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Short Views on Insect Biochemistry and Molecular Biology Vol.(1), October 2014

2014

Section IV
Insect Molecular Genetics

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Short Views
on Vol.
Insect (1)
Biochemistry
and 2014
Molecular Biology Vol.(1), 2014
355 361,

Chapter 15
Insect exuvium extracted DNA marker: a good complementary
molecular taxonomic characteristics with special reference to
mosquitoes
K.J. Dhananjeyan1, R. Paramasivan1, V. Thenmozhi1, R.Chandrasekar2 and B.K. Tyagi1
1

Centre for Research in Medical Entomology (ICMR), 4-Sarojini Street, Chinna Chokkikulam,
Madurai 625002, TN, India
2
Department of Biochemistry and Molecular Biophysics, 238 Burt Hall, Kansas State Univeristy,
Manhattan, KS, USA

Abstract

Many a insect species, particularly sibling species complexes and those sylvatic taxa whose adults
are difficult to catch with information on breeding habitats, pose challenge to their correct
identification. Many genetical and molecular techniques have earlier been attempted on mosquito
species, but without much success in complimenting existing dichotomous keys. Taking cues
from a major molecular taxonomic breakthrough in odonata, we have adopted this technique
refered to as application of genomic DNA extracted from nondestructible structures in
mosquito taxonomy of important Culex, Anopheles and Aedes species.
Key words: Taxonomy, DNA marker, exuvium, cytochrome oxidase, moquito
*For Correspondence (email: abktyagi@gmail.com; abk. tyagi@yahoo.co.in )

1. Introduction
Insects are one of the highly diverged forms of life that exist in the biosphere.
And it has been estimated that only 10 % of the insects have been described and
approximately 10 million species await their discovery. On such grounds, it becomes

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Overview
1. Introduction
2. Advantage and disadvantages of morphology
based identifications.
3. Cytotaxonomy and molecular taxonomy
4. Acknowledgments
5. References

important that identification of individual

biological species.The Biological Species
Concept (BSC) is based on whether
individuals interbreed with one another to
produce viable, fertile off-spring. To establish a biological species it is necessary to
understand its morphological, physiological
and gentic characteristics, thereby its interbreeding capacity could be appreciated.

The field of taxonomy that mainly depends on the morphological characteristics,

as far as insects are concerned is very acute of accommodating and/or positioning the
described species appropriately, as could be inferred from many instances such as the
Ephemeroptera (Mayflies) positioned phylogenetically within the winged (Pterygota)
insects (1); elevation of Ochlerotatus to genus level from subgenus of Aedes (2), etc. In
the field of medical entomology, it is very important that the vector species be
identified precisely, so that the vector control strategies can be designed properly and
can curtail an epidemic. Precise identification of insect species is essential in many
fields such as agriculture (pest management); Forensic Sciences (understanding the
time of crime based on the occurrence of the species and its life stage at the time of
investigation); Conservation of biodiversity (identifying species those are endangered)
etc. Therefore, taxonomy forms the basis for every biological study.

2. Advantages and Disadvantages of morphology based identifications

The morphology based identification technique has many advantages such as:
they can be used at fields; require minimum instrumentations; economical etc. The
disadvantages of morphology based taxonomy are that they require trained and skilful
personnel, specific life stage of the specimen in query; inability to distinguish closely
related and sibling species, species with overlapping characters.

3. Cytotaxonomy and Molecular taxonomy

To overcome the difficulties with morphological techniques, cytotaxonomy was
developed. And with cytotaxonomy many previously undefined species were resolved
such as Anopheles culicifacies C from A & B (3). Even cytotaxonomy has its own
disadvantages such as its inapplicability to all stages and sex of the specimen being
studied, requiring well skilled personnel to run the test as well as to interpret the
readings.

