs. A similar strategy, along
with 12,660 BC1 individuals derived from a cross of PI 124112 and the
gynoecious cultivar “Gynadou”, was implemented to clone the gynoecious
(g) gene (Martin et al. 2009). The analysis located the g locus within a 1.4
kbp no-coding region containing a heavily methylated DNA transposon
element of the hAT family. Subsequent analysis found that this methylation
extends to one neighboring transcription factor gene [C2H2 zinc-finger of
the WIP protein subfamily (designated as CmWIP1)]. Functional analysis
determined that expression of CmWIP1 leads to carpel abortion, resulting
in the development of unisexual male flowers in melon.
In melon, the orange gene (Or; Lu et al. 2006) was mapped to LG IX and
aligned with a major QTL interval associated with the genetic control of
flesh color (Fig. 8-2; Cuevas et al. 2009). Syntenic relationships between this
and other melon maps (Perin et al. 2002; Fukino et al. 2008) indicates that
the Or gene maps to the same genomic region as wf, which conditions flesh
color in melon (Imam et al. 1972). The Or gene regulates the synthesis of
an DnaJ Cysteine-rich-domain protein (i.e., chaperon protein). This protein
is involved in the differentiation of proplastid and/or other non-colored
plastid into chromoplast plastids, which then provides a “metabolic sink”
for carotenoid accumulation (Lu et al. 2006). Such casual associations
must be subjected to rigorous confirmation (i.e., functional analysis and/
or genetic transformation). If confirmed, knowledge of such associations
can improve our knowledge and understanding of the gene-regulated
physiological processes in cucurbits.
LG IX LG IX Thresho LOD
Fukino et al. 2008 F2-3 population ld LOD = LOD =
0.0 CMN53_68 0.0 CMTC47/ CMN22_47
-carotene in
13.1 CMN53_68a -carotene in
16.3 CMN04_19
18.4 ECM150
21.0 ECM66
28.6 CMN04_19
34.2 wf
37.1 ECM180
40.3 Or
70.7 CMMS35_05
Figure 8-2 Colinearity and synteny relationships in Linkage Group IX between melon (Cucumis
melo L.) maps constructed using recombinant inbred lines derived from a cross between “AR
5” (orange-fleshed) x “Harukei 3” (green-fleshed) [Fukino et al. 2008; mapping of white flesh
color (wf; Imam et al. 1972)], and an F2-3 population derived from a cross between Chinese line
“Q 3-2-2” (white-fleshed) and “Top Mark” (orange-fleshed). The latter was used to map Or
gene (Lu et al. 2006) and QTL-associated with β-carotene (i.e., orange flesh) in melon. Figure
adapted with kind permission from Springer Science and Business Media: Cuevas et al. 2009,
Theor Appl Genet 119: 741–756.
Figure 8-3 Comparative analysis of the melon (Cucumis melo L.; n = x = 12) and watermelon
(Citrullus lanatus (Thumb.) Matsum & Nakai; n = x = 11) with the cucumber (Cucumis sativus
L.; n = x = 7) sequence map. The watermelon genetic maps employed in the analysis are
organized into 18 linkage groups. Figure adapted by permission from Macmillan Publishers
Ltd: Nature Genetics; Huang et al. 2009.
Color image of this figure appears in the color plate section at the end of the book.
Figure 8-4 Overview of microsynteny between melon (Cucumis melo L.) BAC 1-21-10 and
regions in the Arabidopsis thaliana, Medicago truncatula, and Populus trichocarpa (not drawn
to scale). Genes are represented by arrows where gene name, number or ID given above or
below the arrow. Homologous genes are illustrated with the same color and indicated by
narrow connecting lines of the corresponding color. Arrows representing genes that have
one or more ESTs are designate with a spot. Genes without homologs are given in black.
Transposable elements are delineated in gray and indicated by Tn. At1g, At2g, At4g referrers
to A. thaliana chromosomes 1, 2 and 4, respectively. Cm11 referrers to C. melo Linkage Group
11, Pt_XI referrers to Populus trichocarpa Linkage Group XI, and Pt_204 referrers to Populus
unmapped scaffold 204. Mt4 referrers to M. truncatula chromosome 4, consists of BAC
clones AC137837, AC153460 and AC144608, and * = End of scaffold. Figure adapted with
kind permission from Springer Science and Business Media: Deleu et al. (2007), Mol Genet
Genomics 278: 611–622.
Color image of this figure appears in the color plate section at the end of the book.
was syntenic to three Arabidopsis (At1g, At3g, and At5g), two Populus (Pt_XIII
and the scaffold Pt_70), and two Medicago (Mt1 and Mt7) genomic regions.
As was the case with the melon genomic region on LG XI, the greatest
synteny was detected in two Populus genomic regions. In fact, the highest
microsynteny values [53.8 (Pt_XI) and 40.9 (Pt_204)%, LG IV; 54.2 (Pt_XIII),
and 59.6 (Pt_70)%, LG XI] were obtained with Populus genome. Although,
the comparative analysis of additional melon sequences are necessary to
verify this putatively high degree of synteny, data currently suggest that the
Populus genome may have utility for genomic analysis (i.e., gene position
and candidate gene) in the Cucurbitaceae.
References
Akashi Y, Fukuda N, Wako T, Masuda M, Kato K (2002) Genetic variation and phylogenetic
relationships in east and south Asian melons, Cucumis melo L., based on the analysis of
five isozymes. Euphytica 125: 385–396.
Albar L, Bangratz-Reyser M, Hebrard E, Ndjiondjop MN, Jones M, Ghesquiere A (2006)
Mutations in the eIF(iso)4G translation initiation factor confer high resistance of rice to
Rice yellow mottle virus. Plant J 47: 417–426.
Al-Faifi S, Meyer JD, Garcia-Mas J, Monforte AJ, Havey MJ (2008) Exploiting synteny in
Cucumis for mapping Psm: a unique locus controlling paternal mitochondrail sorting.
Theor Appl Genet 117: 523–529.
Arumuganathan K, Earle ED (1991) Nuclear DNA content in some important plant species.
Plant Mol Biol Rep 9: 211–215.
Asins MJ (2002) Present and future of quantitaive trait locus analysis in plant breeding. Plant
Breed 121: 281–291.
Atsmon D, Lang A, Light EN (1968) Contents and recovery of gibberellins in monoecious and
gynoecious cucumber plants. Plant Physiol 43: 806–810.
Bang H, Kim S, Leskovar D, King S (2007) Development of a codominant CAPS markers for
allelic selection between canary yellow and red watermelon based on SNP in lycopene
β-cyclase (LCYB) gene. Mol Breed 20: 63–72.
Baudracco-Arnas S, Pitrat M (1996) A genetic map of melon (Cucumis melo L.) with RFLP, RAPD,
isozyme, disease resistance and morphological markers. Theor Appl Genet 93: 57–64.
Bernardo R (2008) Molecular markers and selection for complex traits in plants: learning from
the last 20 years. Crop Sci 48: 1649–1664.
Blanc G, Charcosset A, Mangin B, Gallais A, Moreau L (2006) Connected populations for
detecting quantitative trait loci and testing for epistasis: an application in maize. Theor
Appl Genet 113: 206–224.
Boualem A, Fergany M, Fernandez R, Troadec C, Martin A, et al. (2008) A conserved mutation
in an ethylene biosynthesis enzyme leads to andromonoecy in melons. Science 321:
836–838.
Bradeen JM, Staub JE, Wyse C, Antonise R, Peleman J (2001) Towards an expanded and
intergrated linkage map of cucumber (Cucumis sativus L.). Genome 44: 111–119.
Brotman Y, Silberstein L, Kovalski I, Perin C, Dogimont C, Pitrat M, Klinger J, Thompson
GA, Perl-Treves R (2002) Resistance gene homologies in melon are linked to genetic loci
conferring disease and pest resistance. Theor Appl Genet 104: 1055–1063.
Brown RN, Myers JR (2002) A genetic map of squash (Cucurbita sp.) with randomly amplified
polymorphic DNA markers and morphological markers. J Am Soc Hort Sci 127:
568–575.
Buckler ES, Holland JB, Bradbury PJ, et al. (2009) The genetic architecture of maize flowering
time. Science 325: 714–718.
BurgerY, Katzir N, Tzuri G, Portnoy V, Saar U, Shriber S, Perl-Treves R, Cohen R (2003) Variation
in the response of melon genotypes to Fusarium oxysporum f.sp. melonis race 1 determined
by inoculation test and molecular markers. Plant Pathol 52: 204–211.
Byers RE, Baker LR, Sell HM, Herner RC, Dilley DR (1972) Female flower induction on
androecious cucumber, Cucumis sativus L. J Am Soc Hort Sci 98: 197–199.
Chen JF, Staub JE (1997) Attempts at colchicine doubling of an interspecific hybrid of Cucumis
sativus × C. hystrix. Cucurbit Genet Coop Rep 20: 24–26.
Chen JF, Staub JE, Adelberg JW, Lewis S, Kunkle B (2002) Synthesis and preliminary
characterization of a new species (amphidiploid) in Cucumis. Euphytica 123: 315–322.
Chen JF, Staub JE, Qian CT, Jiang JM, Luo XD, Zhuang FY (2003) Reproduction and cytogenetic
characterization of interspecific hybrids derived from Cucumis hystrix Chakr. × C. sativus
L. Theor Appl Genet 106: 688–695.
Chiba N, Suwabe K, Nunome T, Hirai M (2003) Development of mircosatellite markers in melon
(Cucumis melo L.) and their application to major cucurbit crops. Breed Sci 53: 21–27.
Clayberg CD (1992) Interaction and linkage test of flesh color genes in Cucumis melo L. Cucurbit
Genet Coop Rep 15: 53.
Cohen Y, Cohen S, Eyal H, Thomas CE (1985) Inheritance of resistance to downy mildew in
Cucumis melo PI 124111. Cucurbit Genet Coop Rep 8: 36–38.
Cramer CS, Wehner TC (1998) Fruit yield and yield component means and correlations of four
slicing cucumber populations improved through six to ten cycles of recurrent selection.
J Am Soc Hort Sci 123: 388–395.
Cramer CS, Wehner TC (1999) Little heterosis for yield and yield components in hybrids of
six cucumber inbreds. Euphytica 110: 99–108.
Cramer CS, Wehner TC (2000) Path analysis of the correlation between fruit number and plant
traits of cucumber populations. HortScience 35: 708–711.
Cuevas HE, Staub JE, Simon PW, Zalapa JE, McCreight JD (2008) Mapping of genetic loci that
regulated quantity of β-carotene in fruit of U.S. Western Shipping melon (Cucumis melo
L.). Theor Appl Genet 117: 1345–1359.
Cuevas HE, Staub JE, Simon PW, Zalapa JE (2009) A consensus linkage map identifies genomic
regions controlling fruit maturity and beta-carotene-associated flesh color in melon
(Cucumis melo L.) Theor Appl Genet 119: 741–756.
Dane F, Liu J (2007) Diversity and origin of cultivated and citron type of watermelon. Genet
Resour Crop Evol 54: 1255–1265.
Danin-Poleg Y, Tadmor Y, Tzuri G, Reis N, Hirschberg J, Katzir N (2002) Construction of a
genetic map of melon with molecular markers and horticultural traits, and localization
of genes associated with ZYMV resistance. Euphytica 125: 373–384.
de Ponti OMB (1975) Breeding parthenocarpic pickling cucumbers (Cucumis sativus L.):
Necessity, genetical possibilities, environmental influences and selection criteria.
Euphytica 25: 29–40.
Delannay I (2009) Use of molecular markers to increase genetic diversity of Beit Alpha,
European long, and U.S. processing cucumber market clasess of cucumber (Cucumis
sativus L.) through marker assited selection. PhD Disser, Univ of Wisconsin, Madison,
WI, USA.
Deleu W, González V, Monfort A, Bendahmane A, Puigdoménech P, Arus P, Garcia-Mas J
(2007) Structured of two melon regions reveals high microsynteny with sequenced plant
species. Mol Genet Genom 278: 611–622.
Dijkhuizen A, Staub JE (2003) Effects of environment and genetic background on QTL affecting
yield and fruit quality traits in a wide cross in cucumber [Cucumis sativus L. x Cucumis
hardwickii (R.) Alef.] J New Seeds 4: 1–30.
Dijkhuizen A, Kennard WC, Havey MJ, Staub JE (1996) RFLP variability and genetic
relationships in cultivated cucumber. Euphytica 90: 79–89.
Dogimont C, Leconte L, Périn C, Thabuis A, Lecoq H, Pitrat M (2000) Identification of QTLs
contributing to resistance to different strains of cucumber mosaic cucumovirus in melon.
Acta Hort 510: 391–398.
Dogimont C, Chovelon V, Tual S, Boissot N, Rittener V, Giovinazzo N, Bendahmane A
(2008) Molecular diversity at the Vat/Pm-W resistance locus in melon. In: M Pitrat
(ed) Cucurbitaceae 2008, Proc IX EUCARPIA Meeting on Genetics and Breeding of
Cucurbitaceae 21–24 May, 2008, INRA, Avignon, France, pp 219–227.
Eduardo I, Arus P, Monforte AJ, Obando J, Fernandez-Trujillo JP, Martinez JA, Alarcon AI, Alvarez
JM, van der Knapp E (2007) Estimating the genetic architecture of fruit quality traits in
melon using a genomic library of near isogenic lines. J Am Soc Hort Sci 132: 80–89.
El-Shawaf IIS, Baker LR (1981) Inheritance of parthenocarpic yield in gynoecious pickling
cucumber for once-over mechanical harvest by diallel analysis of six gynoecious lines.
J Am Soc Hort Sci 106: 359–364.
Epinat C, Pitrat M (1994a) Inheritance of resistance to downy mildew (Pseudoperonospora
cubensis) in muskmelon (Cucumis melo). I. Analysis of a 8 × 8 diallel table. Agronomie
14: 239–248.
Epinat C, Pitrat M (1994b) Inheritance of resistance to downy mildew (Pseudoperonospora
cubensis) in muskmelon (Cucumis melo). II. Generation means analysis of 5 genitors.
Agronomie 14: 249–257.
Epinat C, Pitrat M, Bertrand F (1993) Genetic analysis of resistance of five melon lines to
powdery mildews. Euphytica 65: 135–144.
Essafi A, Díaz-Pendón JA, Moriones E, Monforte AJ, Garcia-Mas J, Martín-Hernández AM
(2009) Dissection of the oligogenic resistance to Cucumber mosaic virus in the melon
accession PI 161375. Theor Appl Genet 118: 275–284.
Fan Z, Robbins MD, Staub JE (2006) Population development by phenotypic selection with
subsequent marker assisted selection for line extraction in cucumber (Cucumis sativus
L.). Theor Appl Genet 112: 843–855.
Fanourakis NE, Simon PW (1987) Analysis of genetic linkage in cucumber. J Hered 78:
238–242.
FAO (2004) FAOSTAT Agricultural Database. Food and Agriculture Organization of the United
Nations, Rome, Italy: Accessed online at http: //apps.fao.org
Fazio G (2001) Comparative study of marker-assisted and phenotypic selection and genetic
analysis of yield components in cucumber. PhD Disser, Univ of Wisconsin, Madison,
WI, USA.
Fazio G, Staub JE (2003b) Comparative analysis of response to phenotypic and marker-assisted
selection for multiple lateral branching in cucumber (Cucumis sativus L.). Theor Appl
Genet 107: 875–883.
Fazio G, Staub JE, Chung SM (2002) Development and characterization of PCR markers in
cucumber (Cucumis sativus L.). J Am Soc Hort Sci 127: 545–557.
Fazio G, Staub JE, Stevens ML (2003a) Genetic mapping and QTL analysis of horticultural
traits in cucumber (Cucumis sativus L.) using recombinant inbred lines. Theor Appl
Genet 107: 864–874.
Fernández-Silva I, Eduardo I, Blanca J, Esteras C, Pico B, Nuez F, Arus P, Garcia-Mas J, Monforte
AJ (2008) Bin mapping of genomic and EST-derived SSRs in melon (Cucumis melo L.).
Theor Appl Genet 118: 139–150.
Fredrick LR, Staub JE (1989) Combining ability analysis and evaluation of nearly homozygous
lines derived from Cucumis sativus var. hardwickii (R.) Alef. J Am Soc Hort Sci 114:
332–338.
Fukino N, Sakata Y, Kunihisa M, Matsumoto S (2007) Characterization of novel simple
sequence repeat (SSR) markers for melon (Cucumis melo L.) and their use for genotype
identification. J Hort Sci Biotechnol 82: 330–334.
Fukino N, Ohara T, Monforte A, Sugiyama M, Sakata Y, Kunihisa M, Matsumoto S (2008)
Identification of QTLs for resistance to powdery mildew and SSR markers diagnostic
for powdery mildew resistance genes in melon (Cucumis melo L.). Theor Appl Genet
118: 165–175.
Gale MD, Devos KM (1998) Plant comparative genetics after 10 years. Science 282: 656–659.
Galun E (1959) Effects of gibberellic acid and naphthalene acetic acid on sex expression and
some morphological characters in the cucumber plant. Phyton 13: 1–8.
Galun E (1961) Study of the inheritance of sex expression in the cucumber. The interaction of
major genes with modifying genetic and non-genetic factors. Genetica 32: 134–163.
Gong L, Pachner M, Kalai K, Lelley T (2008a) Microsatellite for the genus Cucurbita and an
SSR-based genetic linkage map of Cucurbita pepo L. Theor Appl Genet 117: 37–48.
Gong L, Pachner M, Kalai K, Lelley T (2008b) SSR-based genetic linkage map of Cucurbita
moschata and its synteny with Cucurbita pepo. Genome 51: 878–887.
Gonzalo MJ, Oliver M, Garcia-Mas J, Monfort A, Dolcet-Sanjuan R, Katzir N, Aroes P, Monforte
AJ (2005) Simple-sequence repeat markers used in merging linkage maps of melon
(Cucumis melo L.). Theor Appl Genet 110: 802–811.
Grummet R, Kabelka E, McQueen S, Wai T, Humphrey R (2000) Characterization of sources of
resistance to the watermelon strain of Papaya ringspot virus in cucumber: allelism and
co-segregation with other portyvirus resistances. Theor Appl Genet 101: 463–472.
Hashizume T, Skimamoto I, Harushima Y, Yui M, Sato T, Imai T, Hirai M (1996) Construction
of a linkage map for watermelon [Citrullus lanatus- (Thunb.) Matsurn & Nakai] using
random amplified polymorphic DNA (RAPD). Euphytica 90: 265–273.
Hashizume T, Shimamoto I, Hirai M (2003) Construction of a linkage map and QTL analysis
of horticultural traits for watermelon [Citrullus lanatus (Thunb.) Matsum & Nakai] using
RAPD, RFLP and ISSR markers. Theor Appl Genet 106: 779–785.
Hawkins LK, Dane F, Kubisiak TL, Rhodes BB, Jarret RL (2001) Linkage mapping in a watermelon
population segregating for fusarium wilt resistance. J Am Soc Hort Sci 126: 344–350.
Helentjaris T, Slocum M, Wright S, Schaefer A, Nienhuis J (1986) Construction of genetic
linkage maps in maize and tomato using restriction fragment length polymorphism.
Theor Appl Genet 72: 761–769.
Hoisington D, Khairallah M, Ribaut JM, Skovmand B, Taba S, Warburton M (1999) Plant
genetic resources: what can they contribute toward increased crop productivity? Proc
Natl Acad Sci USA 99: 8133–8138.
Horejsi T, Staub JE (1999) Genetic variation in cucumber (Cucumis sativus L.) as assessed by
random amplified polymorphic DNA. Genet Resour Crop Evol 46: 337–350.
Horejsi T, Staub JE, Thomas C (2000) Linkage of random amplified polymorphic DNA markers
to downy mildew resistance in cucumber (Cucumis sativus L.). Euphytica 115: 105–113.
Howad W, Yamamoto T, Dirlewanger E, Testolin R, Cosson P, Cipriani G, Monforte AJ, Georgi
L, Abbott AG, Arús P (2005) Mapping with a few plants: using selective mapping for
microsatellite saturation of the Prunus reference map. Genetics 171: 1305–1309.
Huang S, Ruiqiang L, Zhonghua Z, Li L, Xingfang G, Wei F, William JL, et al. (2009) The
genome of cucumber, Cucumis sativus L. Nat Genet 41: 1275–1281.
Hughes MB (1948) The inheritance of two characters of Cucumis melo and their interrelationship.
Proc Am Soc Hort Sci 52: 399–402.
Hulbert SH, Webb GA, Smith SM, Sun Q (2001) Resistance gene complexes: evolution and
utilization. Annu Rev Phytopathol 39: 285–312.
Imam MKL, Abo-Bakr MA, Hanna HY (1972) Inheritance of some economic characters in
crosses between sweet melon and snake cucumber. I. Inheritance of qualitative characters.
Assiut J Ag Sco 3: 363–380.
Jeffrey C (1990) Systematics of the Cucurbitaceae: An overview. In: DM Bates, RW Robisnson,
C Jeffrey (eds) Biology and Utilization of the Cucurbitaceae. Cornell Univ Press, Ithaca,
NY, USA, pp 2–9.
Joobeur T, King JJ, Nolin SJ, Thomas CE, Dean RA (2004) The fusarium wilt resistance locus Fom-2
of melon contains a single resistance gene with complex features. Plant J 39: 283–297.
Kabelka E, Grumet R (1997) Inheritance of resistance to the Moroccan watermelon mosaic
virus in the cucumber line TMG-1 and cosegregation with zucchini yellow mosaic virus
resistance. Euphytica 95: 237–242.
Kabelka E, Ullah Z, Grumet R (1997) Multiple alleles for zucchini yellow mosaic virus resistance
at the zym locus in cucumber. Theor Appl Genet 95: 997–1004.
Kenigsbuch D, Cohen Y (1989) Inheritance of resistance to downy mildew in a gynoecious
muskmelon. Plant Dis 73: 994–996.
Kenigsbuch D, Cohen Y (1990) Inheritance of gynoecy in muskmelon. Genome 33: 317–320.
Kenigsbuch D, Cohen Y (1992a) Inheritance of resistance to downy mildew in Cucumis melo PI
124112 and commonality of resistance genes with PI 124111F. Plant Dis 76: 615–617.
Kenigsbuch D, Cohen Y (1992b) Inheritance and allelism of genes for resistance to races 1 and
2 of Sphaerotheca fuliginea in muskmelon. Plant Dis 76: 626–629.
Kennard WC, Poetter K, Dijkhuizen A, Meglic V, Staub JE, Havey MJ (1994) Linkages among
RFLP, RAPD, isozyme, disease-resistance, and morphological markers in narrow and
wide crosses of cucumber. Theor Appl Genet 89: 42–48.
Kirkbride JH (1993) Biosystematic Monograph of the Genus Cucumis (Cucurbitaceae). Parkway
Publ, Boone, NC, USA.
Knerr LD, Staub JE (1992) Inheritance and linkage relationships of isozyme loci in cucumber
(Cucumis sativus L.). Theor Appl Genet 84: 217–224.
Kong Q, Xiang C, Yu Z, Zhang Z, Liu F, Peng C, Peng X (2007) Mining and charactering
microsatellites in Cucumis melo expressed sequence tags from sequence database. Mol
Ecol Notes 7: 281–283.
Koo D, Choi H, Cho J, Hur Y, Bang J (2005) A high-resolution karyotype of cucumber (Cucumis
sativus L. ‘Winter Long’) revealed by C-banding, pachytene analysis, and RAPD-aided
fluorescence in situ hybridization. Genome 48: 534–540.
Kubicki B (1969) Investigations on sex determination in cucumber (Cucumis sativus L.). Genet
Pol 10: 3–143.
Kupper RS, Staub JE (1988) Combining ability between lines of Cucumis sativus L. and Cucumis
sativus var. hardwickii (R) Alef. Euphytica 38: 197–210.
Lebeda A, Widrlechner MP, Staub J, Ezura H, Zalapa J, Krístová E (2007) Cucurbits
(Curcurbitacea; Cucumis ssp., Cucurbita spp., Citrullus spp.). In: RJ Singh (ed) Genetic
Resources, chromosome engineering, and crop improvement. CRC Press, Boca Raton,
FL, USA, pp 271– 376.
Lecoq H, Pitrat M (1980) Inheritance of resistance to cucumber mosaic virus transmission by
Aphis gossypii in Cucumis melo. Phytopathology 70: 958–961.
Leeuwen H, Monfort A, Zhang H, Puigdoménech P (2003) Identification and characterization
of a melon genomic region containing a resistance gene cluster from a constructed BAC
library. Microcolinearity between Cucumis melo and Arabidopsis thaliana. Plant Mol Biol
51: 703–718.
Leeuwen H, Garcia-Mas J, Coca M, Puigdoménech P, Monfort A (2005) Analysis of the melón
genome in regions encompassing TIR-NBS-LRR resistance genes. Mol Genet Genom
273: 240–251.
Levi A, Thomas CE, Keinath AP, Wehner TC (2000) Estimation of genetic diversity among
Citrullus accessions using RAPD markers. Acta Hort 510: 385–390.
Levi A, Thomas CE, Keinath AP, Wehner TC (2001a). Genetic diversity among watermelon
(Citrullus lanatus and Citrullus colocynthis) accessions. Genet Resour Crop Evol 48:
559–566.
Levi A, Thomas CE, Wehner TC, Zhang X (2001b) Low genetic diversity indicates the need to
broaden the genetic base of cultivated watermelons. HortScience 36: 1096–1101.
Levi A, Thomas CE, Zhang X, Joobeur T, Dean RA, Wehner TC, Carle BR (2001c) A genetic
linkage map for watermelon based on randomly amplified polymorphic DNA markers.
J Am Soc Hort Sci 126: 730–737.
Levi A, Thomas CE, Joobeur T, Zhang X, Davis A (2002) A genetic linkage map for watermelon
derived from a testcross population: (Citrullus lanatus var. citroides x C. lanatus var. lanatus)
× Citrullus colocynthis. Theor Appl Genet 105: 555–563.
Levi A, Thomas CE, Trebitsh T, Salman A, King J, Karalius J, Newman M, Reddy OUK, Xu Y,
Zhang X (2006) An extended linkage map for watermelon based on SRAP, AFLP, SSR,
ISSR, and RAPD markers. J Am Soc Hort Sci 131: 393–402.
Liou PC, Chang YM, Hsu WS, Cheng YH, Chang HR, Hsiao CH (1998) Construction of a
linkage map in Cucumis melo (L.) using random amplified polymorphic DNA markers.
Acta Hort 461: 123–132.
Liu HB (1998) Statistical genomics: linkage, mapping and QTL analysis. CRC Press, Boca
Raton, FL, USA.
Liu L, Kakihara F, Kato M (2004) Characterization of six varities of Cucumis melo L. based
on morphological and physiological characters, including shelf-life of fruit. Euphytica
135: 305–313.
López-Sesé AI, Staub JE, Katzir N, Gómez-Guillamón ML (2002) Estimation of between and
within accession variation in selected Spanish melon germplasm using RAPD and SSR
markers to assess strategies for large collection evaluation. Euphytica 127: 41–51.
López-Sesé AI, Staub JE, Gomez-Guillamon ML (2003) Genetic analysis of Spanish melon
(Cucumis melo L.) germplasm using a standardized molecular marker array and reference
accessions. Theor Appl Genet 108: 41–52.
Lower RL, Edwards MD (1986) Cucumber breeding. In: MJ Basset (ed) Breeding Vegetable
Crops. AVI Publ, Westport, CT, USA, pp 173–207.
Lu S, Van Eck J, Zhou X, Lopez AB, O’Halloran M, Cosman KM, Conlin BJ, Paolillo DJ, Garvin
DF, Vrebalov J, Kochian LV, Küpper H, Earle ED, Cao J, Li L (2006) The cauliflower Or
gene encodes a Dna-J cysteine-rich domain-containing protein that mediates high levels
of β-carotene accumulation. Plant Cell 18: 3594–3605.
Luan F, Delannay I, Staub JE (2008) Melon (Cucumis melo L.) diversity analyses provide
strategies for germplasm curation, genetic improvement, and evidentiary support of
domestication patterns. Euphytica 164: 445–461.
Luo M, Wang YH, Frisch D, Joobeur T, Wing R, Dean RA (2001) Melon BAC library construction
using improved methods and identification of clones linked to the locus conferring
resistance to melon Fusarium wilt (Fom-2). Genome 44: 154–162.
Mackay TFC (2001) The genetic architecture of quantitative traits. Annu Rev Genet 35:
303–339.
Martin A, Troadec C, Boualem A, Rajab M, Fernandez R, Morin H, Pitrat M, Dogimont C,
Bendahmane A (2009) A transposon-induced epigenetic change leads to sex determination
in melon. Nature 461: 1135–1139.
Martyn RD, Gordon TR (1996) Fusarium wilt of melon. In: TA Zitter, DL Hokins, CE Thomas
(eds) Compendium of Cucurbit Diseases. APS Press, Minneapolis, MA, USA, pp
14–15.
McCreight JD (2003) Genes for resistance to powdery mildew races 1 and 2 U.S. in melon PI
313970. HortScience 38: 591–594.
Meglic V, Staub JE (1996) Inheritance and linkage relationships of allozyme and morphological
loci in cucumber (Cucumis sativus L.). Theor Appl Genet 92: 865–872.
Paterson AH, Bowers JE, Burow MD, Draye X, Elsik CG, Jiang C, Katsar CS, Lan T, Lin Y,
Ming R, Wright RJ (2000) Comparative genomics of plant chromosome. Plant Cell 12:
1523–1540.
Pauquet J, Burget E, Hagen L, Chovelon V, Le Menn A, Valot N, Desloire S, Caboche M,
Rousselle P, Pitrat M, Bendahmane A, Dogimont C (2004) Map-based cloning of the Vat
gene from melon conferring resistance to both aphid colonization and aphid transmission
of several viruses. In: A Lebeda, H Paris (eds) Cucurbitaceae 2004, Proc 8th EUCARPIA
Meeting on Cucurbit Genetics and Breeding, Palaky Univ, Olomouc, Czech Republic,
pp 325–329.
Perchepied L, Dogimont C, Pitrat M (2005a) Strain-specific and recessive QTL involved in the
control of partial resistance to Fusarium oxysporum f. sp. melonis race 1.2 in a recombinant
inbred line population of melon. Theor Appl Genet 111: 65–75.
Perchepied L, Mardin M, Dogimont C, Pitrat M (2005b) Relationship between loci conferring
downy mildew and powdery mildew resistance in melon assessed by quantitative trait
loci mapping. Phytopatology 95: 556–565.
Périn C, Hagen LS, de Conto V, Katzir N, Danin-Poleg Y, Portnoy V, Baudracco-Arnas S,
Chadoeuf J, Dogimont C, Pitrat M (2002) A reference map of Cucumis melo based on two
recombinant inbred line populations. Theor Appl Genet 104: 1017–1034.
Pierce LK, Wehner TC (1990) Review of genes and linkage groups in cucumber. HortScience
25: 605–615.
Pitrat M (1984) Linkages studies in muskmelon. Cucurbit Genet Coop Rep 7: 51–63.
Pitrat M (2002) Gene list for melon. Cucurbit Genet Coop Rep 25: 76–93.
Pitrat M (2008) Melon. In: J Prohens, F Nuez (eds) Handbook of Plant Breeding. Springer,
New York, USA, pp 283–315.
Pitrat M, Dogimont C, Bardin M (1998) Resistance to fungal diseases of foliage in melon. In: JD
McCreight (ed) Cucurbitaceae ’98: Evaluation and Enhancement of Cucurbit Germplasm.
ASHS Press, Alexandria, VA, USA, pp 167–173.
Poole CF, Grimbal PC (1939) Inheritance of new sex forms in Cucumis melo L. J Hered 30:
21–25.
Ren Y, Zhang Z, Liu J, Staub JE, Han Y, Cheng Z, Li Z, Lu J, Miao H, Kang H, Xie B, Gu X,
Wang X, Du Y, Jin W, Huang S (2009) An integrated genetic and cytogenetic map of the
cucumber genome. PloS one 4: e5795.
Risch N, Merikangas K (1996) The future of genetic studies of complex human diseases.
Science 273: 1516–1517.
Risser G, Rode JC (1973) Breeding for resistance to Fusarium oxysporum f. melonis. In:
EUCARPIA: La se´lection du melon. Montfavet-Avignon, France, pp 37–39.
Ritchel PS, de Lima-Lins TC, Lourenco-Tristan R, Cortopasi-Buso GS, Buso JA, Ferreiram
ME (2004) Development of microsatellite marker from an enriched genomic library for
genetic analysis of melon (Cucumis melo L.). BMC Plant Biol 4: 9.
Robbins MD, Casler MD, Staub JE (2008) Pyramiding QTL for multiple lateral branching in
cucumber using nearly isogenic lines. Mol Breed 22: 131–139.
Robbins MD, Staub JE (2009) Comparative analysis of marker-assisted and phenotypic selection
for yield components in cucumber. Theor Appl Genet 119: 621–634.
Robinson RW, Whitaker HM, Bohn GW (1976) Genes of the Cucurbitaceae. HortScience 11:
554–568.
Robinson RW, Decker-Walters DS (1997) Cucurbits. Crop Production Science in Horticulture
Series. CAB International, Wallingford, UK.
Roy RP, Saran S (1990) Sex expression in the Cucurbitaceae, In: DM Bates, CH Jeffery (eds)
Biology and Utilization of the Cucurbitaceae. Comstock, Cornell Univ Press, Ithaca, NY,
USA, pp 251–268.
Rudich J, Halevy AH, Kedar N (1972a) Ethylene evolution from cucumber plants as related
to sex expression. Plant Physiol 49: 998–999.
Rudich J, Halevy AH, Kedar N (1972b) The level of phytohormones in monoecious and gynoecious
cucumbers as affected by photoperiod and ethephon. Plant Physiol 50: 585–590.
Salvi S, Tuberosa R (2005) To clone or not to clone plant QTLs: present and future challenges.
Trends Plant Sci 10: 297–304.
Schmidt R (2002) Plant genome evolution: lessons from comparative genomics at the DNA
level. Plant Mol Biol 48: 21–37.
Sensoy S, Buyukalaca S, Abak K (2007) Evaluation of genetic diversity in Turkish melons
(Cucumis melo L.) based on phenotypic characters and RAPD markers. Genet Resour
Crop Evol 54: 1351–1365.
Serquen FC, Bacher J, Staub JE (1997a) Mapping and QTL analysis of a narrow cross in
cucumber (Cucumis sativus L.) using random amplified polymorphic DNA markers.
Mol Breed 3: 257–268.
Serquen FC, Bacher J, Staub JE (1997b) Genetic analysis of yield components in cucumber
(Cucumis sativus L.) at low plant density. J Am Soc Hort Sci 122: 522–528.
Shifriss O (1961) Sex control in cucumbers. J Hered 52: 5–12.
Shimotsuma M (1963) Cytogenetic and evolutionary studies in the genus Citrullus. Seiken
Jiho 15: 24–34.
Silberstein L, Kovalski I, Brotman Y, Perin C, Dogimont C, Pitrat M, Klinger J, Thompson
G, Portnoy V, Katzir N, Perl-Treves R (2003) Linkage map of Cucumis melo including
phenotypic traits and sequence-characterized genes. Genome 46: 761–773.
Smith BD (1997) The initial domestication of Cucurbita pepo in the Americas 10,000 years ago.
Science 276: 932–934.
Staub JE, Serquen FC, Gupta M (1996a) Genetic markers, map construction and their application
in plant breeding. HortScience 31: 729–741.
Staub JE, Bacher J, Poetter K (1996b) Factors affecting the application of random amplified
polymorphic DNAs in cucumber (Cucumis sativus L.). HortScience 31: 262–266.
Staub JE, Serquen FC, McCreight JD (1997) Genetic diversity in cucumber (Cucumis sativus L.):
III. An evaluation of Indian germplasm. Genet Resour Crop Evol 44: 315–326.
Staub JE, Serquen FC, Horejsi T, Chen JF (1999) Genetic diversity in cucumber (Cucumis sativus
L.): IV. An evaluation of Chinese germplasm. Genet Resour Crop Evol 46: 297–310.
Staub JE, Danin-Poleg Y, Fazio G, Horejsi T, Reis N, Katzir N (2000) Comparative analysis of
cultivated melon groups (Cucumis melo L.) using random amplified polymorphic DNA
and single sequence repeat markers. Euphytica 115: 225–241.
Staub JE, Dane F, Reitsma K, Fazio G, López-Sesé AI (2002) The formation of test arrays and
a core collection in (Cucumis sativus L.) using phenotypic and molecular marker data. J
Am Soc Hort Sci 127: 558–567.
Staub JE, Fanourakis N, López-Sesé AI (2004) Genetic diversity in melon (Cucumis melo L.)
landraces from the island of Crete as assessed by random amplified polymorphic DNA
and simple sequence repeat markers. Euphytica 136: 151–166.
Staub JE, Sun Z, Chung SM, Lower RL (2007) Evidence for colinearity among genetic linkage
maps in cucumber. HortScience 42: 20–27.
Staub JE, Robbins MD, Wehner TC (2008) Cucumber. In: J Prohens, F Nuez (eds) Vegetables
I: Asteraceae, Brassicaceae, Chenopodiaceae, and Cucurbitaceae. Springer, New York,
USA, pp 241–282.
Stein N, Perovic D, Kumlehn J, Pellio B, Stracke S, Streng S,Ordon F, Graner A (2005) The
eukaryotic translation initiation factor 4E confers multiallelic recessive Bymovirus
resistance in Hordeum vulgare (L.). Plant J 42: 912–922.
Stepansky A, Kovalski I, Perl-Treves R (1999) Intraspecific classification of melons (Cucumis melo
L.) in view of their phenotypic and molecular variation. Plant Syst Evol 217: 313–332.
Sun Z, Lower RL, Staub JE (2006a) Variance component analysis of parthenocarpy in elite U.S.
processing type cucumber (Cucumis sativus L.) lines. Euphytica 148: 333–341.
Sun Z, Lower RL, Staub JE (2006b) Analysis of generation means and components of variance
for parthenocarpy in cucumber (Cucumis sativus L.). Plant Breed 125: 277–280.
Sun Z, Lower RL, Chung SM, Staub JE (2006c) Identification and comparative analysis of
quantitative trait loci (QTL) associated with parthenocarpy in processing cucumber.
Plant Breed 125: 281–287.
Tatlioglu T (1993) Cucumber Cucumis sativus L. In: G Kalloo, BO Bergh (eds) Genetic
Improvement of Vegetable Crops. Pergamon Press, Tarrytown, NY, USA, pp 197–234.
Trebitsh T, Staub JE, O’Neill SD (1997) Identification of an 1-aminocyclopropane-1-carboxylate
synthase gene linked to the Female gene (F) that determines female sex expression in
cucumber (Cucumis sativus L.). Plant Physiol 113: 987–995.
Vision TJ, Brown DG, Shmoys DB, Durrett RT, Tanksley SD (2000) Selective mapping: a strategy
for optimizing the construction of high-density linkage maps. Genetics 155: 407–420.
Vokalounakis DJ (1992) Heart leaf, a recessive leaf shape marker in cucumber: linkage with
disease resistance and other traits. J Hered 83: 217–221.
Wai T, Grumet R (1995) Inheritance of resistance to watermelon strain of papaya ringspot
virus in the cucumber line ‘TMG-1’. HortScience 30: 338–340.
Wai T, Staub JE, Kabelka E, Grumet R (1997) Linkage analysis of potyvirus resistance alleles
in cucumber. J Hered 88: 454–458.
Wang YH, Thomas CE, Dean RA (1997) A genetic map of melon (Cucumis melo L.) based
on amplified fragment length polymorphism (AFLP) markers. Theor Appl Genet 95:
791–798.
Wang YH, Thomas CE, Dean RA (2000) Genetic mapping of a fusarium wilt resistance gene
(Fom-2) in melon (Cucumis melo L.). Mol Breed 6: 379–389.
Wehner TC (1989) Breeding for improved yield in cucumber. In: J Janick (ed) Plant Breeding
Review, John Wiley & Son, Inc Hoboken, NJ vol 6, pp 323–359.
Wehner TC, Staub JE, Peterson CE (1987) Inheritance of littleleaf and multi-branched plant
habit in cucumber. Cucurbit Genet Coop Rep 10: 33–34.
Yu J, Buckler ES (2006) Genetic association mapping and genome organization of maize. Curr
Opin Biotechnol 17: 155–160.
Yu J, Pressoir G, Briggs WH, Vroh Bi I, Yamasaki M, Doebley JF, McMullen MD, Gaut BS,
Nielsen DM, Holland JB, Kresovich S, Buckler ES (2006) A unified mixed-model method
for association mapping that accounts for multiple levels of relatedness. Nat Genet 38:
203–208.
Yu J, Holland JB, McMullen MD, Buckler ES (2008) Genetic design and statistical power of
nested association mapping in maize. Genetics 178: 539–551.
Yuan XJ, Li XZ, Pan JS, Wang G, Jiang S, Li XH, Deng SL, He HL, Si MX, Lai L, Wu AZ, Zhu
LH, Cai R (2008) Genetic linkage map construction and location of QTL for fruit-related
traits in cucumber. Plant Breed 127: 180–188.
Zalapa JE, Staub JE, McCreight JD, Chung SM, Cuevas HE (2007) Detection of QTL for yield-
related traits using recombinant inbred lines derived from exotic and elite US Western
Shipping melon germplasm. Theor Appl Genet 114: 1185–1201.
Zhang Q, Gabert AC, Baggett JR (1992) Parents and mating systems affect the transfer
of gynoecious flowering to Chinese monoecious cucumbers. J Am Soc Hort Sci 117:
515–517.
Zhang RB, Xu Y, Vi K, Zhang HY, Liu LG, Yi GG, Levi A (2004) A genetic linkage map for
watermelon derived from recombinant inbred lines. J Am Soc Hort Sci 129: 237–243.
Zheng XY, Wolff DW, Baudracco-Arnas S, Pitrat M (1999) Development and utility of cleaved
amplified polymorphic sequences (CAPS) and restriction fragment length polymorphism
(RFLPs) linked to the Fom-2 fusarium wilt resistance gene in melon (Cucumis melo L.).
Theor Appl Genet 99: 453–463.
Zhu C, Gore M, Buckler ES, Yu J (2008) Status and prospects of association mapping in plants.
Plant Genome 1: 5–20.
Zitter TA, Hopkins DL, Thomas CE (1996) Compendium of Cucurbit Diseases. APS Press,
Saint Paul, MN, USA.
Zraide A, Stift G, Pachner M, Shojaeiyan A, Gong L, Lelley T (2007) A consensus map for
Cucurbita pepo. Mol Breed 20: 375–388.
ABSTRACT
Melon is an economically important vegetable crop and belongs to the
Cucurbitaceae family, which includes several other important crops
such as watermelon, cucumber, pumpkin and squash. It has served
as a model system for fruit ripening and sex determination studies. In
recent years, significant progress has been made in melon structural
and functional genomics. Several BAC libraries, a physical map, and
a number of high-density genetic linkage maps have been constructed
for melon and currently its genome is being sequenced by the Spanish
Genomics Initiative using a whole-genome shotgun strategy with the
Roche 454 GS FLX Titanium system. A large collection of ESTs consisting
of approximately ~130,000 ESTs derived from various tissues and
genotypes was created and is publicly available at the International
Cucurbit Genomics Initiative (ICuGI) website (http: //www.icugi.org).
Availability of these ESTs allowed the design of melon microarrays for
transcriptome analysis and the identification of SSR/SNP markers for
breeding programs. Other functional genomic resources and studies in
melon, including proteomics and metabolomics researches and melon
mutant libraries, are also discussed and summarized in this chapter.
Keywords: melon, genome sequencing, expressed sequence tags,
transcriptome profiling, molecular breeding
1
Boyce Thompson Institute for Plant Research, Cornell University, Ithaca, NY 14853, USA.
2
USDA-ARS, Robert W. Holley Center for Agriculture and Health, Ithaca, NY 14853, USA.
a
Current address: College of Medicine, Texas A&M Health Science Center, Temple, TX 76504,
USA; e-mail: lyang@medicine.tamhsc.edu
Corresponding author: zf25@cornell.edu
9.1 Introduction
Melon (cucumis melo L.) belongs to the family Cucurbitaceae, which includes
several other important crops such as watermelon, cucumber, pumpkin
and squash. Melon is one of the most intensively studied species in terms
of fruit ripening and sex determination and is becoming an increasingly
important economic vegetable crop. During 2003–2005, the average global
production of melon reached 60.4 billion pounds, ranking at the 16th
position and sharing 1.3% of the world production of fruits and vegetables
(FAOStat database, 05/2006). In addition, melon is also one of the favorite
fruits for dessert and salad uses because of its unique flavor and aroma.
The average per capita consumption of melon in the US has been increasing
consecutively each decade since the 1960s. During the past few years
(2000–2006), the estimated average US per capita consumption exceeded
12 pounds per year, an 8% rise from 1990–1999 (Vegetables and Melons
outlook/VGS-320/April 19, 2007; Economic Research Service, USDA).
Compared with other major crops in the cucurbit family, melon is
one of the species producing fruits with the greatest amount of genetic
and morphological diversity. Melon includes a wide variety of cultivars
producing fruit deferring in many traits including fruit shape (round,
flat, elongated), size (from 50 g to 15 kg), flesh color (orange, light green,
white), sweetness (high or low sugar content), aroma volatiles and fruit
texture (Nunez-Palenius et al. 2008). In addition, melon fruits also have
significant variations in ripening behavior. Melon fruits can be categorized
as either climacteric or non-climacteric types based on their ripening
related respiration rate and ethylene evolution profiles. Usually fruit
in C. melo var. cantalupensis Naud and C. melo var. reticulatus Naud are
considered as climacteric types, including Cantaloup, Vedrantaise, Noy
Yizre’el and Dulce cultivars. Fruits from C. melo var. inodorus Naud are
generally classified as non-climacteric types, including for example Tam
Dew and Rochet cultivars. In addition to ripening physiology, climacteric
and non-climacteric melons also differ in additional phenotypes. Most of
the climacteric melons have orange flesh, high aroma and quick softening
upon ripening, while non-climacteric melons usually display pale-green
flesh, low level of aroma and slow softening resulting in typically longer
shelf-life than climacteric varieties. It is interesting to note that climacteric
and non-climacteric varieties exist in the same species implying that in at
least the case of melon, these differences are more likely the result of genetic
differences in ethylene synthesis or response and probably do not reflect
major differences in primary ripening programs (Giovannoni 2007).
Extensive molecular and genetic studies have been carried out in
order to better understand regulatory mechanisms underlying these traits
with the aim to improve melon fruit quality and to extend storage time.
than 24 million reads representing 17.6× of the melon genome have been
sequenced and assembled. Additionally, 0.05× of BAC end sequences have
been included in the assembly. The current melon assembly contains 367
Mb (81.5%) of the genome sequence, with an N50 scaffold size of 5.2 Mb
and an N90 index of 77. Anchoring the assembled genome scaffolds onto
melon chromosomes based on genetic and physical maps and annotating
the genome are currently underway (Garcia-Mas, pers. comm.).
indicated that 79% of them could be validated and most of the SFPs (81%)
contained SNPs. The usefulness of these SFPs was further confirmed by
testing them in parents of another mapping population, PI161375 and “Piel
de Sapo” (Ophir et al. 2010). This approach provides an efficient way to
discover markers on a large scale for melon, which further facilitates genetic
mapping and molecular-assisted breeding.
Genetic maps in melon have been generated using different mapping
populations and different types of molecular markers. Nevertheless there
is no unique consensus map with the same name for the linkage groups.
In 2005, the International Cucurbit Genomics Initiative (ICuGI; http: //
www.icugi.org) was founded and one of its main goals was to develop a
consensus melon genetic map by merging the existing melon genetic maps
using SSRs as anchor markers. The consensus map is now available at ICuGI
website (http: //www.icugi.org). The map contains a total of 1,244 markers,
including 544 SSRs, 223 SNPs, 235 RFLPs, 109 AFLPs, 92 RAPDs, 18 ISSRs,
7 INDELs, 9 morphological traits, and 7 other markers. The map has 12
linkage groups and a total length of 1,449.1 cM, with an average distance
of 1.16 cM between points (ICuGI unpublished; http: //www.icugi.org).
This consensus map is significantly more saturated than any of the melon
genetic maps published so far and expected to play a significant role in
melon whole-genome sequencing and melon breeding programs.
~1,800 ESTs from phloem-sap of melon cultivar Hales Best Jumbo (Table
9-1). Comparison of these ESTs to those from other tissues allowed the
identification of a set of phloem-sap specific genes, which were mainly
associated with biotic stimulus, response to stress, and metal-ion binding
(Omid et al. 2007). Recently ICuGI released ~94,000 ESTs (Table 9-1; http: //
www.icugi.org), which represents a significant addition to the current melon
EST and functional genomic resources. These ESTs were generated from
fruits, flowers, and callus of four different genotypes: Dulce, Vedrantais,
PI161375, and “Piel de Sapo” T111, as well as melon necrotic spot virus
(MNSV)-infected leaf, root, and cotyledon of “Piel de Sapo” T111; more
than three-fourths of these ESTs were derived from full-length cDNA clones
(Table 9-1). To date, a total of 129,067 melon ESTs have been generated (Table
9-1). All these ESTs, together with 173 mRNA sequences from GenBank,
were assembled into 24,444 unigenes with an average length of 776.7 bp,
comprising 11,653 contigs with an average length of 972 bp and 12,791
singletons of 598.7 bp. The melon unigenes were extensively annotated by
comparing them to different kinds of sequence databases and by assigning
them gene ontology (GO) terms. The sequences and annotations of all
melon ESTs and unigenes are currently available at ICuGI website (http:
//www.icugi.org) in a searchable manner. Functional classification of the
melon unigenes according to a set of plant-specific GO slims, which are
a list of high-level GO terms providing a broad overview of the ontology
content (http: //www.geneontology.org/GO.slims.shtml), is shown in Fig.
9-1. A number of genes that are potentially involved in fruit and flower
development, fruit ripening, and responses to different biotic/abiotic
stresses can be identified from the melon EST collection (Fig. 9-1; ICuGI
unpublished). As shown in Table 9-1, melon ESTs were generated from
more than 10 different genotypes which show high genetic diversity, thus
SNPs were expected to be enriched in the EST collection. We were able to
discover a total of 3,073 high-confidence SNPs, among which were 1,972
transitions, 976 transversions, and 125 single-base indels. Furthermore, over
4,000 SSRs were also identified from the melon EST collection (ICuGI, http:
//www.icugi.org). These SNPs and SSRs are potential valuable markers
for melon breeding programs and part of them have been used to construct
high density genetic maps (Deleu et al. 2009; Harel-Beja et al. 2010).
growth 222
pollination 104
abscission 21
ripening 78
tropism 76
transport 2400
translation 870
transcription 1746
reproduction 795
Figure 9-1 Functional classification of unigenes derived from a collection of ~130,000 melon
ESTs.
the last decade. Currently there are two microarray platforms available for
melon. The first is a cDNA array, which was designed based on the early
collection of melon ESTs. The array contains 3,068 unique cDNA clones
with each clone printed in triplicate on the array, as well as 12 negative
controls. The array is available through the ICuGI website (http: //www.
icugi.org). The second melon array is an oligo-nucleotide array constructed
using the Nimblegen Maskless Array Synthesis technology and currently
is commercially available at the Nimblegen Company. The array was
cultivar (Dulce) during fruit ripening (Liu, Fei, Katzir and Giovannoni
unpubl.data). Mascarell-Creus et al. (2009) reported using the melon
Nimblegen oligo-nucleotide array to characterize global gene expression
profiles during fruit ripening of a non-climacteric melon, “Piel de Sapo”
T111, and in response to viral and fungal infections in melon roots and
cotyledons. They found that fruit ripening of “Piel de Sapo” T111 involved
down-regulation of ethylene biosynthesis genes and differential regulation
of sugar metabolism and cell wall-loosening enzymes, while in cultivar
agrestis Pat 81, responses of roots to fungi Monosporascus cannonballus
involved down-regulation of signal transduction pathways and cell wall
and cytoskeleton rearrangements, and in cultivar “Piel de Sapo” tendril,
cotyledons infected with cucumber mosaic virus (CMV) induced structural
cell-cycle deregulation (Mascarell-Creus et al. 2009).
It has been proved that the occurrence of ESTs from large unbiased
(non-normalized and non-subtracted) cDNA libraries represents the
relative expression of genes from which the ESTs are derived. Using ESTs
as an approach of expression profiling has been reported in several plant
species including tomato (Fei et al. 2004), grape (da Silva et al. 2005),
and apple (Park et al. 2006). As shown in Table 9-1, the current melon
EST collection includes ESTs derived from unbiased libraries (especially
those by ICuGI) of different tissues, development stages, and conditions.
These ESTs provided a valuable and rich source for identification of tissue
specific genes, highly expressed genes, and differentially expressed genes.
However, the overall effort and cost of EST sequencing are major hurdles
of this approach. This situation has changed during the past several years
due to rapid advances of next-generation sequencing technologies, led by
Illumina/Solexa and Roche/454 sequencing by synthesis technologies,
which can generate million to 10s of millions short tags (40–400 bp)
efficiently and cost effectively. In addition, these technologies do not require
labor intensive cloning effort and the cost can be reduced significantly.
RNA-seq, an emerging technology for large-scale gene expression analysis
through sequencing the whole RNA population using high-throughput
sequencing technologies (Wang et al. 2009), is expected to eventually phase
out microarrays for transcriptomic studies. Microarray is a hybridization-
based technology, thus it is prone to suffer from hybridization artifacts.
Cross-hybridization of closely related members in the same gene family
or short stretches of nucleotide homologies has been observed in cDNA
and oligo-nucleotide arrays (Kachalo et al. 2002), respectively. Stable
probe secondary structures (Southern et al. 1999) may interfere with
array hybridization. In addition, problems such as high background (e.g.,
nonspecific hybridization) and limited dynamic range (e.g., nonlinear and
saturable hybridization kinetics) have been well documented (Velculescu
and Kinzler 2007). RNA-seq approaches bypass the longstanding technical
References
Adams MD, Kelley JM, Gocayne JD, Dubnick M, Polymeropoulos MH, Xiao H, Merril CR, Wu
A, Olde B, Moreno RF, Kerlavage AR, Mccombie WR, Venter JC (1991) Complementary-
DNA sequencing—expressed sequence tags and human genome project. Science 252:
1651–1656.
Allwood JW, Erban A, de Koning S, Dunn WB, Luedemann A, Lommen A, Kay L, Löscher R,
Kopka J, Goodacre R (2009) Inter-laboratory reproducibility of fast gas chromatography–
electron impact–time of flight mass spectrometry (GC-EI-TOF/MS) based plant
metabolomics. Metabolomics 5: 479–496.
Arabidopsis Genome Initiative (2000) Analysis of the genome sequence of the flowering plant
Arabidopsis thaliana. Nature 408: 796–815.
Arumuganathan K, Earle E (1991) Nuclear DNA content of some important plant species.
Plant Mol Biol Rep 9: 210–220.
Ayub R, Guis M, Ben Amor M, Gillot L, Roustan JP, Latché A, Bouzayen M, Pech JC (1996)
Expression of ACC oxidase antisense gene inhibits ripening of cantaloupe melon fruits.
Nat Biotechnol 14: 862–866.
Bauchot AD, Mottram DS, Dodson AT, John P (1998) Effect of ACO antisense genes on
the formation of volatile esters in cataloup Charentais melon. J Agric Food Chem 46:
4787–4792.
Baudracco-Arnas S, Pitrat M (1996) A genetic map of melon (Cucumis melo L.) with RFLP, RAPD,
isozyme, disease resistance and morphological markers. Theor Appl Genet 93: 57–64.
Biais B, Allwood JW, Deborde C, Xu Y, Maucourt M, Beauvoit B, Dunn WB, Jacob D, Goodacre
R, Rolin D, Moing A (2009) 1H NMR, GC-EI-TOFMS, and data set correlation for fruit
metabolomics: application to spatial metabolite analysis in melon. Anal Chem 81:
2884–2894.
Biais B, Beauvoit B, William Allwood J, Deborde C, Maucourt M, Goodacre R, Rolin D, Moing
A (2010) Metabolic acclimation to hypoxia revealed by metabolite gradients in melon
fruit. J Plant Physiol 167: 242–245.
Boualem A, Fergany M, Fernandez R, Troadec C, Martin A, Morin H, Sari MA, Collin F,
Flowers JM, Pitrat M, Purugganan MD, Dogimont C, Bendahmane A (2008) A conserved
mutation in an ethylene biosynthesis enzyme leads to andromonoecy in melons. Science
321: 836–838.
Chan AP, Crabtree J, Zhao Q, Lorenzi H, Orvis J, Puiu D, Melake-Berhan A, Jones KM, Redman
J, Chen G, Cahoon EB, Gedil M, Stanke M, Haas BJ, Wortman JR, Fraser-Liggett CM,
Ravel J, Rabinowicz PD (2010) Draft genome sequence of the oilseed species Ricinus
communis. Nat Biotechnol 28: 951–956.
Cuevas HE, Staub JE, Simon PW, Zalapa JE, McCreight JD (2008) Mapping of genetic loci that
regulate quantity of beta-carotene in fruit of US Western Shipping melon (Cucumis melo
L.). Theor Appl Genet 117: 1345–1359.
Cuevas HE, Staub JE, Simon PW, Zalapa JE (2009) A consensus linkage map identifies genomic
regions controlling fruit maturity and beta-carotene-associated flesh color in melon
(Cucumis melo L.). Theor Appl Genet 119: 741–756.
Danin-Poleg Y, Tadmor Y, Tzuri G, Reis N, Hirschberg J, Katzir N (2002) Construction of a
genetic map of melon with molecular markers and horticultural traits, and localization
of genes associated with ZYMV resistance. Euphytica 125: 373–384.
Deleu W, Esteras C, Roig C, Gonzalez-To M, Fernandez-Silva I, Gonzalez-Ibeas D, Blanca J,
Aranda MA, Arus P, Nuez F, Monforte AJ, Pico MB, Garcia-Mas J (2009) A set of EST-SNPs
for map saturation and cultivar identification in melon. BMC Plant Biol 9: 90.
Du CX, Fan HF, Guo SR, Tezuka T, Li J (2010) Proteomic analysis of cucumber seedling roots
subjected to salt stress. Phytochemistry 71: 1450–1459.
Ezura H, Fukino N (2009) Research tools for functional genomics in melon (Cucumis melo L.):
Current status and prospects. Plant Biotechnol 26: 359–368.
Fei Z, Tang X, Alba RM, White JA, Ronning CM, Martin GB, Tanksley SD, Giovannoni JJ (2004)
Comprehensive EST analysis of tomato and comparative genomics of fruit ripening.
Plant J 40: 47–59.
Fernandez-Silva I, Eduardo I, Blanca J, Esteras C, Pico B, Nuez F, Arus P, Garcia-Mas J, Monforte
AJ (2008) Bin mapping of genomic and EST-derived SSRs in melon (Cucumis melo L.).
Theor Appl Genet 118: 139–150.
Fiehn O, Kopka J, Dormann P, Altmann T, Trethewey RN, Willmitzer L (2000) Metabolite
profiling for plant functional genomics. Nat Biotechnol 18: 1157–1161.
Flores F, El Yahyaoui F, de Billerbeck G, Romojaro F, Latché A, Bouzayen M, Pech JC, Ambid
C (2002) Role of ethylene in the biosynthetic pathway of aliphatic ester aroma volatiles
in Charentais Cantaloupe melons. J Exp Bot 53: 201–206.
Fukino N, Sugiyama M, Ohara T, Sainoki H, Kubo N, Hirai M, Matsumoto S, Sakata Y
(2008a) Detection of quantitative trait loci affecting lateral branching in Cucumis melo.
In: Cucurbitacea 2008. Proc IXth EUCARPIA Meeting on Genetics and Breeding of
Cucurbitaceae. Avignon, France, pp 505–509.
Fukino N, Ohara T, Monforte AJ, Sugiyama M, Sakata Y, Kunihisa M, Matsumoto S (2008b)
Identification of QTLs for resistance to powdery mildew and SSR markers diagnostic
for powdery mildew resistance genes in melon (Cucumis melo L.). Theor Appl Genet
118: 165–175.
Giovannoni JJ (2007) Fruit ripening mutants yield insights into ripening control. Curr Opin
Plant Biol 10: 283–289.
Gonzalez VM, Garcia-Mas J, Arus P, Puigdomenech P (2010) Generation of a BAC-based
physical map of the melon genome. BMC Genom 11: 339.
Gonzalez-Ibeas D, Blanca J, Roig C, González-To M, Picó B, Truniger V, Gómez P, Deleu W,
Caño-Delgado A, Arús P, Nuez F, Garcia-Mas J, Puigdomènech P, Aranda MA (2007)
MELOGEN: an EST database for melon functional genomics. BMC Genom 8: 306.
Gonzalo MJ, Oliver M, Garcia-Mas J, Monfort A, Dolcet-Sanjuan R, Katzir N, Arús P, Monforte
AJ (2005) Simple-sequence repeat markers used in merging linkage maps of melon
(Cucumis melo L.). Theor Appl Genet 110: 802–811.
Harel-Beja R, Tzuri G, Portnoy V, Lotan-Pompan M, Lev S, Cohen S, Dai N, Yeselson L, Meir A,
Libhaber SE, Avisar E, Melame T, van Koert P, Verbakel H, Hofstede R, Volpin H, Oliver
M, Fougedoire A, Stalh C, Fauve J, Copes B, Fei Z, Giovannoni J, Ori N, Lewinsohn E,
Sherman A, Burger J, Tadmor Y, Schaffer AA, Katzir N (2010) A genetic map of melon
highly enriched with fruit quality QTLs and EST markers, including sugar and carotenoid
metabolism genes. Theor Appl Genet 121: 511–533.
Huang S, Li R, Zhang Z, Li L, Gu X, Fan W, Lucas WJ, Wang X, Xie B, Ni P, Ren Y, Zhu H, Li
J, Lin K, Jin W, Fei Z, Li G, Staub J, Kilian A, van der Vossen EA, Wu Y, Guo J, He J, Jia
Z, Ren Y, Tian G, Lu Y, Ruan J, Qian W, Wang M, Huang Q, Li B, Xuan Z, Cao J, Asan,
Wu Z, Zhang J, Cai Q, Bai Y, Zhao B, Han Y, Li Y, Li X, Wang S, Shi Q, Liu S, Cho WK,
Kim JY, Xu Y, Heller-Uszynska K, Miao H, Cheng Z, Zhang S, Wu J, Yang Y, Kang H,
Li M, Liang H, Ren X, Shi Z, Wen M, Jian M, Yang H, Zhang G, Yang Z, Chen R, Liu S,
Li J, Ma L, Liu H, Zhou Y, Zhao J, Fang X, Li G, Fang L, Li Y, Liu D, Zheng H, Zhang Y,
Qin N, Li Z, Yang G, Yang S, Bolund L, Kristiansen K, Zheng H, Li S, Zhang X, Yang H,
Wang J, Sun R, Zhang B, Jiang S, Wang J, Du Y, Li S (2009) The genome of the cucumber,
Cucumis sativus L. Nat Genet 41: 1275–1281.
International Brachypodium Initiative (2010) Genome sequencing and analysis of the model
grass Brachypodium distachyon. Nature 463: 763–768.
International Rice Genome Sequencing Project (2005) The map-based sequence of the rice
genome. Nature 436: 793–800.
Jaillon O, Aury JM, Noel B, Policriti A, Clepet C, Casagrande A, Choisne N, Aubourg S,
Vitulo N, Jubin C, Vezzi A, Legeai F, Hugueney P, Dasilva C, Horner D, Mica E, Jublot
D, Poulain J, Bruyère C, Billault A, Segurens B, Gouyvenoux M, Ugarte E, Cattonaro F,
Anthouard V, Vico V, Del Fabbro C, Alaux M, Di Gaspero G, Dumas V, Felice N, Paillard
S, Juman I, Moroldo M, Scalabrin S, Canaguier A, Le Clainche I, Malacrida G, Durand
E, Pesole G, Laucou V, Chatelet P, Merdinoglu D, Delledonne M, Pezzotti M, Lecharny
A, Scarpelli C, Artiguenave F, Pè ME, Valle G, Morgante M, Caboche M, Adam-Blondon
AF, Weissenbach J, Quétier F, Wincker P; French-Italian Public Consortium for Grapevine
Genome Characterization (2007) The grapevine genome sequence suggests ancestral
hexaploidization in major angiosperm phyla. Nature 449: 463–467.
Joobeur T, King JJ, Nolin SJ, Thomas CE, Dean RA (2004) The Fusarium wilt resistance locus
Fom-2 of melon contains a single resistance gene with complex features. Plant J 39:
283–297.
Kachalo S, Arbieva Z, Liang J (2002) Assessing effect of cross hybridization on oligonucleotide
microarrays. In: Critical Assessment of Microarray Data Analysis (CAMDA02).
Kater MM, Franken J, Carney KJ, Colombo L, Angenent GC (2001) Sex determination in
the monoecious species cucumber is confined to specific floral whorls. Plant Cell 13:
481–493.
Kenigsbuch D, Cohen Y (1990) The inheritance of gynoecy in muskmelon. Genome 33:
317–320.
van Leeuwen H, Monfort A, Zhang H-B, Puigdoménech P (2003) Identification and
characterisation of a melon genomic region containing a resistance gene cluster from a
constructed BAC library. Microcolinearity between Cucumis melo and Arabidopsis thaliana.
Plant Mol Biol 51: 703–718.
Lin MK, Lee YJ, Lough TJ, Phinney BS, Lucas WJ (2009) Analysis of the pumpkin phloem
proteome provides insights into angiosperm sieve tube function. Mol Cell Proteom 8:
343–356.
Luo MZ, Wang YH, Frisch D, Joobeur T, Wing RA, Dean RA (2001) Melon bacterial artificial
chromosome (BAC) library construction using improved methods and identification of
clones linked to the locus conferring resistance to melon Fusarium wilt (Fom-2). Genome
44: 154–162.
Martin A, Troadec C, Boualem A, Rajab M, Fernandez R, Morin H, Pitrat M, Dogimont C,
Bendahmane A (2009) A transposon-induced epigenetic change leads to sex determination
in melon. Nature 461: 1135–1138.
Mascarell-Creus A, Cañizares J, Vilarrasa-Blasi J, Mora-García S, Blanca J, Gonzalez-Ibeas D,
Saladié M, Roig C, Deleu W, Picó-Silvent B, López-Bigas N, Aranda MA, Garcia-Mas
J, Nuez F, Puigdomènech P, Caño-Delgado AI (2009) An oligo-based microarray offers
novel transcriptomic approaches for the analysis of pathogen resistance and fruit quality
traits in melon (Cucumis melo L.). BMC Genom 10: 467.
Ming R, Hou S, Feng Y, Yu Q, Dionne-Laporte A, Saw JH, Senin P, Wang W, Ly BV, Lewis KL,
Salzberg SL, Feng L, Jones MR, Skelton RL, Murray JE, Chen C, Qian W, Shen J, Du P,
Eustice M, Tong E, Tang H, Lyons E, Paull RE, Michael TP, Wall K, Rice DW, Albert H,
Wang ML, Zhu YJ, Schatz M, Nagarajan N, Acob RA, Guan P, Blas A, Wai CM, Ackerman
CM, Ren Y, Liu C, Wang J, Wang J, Na JK, Shakirov EV, Haas B, Thimmapuram J, Nelson
D, Wang X, Bowers JE, Gschwend AR, Delcher AL, Singh R, Suzuki JY, Tripathi S, Neupane
K, Wei H, Irikura B, Paidi M, Jiang N, Zhang W, Presting G, Windsor A, Navajas-Pérez
R, Torres MJ, Feltus FA, Porter B, Li Y, Burroughs AM, Luo MC, Liu L, Christopher DA,
Mount SM, Moore PH, Sugimura T, Jiang J, Schuler MA, Friedman V, Mitchell-Olds T,
Shippen DE, de Pamphilis CW, Palmer JD, Freeling M, Paterson AH, Gonsalves D, Wang
L, Alam M (2008) The draft genome of the transgenic tropical fruit tree papaya (Carica
papaya Linnaeus). Nature 452: 991–996.
Niemirowiczszczytt K, Kubicki B (1979) Cross fertilization between cultivated species of genera
Cucumis L. and Cucurbita L. Genet Polon 20: 117–124.
Nieto C, Morales M, Orjeda G, Clepet C, Monfort A, Sturbois B, Puigdomènech P, Pitrat M,
Caboche M, Dogimont C, Garcia-Mas J, Aranda MA, Bendahmane A (2006) An eIF4E
allele confers resistance to an uncapped and non-polyadenylated RNA virus in melon.
Plant J 48: 452–462.
Nieto C, Piron F, Dalmais M, Marco CF, Moriones E, Gomez-Guillamon ML, Truniger V, Gomez
P, Garcias-Mas J, Aranda MA, Bendahmane A (2007) EcoTILLING for the identification
of allelic variants of melon eIF4E, a factor that controls virus susceptibility. BMC Plant
Biol 7: 34.
Nunez-Palenius HG, Huber DJ, Klee HJ, Cantliffe DJ (2007) Fruit ripening characteristics
in transgenic “Galia” male parental muskmelon line. Poastharvest Biol Technol 44:
95–100.
Nunez-Palenius HG, Gomez-Lim M, Ochoa-Alejo N, Grumet R, Lester G, Cantliffe DJ
(2008) Melon fruits: genetic diversity, physiology, and biotechnology features. Crit Rev
Biotechnol 28: 13–55.
Oliver M, Garcia-Mas J, Cardús M, Pueyo N, López-Sesé AI, Arroyo M, Gómez-Paniagua H,
Arús P, de Vicente C (2001) Construction of a reference linkage map of melon. Genome
44: 836–845.
Omid A, Keilin T, Glass A, Leshkowitz D, Wolf S (2007) Characterization of phloem-sap
transcription profile in melon plants. J Exp Bot 58: 3645–3656.
Ophir R, Eshed R, Harel-Beja R, Tzuri G, Portnoy V, Burger Y, Uliel S, Katzir N, Sherman
A (2010) High-throughput marker discovery in melon using a self-designed oligo
microarray. BMC Genom 11: 269.
Park S, Sugimoto N, Larson MD, Beaudry R, van Nocker S (2006) Identification of genes with
potential roles in apple fruit development and biochemistry through large-scale statistical
analysis of expressed sequence tags. Plant Physiol 141: 811–824.
Paterson AH, Bowers JE, Bruggmann R, Dubchak I, Grimwood J, Gundlach H, Haberer G,
Hellsten U, Mitros T, Poliakov A, Schmutz J, Spannagl M, Tang H, Wang X, Wicker T,
Bharti AK, Chapman J, Feltus FA, Gowik U, Grigoriev IV, Lyons E, Maher CA, Martis
M, Narechania A, Otillar RP, Penning BW, Salamov AA, Wang Y, Zhang L, Carpita NC,
Freeling M, Gingle AR, Hash CT, Keller B, Klein P, Kresovich S, McCann MC, Ming R,
Peterson DG, Mehboob-ur-Rahman, Ware D, Westhoff P, Mayer KF, Messing J, Rokhsar
DS (2009) The Sorghum bicolor genome and the diversification of grasses. Nature 457:
551–556.
Perchepied L, Bardin M, Dogimont C, Pitrat M (2005a) Relationship between loci conferring
downy mildew and powdery mildew resistance in melon assessed by QTL mapping.
Phytopathology 95: 556–565.
Perchepied L, Dogimont C, Pitrat M (2005b) Strain-specific and recessive QTLs involved in
control of partial resistance to Fusarium oxysporum f. sp. melonis race 1.2 in a recombinant
inbred line population of melon. Theor Appl Genet 111: 65–74.
Périn C, Hagen LS, de Conto V, Katzir N, Danin-Poleg Y, Portnoy V, Baudracco-Arnas S,
Chadoeuf J, Dogimont C, Pitrat M (2002) A reference map of Cucumis melo based on two
recombinant inbred line populations. Theor Appl Genet 104: 1017–1034.
Pitrat M, Chauvet M, Foury C (1999) Diversity, history and productivity of cultivated cucurbits.
In: K Abak, S Buyukalaca (eds) 1st Int Symp on Cucurbtis. ISHS Adana, Turkey, pp
21–28.
Poole CF, Grimball PC (1939) Inheritance of new sex forms in Cucumis melo L. J Hered 30:
21–25.
Puigdomènech P, Martínez-Izquierdo JA, Arús P, Garcia-Mas J, Monforte AJ, Nuez F, Picó B,
Blanca J, Aranda M, Arnau V, Robles A (2007) The Spanish melon genomics initiative.
Acta Hort 731: 47–54.
Rudd S (2003) Expressed sequence tags: alternative or complement to whole genome
sequences? Trends Plant Sci 8: 321–329.
Samuel P (2008) Transcriptional profiling of aphid resistant and susceptible melon (Cucumis
melo) following cotton-melon aphid (Aphis gossypii) feeding. PhD thesis. Univ of Arkansas,
AK, USA.
Schmutz J, Cannon SB, Schlueter J, Ma J, Mitros T, Nelson W, Hyten DL, Song Q, Thelen JJ,
Cheng J, Xu D, Hellsten U, May GD, Yu Y, Sakurai T, Umezawa T, Bhattacharyya MK,
Sandhu D, Valliyodan B, Lindquist E, Peto M, Grant D, Shu S, Goodstein D, Barry K,
Futrell-Griggs M, Abernathy B, Du J, Tian Z, Zhu L, Gill N, Joshi T, Libault M, Sethuraman
A, Zhang XC, Shinozaki K, Nguyen HT, Wing RA, Cregan P, Specht J, Grimwood
J, Rokhsar D, Stacey G, Shoemaker RC, Jackson SA (2010) Genome sequence of the
palaeopolyploid soybean. Nature 463: 178–183.
Schnable PS, Ware D, Fulton RS, Stein JC, Wei F, Pasternak S, Liang C, Zhang J, Fulton L, Graves
TA, Minx P, Reily AD, Courtney L, Kruchowski SS, Tomlinson C, Strong C, Delehaunty K,
Fronick C, Courtney B, Rock SM, Belter E, Du F, Kim K, Abbott RM, Cotton M, Levy A,
Marchetto P, Ochoa K, Jackson SM, Gillam B, Chen W, Yan L, Higginbotham J, Cardenas
M, Waligorski J, Applebaum E, Phelps L, Falcone J, Kanchi K, Thane T, Scimone A, Thane
N, Henke J, Wang T, Ruppert J, Shah N, Rotter K, Hodges J, Ingenthron E, Cordes M,
Kohlberg S, Sgro J, Delgado B, Mead K, Chinwalla A, Leonard S, Crouse K, Collura K,
Kudrna D, Currie J, He R, Angelova A, Rajasekar S, Mueller T, Lomeli R, Scara G, Ko A,
Delaney K, Wissotski M, Lopez G, Campos D, Braidotti M, Ashley E, Golser W, Kim H,
Lee S, Lin J, Dujmic Z, Kim W, Talag J, Zuccolo A, Fan C, Sebastian A, Kramer M, Spiegel
L, Nascimento L, Zutavern T, Miller B, Ambroise C, Muller S, Spooner W, Narechania A,
Ren L, Wei S, Kumari S, Faga B, Levy MJ, McMahan L, Van Buren P, Vaughn MW, Ying K,
Yeh CT, Emrich SJ, Jia Y, Kalyanaraman A, Hsia AP, Barbazuk WB, Baucom RS, Brutnell
TP, Carpita NC, Chaparro C, Chia JM, Deragon JM, Estill JC, Fu Y, Jeddeloh JA, Han Y,
Lee H, Li P, Lisch DR, Liu S, Liu Z, Nagel DH, McCann MC, SanMiguel P, Myers AM,
Nettleton D, Nguyen J, Penning BW, Ponnala L, Schneider KL, Schwartz DC, Sharma
A, Soderlund C, Springer NM, Sun Q, Wang H, Waterman M, Westerman R, Wolfgruber
TK, Yang L, Yu Y, Zhang L, Zhou S, Zhu Q, Bennetzen JL, Dawe RK, Jiang J, Jiang N,
Presting GG, Wessler SR, Aluru S, Martienssen RA, Clifton SW, McCombie WR, Wing
RA, Wilson RK (2009) The B73 maize genome: complexity, diversity, and dynamics.
Science 326: 1112–1115.
Segarra G, Casanova E, Bellido D, Odena MA, Oliveira E, Trillas I (2007) Proteome, salicylic
acid, and jasmonic acid changes in cucumber plants inoculated with Trichoderma asperellum
strain T34. Proteomics 7: 3943–3952.
Shattuck-Eidens DM, Bell RN, Neuhausen SL, Helentjaris T (1990) DNA sequence variation
within maize and melon: observations from polymerase chain reaction amplification and
direct sequencing. Genetics 126: 207–217.
Silberstein L, Kovalski I, Brotman Y, Périn C, Dogimont C, Pitrat M, Klingler J, Thompson
G, Portnoy V, Katzir N, Perl-Treves R (2003) Linkage map of Cucumis melo including
phenotypic traits and sequence-characterized genes. Genome 46: 761–773.
da Silva FG, Iandolino A, Al-Kayal F, Bohlmann MC, Cushman MA, Lim H, Ergul A, Figueroa
R, Kabuloglu EK, Osborne C, Rowe J, Tattersall E, Leslie A, Xu J, Baek J, Cramer GR,
Cushman JC, Cook DR (2005) Characterizing the grape transcriptome. Analysis of
expressed sequence tags from multiple Vitis species and development of a compendium
of gene expression during berry development. Plant Physiol 139: 574–597.
Southern E, Mir K, Shchepinov M (1999) Molecular interactions on microarrays. Nat Genet
21: S5–9
Tadmor Y, Katzir N, Meir A, Yaniv-Yaakov A, Sa’ar U, Baumkoler F, Lavee T, Lewinsohn E,
Schaffer A, Burger J (2007) Induced mutagenesis to augment the natural genetic variability
of melon (Cucumis melo L.). Plant Sci 55: 159–169.
Tanurdzic M, Banks JA (2004) Sex-determining mechanisms in land plants. Plant Cell 16:
S61–71.
Tezuka T, Waki K, Yashiro K, Kuzuya M, Ishikawa T, Takatsu Y, Mijagi M (2009) Construction
of a linkage map and identification of DNA markers linked to Fom-1, a gene conferring
resistance to Fusarium oxysporum f.sp. melonis race 2 in melon. Euphytica 168: 177–188.
Tuskan GA, Difazio S, Jansson S, Bohlmann J, Grigoriev I, Hellsten U, Putnam N, Ralph S,
Rombauts S, Salamov A, Schein J, Sterck L, Aerts A, Bhalerao RR, Bhalerao RP, Blaudez
D, Boerjan W, Brun A, Brunner A, Busov V, Campbell M, Carlson J, Chalot M, Chapman
J, Chen GL, Cooper D, Coutinho PM, Couturier J, Covert S, Cronk Q, Cunningham R,
Davis J, Degroeve S, Déjardin A, Depamphilis C, Detter J, Dirks B, Dubchak I, Duplessis S,
Ehlting J, Ellis B, Gendler K, Goodstein D, Gribskov M, Grimwood J, Groover A, Gunter
L, Hamberger B, Heinze B, Helariutta Y, Henrissat B, Holligan D, Holt R, Huang W,
Islam-Faridi N, Jones S, Jones-Rhoades M, Jorgensen R, Joshi C, Kangasjärvi J, Karlsson
J, Kelleher C, Kirkpatrick R, Kirst M, Kohler A, Kalluri U, Larimer F, Leebens-Mack
J, Leplé JC, Locascio P, Lou Y, Lucas S, Martin F, Montanini B, Napoli C, Nelson DR,
Nelson C, Nieminen K, Nilsson O, Pereda V, Peter G, Philippe R, Pilate G, Poliakov A,
Razumovskaya J, Richardson P, Rinaldi C, Ritland K, Rouzé P, Ryaboy D, Schmutz J,
Schrader J, Segerman B, Shin H, Siddiqui A, Sterky F, Terry A, Tsai CJ, Uberbacher E,
Unneberg P, Vahala J, Wall K, Wessler S, Yang G, Yin T, Douglas C, Marra M, Sandberg
G, Van de Peer Y, Rokhsar D (2006) The genome of black cottonwood, Populus trichocarpa
(Torr. & Gray). Science 313: 1596–604.
Vaucheret H (2006) Post-transcriptional small RNA pathways in plants: mechanisms and
regulations. Genes Dev 20: 759–771.
Velasco R, Zharkikh A, Affourtit J, Dhingra A, Cestaro A, Kalyanaraman A, Fontana P,
Bhatnagar SK, Troggio M, Pruss D, Salvi S, Pindo M, Baldi P, Castelletti S, Cavaiuolo M,
Coppola G, Costa F, Cova V, Dal Ri A, Goremykin V, Komjanc M, Longhi S, Magnago P,
Malacarne G, Malnoy M, Micheletti D, Moretto M, Perazzolli M, Si-Ammour A, Vezzulli S,
Zini E, Eldredge G, Fitzgerald LM, Gutin N, Lanchbury J, Macalma T, Mitchell JT, Reid J,
ABSTRACT
This chapter elucidates the challenges in genetics, breeding, and
genomics of watermelon, dealing with crop origin and major diseases
and pests. It also delineates challenges in enhancing watermelon
cultivars for disease and pest resistance, genetic mapping and the
identification of disease resistance genes. It describes the recent
advances in genomics of watermelon and the construction of a physical
map and genetic maps for watermelon. At present, about 80% of the
watermelon (Citrullus lanatus var. lanatus) genome has been sequenced
and assembled using the advanced 454 and Solexa technologies. There
are two major genomic projects for watermelon. The first is in China
using the elite Chinese watermelon line 97103 for whole genome
sequencing. The second sequencing project is in the USA, using the
heirloom cultivar Charleston Gary genome for sequencing. Alongwith
the ongoing genomic project, there is a great need to phyenotype
and identify genes associated with disease or pest resistance in wild
watermelon and incorporate these genes into watermelon cultivars
without compromising fruit quality. The genomic and genetic studies
described here should be a useful platform for further studies and gene
discovery in watermelon.
Keywords: Citrullus, genomics, genetic map, resistance, genome
sequencing
1
USDA-ARS, U.S. Vegetable Laboratory, 2700 Savannah Highway, Charleston, SC, 29414,
USA.
2
Department of Biology, West Virginia State University, Institute, WV 25112, USA.
3
National Engineering Research Center for Vegetables Banjing, 100097 Beijing, PO Box 2443,
PR China.
4
Syngenta Seeds, 21435 Road 98, Woodland, CA 95695, USA.
*Corresponding author: Amnon-Levi@usda.ars.gov
Citrullus colocynthis
Figure 10-1 A dendogram showing genetic relationships among 31 Citrullus spp. based on
the neighbor-joining (NJ) method (Saitou and Nei 1987) using amplified fragment length
polymorphism (AFLP) and simple sequence repeat (SSR) fragments. Numbers shown at
different nodes represent percentage confidence limits obtained in the bootstrap analysis.
1 2 3 4 5 6 7 8 9 10 11 12
p8/Dra1
–1.8 Kb
Figure 10-2 Autoradiogram (chloroplast DNA P8 fragment to DNA digested with restriction
enzyme DraI), showing close affinity of the chloroplast genome of C. colocynthis (PI 386016, PI
388770, and PI 432337; lanes 10–12) to those of C. lanatus subsp. citroides (PI 271778, PI 296341,
and PI 500332; lanes 7–9), but not to those of Citrullus lanatus subsp. lanatus accessions (PI
162667, PI 169289, and PI 185635; lanes 4–6) or watermelon cultivars ( “Allsweet”, “Charleston
Gray” and “Black Diamond”; lanes 1–3).
Watermelon has been cultivated in Africa and in the Middle East for
thousands of years, and in China before 900 AD, and was brought from
Africa to the New World in the 1500s. Although watermelon was first
domesticated near the center of origin in Africa and is popular in most African
countries, Asia is the leading continent in production and consumption of
this important vegetable crop. Over 75% of the world watermelons are
produced in Asia. China is the leading producer, growing 76 million tons
of watermelon per year (about 73% of the total world production), while
the United States produces 3.8 million tons of watermelon per year (about
2.5% of total world production) (http://www.fao.org). Watermelon is
grown in 44 states in the US, Florida, Georgia, California, and Texas,
having long warm growing seasons are the major producing states. In the
US, watermelon production has increased from 1.2 M tons in 1980 to 3.8 M
tons in 2009, with an annual farm value of $470 million (US Department of
Agriculture, Agricultural Statistics, 2009). In recent years, there has been
an increase in consumer demand for seedless watermelons and production
of seedless watermelon has increased significantly. Today, over 80% of the
watermelons produced in the US are triploid seedless watermelons (US
National Watermelon Promotion Board; www.watermelon.org ).
Watermelon needs high temperatures and intense light to promote
flowering, and fruit set and development. Watermelon plants have trailing
vines with branched tendrils at each node, and their leaves are divided
into three or four pairs of lobes. In the field, the vines of certain wild
type accessions can reach up to 8–9 meter long. However, there are dwarf
watermelon accessions with short and/or less-branched stems controlled
by two recessive genes (dw-1 and dw-2). Roots of wild type watermelons
that grow in the African deserts are extensive, penetrating deep into the soil
layers. However, in commercial fields, with drip irrigation, most of the root
system of watermelon cultivars resides in the upper soil surface.
Watermelon has small flowers (about 0.5–0.6” and 0.6–1.0” in diameter in
C. colocynthis and C. lanatus, respectively) compared with other major cucurbit
crops, including Benincasa hispida (Thunb.) Cogn, Lagenaria siceraria (Mol.)
Standl. and Cucurbita spp.(Levi et al. 2010). Like most cucurbit species, the
flowers of watermelon have five yellow petals. Most watermelon cultivars are
monoecious, having separate male (staminate) and female (pistillate) flowers
(that are typically formed in this order) on the same plant. Many of the wild
type watermelon accessions are andromonoecious, producing both male and
perfect (hermaphroditic) flowers on the same plant. The female flowers have
an inferior ovary, and the size and shape of the ovary is correlated with final
fruit size and shape. In many cultivars, the male flowers are positioned at
most nodes while the female or perfect flowers are positioned at every seventh
node. The male:female flower ratio of typical watermelon cultivars ranges
from 3:1 (Extra Early Sugar Baby) to 7:1 (Black Diamond) to 11:1 (Charleston
Gray) and to 15:1 (C. lanatus subsp. citroides PI 500331). Watermelon plants
start to form flower at the 5–7th internodes, at about 4 weeks after planting.
Ethylene is known to play a critical role in floral sex determination of cucurbit
species. Ethylene promotes female flowers in cucumber and melon. However,
in watermelon it is possible that ethylene promotes the formation of male
flowers (Salman-Minkov et al. 2008).
The phylogenetic relationships among Citrullus species and subspecies
were examined using isozymes (Zamir et al. 1984; Navot and Zamir 1987),
genomic DNA markers (Jarret et al. 1997; Levi et al. 2001a, b) and organelle
DNA markers (Dane and Lang 2004; Levi and Thomas 2005). These genetic
studies (Navot and Zamir 1987; Levi et al. 2001a, b) indicated that although
possessing a wide phenotypic diversity, a narrow genetic base exists
among watermelon cultivars. In these studies, most isozymes produced
monomorphic patterns among cultivars (Zamir et al. 1984; Biles et al. 1989).
RAPD analysis produced low polymorphism, but provided informative
markers that could distinguish among watermelon cultivars that share a
narrow genetic base (Hashizume et al. 1993; Zhang et al. 2004). This narrow
genetic base is a result of many years of cultivation and continuous selection
for a set of desirable qualities, suitable to the needs of growers, shippers,
and consumers.
Over 314 American heirloom cultivars are maintained by the USDA, ARS,
Regional Plant Introduction Station, Griffin, Georgia and are considered an
essential germplasm resource for watermelon breeding programs. Among
them are “Allsweet”, “Au-Producer”, “Charleston Gray”, “Crimson Sweet”,
“Jubilee”, and “Peacock”. These heirloom cultivars are grown throughout
the world and are widely used as parents for many F1 hybrid lines. There
is incomplete information regarding the ancestries of many American
watermelon cultivars developed during the 19th and early 20th centuries.
For a long time, the identification of watermelon cultivars relied mainly
on fruit characteristics. Today, molecular markers are extremely useful in
determining genetic relatedness and genetic purity of cultivars. Overall, the
watermelon cultivars do not show any inbreeding depression, and inbred
Figure 10-3 Root systems of Citrullus colocynthis, C. lanatus subsp. lanatus “Charleston Gray”
heavily infected with peanut root-knot nematode (RKN), Meloidogyne arenaria race 1, versus
C. lanatus subsp. citroides showing resistance to RKN.
Color image of this figure appears in the color plate section at the end of the book.
maturing, and ripe stages (Davis et al. 2006), and constructed normalized
cDNA libraries that were subtracted by hybridization with leaf cDNA. Eight
thousand eight hundred cDNA clones of the watermelon flesh subtraction
library were sequenced and ESTs associated with fruit setting, development,
and ripening were identified. These 8,800 ESTs were assembled into 4,770
EST-unigenes. These EST-unigenes can be found at the International Cucurbit
Genomics Initiative (ICuGI) website (http://www.icugi.org). About 29%
of the 4,770 watermelon EST-unigenes had no detectable homologous gene
sequences to any other genomes or protein sequences in GenBank (Fig. 10-4)
(Levi et al. 2006; Wechter et al. 2008). These results indicate that watermelon
may offer a large number of unique genes and metabolites that may not exist
in other plant species. Also, about 20% of the EST-unigenes correspond to
genes with unknown function, whereas about 50% correspond to genes with
known function in other plant species. These “EST-unigenes” are mainly
Figure 10-4 Classification of 880 EST-unigenes Illini Red watermelon fruit based on homology
of 880 EST-unigenes to gene sequences in other plant species.
Color image of this figure appears in the color plate section at the end of the book.
population derived from a cross between PI 296341-FR and the elite Chinese
breeding line 97103 (Xu et al. 2010). Also, the expression of 832 EST-unigenes
was examined using microarray and quantitative real-time PCR approach
to elucidate the genetic flow of events associated with fruit development
and ripening in watermelon fruit (Wechter et al. 2008). The microarray
analysis identified 335 unique ESTs that are differentially modulated by
at least two-fold in watermelon fruit during the early, ripening, or mature
stage when compared to leaf. Primers for the differentially expressed genes
have been developed and are being used to determine their association with
fruit quality. This microarray study also elucidated the role of ethylene in
fruit development in watermelon and in non-climacteric fruits (Salman-
Minkov et al. 2008), showing that ethylene levels are highest during the
early development stages and decrease during maturation and ripening of
the watermelon fruit. These genomic and metabolomic profiles provided
valuable information on genes and metabolites that affect watermelon fruit
quality (Wechter et al. 2008).
In the US, watermelon is a major specialty crop, which provides
an important source of income for farmers and a vital lifeline to the
surrounding rural communities in the southern states. However, in recent
years, watermelon growers in the US have faced new challenges related to
heightened competition with vegetables from Central and South America;
increased production costs due to the significant increase in the cost of oil
and its byproducts; and increased disease, pest, and environmental pressures
(urbanization and reduction in agricultural land, poor soil conditions,
increased salinity and poor water quality, and intense use of pesticides and
other chemicals), that reduce watermelon yield and quality.
Fon race 0 and 1, but not to race 2. Fon is a seed-borne pathogen and can
rapidly spread in watermelon production areas. An RIL population (Zhang
et al. 2004) derived from a cross between the Fon race 2 resistant PI 296341-
FR (C. lanatus var. citroides) and the high quality inbred watermelon line
97103 (C. lanatus var. lanatus) has been used in genetic studies to identify
markers associated with Fusarium wilt resistance. Xu et al. (2010) have
constructed a suppression substractive hybridization (SSH) library from
root tissues of PI296341-FR that was infected with Fusarium wilt race
1. They sequenced approximately 3,895 cDNA clones. Their sequence
analysis identified a large number of the EST-unigenes that are putatively
associated with the disease-defense response. They analyzed the gene
expression of the root tissue infected by F. oxysporum race 1, using 32
Agilent 8 × 15K microarray chips designed on the watermelon ESTs and
public databases. This gene expression study identified 12 putative genes
in the Glycosphingolipid metabolic pathway that may be associated with
resistance to F. oxysporum race 1.
Root-knot nematodes (RKN) are among the most important pests of
cucurbit crops worldwide. Three primary species, Meloidogyne incognita,
M. arenaria, and M. javanica, cause substantial damage to watermelon
throughout the southern US (Thomason and McKinney 1959; Winstead
and Riggs 1959; Sumner and Johnson 1973; Thies 1996). The RKN invade
watermelon roots, inducing root galls that damage the vascular system and
interfere with the uptake of water and translocation of minerals, resulting in
stunted plants that produce poor or no yields (Williamson and Hussey 1996).
Root-knot nematodes also increase the occurrence and severity of Fusarium
wilt in watermelon and could reduce resistance in Fusarium wilt-resistant
varieties (Sumner and Johnson 1973). The RKN have been controlled
in watermelon by pre-plant fumigation with methyl bromide or other
nematicide treatments. The exclusion of methyl bromide from pre-plant soil
fumigation is projected to result in annual yield losses of at least 15 to 20% for
watermelon in Georgia and Florida (Lynch and Carpenter 1999). Thies and
Levi (2003, 2007) screened a watermelon germplasm and identified several
C. lanatus subsp. citroides PIs that contain resistance to root-knot nematodes
(Fig. 10-3). These C. lanatus subsp. citroides PIs are being used in breeding
programs and in developing rootstocks for grafting experiments in fields
infected with root-knot nematodes (Thies et al. 2010). Watermelon has also
suffered major losses from aphid and whitefly-transmitted viruses. The
aphid-transmitted potyviruses, including ZYMV, WMV, PRSV-Watermelon
(W), and cucumber mosaic virus (CMV) are considered the most important
viruses of watermelon (Provvidenti 1996; Lecoq et al. 1998). Recently, the
whitefly transmitted squash vein yellowing virus (SqVYV) was identified
as the causal agent of watermelon vine decline (WVD) that has caused
severe damage to the watermelon industry in Southwest Florida, Indiana,
and South Carolina (Adkins et al. 2007; Egel and Adkins 2007; Kousik et al.
2009). These viral diseases are difficult to control due to their transmission
by insect vectors (aphid or whitefly). Several mechanisms for resistance
to potyviruses in cucurbit species were elucidated in recent studies (Ling
et al. 2009). Recently, Ling et al. (2009) and Harris et al. (2009b) identified
markers associated with ZYMV resistance in the C. lanatus var. lanatus PI
595203. The markers are on the “eIF4e” gene sequence, which is known to
be one of the eukaryotic elongation factor genes that are associated with
resistance to poyviruses in plants (Ling et al. 2009). These markers (Fig.
10-5) are being used by seed company breeders to incorporate ZYMV
resistance into watermelon. However, our experiments indicated that the
marker is not useful for incorporating resistance to all ZYMV strains, and
in addition to the “eIF4e” gene a modifier gene(s) might be associated with
the resistance (A. Levi unpubl. data).
a) Line: P N F1 F2
Phenotype: R S S R S R S S S S S M
Genotype:
N
P
Figure 10-5 A restriction fragment length polymorphism generated using CAPS-1 marker for
the eukaryotic elongation factor “eIF4E” gene sequence on randomly selected F2 progenies
along with F1 and parental materials (P: PI 595203, N: New Hampshire Midget). Phenotype
on virus susceptibility was determined through seedling inoculation with ZYMV-FL (R:
resistance, S: susceptible). Genotype was determined through comparisons to RFLP patterns
in parents, P: PI 595203 (106 bp); N: New Hampshire Midget (131 bp). M is 1 kb plus molecular
weight marker.
2 3 4 5
1
0 WMN73C12 WMN11H07 SSR20354 WSSH387 WMN42A02 WMU799
WMN74D10 WSSH765 0 Bac_SSR33 0 WSSH970 0 WMG00426
0 wep33 4 WMG00362 WMG00327
7 wep32 3 WMG00222 WMG00653 5 WMN54F06 WMN04G10 WMN04F04 WSSH163
WMG00654 WMG00007 WMU567 wep30 SSR17818 WMN07G11 WMG01643 WMF00056
9 wep22 13 9 6 WMG01099
11 WMG01836 WMG01835 WMG01156 WMG01426 WMU617 WSSH1194 WSSH1004
11 WMG01634 WMG00863 15 Bac_SSR8 WSSH1015 WSSH1196 WMI00092 WMG00955
16 WSSH984 8 WMG00769 WMG01557
18 WMG00034 WMF00190 WMG00864 16 WMG00131 WMG00207 WMG00208
18 WMG01413 12 WMG00124 WMF00087
WMG00291 SSR10055 12 WMG00344
WMG00627 WMG00751 19 wep18 16 WMG00326 WMG00316 WMG00482
20 SSR10042 WMF00091 WMG00652 WMF00041
WMG01166 WMG01617 WMG01324 WMG00958 WMN03E10 19 SSR15737 10
20 WMG00658 WMG00455
25 WMG01168 WMG01379 WMG01417 WSSH497 20 WSSH410
WMG01303 WMG00949 22 WMN36H04 24 WMN30B12 WMI00015
28 SSR01859 13 WMG01433 WMG00651
30 WMI00072 14 WMG00003 WMG01281 24 SSR04992 WSSH572 30 WMG00102
WMG01568 WMG01911 25 wep16 wep48 32 WMG01592 WMG00354 WMG00971
35 WMG01114 WSSH531 WMG00464 WMG01374
37 WMG00255 WMG00166 WMG01912 WMF00243 WMG00676 WMG01691 36 WMN30E07 WSSH399 14
WMF00244 WMG01534 WMG01212 WMG00911 37 WMU880 WMF00038 WMG01752
WSSH322 WMN22H02 26 WMF00184
44 WMI00043 WMI00086 WMG00805 WMF00020 WMG01521 WMF00110
wep3 18 WMG00419
WMG00602 WMG00845 18 WMG00183 WMG00085 WMF00070 WMI00046 WMG01034 WMG00933
WMG01451 WMG00566 WMI00054 WMG00998 WMG01183 WMG01427 21 WMG01455 WMG00854
45 WMG01041 WMG01404 21 22 WMG01163
WMG01576 WMG01771 WMG00681 WMG01553 28 WMG01458 WMG00453 WMG00564 WSSH1133
38 24 WMG01686
SSR01534 SSR07284 22 WMI00013 WMG00454 WMF00129 WMG00808
WMG00328 WMG01096 29 WMG01276 WMG00807 WMG00898 25 WMG01068 WMG01067
50 WMG01487 WMG00230 WSSH752 WSSH321
WMG00846 27 WMF00144 WMG01097 WMI00018 WMI00017 WMG01144 WMG00589 27
WMG01393 WMG00765 WMG01076 WMG00660 WMI00091 MCPI_14
52 WMG01852 WMG00985 30 31 WMI00021
53 WMG00582 WMG01351 31 WMG00314 WMG01602 WMG01623 WMG01047 45 WMG00405
34 WMG00221 WMG01454 48 WMI00069 38 WMI00076
54 WMU226 wep65 WSSH1125
WMG00071 WMG01369 WMG00009 WMF00101 WMG00486 WMG00485 53 WMG00458
35 31 WMG01151 WMG01685 WMG01192 WMG00410 41 WSSH1113 WSSH1199
WMG00708 WMG00281 WMG00010
45 MCPI_15 MCPI_11 WMG01199 54 WMG00457 WMG01750 WMU328 WMN39F06
WMF00009 WMG00546
WMG01126 WMG01758 46 WSSH740 wep62 32 WMG01728 WMG01708
56 WMG00080 WMG01743 47 WMG00058 WMG00114 WMG00874 WMG01383
WMG01007 WMG01601 49 WSSH786 WMG00226 WMG01335
WMG01032 WMG00018 52 WMU656 33 WMG00873 WMG00038
WMG00948 WMG00515 55 WMN08D09 EIF4e_SSR12 WMG01296
WSSH820 56 WMN23G09 WMG00337 EIF4e_SSR7
58 WMU149
61 WMG00621
WMG00201 WMG00029 6
68 WSSH872 63
WMG00084 WMG00813 WMG00028
WMG01119 WMG01113 WMU157 WSSH313 SSR20083 WMU597
67 WSSH573 0
WMG00126 WMG01577 WSSH892
WMF00096 WMG01346 73 WMU316 WSSH14 WSSH15
78 WMN09D12 6 WSSH398
WMG00480 WMG00795
70 MCPI_47 MCPI_27 11 WMN16F06
WMG01139 WMG01362
WMG00554 WMG00288 MCPI_29 MCPI_13 12 WMU512
81
WMG00667 WMG00668 WSSH705 SSR04385 15 WSSH807 WMN77H08
WMG01357 WMG00462 MCPI_07 19 WMG01309 WMF00048
WMF00002 WMG00040 20 WMG00133
71 MCPI_41 WMG00246 WMG01776 WMI00090
WMG00592 WMG00365 WMI00020 WMI00031
72 24 WMG01790 WMG01718
WMG00976
75 WMG01107 WMG01358 WMG00287
76 WMG01824 WMG00647 WMG00544
79 WMG00076 9
80 WMG00075 10
wep24 WSSH891
SSR11439 WMU400 WSSH698 wep64
84 WMU297 MCPI_21 0 SSR20063 SSR33278 0 SSR12763 WMU307
WMN23E12 wep19 3 WMI00067
WMN17F06 wep74 1 WMN74G08 7 SSR21065
87 3 WMN55B08 11 WMG00269
WMU995
13 WMG01478
91 SSR15203 8 5 WMU525 WMN73E02
16 WMG01136
92 WMG00228 WMG00165
WMG00383 WMG00013
7 13
wep63 SSR11741
SSR00048 23 WMU227 WMN54E05
99 WMG00014 WMI00050 WMG00369 WMG00049 16 WMG01133 26 wep14
0 22 WSSH422 MCPI_37 MCPI_46
104 WMG00801 WMI00087 WMI00034 WMG00050 27
107 WMG01709 0 WMG01397 WMG00434 4 WMG00329 WMG00265 26 WMG00212 WMG00567 MCPI_25 MCPI_10
WMG00304 WMG00324 6 WMF00044 WMG00969 WMG00483 29 WMG00079
WMG01507 WMG01859 SSR11343 SF2SR10 29 WMG00635 WMI00029 WMG00712 WMG01407
2 31 WMG00711
WMG00345 WMN24D04 WMU398 WMG00019
3 WMI00055 WMF00097 WMG00373 WMU529 WMG00016 34 WMG01881
5 WMG01045 WMG00374 WMI00041 WMG01523 WMG00959 36 WSSH405 WMG01035
7 WMI00048 WMG01714 WMG01491 WMG00926 WMG00883 38 WSSH307 WMG00903
WMG01005 WMG01436 WMG01461 WMG00037 WMG00884 WMG01887 WMG00618 WMG00932
8 WMG01103 WMG01883 WMG00609 WMG00571 WMG00851 WMG01722 40 WMG00844 WMG01039
WMG01701 WMI00084 12 WMG00333 WMG01314 WMG01178
WMG01292 WMG00610
WMG00353 WMG01639 WMG01001 WMG00886 30 WMG00508 WMG01512 41 WMG00686
10 43 WMG00819
WMG01094 WMG01724 WMG01287 WMG00522 WMG01387 WMG00593
11 WMF00003 WMI00060 WMG00528 WMG01209 WMG00688 WMG01731 46 WMG01442
14 WMG00612 WMG01589 WMG00620 WMG00861 WMG01256 49 WSSH306
15 WMG00611 WMG01147 WMG01698 WMG00826 WMG00507 MCPI_33 50 WSSH1200
18 WMG01149 WMG00371 WMU97 WSSH799 WMU56 WMU155_2 WMG00257
51
27 WSSH1110 WMG00252 WMG00992 WMG01838 WMG00671 WMG00435
31 wep79 SSR07236 WMG00991 WMG00726 WMI00093 WSSH889 WMG00867 WMG00850
33 55 WMG00262
38 wep59 WMG01402 WMF00251 MCPI_23
WMN12E07 SSR14498 19 WMF00071 WMF00250 36 WMG00001 56 SSR02008 WMN20D05
39 WSSH917 WSSH192 SSR20270
WMG00738 WSSH837 38 WMN07D02 SSR22259
WMU3 41 wep50 57 WMU155_1 WMN09C06
21 WMN09B07 WSSH434 43 WMG00068 WSSH341 WSSH711
48 WMG00030
11 12 13 14
WMG01161 WMG00090
0 WSSH1136 WMG01636
WMI00078 WMG00134 0 0 SSR13292 MCPI_26 0 Fo_SSR23 Fo_SSR13
2 wep55 WMN44B03 WSSH791
4 WMI00080 10 WMU758 SSR00842 10
WSSH118 WSSH1151 Fo_SSR44 Fo_SSR16
Fo_SSR26 Fo_SSR12
15
11 WMU548 MCPI_24 WMU430 3
5 WMI00081 WMN65D12 WSSH1174 MCPI_09 Fo_SSR27 Fo_SSR29
WMN19D04 SSR23148 14 WMU626 15 WMN12A04 WMN28B09
6 16 SSR07461 WMU134 Fo_SSR36 Fo_SSR37 0
WMI00065 21 WSSH28 WMN37G11 WMN20A02 WMN76F12
9 C31 C80 20 WMN67H04 WSSH331 SSR20704
22 WMN18B09 SSR02237 WMN29C04 WSSH1201 9 wep13 WMN02H02
25 WMU503 SF12SR5 WMN15E04 5 SSR05195 SSR15802
SSR23265 14 Fo_SSR6 WSSH195
28 WMN41G05 WMN19F06 17 WSSH326
WSSH214 WSSH215
20 wep21
Figure 10-6 Genetic linkage map for watermelon based on 103 RILs derived from a cross
between the Fusarium wilt resistant PI 296341-FR and high quality Chinese watermelon line
97103. Most of the markers in this linkage map represent SSRs and EST-SSRs that exist in
EST-unigenes of watermelon (Levi et al. 2006).
on several linkage groups and did not cover all regions of the watermelon
genome (Levi et al. 2006). Xu et al. (2010) have constructed an extensive
map, based on 555 SSR and EST-SSR markers that have proven quite useful
as a watermelon genome representation. This map consists of 11 linkage
groups with a total coverage of 739 cM and the average distance between
markers of 1.33 cM. Many markers are still required for the construction
of a dense map that can be used extensively in watermelon breeding
programs and to isolate genes that control fruit quality or confer resistance
to diseases and pests. Recently, they implemented a simple procedure for
developing and using a new type of PCR primers, named “high frequency
oligonucleotides—targeting active genes (HFO-TAG)”. These primers proved
useful in producing polymorphic markers in watermelon cultivars and in
genetic mapping of watermelon (Levi et al. 2010). The HFO-TAG primers
are constructed by first using a “practical extraction and report language
(Perl)” script to identify short oligonucleotides (8-, 9-, and 10-base) that exist
in high frequency in a 4,700 EST-unigene watermelon fruit library (Levi
et al. 2006; Wechter et al. 2008). This computer-based screening yielded
3,200 oligonucleotides that exist 32 to 335 times in the 4,700 EST-unigenes
constructed for watermelon. Of these, 192 HFO-TAG primers (present 51-269
times in the 4,700 EST-unigenes) were used to amplify DNA from closely
related watermelon cultivars. The average number of DNA fragments
produced by a single HFO-TAG primer among the watermelon cultivars was
considerably higher than the number of fragments produced by inter-simple
sequence repeat (ISSR) or randomly amplified polymorphic DNA (RAPD)
primers. Also, the HFO-TAG primers produced considerably more fragments
than the ISSR or RAPD primers from a watermelon cDNA library that was
used as a template. These results suggest that the HFO-TAG primers should
be more specific in targeting active gene loci. Xu et al. (2010) have sequenced
the genome of the elite Chinese watermelon line 97103. Additionally, they
produced extensive EST data for watermelon. A sequencing project for the
genome of the heirloom cultivar Charleston Gray has been conducted by Levi
et al. (2011). These extensive EST data should be useful for developing HFO-
TAG primers that can be utilized in genetic mapping and targeting of gene loci
of watermelon. Several linkage maps were constructed for watermelon using
a BC1 population (Levi et al. 2001c), a testcross population (Levi et al. 2002,
2006), an F2 population (Hashizume et al. 2003) and a recombinant inbred
line (RIL) population (Zhang et al. 2004; Xu et al. 2010). The genetic maps
and mapping populations (Levi et al. 2006) have been useful for mapping
the eukaryotic elongation factor “eIF4E” gene linked to ZYMV resistance in
watermelon and for identification of markers linked to this resistance (Ling
et al. 2009; Harris et al. 2010a). Also these maps have been used for genetic
mapping of NBS-LRR genes (possible resistance-gene analogs) that we
recently identified in watermelon (Harris et al. 2009b).
watermelon lines with different fruit qualities and resistance to diseases and
pests were selected and used for shallow sequencing and SNP discovery
(Tables 10-1 and 10-2; Fig. 10-7). As in previous studies using DNA markers
(Jarret et al. 1997; Levi et al. 2001b, 2001c), a large number of SNPs was
discovered among the C. lanatus var. lanatus (watermelon cultivars) versus
the wild type C. lanatus var. citroides (cow watermelon) while lower genetic
diversity existed within the subspecies.
A special assembling platform was constructed for the Solexa sequence
data assembly. The total length of the assembled genome sequence was
360.3 Mb, which is about 83.8% of the genome and 16.2% smaller than the
estimated genome size 430 Mb. The size of scaffold N50 is 2.51 Mb and
Figure 10-7 The genetic diversity estimation based on number of single nucleotide
polymorphisms (SNPs) among 17 sequenced genomes.
Table 10-1 Seventeen watermelon genotypes that were sequenced and of which genomes
were assembled and used for SNP discovery.
GS17 Sy-904304 C. lanatus var. lanatus Allsweet type, resistant to Fusarium wilt
race 1 & anthracnose
the number of them is 41. The coverage of the watermelon genome by this
assembly was confirmed using the available EST and BAC sequences. The
assembly contains 99.6% of the 587,291 watermelon ESTs and 92.9% of the
three finished BAC sequences.
SSR, InDel and SV markers generated based on the WWGS and
BAC end sequences, EST-SSR generated based on the SSH cDNA library
sequences, and EST sequences of watermelon fruit development were used
to construct a high-density genetic map using 103 RILs derived from the
cross of PI296341-FR and 97103. Presently, the genetic map consists of 609
SSR markers with 15 linkage groups. The total coverage is 589.1 cM with
an average distance of 0.97 cM. Using this map, Xu et al. (2010) were able
to anchor 189 of the 201 N90 scaffold, which represent 89.04% assembled
sequence, onto 15 chromosomes.
Three gene-prediction methods (cDNA-EST, homology based, and ab
initio) were used to identify protein-coding genes and then built a consensus
gene set by merging all of the results. Xu et al. (2010) predicted 23,738 genes,
with a mean coding sequence size of 1,102 bp and an average of 4.61 exons
per gene Forty-five NBS R-genes were observed. Under an 80% sequence
overlap threshold, they found that 76.14% of the genes were supported by
GS1 GS10 GS11 GS12 GS13 GS14 GS15 GS16 GS2 GS3 GS4 GS5 GS6 GS7 GS8 GS17 GS9
GS1
GS10 0.46
GS11 7.14 7.21
GS12 0.95 1.02 7.04
GS13 5.36 5.41 4.77 5.38
GS14 0.84 0.92 7.05 1.05 5.38
GS15 7.18 7.25 1.45 7.08 4.9 7.07
GS16 7.21 7.28 1.14 7.12 4.88 7.12 1.3
GS2 0.38 0.45 7.22 1.01 5.42 0.9 7.26 7.29
GS3 0.32 0.51 7.19 1.01 5.41 0.9 7.23 7.26 0.42
GS4 0.38 0.44 7.18 1.03 5.41 0.93 7.23 7.26 0.38 0.43
GS5 0.25 0.53 7.16 1.02 5.39 0.91 7.2 7.24 0.49 0.35 0.49
GS6 0.34 0.54 7.17 1.03 5.4 0.93 7.22 7.25 0.51 0.38 0.5 0.41
GS7 0.34 0.55 7.17 1.01 5.4 0.9 7.22 7.25 0.51 0.39 0.5 0.4 0.39
GS8 0.36 0.51 7.21 1.01 5.41 0.9 7.25 7.29 0.45 0.23 0.48 0.4 0.42 0.42
GS17 0.4 0.47 7.2 1.03 5.41 0.92 7.24 7.28 0.34 0.47 0.38 0.53 0.55 0.55 0.51
GS9 0.43 0.45 7.19 1.04 5.41 0.93 7.23 7.26 0.45 0.51 0.42 0.55 0.57 0.57 0.53 0.43
Watermelon 327
© 2012 by Taylor & Francis Group, LLC
328 Genetics, Genomics and Breeding of Cucurbits
multiple gene finders. About 83.21% of the genes have homologs in the
TrEMBL protein database, and 68.37% can be classified by InterPro. In total,
83.56% of the genes have either known homologs or can be functionally
classified. On the basis of pair-wise protein sequence similarities, they
carried out a gene family clustering analysis on all genes in sequenced
plants. The watermelon genes consist of 15,460 families. Of these, 3,099 are
watermelon unique families, among which 2,543 are single-gene families.
Comparing the gene families of watermelon with that of cucumber and
Arabidopsis, they found 12,216 gene families that were shared by at least
two species, and 9,916 clusters that were shared by all three species. There
were 868 watermelon-specific gene families (Xu et al. 2010).
Additional 16 watermelon lines were resequenced to discover the
diversity and the key agricultural traits and useful genes. Total base
pairs are 65 Gb in length. Resequencing of 16 watermelon germplasm
materials was completed with a sequencing depth of 6.30–18.79x. After
the preliminary analysis, the SNP density of the 16 watermelon inbred
lines was 0.023%–0.729%. The genetic diversity of these 16 genotypes and
watermelon line 97103 were analyzed based on these SNPs. The genetic
diversity of these genotypes is in agreement with the results derived from
the 30 SSR markers.
information developed here will not only provide extensive genetic and
physical mapping of the watermelon genome, but will also be incorporating
useful genes into watermelon cultivars. The striking differences between
wild and cultivated watermelon in terms of resistance to disease and pests
are evident by studies on Fusarium wilt (Netzer and Martyn 1989), root-knot
nematodes (Thies and Levi 2003, 2007), potyviruses (Ling et al. 2009; Harris
et al. 2009a), and whiteflies (Simmons and Levi 2002). Extensive genome
sequencing, as well as fine-mapping of this important crop, with particular
focus on the resistance genes, will greatly contribute to the breeder’s ability
to generate resistant cultivars. Hongbin Zhang and his team (Texas A&M
University) have constructed a binary large-insert, plant-transformation-
competent, BIBAC-library from the DNA of watermelon cultivar Charleston
Gray. The library contains a total of 26,112 clones arrayed as individual clones
in 68 microplates (384-well in each). Analysis of a sample of random clones
showed that the library has an average insert size of 170 kb, representing a
10x coverage of the watermelon haploid genome, thus providing greater than
99% probability of obtaining at least one positive clone from the library with a
single-copy probe. The library was constructed in a binary vector that could be
directly transformed into plants via either Agrobacterium-mediated (Hamilton
et al. 1996; Liu et al. 1999; Tao et al. 2001) or particle bombardment (Zhang
and Chang 2009) method. It could be used for genome-wide or large-scale
functional analysis, molecular breeding, and molecular pharming. The BIBAC
library developed for “Charleston Gray” by Hongbin Zhang (pers. comm.) is
well suited for the development of a physical map (Ren et al. 2005). A total of
~30,000 (~11x) clones (Ren et al. 2005), ~15,000 clones from each library, can
be selected, sequenced, and fingerprinted (Zhang and Chang 2009).
References
Adkins ST, Webb S, Achor D, Roberts P, Baker C (2007) Identification and characterization of
a novel whitefly-transmitted member of the family potyviridae isolated from cucurbits
in Florida. Phytopathology 97: 145–154.
Agarwal S, Rao AV (2000) Tomato lycopene and its role in human health and chronic diseases.
J Can Med Assoc 163: 739–744.
Bang H, Kim S, Leskovar D, King S (2007) Development of a codominant CAPS marker for
allelic selection between canary yellow and red watermelon based on SNP in lycopene
beta-cyclase (LCYB) gene. Mol Breed 20: 63–72.
Bates DM, Robinson RW (1995) Cucumbers, melons and water-melons. In: J Smartt, NW
Simmonds (eds) Evolution of Crop Plants. Longman, London, UK, pp 89–96.
Belkhadir Y, Nimchuk Z, Hubert DA, Mackey D, Dangl JL (2004a) Arabidopsis RIN4 negatively
regulates disease resistance mediated by RPS2 and RPM1 downstream or independent of
the NDR1 signal modulator, and is not required for the virulence functions of bacterial
type III effectors AvrRpt2 or AvrRpm1. Plant Cell 16: 2822–2835.
Belkhadir Y, Subramaniam R, Dangl JL (2004b) Plant disease resistance protein signaling:
NBS-LRR proteins and their partners. Curr Opin Plant Biol 7: 391–399.
Biles CL, Martyn RD, Wilson HD (1989) Isoenzymes and general proteins from various
watermelon cultivars and tissue types. HortScience 24: 810–812.
Brotman Y, Silberstein L, Kovalski L, Perin C, Dogimont C, Pitrat M, Klingler J, Thompson
G, Perl-Treves R (2002) Resistance gene homologues in melon are linked to genetic loci
conferring disease and pest resistance. Theor Appl Genet 104: 1055–1063.
Buckler ES, 4th, Phelps-Durr TL, Buckler CSK, Dawe RK, Doebley JF, Holtsford TP (1999) Meiotic
drive of chromosomal knobs reshaped the maize genome. Genetics 153: 415–426.
Burkill HM (1985) The Useful Plants of West Tropical Africa, vol 1. 2nd edn. Royal Botanic
Gardens, Kew, UK, 960 pp.
Chang Y-L, Tao Q, Scheuring C, Ding K, Meksem K, Zhang H-B (2001) An integrated map of
Arabidopsis thaliana for functional analysis of its genome sequence. Genetics 159: 1231–1242.
Chen M, Presting G, Barbazuk WG, Goicoechea JL, Blackmon B, Fang G, Kim H, Frisch D,
Yu, Y, Sun S, Higingbottom S, Phimphilai J, Phimphilai D, Thurmond S, Gaudette B,
Li P, Liu J, Hatfield J, Main D, Farrar K, Henderson C, Barnett L, Costa R, Williams B,
Walser S, Atkins M, Hall C, Budiman MA, Tomkins JP, Luo M, Bancroft I, Salse J, Regad F,
Mohapatra T, Singh K, Tyagi AK, Soderlund C, Dean RA, Wing RA (2002). An integrated
physical and genetic map of the rice genome. Plant Cell 14: 537–545.
Dane F, Hawkins LK, Norton JD (1998) New resistance to race 2 of Fusarium oxysporum f. sp.
niveum in watermelon. Cucurbit Genet Coop Re 21: 37–39.
De Winter B (1990) A new species of Citrullus (Benincaseae) from the Namib desert, Namibia.
Bothalia 20: 209–211.
Dane F, Lang P (2004) Sequence variation at cpDNA regions of watermelon and related wild
species: implications for the evolution of Citrullus haplotypes. Am J Bot 91: 1922–1929.
Dane F, Lang P, Bakhtiyarova R (2004) Comparative analysis of chloroplast DNA variability
in wild and cultivated Citrullus species. Theor Appl Genet 108: 958–966.
Davis AR, Levi A, Kim S, Hernandez A, King SR (2006) RNA extraction method from fruit
tissue high in water and sugar. HortScience 41: 1292–1294.
Deng Z, Huang S, Ling P, Chen C, Yu C, Weber CA, Moore GA, Gmiter FG, JR (2000) Cloning
and characterization of NBS-LRR class resistance-gene candidate sequences in citrus.
Theor Appl Genet 101: 814–822.
De Winter B (1990) A new species of Citrullus (Benincaseae) from the Namib desert, Namibia.
Bothalia 20: 209–211.
DeYoung BJ, Innes RW (2006) Plant NBS-LRR proteins in pathogen sensing and host defense.
Nat Immunol 7: 1243–1249.
Durham RE, Liou PC, Gmitter FG Jr, Moore GA (1992) Linkage of restriction length
polymorphisms and isozymes in Citrus. Theor Appl Genet 84: 39–48.
Egel DS, Adkins S (2007) Squash vein yellowing virus identified in Watermelon (Citrullus
lanatus) in Indiana. Plant Dis 91: 1056.
FAO (1995) Production Year Book for 1994. Food and Agricultural Organization of the United
Nations, Rome, Italy vol. 48, No 125.
Fei Z, Tang X, Alba RM, White JA, Ronning CM, Martin GB, Tanksley SD, Giovannoni J (2004)
Comprehensive EST analysis of tomato and comparative genomics of fruit ripening.
Plant J 40: 47–59.
Fraser PD, Bramley PM (2004) The biosynthesis and nutritional uses of carotenoids. Progr
Lipid Res 43: 228–265.
Giovannucci E, Ascherio A, Rimm EB, Stampfer MJ, Colditz GA, Willett WC (1995) Intake
of carotenoids and retinol in relation to risk of prostate cancer. J Natl Cancer Inst 87:
1767–1776.
Giovannucci E, Willett WC, Stampfer MJ, Liu Y, Rimm EB (2002) A prospective study of tomato
products, lycopene, and prostate cancer risk. J Natl Cancer Inst 94: 391–396.
Gusmini G, Song R, Wehner TC (2005) New Sources of Resistance to Gummy Stem Blight in
Watermelon. Crop Sci 45: 582–588.
Guzman P, Sudarshana MR, Seo YS, Rojas MR, Natwick E, Turini T, Mayberry K, Gilbertson
RL (2000) A new bipartite geminivirus (Begomovirus) causing leaf curl and crumpling
in cucurbits in the Imperial Valley of California. Plant Dis 84: 488.
Harris KR, Ling KS, Wechter WP, Levi A (2009a) A Diagnostic SNP marker for selection
of zucchini yellow mosaic virus resistance gene in watermelon breeding. J Am Soc
HortScience 134: 529–534.
Harris KR, Wechter WP, Levi A (2009b) Isolation, sequence analysis, and linkage mapping of NBS-
LRR disease resistance gene homologs in watermelon. J. Am Soc Hort Sci 134: 649–657.
Hamilton CM, Frary A, Lewis C, Tanksley SD (1996) Stable transfer of intact high molecular
weight DNA into plant chromosomes. Proc Natl Acad Sci USA 93: 9975–9979.
Hashizume T, Sato T, Hirai M (1993) Determination of genetic purity of hybrid seed in
watermelon (Citrullus lanatus) and tomato (Lycopersicon esculentum) using random
amplified polymorphic DNA (RAPD). Jpn J Breed 43: 367–375.
Hashizume T, Shimamoto I, Harushima Y, Yui M, Sato T, Imai T, Hirai M (1996) Construction
of a linkage map for watermelon (Citrullus lanatus (Thunb.) Matsum & Nakai) using
random amplified polymorphic DNA (RAPD). Euphytica 90: 265–273.
Hashizume T, Shimamoto I, Hirai M (2003) Construction of a linkage map and QTL analysis
of horticultural traits for watermelon [Citrullus lanatus (THUNB.) MATSUM & NAKAI]
using RAPD, RFLP and ISSR markers. Theoretical and applied genetics 106: 779–785.
Hawkins LK, Dane F, Kubisiak TL, Rhodes B, Jarret BL (2001) Linkage mapping in a watermelon
population segregating for fusarium wilt resistance. J Am Soc Hort Sci 126: 344–350.
Hong Y, Levay K, Murphy JF, Klein PG, Shaw JG, Hunt AG (1995) A potyvirus polymerase
interacts with the viral coat protein and VPg in yeast cells. Virology 214: 159–166.
Hoskins RA, Nelson CR, Berman BP, Laverty TR, George RA, Ciesiolka L, Naeemuddin
M, Arenson AD, Durbin J, David RG, Tabor PE, Bailey MR, DeShazo DR, Catanese J,
Mammoser A, Osoegawa K, Jong PJ De, Celniker SE, Gibbs RA, Rubin GM, Scherer SE
(2000) A BAC-based physical map of the major autosomes of Drosophila melanogaster.
Science 287: 2271–2274.
Jarret RL, Newman M (2000) Phylogenetic relationships among species of Citrullus and the
placement of C. rehmii De Winter as determined by Internal Transcribed Spacer (ITS)
sequence heterogeneity. Genet Resour Crop Evol 47: 215–222.
Jarret RL, Merrick LC, Holms T, Evans J, Aradhya MK (1997) Simple sequence repeats in
watermelon [Citrullus lanatus (Thunb.) Matsum. & Nakai]. Genome 40: 433–441.
Jeffrey C (1975) Further notes on Cucurbitaceae: III. Some African taxa. Kew Bull 30:
475–493.
Jeffrey C (1978) Cucurbitaceae. Flora Zambesiaca 4: 433–434.
Jeffrey C (1990) Appendix, an outline classification of the Cucurbitaceae. In: DM Bates, RW
Robinson, C Jeffrey (eds) Biology and Utilization of the Cucurbitaceae. Cornell Univ,
Ithaca, NY, USA, p 449.
Jeffrey C (2001) Cucurbitaceae. In: P Hanelt (ed) Institute of Plant Genetics and Crop Plant
Research Mansfeld’s Encyclopedia of Agricultural and Horticultural crops, vol 3. Springer,
Berlin, Germany, pp 1510–1557.
Jia Y, McAdams SA, Bryan GT, Hershey HP, Valent B (2000) Direct interaction of resistance
gene and avirulence gene products confers rice blast resistance. EMBO J 19: 4004–4014.
ABSTRACT
Cucumber is the first cucurbit completely sequenced. This chapter
covers genetic and genomic resources, genome features, comparative
analysis among cucurbits, resistance genes in cucumber genome. As of
the end of 2009, a total of 8,113 expressed sequence tag (EST) sequences
were deposited in GenBank dbEST and an additional 359,108 EST
sequences were obtained. These EST sequences had been assembled
into 81,401 unigenes, which can be accessed at www.icugi.org. A fosmid
library with the insert size of 35–40 Kb was constructed using the
sequenced genotype 9930 and a BAC library with an average insert size
of 101 Kb was also constructed using “Chinese long” inbred line 228.
The sequenced genome of “Chinese long” inbred line 9930 was 243.5
Mb covering more than 96% of the genic regions and contained 26,682
protein-coding and 292 rRNA fragments, 699 tRNA, 238 snoRNA, 192
snRNA, and 171 miRNA genes. Total repeats account for 24% of the
genome, including 10.4% retrotransposon sequences. Chromosome
karyotyping and generation of markers based on the completed
sequence are also discussed.
Keywords: cucumber, BAC library, EST, genome sequencing, SSR
marker
This chapter includes: 1) Genetic and genomic resources for the cucumber
genome project; 2) Whole genome features of cucumber; 3) Comparative
analysis among cucurbits; 4) Pathogen resistance and expanded genes in
cucumber genome; 5) Cucumber as a mode genome for vascular biology.
1
Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing,
100081, China.
*Corresponding author: huangsanwen@caas.net.cn
Figure 11-1 Ideogram showing the position and intensity of Type I/II, Type III, Type IV,
45S rDNA/CsRP2 and 5S rDNA on cucumber metaphase chromosomes (Han et al. 2008).
Chromosome nomenclature follows Koo et al. (2005).
Color image of this figure appears in the color plate section at the end of the book.
Table 11-1 Summary of the cucumber genetic map with RIL mapping population from the
inter-subspecific cross between Gy14 and PI 183967 (Ren et al. 2009).
The above SSR markers were used to examine the genetic affinity of
diverse cucumber inbred lines and evaluate their potential in marker-
assisted selection (MAS). Approximately 65% of the 995 SSRs examined
were polymorphic in these 11 lines. The polymorphism information
content (PIC) values ranged from 0.17 to 0.84, with an average value of
0.44 indicating that the SSR markers employed provided for a robust
discrimination among this germplasm array. Moreover, the SSRs with PIC
values from 0.4 to 0.6 were most common, and ~250 SSRs with PIC value
> 0.5 were highly polymorphic. Therefore, these highly informative SSR
markers would most likely be useful in tracing economically important
traits in breeding populations.
This genentic map might have a broader scope of deployment and use
in cucurbit breeding. An appreciable number of the SSR markers were able
to amplify products in melon [487 (48.9%)], watermelon [258 (25.9%)] and
pumpkin [221 (22.2%)]. Moreover, these SSR markers detected relatively
high levels of polymorphism in these species (melon, 39.6%, watermelon,
46.5%, and pumpkin, 54.8%). Thus, these markers are also potentially useful
in these related crop species.
26 1 21.7 26 1 21.0
27 8 19.6 27 4 24.6
28 3 23.4 28 2 26.9 28 2 25.3
29 1 27.6 29 1 23.8 29 1 26.0
30 2 28.6 30 3 23.0 30 1 24.9
31 1 27.8
32 1 26.3 32 2 26.5 32 2 26.6 32 3 29.1
33 2 25.8 33 1 27.3 33 2 28.0
34 1 28.1
35 1 28.1 35 3 33.6 35 1 31.5
36 2 32.3 36 1 30.4 36 1 32.2
37 3 29.3 37 2 36.0 37 2 29.1 37 6 34.4 37 3 31.3
38 2 30.0 L 38 34 35.1
39 1 37.5 39 1 31.0 39 3 33.3 39 2 35.6
40 1 32.3 40 2 31.7 40 2 33.7
41 1 37.3 41 3 35.6
42 4 33.5 42 1 39.8 42 1 33.0 42 5 36.3 42 4 35.6
43 1 34.5 43 1 41.1 43 1 38.3 43 1 39.0
44 1 41.8 44 2 39.0 44 2 37.2 44 1 39.6
45 1 35.7 45 3 42.9
46 7 39.6
47 1 37.1 47 3 44.9
48 1 37.0
49 6 38.5 49 3 46.2 49 4 43.2
50 4 39.4
51 1 41.3 51 2 48.4 51 2 39.0 51 3 46.0
52 1 43.6
53 5 40.7 53 1 48.4
54 1 48.9 54 1 45.3
55 3 43.3 55 1 49.6 55 4 48.6
56 3 44.4 56 1 52.8 56 1 42.1 56 2 50.3 56 3 46.7 56 1 50.9
57 7 47.4 57 1 51.6
58 1 46.1 58 1 43.9 58 4 47.9
59 5 48.7 59 4 52.9
60 7 57.5 60 1 45.4 60 3 53.1 60 4 49.7
61 1 48.3 61 6 58.2 61 3 53.8 61 6 50.5 SDR
62 1 59.3 L 62 4 54.5 62 2 55.0
63 13 60.8 63 2 47.0 63 1 51.6
64 1 50.3 64 1 52.7 64 3 56.4
65 2 48.7 65 3 53.4
66 1 61.5 66 5 49.4 66 3 54.3 66 1 58.5
67 2 59.3
68 1 51.1 68 2 55.7 L 68 9 60.1
69 2 52.4 69 1 62.9 69 1 52.7 69 1 56.4
70 2 64.1 70 1 59.9 70 6 57.9
71 1 53.3 71 4 66.3 71 2 54.0 71 1 58.6
L 125 4 106.5
127 1 98.6
128 3 99.8
130 1 101.1
131 1 101.8
132 1 102.7
133 2 103.7
134 1 104.0
L 135 12 105.1
136 3 105.2
137 2 107.7
139 2 109.4
143 1 112.7
Figure 11-2 Cucumber SSR linkage map (Ren et al. 2009). The bin names and genetic distances
in cM are respectively listed on the left and right of the chromosomes. The number of SSR
markers in each filled bin is indicated in the boxes. White boxes indicated a recombination
event with no markers. The short and long arms are indicated with S and L, respectively. SDR
= segregation distortion region.
Figure 11-3 Integration of the seven linkage groups of cucumber with individual chromosomes
(Ren et al. 2009). (A1) Distribution of Type I/II (green) and Type III (red) repeats on cucumber
chromosomes. (A2) DAPI staining was converted to black and white images. (A3) Localization
of chromosome-specific fosmid clones on both arms of individual chromosomes, genetic
location of arm-specific fosmid clones are indicated in Fig. 11-2. (B) Localization of fosmids 4S
(red) and 4L (green) together with Type III (red) and 45S rDNA (green) repeats on the mitotic
chromosomes. Bar = 2.5 µm.
Color image of this figure appears in the color plate section at the end of the book.
depth distribution of sequenced reads (350 Mb) (Huang et al. 2009). The
majority of the remaining 30% unassembled regions of the genome are
likely heterochromatic satellite or rRNA sequences. By aligning the EST
sequences against the assembly, it was estimated that more than 96% of
the genic regions were included in the assembled genome.
Using the above mentioned map, 72.8% of the assembled sequences
can be anchored onto the seven chromosomes. Among the 1,885 markers,
1,763 (93.5%) were uniquely aligned and used for constructing the
pseudochromosomes. The majority (98.7%) of the markers were collinear
with the sequence assembly (Fig. 11-4a). Comparison of the genetic and
physical distances between markers showed recombination suppression
of two 10 Mb regions at either end of chromosome 4, a 20 Mb region
on chromosome 5, and an 8 Mb region on chromosome 7. Using FISH,
segmental inversion within the suppression region on chromosome 5
between Gy14 and PI183967 was detected (Fig. 11-4b) that provides an
explanation for recombination suppression in these regions. These regions
of recombination suppression are additionally useful for studying cucumber
evolution during domestication.
Figure 11-4 Integrated genetic/physical map of cucumber (Huang et al. 2009). (a) Genetic
versus physical distance map of the seven cucumber chromosomes. The genetic map was
constructed using an RIL mapping population from the inter-subspecific cross between Gy14
(domestic cucumber) and PI183967 (wild cucumber). (b) The segmental inversion between the
domestic cultivar Gy14 and the wild accession PI183967 on cucumber chromosome 5 detected
by FISH. 12-2 and 12-7 denote individual fosmid clones. (Scale bars, 1 µm)
Color image of this figure appears in the color plate section at the end of the book.
Table 11-3 Repeat content in the assembled cucumber genome (Huang et al. 2009).
and 6 were colinear to melon chromosomes 2+12, 3+5, 4+6, 9+10, and 8+11,
respectively, indicating that, after speciation, these cucumber chromosomes
each resulted from a fusion of two ancient chromosomes. In addition to
chromosome fusion, the comparison also showed the occurrence of several
inter-chromosome and intra-chromosome rearrangements (Fig. 11-5a).
Figure 11-5 Comparative genomic analysis of cucurbits (Huang et al. 2009). (a) Comparative
analysis of the melon and watermelon genetic maps with the cucumber sequence map. (b)
Syntenic blocks between the cucumber genome (scaffold000089) and a melon BAC sequence
(accession: EF188258.1). Genes are drawn as black arrows with the orientation indicated on
the sequence. Transposable elements (TEs) are illustrated as rectangles; retrotransposable
elements are in red, DNA transposons are in blue and unclassified TEs are in green. Orthologous
sequence regions between the two genomes are displayed.
Color image of this figure appears in the color plate section at the end of the book.
Using the annotated genes in the four melon BACs, it was estimated
that cucumber and melon diverged about 4~7 million years ago (Mya) based
on the divergence age of Arabidopsis and papaya (54~90 Mya) (Huang et
al. 2009).
6-1
6-2 7-1
Cen
6-3 7-2
6-4
Cen
6-5/6-6
Cen 7-3
6-6
6-10
7-7
6-11
7-8
6-12
Cucumber Melon Cucumber Melon
chromosome 6 chromosome I chromosome 7 chromosome II
Figure 11-6 Diagrammatic illustration of the marker orders and centromere positions of two
pairs of cucumber and melon chromosomes (Han et al. 2009).
Color image of this figure appears in the color plate section at the end of the book.
Figure 11-7 Genomic locations of R genes on the cucumber chromosomes (Huang et al.
2009). Three R genes could not be anchored on specific chromosome.
Color image of this figure appears in the color plate section at the end of the book.
Figure 11-8 Lineage-specific expansion of the lipoxygenase (LOX) family in the cucumber
genome (Huang et al. 2009). The LOX family was divided into two groups, “Type I” and “Type
II”. The two tandem duplicated gene clusters were ordered and displayed on chromosomes
2 and 4, plus one unmapped scaffold of the cucumber genome.
Color image of this figure appears in the color plate section at the end of the book.
harboring six copies. Fourteen of the LOX genes are specific to the cucumber
lineage. The volatile (E, Z)-2, 6-nonadienal (NDE) gives cucumber its “fresh
green” flavor (Buescher and Buescher 2001) and confers resistance to some
bacteria and fungi (Cho et al. 2004). LOX and one type of hydroperoxide
lyase (9-HPL) synthesize NDE from linolenic acid precursors. Genes
with 9-HPL activity are rarely found in other plants (Matsui et al. 2000).
However, cucumber contains two tandem HPL genes, one of which has
been experimentally confirmed as having 9-HPL activity (Matsui et al. 2000).
The expansion of the LOX gene family and the duplicated HPL genes may
relate to the high level of NDE synthesis in cucumber.
Expansins are cell-wall loosening proteins in plants (Cosgrove 2000).
In cucumber, the expansin subfamily, EXLA, has undergone significant
expansion via tandem duplication (8 genes in cucumber as compared to 1
to 3 in other sequenced genomes); this event may well have contributed to
the development of tendril coiling in cucumber.
Among the sugar transporter proteins, polyol transorter (PLT) gene
family was expanded significantly in cucumber compared with other plants.
A specific gene cluster resulted from tandem duplication contributed mostly
to the expansion.
(Illumina GA). This makes it possible to carry out rapid and low-cost
sequencing for other important plant species. Equipped with this reference
genome, many diverse cucumber lines can be rapidly sequenced at low cost.
Using these resequenced sequences, many genes or signatures related to
the domestication processes and adaptation to diverse environments can be
detected. These genes or signatures will enable marker-assisted breeding of
high-yielding, disease-resistant, and fresh green-scented cucumber.
References
Arumuganathan K, Earle E (1991) Nuclear DNA content of some important plant species.
Plant Mol Biol Rep 9: 208–218.
Bowers JE, Chapman BA, Rong J, Paterson AH (2003) Unravelling angiosperm genome
evolution by phylogenetic analysis of chromosomal duplication events. Nature 422:
433–438.
Buescher RH, Buescher RW (2001) Production and stability of (E, Z)-2, 6-nonadienal, the major
flavor volatile of cucumbers. J Food Sci 66: 357–361.
Cho MJ, Buescher RW, Johnson M, Janes M (2004) Inactivation of Pathogenic Bacteria
by Cucumber Volatiles (E, Z)-2, 6-Nonadienal and (E)-2-Nonenal. J Food Protec 67:
1014–1016.
Cosgrove DJ (2000) Loosening of plant cell walls by expansins. Nature 407: 321–326.
Danin-Poleg Y, Reis N, Baudracco-Arnas S, Pitrat M, Staub JE, Oliver M, Arus P, deVicente
CM, Katzir N (2000) Simple sequence repeats in Cucumis mapping and map merging.
Genome 43: 963–974.
Deleu W, Esteras C, Roig C, Gonzalez-To M, Fernandez-Silva I, Gonzalez-Ibeas D, Blanca J,
Aranda M, Arus P, Nuez F, Monforte A, Pico M, Garcia-Mas J (2009) A set of EST-SNPs
for map saturation and cultivar identification in melon. BMC Plant Biol 9: 90.
Fazio G, Staub JE, Stevens MR (2003) Genetic mapping and QTL analysis of horticultural
traits in cucumber (Cucumis sativus L.) using recombinant inbred lines. Theor Appl
Genet 107: 864–874.
Fernandez-Silva I, Eduardo I, Blanca J, Esteras C, Picó B, Nuez F, Arús P, Garcia-Mas J, Monforte
A (2008) Bin mapping of genomic and EST-derived SSRs in melon ( Cucumis melo L.).
Theor Appl Genet 118: 139.
Ganal M, Hemleben V (1988) Insertion and amplification of a DNA sequence in satellite DNA
of Cucumis sativus L. (cucumber). Theor Appl Genet 75: 357–361.
Ganal M, Riede I, Hemleben V (1986) Organization and sequence analysis of two related
satellite DNAs in cucumber (Cucumis sativus L.). J Mol Evol 23: 23–30.
Han YH, Zhang ZH, Liu JH, Lu JY, Huang SW, Jin WW (2008) Distribution of the tandem
repeat sequences and karyotyping in cucumber (Cucumis sativus L.) by fluorescence in
situ hybridization. Cytogenet Genome Res 122: 80–88.
Han Y, Zhang Z, Liu C, Liu J, Huang S, Jiang J, Jin W (2009) Centromere repositioning in
cucurbit species: Implication of the genomic impact from centromere activation and
inactivation. Proc Natl Acad Sci USA 106: 14937–14941.
Huang S, Li R, Zhang Z, Li L, Gu X, Fan W, Lucas WJ, Wang X, Xie B, Ni P, Ren Y, Zhu H, Li
J, Lin K, Jin W, Fei Z, Li G, Staub J, Kilian A, van der Vossen EA, Wu Y, Guo J, He J, Jia
Z, Ren Y, Tian G, Lu Y, Ruan J, Qian W, Wang M, Huang Q, Li B, Xuan Z, Cao J, Asan,
Wu Z, Zhang J, Cai Q, Bai Y, Zhao B, Han Y, Li Y, Li X, Wang S, Shi Q, Liu S, Cho WK,
Kim JY, Xu Y, Heller-Uszynska K, Miao H, Cheng Z, Zhang S, Wu J, Yang Y, Kang H,
Li M, Liang H, Ren X, Shi Z, Wen M, Jian M, Yang H, Zhang G, Yang Z, Chen R, Liu S,
Li J, Ma L, Liu H, Zhou Y, Zhao J, Fang X, Li G, Fang L, Li Y, Liu D, Zheng H, Zhang Y,
Qin N, Li Z, Yang G, Yang S, Bolund L, Kristiansen K, Zheng H, Li S, Zhang X, Yang H,
Wang J, Sun R, Zhang B, Jiang S, Wang J, Du Y, Li S (2009) The genome of the cucumber,
Cucumis sativus L. Nat Genet 41: 1275–1281.
Jaillon O, Aury JM, Noel B, Policriti A, Clepet C, Casagrande A, Choisne N, Aubourg S,
Vitulo N, Jubin C, Vezzi A, Legeai F, Hugueney P, Dasilva C, Horner D, Mica E, Jublot
D, Poulain J, Bruyère C, Billault A, Segurens B, Gouyvenoux M, Ugarte E, Cattonaro F,
Anthouard V, Vico V, Del Fabbro C, Alaux M, Di Gaspero G, Dumas V, Felice N, Paillard
S, Juman I, Moroldo M, Scalabrin S, Canaguier A, Le Clainche I, Malacrida G, Durand
E, Pesole G, Laucou V, Chatelet P, Merdinoglu D, Delledonne M, Pezzotti M, Lecharny
A, Scarpelli C, Artiguenave F, Pè ME, Valle G, Morgante M, Caboche M, Adam-Blondon
AF, Weissenbach J, Quétier F, Wincker P (2007) The grapevine genome sequence suggests
ancestral hexaploidization in major angiosperm phyla. Nature 449: 463–467.
Kong Q, Xiang C, Yu Z (2006) Development of EST-SSRs in Cucumis sativus from sequence
database. Mol Ecol Notes 6: 1234–1236.
Koo DH, Choi HW, Cho J, Hur Y, Bang JW (2005) A high-resolution karyotype of cucumber
(Cucumis sativus L. ‘Winter Long’) revealed by C-banding, pachytene analysis, and
RAPD-aided fluorescence in situ hybridization. Genome 48: 534–540.
Liavonchanka A, Feussner I (2006) Lipoxygenases: occurrence, functions and catalysis. J Plant
Physiol 163: 348–357.
Lin M-K, Lee Y-J, Lough TJ, Phinney BS, Lucas WJ (2009) Analysis of the Pumpkin Phloem
Proteome Provides Insights into Angiosperm Sieve Tube Function. Mol Cell Proteom
8: 343–356.
Lough TJ, Lucas WJ (2006) INTEGRATIVE PLANT BIOLOGY: Role of Phloem Long-Distance
Macromolecular Trafficking. Annu Rev Plant Biol 57: 203–232.
Matsui K, Ujita C, Fujimoto S, Wilkinson J, Hiatt B, Knauf V, Kajiwara T, Feussner I (2000) Fatty
acid 9- and 13-hydroperoxide lyases from cucumber. FEBS Lett 481: 183–188.
Ming R, Hou S, Feng Y, Yu Q, Dionne-Laporte A, Saw JH, Senin P, Wang W, Ly BV, Lewis KL,
Salzberg SL, Feng L, Jones MR, Skelton RL, Murray JE, Chen C, Qian W, Shen J, Du P,
Eustice M, Tong E, Tang H, Lyons E, Paull RE, Michael TP, Wall K, Rice DW, Albert H,
Wang ML, Zhu YJ, Schatz M, Nagarajan N, Acob RA, Guan P, Blas A, Wai CM, Ackerman
CM, Ren Y, Liu C, Wang J, Wang J, Na JK, Shakirov EV, Haas B, Thimmapuram J, Nelson
D, Wang X, Bowers JE, Gschwend AR, Delcher AL, Singh R, Suzuki JY, Tripathi S, Neupane
K, Wei H, Irikura B, Paidi M, Jiang N, Zhang W, Presting G, Windsor A, Navajas-Pérez
R, Torres MJ, Feltus FA, Porter B, Li Y, Burroughs AM, Luo MC, Liu L, Christopher DA,
Mount SM, Moore PH, Sugimura T, Jiang J, Schuler MA, Friedman V, Mitchell-Olds T,
Shippen DE, dePamphilis CW, Palmer JD, Freeling M, Paterson AH, Gonsalves D, Wang
L, Alam M (2008) The draft genome of the transgenic tropical fruit tree papaya (Carica
papaya Linnaeus). Nature 452: 991–996.
Nieto C, Morales M, Orjeda G, Clepet C, Monfort A, Sturbois B, Puigdomènech P, Pitrat M,
Caboche M, Dogimont C, Garcia-Mas J, Aranda MA, Bendahmane A (2006) An eIF4E
allele confers resistance to an uncapped and non-polyadenylated RNA virus in melon.
Plant J 48: 452–462.
Ren Y, Zhang Z, Liu J, Staub JE, Han Y, Cheng Z, Li X, Lu J, Miao H, Kang H, Xie B, Gu X,
Wang X, Du Y, Jin W, Huang S (2009) An integrated genetic and cytogenetic map of the
cucumber genome. PLoS ONE 4: e5795.
Staub JE, Serquen FC, Horejsi T, Chen J-F (1999) Genetic diversity in cucumber (Cucumis sativus
L.): IV. An evaluation of Chinese germplasm1. Genet Resour Crop Evol 46: 297–310.
Taler D, Galperin M, Benjamin I, Cohen Y, Kenigsbuch D (2004) Plant eR Genes That Encode
Photorespiratory Enzymes Confer Resistance against Disease. Plant Cell 16: 172–184.
Tuskan GA, Difazio S, Jansson S, Bohlmann J, Grigoriev I, Hellsten U, Putnam N, Ralph S,
Rombauts S, Salamov A, Schein J, Sterck L, Aerts A, Bhalerao RR, Bhalerao RP, Blaudez
D, Boerjan W, Brun A, Brunner A, Busov V, Campbell M, Carlson J, Chalot M, Chapman
J, Chen GL, Cooper D, Coutinho PM, Couturier J, Covert S, Cronk Q, Cunningham R,
Davis J, Degroeve S, Déjardin A, Depamphilis C, Detter J, Dirks B, Dubchak I, Duplessis S,
Ehlting J, Ellis B, Gendler K, Goodstein D, Gribskov M, Grimwood J, Groover A, Gunter
ABSTRACT
The Cucurbitaceae is one of only a handful of plant families
characterized by predominantly unisexual flower production. Sex types
are highly variable, and a given genus can include both monoecious
and dioecious species. Dioecy is codified by divergent chromosomes
or gene complexes, while monoecy is conferred by a small number of
independently segregating genes. Microscopic analysis of cucumber
(Cucumis sativus) and melon (C. melo) indicates that unisexuality
results from specific suppression of either stamen or carpel primordia
subsequent to initial whorl formation. Sexual identity of a given bud
is caused by hormone balance, primarily ethylene level, which is,
in turn, influenced by genetic, developmental, and environmental
factors. Recently great progress has been made to elucidate underlying
molecular bases for sex determination. Primary sex determination
loci, A and M from melon and cucumber respectively, which cause
suppression of stamens, have been cloned and found to encode a
key enzyme for ethylene biosynthesis, ACS (1-amino-1-cyclopropane
carboxylate synthase). Similarly, the dominant F locus from cucumber,
which prevents suppression of carpels, also encodes an ACS gene. The
dominant G locus from melon, which causes carpel suppression, also
has been cloned and found to encode a WIP family transcription factor.
Further insight has been gained from studies with transgenic melon
expressing ethylene biosynthesis or perception genes, demonstrating
the essential role of ethylene perception and indicating that cross-
talk between the floral whorls is important for the sex determination
process.
Keywords: sex determination, floral development, ethylene, monoecy,
cucumber, melon, Cucumis
1
Department of Horticulture and Graduate Program in Genetics, Michigan State University,
East Lansing, MI 48824, USA; e-mail: taftjess@msu.edu
*Corresponding author: grumet@msu.edu
Petal
Petal
Nectary
Anther
Ovary
Nectary
Ovule
Figure 12-1 Sexual differentiation of cucumber flower buds. Longitudinal sections of cucumber
buds under dissection microscope. A. Male bud at stage 11. B. Female bud at stage 12 (anthesis).
Bud development stages are assigned as per Bai et al. (2004).
Color image of this figure appears in the color plate section at the end of the book.
Figure 12-2 The ABC model of floral development. Three sets of MADS box transcription
factors interact in different combinations to specify sequential development of the four floral
organ whorls.
Table 12-2 Cucumber floral bud development stages based on Bai et al. (2004).
Non sex-specific Sex specific
development development
Stage Male Female
1 Inflorescence initiation
2 Sepal whorl initiation
3 Petal whorl initiation
4 Stamen whorl initiation
5 Carpel whorl initiation
ACS and ACO are generally encoded by multigene families whose members
are differentially regulated and can be induced by a variety of internal
signals, including other hormones and ethylene itself (Rottman et al. 1991;
Bleecker and Kende 2000; Salmon-Minkov et al. 2008).
A key breakthrough in the understanding of the mechanism of sex
determination in cucumber came from the discovery that the dominant F
allele, which allows for constitutive production of carpels (i.e., gynoecious
or hermaphrodite plants), completely co-segregated with an additional
copy of an ACS gene, CsACS1G (Trebitsch et al. 1997). CsACS1G is absent in
near-isogenic, monoecious lines and expression of CsACS1G transcript was
observed in apices of gynoecious, but not isogenic monoecious cucumbers
(Trebitsch et al. 1997; Kamachi et al. 2000). This led to the clear implication
that the F locus increases femaleness by regulating endogenous ethylene
production, which in turn promotes carpel development. CsACS1G is located
in tandem with CsACS1, and appears to be the result of recombination
between CsACS1 and a branched chain amino acid transferase gene
(Mibus and Tatlioglu 2004; Knopf and Trebitsh 2006). CsACS1 and 1G show
complete homology in the coding region, 400 bp of the proximal promoter,
and the 3’ untranslated regions, suggesting a recent gene duplication event
(Mibus and Tatlioglu 2004; Knopf and Trebitsh 2006). Thus, the feminizing
effect of F may be a gene dosage effect leading to the increased ethylene
production that has been observed in gynoecious lines and/or differential
regulation resulting from differences in the distal promoter region (Knopf
and Trebitsh 2006).
Two other ACS genes, CsACS2 and CsACS4 also exhibit increased
expression with the transition to the female phase (Kamachi et al. 1997;
Yamasaki et al. 2001, 2003a, b). CsACS2 transcript was not detected prior
to the formation of pistil primordia, after which it was localized to the
ovary, just below the pistil primordia (Yamasaki et al. 2003a; Saito et al.
2007). Timing and level of expression of the CsACS2 gene in the apices of
monoecious and gynoecious cucumbers also coincided with the action of
ethylene in the induction of the first female bud (Kamachi et al. 1997, 2000).
It was suggested that CsACS2 is critical for female flower development,
and that CsACS1 (the F locus) may act to accelerate the timing and increase
levels of CsACS2 via higher ethylene production (Kamachi et al. 2000).
Sex- and ethylene-related differential expression also has been observed
for ACO genes (Kahana et al. 1999). Application of the ethylene releasing
compound, ethrel, caused different effects on distinct ACO family members
and in different sex types, indicating likely functional differentiation among
the ACO genes. ACO3 was expressed more strongly in developing stamens
and pistils of female buds than male. On the other hand, ACO2 was not
expressed in gynoecious apices and levels decreased in apices of monoecious
plants when they reached the stage of female flower production. ACO2
Acknowledgments
We thank Drs. David Dilley and Holly Little for helpful reviews of this
chapter. This work was in part supported by research grant US-3735-
05C from the United States-Israel Binational Agricultural Research and
Development (BARD) Fund.
References
Ainsworth C (2000) Boys and girls come out to play: the molecular biology of dioecious plants.
Ann Bot 86: 211–221.
Akimoto J, Fukuhara T, Kikuzawa K (1999) Sex ratios and genetic variation in a functionally
androdioecious species, Schizopepon bryoniaefolius (Cucurbitaceae). Am J Bot 86:
880–886.
Ando S, Sato Y, Kamachi S, Dakai S (2001) Isolation of a MADS-box gene (ERA17) and
correlation of its expression with the induction of formation of female flowers by ethylene
in cucumber plants (Cucumis sativus L.). Planta 213: 943–952.
Atsmon D, Galon E (1960) A morphological study of staminate, pistillate, and hermaphrodite
flowers in Cucumis sativus L. Phytomorphology 10: 110–115.
Atsmon D, Lang A, Light EN (1968) Contents and recovery of gibberellins in monoecious and
gynoecious cucumber plants. Plant Physiol 43: 806–810.
Augustine JJ, Bakier LR, Sell HM (1973) Female flower induction on androecious cucumber,
Cucumis sativus L. J Am Soc Hort Sci 98: 197–199.
Hao YJ, Wang DH, Peng YB, Bai SL, Xu LY, Li YQ, Zu ZH, Bai SN (2003) DNA damage in the
early primordial anther is closely correlated with stamen arrest in the female flower of
cucumber (Cucumis sativus L.) Planta 217: 888–895.
Huai Q, Xia Y, Chen Y, Callahan B, Li N, Ke H (2001) Crystal structure of 1-aminocyclopropane-
1-carboxylate (ACC) synthase in complex with aminoethoxyvinylglycine and
pyridoxal-5’-phosphate provide new insight into catalytic mechanisms. J Biol Chem
276: 38210–38216.
Huang S, Li R, Zhang Z, Li L, Gu X, Fan W, Lucas WJ, Wang X, Xie B, Ni P, Ren Y, Zhu H, Li
J, Lin K, Jin W, Fei Z, Li G, Staub J, Kilian A, van der Vossen EAG, Wu Y, Guo J, He J, Jia
J, Ren Y, Tan G, Lu Y, Ruan J, Qian W, Wang M, Huang Q Li B, Xuan Z, Cao J, Asan, Wu
Z, Ahang J, Cai Q, Bai Y, Zho B, Han Y, Li Y, Li X, Wang S, Shi Q Liu S, Cho WK, Kim
JY, Xu Y, Heller-Uszynska K, Miao H, Cheng Z, Zhang S, Wu J, Yang Y, Kang H, Li M,
Liang H, Ren X, Shi A, Wen M, Jian M, Yang H, Zhang G, Yang Z, Chen R, Liu S, Li J, Ma
L, Liu H, Zhou Y, Zhao J, Fang X, Li G, Fang L, Li Y, Liu D, Zheng H, Zhang Y, Qin N,
Li Z, Yang G, Yang S, Bolund L, Kristiansen K, Zheng H, Li S, Zhang X, Yang H, Wang J,
Sun R, Zhang B, Jiang S, Wang J, Du Y, Li S (2009) The genome of the cucumber, Cucumis
sativus L. Nat Genet: doi: 10.1038/ng.475.
Irish VF, Yamamoto YT (1995) Conservation of floral homeotic gene function between
Arabidopsis and Antirrhinum. Plant Cell 7: 1635–1644.
Iwahori S, Lyons JM, Smith OE (1970) Sex expression in cucumber plants as affected by
2-chloroethylphosphonic acid, ethyene, and growth regulators. Plant Physiol 46:
412–415.
Johnson PR, Ecker JR (1998) The ethylene gas signal transduction pathway: A molecular
perspective. Annu. Rev Genet 32: 227–254.
Kahana A, Silberstein L, Kessler N, Goldstein RS, Perl-Treves R (1999) Expression of ACC
oxidase genes differs among sex genotypes and sex phases in cucumber. Plant Mol Biol
41: 517–528.
Kamachi S, Sekimoto H, Kondo N, Sakai S (1997) Cloning of a cDNA for a 1-aminocyclopropane-
1-carboxylate synthase that is expressed during development of female flowers at the
apices of Cucumis sativus L. Plant Cell Physiol 38: 1197–1206.
Kamachi S, Mizusawa H, Matsura S, Sakai S (2000) Expression of two 1-aminocyclopropane-
1-carboxylate synthase genes, CS-ACS1 and CS-ACS2, correlated with sex phenotypes
in cucumber plants (Cucumis sativus L). Plant Biotechnol 17: 69–74.
Karchi Z (1970) Effects of 2-chloroethanephosphonic acid on flower types and flowering
sequences in muskmelon. J Am Soc Hort Sci 95: 575–578.
Kater MM, Franken J, Carney KJ, Clombo L, Angenent GC (2001) Sex determination in
the monoecious species cucumber in confined to specific floral whorls. Plant Cell 13
481–493.
Kenigsbuch D, Cohen Y (1990) The inheritance of gynoecy in muskmelon. Genome 33:
317–320.
Knopf R, Trebish T (2006) The female specific CsACS1G gene of cucumber. A case of gene
duplication and recombination between the non-sex specific 1-aminocyclopropane-1-
carboxylate synthase gene and a branched chain amino acid transaminase gene. Plant
Cell Physiol 47: 1217–1228.
Krupnick GA, Brown K, Stephenson AG (1999) The influence of fruit on the regulation of
internal ethylene concentrations and sex expression in Cucurbita texana. Int. J Plant Sci
160: 321–330.
Kubicki B (1969a) Sex determination in muskmelon (Cucumis melo L.). Genet Polan 10:
145–163.
Kubicki B (1969b) Investigations on sex determination in cucumber (Cucumis sativus L.) VI.
Androecium. Genet Polan 10: 87–99.
Li Z, Pan JS, Guan Y, Tao QY, He HL, Si LT, Cai R (2008) Development and fine mapping of
three co-dominant SCAR markers linked to the M/m gene in the cucumber plant (Cucumis
sativus L.). Theor Appl Genet 117: 1253–1260.
Li Z, Huang S, Liu S, Pan J, Zhang Z, Tao Q, Shi Q, Jia Z, Zhang W, Chen H, Si L, Zhu Z,
Cai R (2009) Molecular isolation of the M gene suggests that a conserved-residue
conversion induces the formation of bisexual flowers in cucumber plants. Genetics 182:
1381–1385.
Little HA, Papadopoulou E, Hammar SA, Grumet R (2007) The influence of ethylene perception
on sex expression in transgenic melon as assessed by expression of the mutant ethylene
receptor gene, At-etr1-1, under control of constitutive and floral-targeted promoters. Sex
Plant Reprod 20: 123–136.
Lower RL, Nienhuis J (1990) Prospects for increasing yields of cucumbers via Cucumis sativus
var. hardwickii germplasm. In: DM Bates, RW Robinson, C Jeffrey (eds) Biology and
Utilization of the Cucurbitaceae. Cornell Univ Press, Ithaca, NY, USA, pp 397–405.
Magdum MB, Shinde NN, Seshadri VS (1982) Androecious sex form in muskmelon. Cucurbit
Genet Coop 5: 24–25.
Makus DJ, Pharr DM, Lower RL (1975) Some morphogenic differences between monoecious
and gynoecious cucumber seedlings as related to ethylene production. Plant Physiol
55: 352–355.
Martin A, Troadec C, Boualem A, Mazen R, Fernandez R, Morin H, Pitrat M, Dogimont C,
Bendahmane A (2009) A transposon-induced epigenetic change leads to sex determination
in melon. Nature 461: 1135–1139.
McMurray AL, Miller CH (1968) Cucumber sex expression modified by 2-chloroethanephosphonic
acid. Science 162: 1397–1398.
Mibus H, Tatlioglu T (2004) Molecular characterization and isolation of the F/f gene for
femaleness in cucumber (Cucumis sativus L.). Theor Appl Genet 109: 1669–1676.
Ming R, Wang JP, Moore PH, Patterson AH (2007) Sex chromosomes in flowering plants. Am
J Bot 94: 141–150.
Mockaitis JM, Kivilaan A (1964) Graft-induced sex changes in Cucumis melo L. Nature 202:
216.
Ngouajio M, Mennan H (2005) Weed populations and pickling cucumber (Cucumis sativus)
yield under summer and winter cover crop systems. Crop Protec 24: 421–526.
Owens KW, Peterson CE, Tolla GE (1980) Production of hermaphrodite flowers on gynoecious
muskmelon by silver nitrate and aminoethoxyvinylglycine. HortScience 15: 654–655.
Oyama RK, Voltz SM, Renner SS (2009) A sex-linked SCAR marker in Bryonia dioica
(Cucurbitaceae), a dioecious species with XY sex-determination and homorophic sex
chromosomes. J Evol Biol 22: 214–224.
Papadopoulou E, Grumet R (2005) Brassinosteroid-induced femaleness in cucumber and
relationship to ethylene production. HortScience 40: 1763–1767.
Papadopoulou E, Little HA, Hammar SA, Grumet R (2005) Effect of modified endogenous
ethylene production on sex expression, bisexual flower development, and fruit production
in melon (Cucumis melo L. ). Sex Plant Reprod 18: 131–142.
Perl-Treves R (1999) Male to female conversion along the cucumber shoot: approaches to
studying sex genes and floral development in Cucumis sativus. In: CC Ainsworth (ed)
Sex Determination in Plants. Bios Scientific Publ, Oxford, UK, pp 189–215.
Perl-Treves R, Kahana A, Rosenman N, Xiang Y, Silberstein L (1998) Expression of multiple
AGAMOUS-like genes in male and female flower of cucumber (Cucumis sativus L.). Plant
Cell Physiol 39: 701–710.
Pike LM, Peterson CE (1969) Gibberellin A4/A7 for induction of staminate flowers on the
gynoecious cucumber (Cucumis sativus L.). Euphytica 18: 106–109.
Renner SS, Ricklefs RE (1995) Dioecy and its correlates in the flowering plants. Am J Bot 82:
596–606.
Robinson RW, Decker-Walters DS (eds) (1997) Cucurbits. CAB International. New York, NY,
USA, pp 9–10; 17–19; 195–198.
Robinson RW, Shannon S, La Guardia MD (1969) Regulation of sex expression in the cucumber.
BioScience 19: 141–142.
Yamasaki S, Fujii N, Matsura S, Mizusawa H, Takahashi H (2001) The M locus and ethylene-
controlled sex determination in andromonoecious cucumber plants. Plant Cell Physiol
42: 608–619.
Yamasaki S, Fujii N, Takahashi H (2003a) Photoperiodic regulation of CS-ACS2, CS-ACS4 and
CS-ERS gene expresion contributes to the femaleness of cucumber flowers through diurnal
ethylene production under short day conditions. Plant Cell Env 26: 537–546.
Yamasaki S, Fujii N, Takahashi H (2003b) Characterization of ethylene effects on sex
determination of cucumber plants. Sex Plant Reprod 16: 103–111.
Yin T, Quinn JA (1995) Tests of a mechanistic model of one hormone regulating both sexes in
Cucumis sativus (Cucurbitaceae). Am J Bot 82: 1537–1546.
Zhang LB, Simmons MP, Kocyan A, Renner SS (2006) Phylogeny of the Cucurbitales based
on DNA sequences of nine loci from three genomes: implications for morphological and
sexual system evolution. Mol Phylogenet Evol 39: 305–322.
ABSTRACT
Cucurbit plants are important not only as crops for world food
production but also as model plants for elucidating significant traits
in plant development and responses, such as sex determination and
signaling through the phloem sap. To achieve sustainable production
and to extend our understanding of development and environmental
responses in cucurbit plants, continuous research and development
(R&D) is needed. As described in previous chapters of this book,
significant amounts of knowledge in terms of conventional and
molecular genetics and breeding are currently available for this plant
species. In addition, emerging technologies, such as high-throughput
sequencing technology and high-resolution metabolomics, have
generated a very large amount of data on cucurbit plants. New resources
such as induced mutant lines have also been generated for this plant
species. Effective and efficient integration of these technologies and
resources into conventional genetic and molecular tools is crucial for
achieving successful R&D in cucurbit plants. In this chapter, future
aspects of R&D useful for extending our knowledge and promoting
the use and breeding of cucurbit plants will be discussed.
Keywords: parthenocarpy, omics, metabolome, phenome, high-
throughput sequencing technology
Gene Research Center, Graduate School of Life and Environmental Sciences, University of
Tsukuba, Ten-nodai 1-1-1, Tsukuba 305-8572, JAPAN; e-mail: ezura@gene.tsukuba.ac.jp
is the nitrogenous substrate used in the synthesis of nitric oxide and plays an
essential role in cardiovascular and immune functions (Collins et al. 2007).
Melon contains a higher amount of gamma-aminobutyric acid (GABA), a
compound with antihypertensive effects (Yoshimura et al. 2010), in its fruits
(Ezura et al. unpubl. result). Cucumisin is a thermostable alkaline serine
protease found in the juice of melon fruits (Cucumis melo L.) (Kaneko and
Tominaga 1975). Its complete nucleotide sequence and deduced amino acid
sequence have been determined (Yamagata et al. 1994). Cucumisin supports
daily food life by promoting the digestion of meats, but it is reported to
be a major allergen in melon fruits (Cuesta-Herranz et al. 2003). Recently,
the preventive effects of melon extracts, which are rich in superoxide
scavenging activity, on abdominal and liver fat and adipokine imbalance
in high-fat-fed hamsters have been reported (Decorde et al. 2009). Melon
extracts prevented aortic lipids and liver steatosis in a diet-induced model
of atherosclerosis (Decorde et al. 2010). Both reports indicate the benefits
of melon for human health.
Minor cucurbitaceae plants include pharmaceutical ingredients, which
may contribute to human health. Momordica charantia Linn, belonging to
the family of Cucurbitaceae, is a useful medicinal and vegetable plant for
human health and one of the most promising plants for diabetes treatment
(Lee et al. 2009). Cucurbitane-type triterpenoids are the main active
constituents of M. charantia and have a number of potential biological and
pharmacological applications because of their antidiabetic, anti-obesity,
anticancer, anti-HIV, antifeedant and antioviposition activities. Since
the early 1960s, the constituents of bitter melon have been investigated,
and several classes of secondary metabolites, including cucurbitane-type
triterpenoids, have been isolated. Charantin, an anti-diabetic compound,
is a typical cucurbitane-type triterpenoid in M. charantia and a potential
and promising substance for the treatment of diabetes. Citrullus colocynthis
Schrad., endemic in southern Tunisia, is used in folk medicine to treat many
inflammatory diseases (Marzouk et al. 2010). After identification and acute
toxicity assays, C. colocynthis aqueous extracts were screened for analgesic
and anti-inflammatory activities using the acetic acid writhing test in mice
and the carrageenan-induced paw edema assay in rats, respectively. All
extracts displayed analgesic and anti-inflammatory activities at different
doses without inducing acute toxicity. Topical results were obtained with
immature fruits followed by seeds. The stem and root extracts were shown
to possess less significant inhibitory activity than analgesic and anti-
inflammatory models. Based on this study, C. colocynthis is a potentially
useful drug suitable for further evaluation for rheumatoid arthritis, and
its folk medicinal use as an analgesic and anti-inflammatory agent is
validated.
13.7 Conclusion
Cucurbit plants are commonly grown in the world. To maintain or further
extend their sustainable production and use, continuous R&D of key
technologies such as an induction of parthenocarpy traits, metabolomic
profiling of functional ingredients for human health, finding new uses such
as phytoremediation, comprehensive use of high-throughput sequencing
technology and informatics, the integration of omics datasets into phenome
data and high-throughput genetic transformation are required. Sharing
bioresources and the information generated by the technologies mentioned
above are also significant means to enhance cucurbit production and
research.
References
Acciarri N, Restaino F, Vitelli G, Perrone D, Zottini M, Pandolfini T, Spena A, Rotino G (2002)
Genetically modified parthenocarpic eggplants: improved fruit productivity under both
greenhouse and open field cultivation. BMC Biotechnol 2: 4.
Alonso JM, Ecker JR (2006) Moving forward in reverse: genetic technologies to enable genome-
wide phenomic screens in Arabidopsis. Nat Rev Genet 7: 524–536.
Arbona V, Iglesias DJ, Talón M, Gómez-Cadenas A (2009) Plant phenotype demarcation
using nontargeted LC-MS and GC-MS metabolite profiling. J Agric Food Chem 57:
7338–7347.
Azevedo-Meleiro CH, Rodriguez-Amaya DB (2007) Qualitative and quantitative differences
in carotenoid composition among Cucurbita moschata, Cucurbita maxima, and Cucurbita
pepo. J Agric Food Chem 55: 4027–33.
Biais B, Allwood JW, Deborde C, Xu Y, Maucourt M, Beauvoit B, Dunn WB, Jacob D, Goodacre
R, Rolin D, Moing A (2009) 1H NMR, GC-EI-TOFMS, and data set correlation for fruit
metabolomics: application to spatial metabolite analysis in melon. Anal Chem 81:
2884–2894.
Britton G, Liaaen-Jensen S, Pfander H (2004) Carotenoids Handbook, Brikhauser Verlag,
Basel, Switzerland.
Campbell S, Arakaki AS, Li QX (2009) Phytoremediation of heptachlor and heptachlor epoxide
in soil by Cucurbitaceae. Int J Phytoremed 11: 28–38.
Collins JK, Wu G, Perkins-Veazie P, Spears K, Claypool PL, Baker RA, Clevidence BA (2007)
Watermelon consumption increases plasma arginine concentrations in adults. Nutrition
23: 261–266.
Cuesta-Herranz J, Pastor C, Figueredo E, Vidarte L, De las Heras M, Durán C, Fernández-Caldas
E, de Miguel J, Vivanco F (2003) Identification of Cucumisin (Cuc m 1), a subtilisin-like
endopeptidase, as the major allergen of melon fruit. Clin Exp Allergy 33: 827–33.
Decorde K, Agne A, Lacan D, Ramos J, Fouret G, Ventura E, Feillet-Coudray C, Cristol JP,
Rouanet JM (2009) Preventive effect of a melon extract rich in superoxide scavenging
activity on abdominal and liver fat and adipokine imbalance in high-fat-fed hamsters. J
Agric Food Chem 57: 6461–6467.
Decorde K, Ventura E, Lacan D, Ramos J, Cristol JP, Rouanet JM (2010) An SOD rich melon
extract Extramel prevents aortic lipids and liver steatosis in diet-induced model of
atherosclerosis. Nutr Metab Cardiovasc Dis 20: 301–7.
Ezura H, Fukino N (2009) Research tools for functional genomics in melon (Cucumis melo L.):
Current status and prospects. Plant Biotechnol 26: 359–368.
Ezura H, Yuhashi KI, Yasuta T, Minamisawa K (2000) Effect of ethylene on Agrobacterium
tumefaciens-mediated gene transfer to melon. Plant Breed 119: 75–79.
Heuberger AL, Lewis MR, Chen MH, Brick MA, Leach JE, Ryan EP. (2010) Metabolomic and
functional genomic analyses reveal varietal differences in bioactive compounds of cooked
rice. PLoS One 5: e12915.
Huitron MV, Diaz M, Dianez F, Camacho F, Valverde A (2007) Effect of 2, 4-D and CPPU on
triploid watermelon production and quality. Hortscience 42: 559–564.
Huang S, Li R, Zhang Z, Li L, Gu X, Fan W, Lucas WJ, Wang X, Xie B, Ni P, Ren Y, Zhu H, Li
J, Lin K, Jin W, Fei Z, Li G, Staub J, Kilian A, van der Vossen EA, Wu Y, Guo J, He J, Jia
Z, Ren Y, Tian G, Lu Y, Ruan J, Qian W, Wang M, Huang Q, Li B, Xuan Z, Cao J, Asan,
Wu Z, Zhang J, Cai Q, Bai Y, Zhao B, Han Y, Li Y, Li X, Wang S, Shi Q, Liu S, Cho WK,
Kim JY, Xu Y, Heller-Uszynska K, Miao H, Cheng Z, Zhang S, Wu J, Yang Y, Kang H,
Li M, Liang H, Ren X, Shi Z, Wen M, Jian M, Yang H, Zhang G, Yang Z, Chen R, Liu S,
Li J, Ma L, Liu H, Zhou Y, Zhao J, Fang X, Li G, Fang L, Li Y, Liu D, Zheng H, Zhang Y,
Qin N, Li Z, Yang G, Yang S, Bolund L, Kristiansen K, Zheng H, Li S, Zhang X, Yang H,
Wang J, Sun R, Zhang B, Jiang S, Wang J, Du Y, Li S (2009) The genome of the cucumber,
Cucumis sativus L. Nat Genet 41: 1275–1281.
Kaneko M, Tominaga N (1975) Isolation and characterization of a proteinase from the sarcocarp
of melon fruit. J Biochem 78: 1287–1296.
Kuromori T, Takahashi S, Kondou Y, Shinozaki K, Matsui M (2009) Phenome analysis in
plant species using loss-of-function and gain-of-function mutants. Plant Cell Physiol
50: 1215–1231.
Lee SY, Eom SH, Kim YK, Park NI, Park SU (2009) Cucurbitane-type triterpenoids in Momordica
charantia Linn. J Med Plant Res 3: 1261–1269.
Martinelli F, Uratsu SL, Reagan RL, Chen Y, Tricoli D, Fiehn O, Rocke DM, Gasser CS, Dandekar
AM (2009) Gene regulation in parthenocarpic tomato fruit. J Exp Bot 60: 3873–3890.
Marzouk B, Marzouk Z, Haloui E, Fenina N, Bouraoui A, Aouni M (2010) Screening of
analgesic and anti-inflammatory activities of Citrullus colocynthis from southern Tunisia.
J Ethnopharma 121: 15–19.
Menezes CB, Maluf WR, Azevedo SM, Faria MV, Nascimento IR, Nogueira DW, Gomes LA,
Bearzoti E (2005) Inheritance of parthenocarpy in summer squash (Cucurbita pepo L.).
Genet Mol Res 4: 39–46.
Nonaka S, Yuhashi K, Takada K, Sugawara M, Minamisawa K, Ezura H (2008a) Ethylene
production in plants during gene transformation suppresses vir gene expression in
Agrobacterium tumefaciens. New Phytologist 178: 647–656.
Nonaka S, Sugawara M, Minamisawa K, Yuhashi KI, Ezura H (2008b) 1-aminocyclopropane-1-
carboxylate deaminase-producing Agrobacterium confers higher ability for gene transfer
into plant cells. Appl Environ Microbiol 74: 2526–2528.
Ntui VO, Khan RS, Chin DP, Nakamura I, Mii M (2010) An efficient Agrobacterium tumefaciens-
mediated genetic transformation of “Egusi” melon (Colocynthis citrullus L.) Plant Cell
Tiss Org Cult 13: 15–22.
Otani T, Seike N, Sakata Y (2007) Differential uptake of dieldrin and endrin from soil by several
plant families and Cucurbita genera. Soil Sci Plant Nutr 53: 86–94.
Puigdomènech P, Martínez-Izquierdo JA, Arús P, Garcia-Mas J, Monforte AJ, et al. (2007) The
Spanish melon genomics initiative. Acta Horticult 731: 47–54.
Ren Y, Zhang Z, Liu J, Staub JE, Han Y, Cheng Z, Li X, Lu J, Miao H, Kang H, Xie B, Gu X,
Wang X, Du Y, Jin W, Huang S (2009) An integrated genetic and cytogenetic map of the
cucumber genome. PLoS One. 4(6): e5795.
Robinson RW, Reiners S (1999) Parthenocarpy in summer squash. Hortscience 34: 715–717.
Rotino GL, Perri E, Zottini M, Sommer H, Spena A (1997) Genetic engineering of parthenocarpic
plants. Nat Biotechnol 15: 1398–13401.
Rotino GL, Acciarri N, Sabatini E, Mennella G, Lo Scalzo R, Maestrelli A, Molesini B,
Pandolfini T, Scalzo J, Mezzetti B, Spena A (2005) Open field trial of genetically modified
parthenocarpic tomato: seedlessness and fruit quality. BMC Biotechnol 5: 32.
Sobolev AP, Testone G, Santoro F, Nicolodi C, Iannelli MA, Amato ME, Ianniello A, Brosio
E, Giannino D, Mannina L (2010) Quality traits of conventional and transgenic lettuce
(Lactuca sativa L.) at harvesting by NMR metabolic profiling. J Agric Food Chem 58:
6928–6936.
Sugiyama K, Morishita M, Nishino E (2002) Seedless watermelons produced via soft-X-
irradiated pollen. Hortscience 37: 292–295.
Sun Z, Lower RL, Staub JE (2006) Analysis of generation means and components of variance
for parthenocarpy in cucumber (Cucumis sativus L.). Plant Breed 125: 277–280.
Tadmor Y, Katzir N, Meir A, Yaniv-Yaakov A, Sa’ar U, Baumkoler F, Lavee T, Lewinsohn E,
Schaffer A, Burger J (2007) Induced mutagenesis to augment the natural genetic variability
of melon (Cucumis melo L.). Israel J Plant Sci 55: 159–169.
Wada M (1930) Über Citrullin, eine neue Aminosäure im Presssaft der Wassermelone, Citrullus
vulgaris Schrad. Biochem Zeit 224: 420.
Xu Y, Guo S, Zhang H, Ren Y, Zhao H, Lv G, Gong G, Fei Z, Kou Q, Zou X, Wang H, Hou
W (2010) International Watermelon Genomics Initiative (IWGI) In: J Thies, A Levi, C
Kousik (eds) Advance and Orientation. Proceeding of Cucurbitacaea 2010, Charleston,
South Carolina.
Yamagata H, Masuzawa T, Nagaoka Y, Ohnishi T, Iwasaki T (1994) Cucumisin, a serine protease
from melon fruits, shares structural homology with subtilisin and is generated from a
large precursor. J Biol Chem 269: 32725–32731.
Yamamoto T, Nagasaki H, Yonemaru J, Ebana K, Nakajima M, Shibaya T, Yano M (2010) Fine
definition of the pedigree haplotypes of closely related rice cultivars by means of genome-
wide discovery of single-nucleotide polymorphisms. BMC Genomics. 2010: 11: 267.
Yoshimura M, Toyoshi T, Sano A, Izumi T, Fujii T, Konishi C, Inai S, Matsukura C, Fukuda
N, Ezura H, Obata A (2010) Antihypertensive effect of a gamma-aminobutyric acid-rich
tomato cultivar ‘DG03-9’ in spontaneously hypertensive rats.
C. melo 142–145, 147, 156, 159, 182 diversity array technology (DArT) 338
C. metuliferus 142, 143, 159 DNA marker 238, 240
C. moschata 140, 167, 168, 173–177, 179, double haploid lines (DHL) 244
180 dry matter 93, 106, 107, 109, 111, 115–117,
C. okeechobeensis 110, 126 120–124
C. pepo 140, 167–171, 173, 176, 177, 179
C. rehmii 8 E
C. sativus 142, 143, 156, 157, 159
C. sororia 8 Egusi 310, 316
C. trigonus 21 ethylene 313, 318
classification 93–95, 103, 114, 121 European Central Cucurbits Database 10,
cleaved amplified polymorphic sequence 11
240 evolution 287
climacteric 287, 288, 298, 299 expressed sequence tag (EST) 245, 254
Cm-eIF4E proteins 218 -unigenes 317–319, 322, 323
coat protein (CP) genes 129
F
Coniandreae 141
Conomon 145–147, 151–154 femaleness 4
Consultative Group on International finger printed contigs 292
Agricultural Research 10 flanking markers 216, 218
Corynespora blight 65 flesh bitterness 69
Crookneck 96, 100, 104, 106, 122, 129 flesh color 66, 118
crown rot 67 flesh thickness 118
cucumber 1–7, 9, 12 flexuosus 148
cucumber beetles 127, 128 flowering and fruiting patterns 116
cucumber genome initiative 336 fluorescence in situ hybridization 9, 336
cucumber mosaic virus 65, 67, 79, 107, 108, foliar blights 126
125 fosmid libraries 340
Cucumis 1, 2, 4–7, 9, 11, 12 fruit fly 66, 67
C. anguria 2 fruit quality 63, 64, 66, 67, 79
C. dudaim 145, 146, 149, 151, 154, 155 fruit rot 126
C. melo 2, 5 Fursa 310
C. metuliferus 2 Fusarium wilt 65, 66, 74, 75, 77, 78, 80, 227,
C. picrocarpus 7 232
C. sativus 2, 5
cucumisin 379 G
cucurbit leaf crumple virus 320
Cucurbita 2, 5, 8, 11, 12 gamma-aminobutyric acid 379
C. ficifolia 3, 8 GC-EI-TOFMS 301
C. mixta 2, 3, 8 gene 4, 8, 12, 13
C. maxima 2, 5 genetic diversity 238, 239, 241, 248, 252,
C. maxima X C. moschata 108, 109 253, 262, 271, 276
C. maxima X C. pepo 93, 97, 108 genetic map 9, 199–201, 206, 208, 219
C. moschata 2, 5, 106 genetically-modified organism (GMO) 83
C. moschata X C. martinezii 108 genic male-sterility 69
C. pepo 2, 5 genome 4, 5, 9
Cucurbitaceae 1, 4–6, 9, 140–142, 144, 160, 173 genotyping 240, 246, 270, 271
cucurbitacins 64 geographically-based 241, 253
germplasm 313, 314, 319, 328
D Germplasm Resources Information
Network (GRIN) 10
dioecy 353–357 gibberellins 360
disease resistance 64, 66, 67, 78, 82, 83, 107, Golden Delicious 105
110, 122, 124, 125 gummy stem blight 66, 78, 82, 126, 127
gynoecious 4, 64, 66, 68, 70, 79, 80, 288, 292, linked marker 225, 227, 230, 232
302 Lipoxygenase 348
gynogenesis 130 long terminal repeat 342
gynomonoecious 65, 80 Luffa 17–19, 21, 25, 26, 28, 30, 36, 37, 46, 47
L. abyssinica 166
H L. acutangula 17–19, 25, 47
L. aegyptiaca 26, 30, 36, 37, 47
hand-pollination 100, 101
L. astorii 36, 37
haploid 130
L. breviflora 166
hermaphrodite 65, 288
L. cylindica 17
herpetospermae 141
L. forskalii 30
heteromorphic 356
L. graveolans 18
heterosis 118, 123–125
L. graveolens 36, 37
high performance liquid chromatography
L. hermaphrodita 18, 47
249
L. operculata 36, 37
high-information-content fingerprinting
L. quinquefida 36, 37
292
L. sphaerica 166
HindIII 291, 292
L. umbellate 30
homomorphic 356
luffeae 140, 141, 181
Hubbard 105, 108, 114, 119
luffinS1 26
hull-less oil seed pumpkins 111
luffinS2 26
hybrid cultivar 112, 118, 123
luffinS3 26
hypoglycemic 23
lutein 378
lycopene 315, 316
I
lycopene β-cyclase 267, 316
immortalized mapping population 244,
264, 275 M
Immunomodulation 23
MADS box 357, 358
inbred backcross lines 270, 271
male-sterile 65, 66, 69, 71, 75
inbreeding depression 93, 123
map-based cloning 199, 200, 207, 216–218
inheritance of traits 62
marker-assisted selection (MAS) 43, 226
inodorus 145, 146, 149–155
Medicago 274, 275
insect resistance 64, 125, 127
M. truncatula 289
internal transcribed spacer (ITS) 143, 146
meiotic drive 321, 322
International Cucurbit Genomic Initiative
melon 1–7, 9, 12
(ICuGI) 247
melon necrotic spot virus (MNSV) 217
International Watermelon Genomics
Melothria medraspatana 68
Initiative 381
metabolic sink 268
inter simple sequence repeat 146, 240
metabolomics 286, 300, 301
Interspecific hybridization 128
microarrays 286, 299, 300
intestinal antiparasitia 23
microsynteny 266, 272–275, 345
isozyme marker 200
mitochondrial 4, 8
modifying genes 255, 258
J
molecular marker 199–201, 204, 205, 207
Joliffieae 140, 141, 180 molecular techniques 128
Momordica 140–142, 145–147, 151–154, 180,
K 181
M. balsamina 18, 28, 29
Kabocha 95, 106, 114, 119–121 M. charantia var. charantia 28, 35, 45
M. charantia 17, 18, 20, 21, 24, 29
L M. charantia var. minima 29
Lagenaria siceraria 140, 166 M. charantia var. muricata 28, 35, 45
leucine-rich repeat (LRR) 250 M. cochinchinensis 18, 20, 28
N R
Figure 1-1 Major and minor cucurbit phylogenetic tree. Chromosome numbers and common
names follow each species name (when available). Molecular clock in million years ago,
if available, was shown on branching points. The tribe to which the species belongs was
shown to the right of vertical bars. Geographical occurrence of species: Green—America;
Black—mainland African; Red—Asia; Blue—Australia. The tree was redrawn after Schaefer
et al. (2009).
Chapter 4
Figure 4-1 Ilustration of the three major squash types grown in North America: acorn (C.
pepo, A) kabocha/buttercup (C. maxima, B) and butternut (C. moschata, C).
Figure 4-2 Pistillate (left) and staminate (right) flowers of monoecious C. maxima.
Figure 4-3 Flower morphology of C. maxima (left), C. pepo (middle) and C. moschata (right),
represented by pistillate flowers.
Figure 4-4 Ilustration of peduncle types in C. maxima (bottom laft), C. moschata (bottom right),
C. pepo subsp. ovifera (top laft), C. pepo subsp. pepo (top right). See text for explanation.
Figure 4-5 Pistillate flower (left) tied off with a Twist-ems tie one day prior to anthesis; pistil-
late flower retied and tagged after pollination.
Chapter 5
Figure 5-1 Distribution map of Cucumis spp. Principal diversity centers based on Kirkbride
(1993).
C. sativus L. and C. sativus L. var. hardwickii (Royle) Alef.
C. hystrix Chakravarty
C. myriocarpus Naudin ▲
C. africanus L.
C. anguria L.
C. dipsaceus Ehrenberg ex Spach
C. zeyheri Sonder
C. ficifolius A. Richard
C. metuliferus E. Meyer ex Naudin
♦ ♦
C. melo L. ssp. Melo and C. melo L. ssp. agrestis (Naudin) Pangalo
C. sagittatus Peyritsch
Figure 5-2 Distribution map of Citrullus spp. Principal diversity centers based on Jeffrey
(2001).
C. colocynthis (L.) Schrad.
C. ecirrhosus Cogn.
C. lanatus ((Thunb.) Matsum & Nakai) var. lanatus ▲ and var. citroides ▲
C. rehmii De Winter ♦
Figure 5-3 Distribution map of Cucurbita spp. Principal diversity centers based on Lira-Saade
(1995) and Sanjur et al. (2002).
C. argyrosperma C. Huber
C. ficifolia Bouché
C. maxima Duchesne ▲
C. moschata Duchesne
C. pepo L.
C. ecuadorensis
C. okeechobeensis (J.K. Small) L.H. Bailey
♦
C. lundelliana L.H. Bailey
C. digitata A. Gray, C. cylindrata L.H. Bailey and C. palmata S. Watson
C. foetidissima Kunth
CHINENSIS
MOMORDICA
ACIDULUS
TIBISH
Color Plate Section 401
CHATE Yes
sometimes
green
CANTALUPENSIS
usually
sometimes
Genetics, Genomics and Breeding of Cucurbits
with warts
RETICULATUS
AMERI
Color Plate Section 403
INODORUS
DUDAIM
thick, lignified
non-lignified
lignified
Chapter 6
Figure 6-1 A genetic map of Cucurbita pepo. The new map contains 659 loci: 178 SSR, 244
AFLP, 230 RAPD, five SCAR markers, and two morphological traits (h and B) (From Gong
et al. 2008b).
Chapter 8
Figure 8-3 Comparative analysis of the melon (Cucumis melo L.; n = x = 12) and watermelon
(Citrullus lanatus (Thumb.) Matsum & Nakai; n = x = 11) with the cucumber (Cucumis sativus
L.; n = x = 7) sequence map. The watermelon genetic maps employed in the analysis are
organized into 18 linkage groups. Figure adapted by permission from Macmillan Publishers
Ltd: Nature Genetics; Huang et al. 2009.
Figure 8-4 Overview of microsynteny between melon (Cucumis melo L.) BAC 1-21-10 and
regions in the Arabidopsis thaliana, Medicago truncatula, and Populus trichocarpa (not drawn
to scale). Genes are represented by arrows where gene name, number or ID given above or
below the arrow. Homologous genes are illustrated with the same color and indicated by
narrow connecting lines of the corresponding color. Arrows representing genes that have
one or more ESTs are designate with a spot. Genes without homologs are given in black.
Transposable elements are delineated in gray and indicated by Tn. At1g, At2g, At4g referrers
to A. thaliana chromosomes 1, 2 and 4, respectively. Cm11 referrers to C. melo Linkage Group
11, Pt_XI referrers to Populus trichocarpa Linkage Group XI, and Pt_204 referrers to Populus
unmapped scaffold 204. Mt4 referrers to M. truncatula chromosome 4, consists of BAC
clones AC137837, AC153460 and AC144608, and * = End of scaffold. Figure adapted with
kind permission from Springer Science and Business Media: Deleu et al. (2007), Mol Genet
Genomics 278: 611–622.
Chapter 10
Figure 10-3 Root systems of Citrullus colocynthis, C. lanatus subsp. lanatus “Charleston Gray”
heavily infected with peanut root-knot nematode (RKN), Meloidogyne arenaria race 1, versus
C. lanatus subsp. citroides showing resistance to RKN.
Figure 10-4 Classification of 880 EST-unigenes Illini Red watermelon fruit based on homology
of 880 EST-unigenes to gene sequences in other plant species.
Chapter 11
Figure 11-1 Ideogram showing the position and intensity of Type I/II, Type III, Type IV,
45S rDNA/CsRP2 and 5S rDNA on cucumber metaphase chromosomes (Han et al. 2008).
Chromosome nomenclature follows Koo et al. (2005).
Figure 11-3 Integration of the seven linkage groups of cucumber with individual chromosomes
(Ren et al. 2009). (A1) Distribution of Type I/II (green) and Type III (red) repeats on cucumber
chromosomes. (A2) DAPI staining was converted to black and white images. (A3) Localization
of chromosome-specific fosmid clones on both arms of individual chromosomes, genetic
location of arm-specific fosmid clones are indicated in Figure 11-2. (B) Localization of fosmids
4S (red) and 4L (green) together with Type III (red) and 45S rDNA (green) repeats on the
mitotic chromosomes. Bar = 2.5 µm.
Figure 11-4 Integrated genetic/physical map of cucumber (Huang et al. 2009). (a) Genetic
versus physical distance map of the seven cucumber chromosomes. The genetic map was
constructed using an RIL mapping population from the inter-subspecific cross between Gy14
(domestic cucumber) and PI183967 (wild cucumber). (b) The segmental inversion between the
domestic cultivar Gy14 and the wild accession PI183967 on cucumber chromosome 5 detected
by FISH. 12-2 and 12-7 denote individual fosmid clones. (Scale bars, 1 µm)
Figure 11-5 Comparative genomic analysis of cucurbits (Huang et al. 2009). (a) Comparative
analysis of the melon and watermelon genetic maps with the cucumber sequence map. (b)
Syntenic blocks between the cucumber genome (scaffold000089) and a melon BAC sequence
(accession: EF188258.1). Genes are drawn as black arrows with the orientation indicated on
the sequence. Transposable elements (TEs) are illustrated as rectangles; retrotransposable
elements are in red, DNA transposons are in blue and unclassified TEs are in green. Orthologous
sequence regions between the two genomes are displayed.
6-1
6-2 7-1
Cen
6-3 7-2
6-4
Cen
6-5/6-6
Cen 7-3
6-6
6-10
7-7
6-11
7-8
6-12
Cucumber Melon Cucumber Melon
chromosome 6 chromosome I chromosome 7 chromosome II
Figure 11-6 Diagrammatic illustration of the marker orders and centromere positions of two
pairs of cucumber and melon chromosomes (Han et al. 2009).
Figure 11-7 Genomic locations of R genes on the cucumber chromosomes (Huang et al. 2009).
Three R genes could not be anchored on specific chromosome.
Figure 11-8 Lineage-specific expansion of the lipoxygenase (LOX) family in the cucumber
genome (Huang et al. 2009). The LOX family was divided into two groups, “Type I” and “Type
II”. The two tandem duplicated gene clusters were ordered and displayed on chromosomes
2 and 4, plus one unmapped scaffold of the cucumber genome.
Chapter 12
Petal
Petal
Nectary
Anther
Ovary
Nectary
Ovule
Figure 12-1 Sexual differentiation of cucumber flower buds. Longitudinal sections of cucumber
buds under dissection microscope. A. Male bud at stage 11. B. Female bud at stage 12 (anthesis).
Bud development stages are assigned as per Bai et al. (2004).