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All these drawbacks are overcome by the DNA based molecular taxonomy as it
could be used for specimens of any stage, sex and form. This is because of the fact that
genomic DNA could be isolated from wide variety of specimens such as immature,
adult, fresh, ethanol preserved, frozen, physically damaged during collection or
handling, museum specimens.
Application of this DNA based molecular taxonomic technique has been very
helpful in discriminating many sibling species belonging to a cryptic species complex.
Genomic DNA markers such as nuclear ribosomal internal transcribed spacers (ITSs),
microstallite sequences, mitochondrial cytochrome oxidase c subunits I & II (mtCOI &
II), NADPH, etc., are used. For example, the ITS markers have been used to identify
sibling species of several Anopheline mosquito complexes such as, Anopheles
maculipennis complex (4), An. quadrimaculatus complex (5) and An. culicifacies
complex (6); Culex vishnui subgroup mosquitoes (7,8) etc. Similarly the mitochondrial
DNA markers have been explored in differentiating the Aedes species mosquitoes (8,9)
For all these molecular taxonomy, the foremost important step and a prerequisite
is good quality of genomic DNA isolated from the specimen, which means that the
specimen, either whole or part, is to be sacrificed. This loss of specimen becomes
devastation in cases such as selection of traits in economically important insects (eg,
Honey bee, silk worm); endangered species (dragon flies, butterflies); loss of voucher
specimen (disease vectors, pests). To minimize the loss, many alternate sources were
used for the isolation of genomic DNA such as legs on one side of the specimens body,
wings or wing clippings and body segments. Even these sources for genomic DNA
isolation were found to cause difficulties. In the case cases of trait selection in honey
bee and silk worm, removal of legs or clipping the wings from the trait selected female
leaves them under-competitive in reproduction, ultimately making the selection
process a failure. In case of endangered species, leg removal or wing clipping renders
the individual weakness; and in case of disease vectors (Fig.1) and pests, loss of
specific discriminating morphological characters.
Recently use of non-destructive materials for the isolation of genomic DNA for
molecular taxonomic works and other molecular studies has been explored. These
non-destructive materials include larval & pupal exuviae and the egg shells. Studies
have revealed that these alternate non-destructive materials successfully yield genomic
DNA that could be used in molecular studies. Exuviae are the insect exoskeleton those
are shed by the immatures when moulting from one instar to the next. Exoskeletons are
biochemically chitin, a polymer of N-acetylglucosamine and do not contain any
nucleic acids. In spite of this nature the genomic DNA yield from exuviae could be
attributed to as then when moulting occurs, the epithelial cells lining the foregut, hind
gut and tracheae are pulled out along with the exoskeleton (10).

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Exuviae of honey bee, Apis melifera, have been successfully used by Gregory &
Rinderer (11) to amplify microsatellite DNA and thereby selecting a better trait of
queen bee to initiate a new colony. The genotypes of the fanning worker bees were also
determined using DNA from the exuviae without hurting or killing the organisms. The
emerging workers were marked with coloured, numbered tags to enable behavioural

Fig.1. Plate showing vector mosquitoes.

a. Aedes aegypti; b. Ae. Albopictus; c. Culex tritaeniorhynchus; d. Cx. vishnui; e. Cx. pseudovishnui

observations over their entire life. Using this new method, Su et al., (12) determined 20
patrilines in a naturally mated queen colony, and discovered that the patriline
composition of bees exhibiting fanning behaviour was significantly different from the
patriline composition of the whole colony. The results confirmed that the genetic
structure of a natural insect society plays a fundamental role in the division of labour.
(11).
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Similarly, genomic DNA from exuviae of odonates has been utilized for studying
the population structure of the odonates by Watts and others (13). In the same way, the
DNA sequences of butter flies were analysed by Feinstein (14) that was isolated from
exuviae. The pupal exuviae of Chironomides were also been used to identify the
specimens to the species level following non-destructive method of genomic DNA
isolation. The work had yield very promising results by making use of the pupal
exuvium for DNA isolation without damaging the skin itself which further was
vouched for morphological identification purpose (15).
In a study by Dhananjeyan et al., in 2010, the use of egg shells along with larval
and pupal exuviae of the vector mosquitoes have been extensively studied. The exuviae
of different species of vector mosquitoes viz., Culex tritaeniorhynchus, Cx. vishnui,
Cx. pseudovishnui, (Fig. 2) Aedes aegypti and Ae. albopictus (Fig. 3) have been studied
for their use in molecular taxonomy
and all the species have provided
genomic DNA usable in species
differentiating PCR reactions. The
left out parts of mosquitoes are as
informative as the adult mosquitoes,
when
molecular
studies
are
concerned. These left-out materials
were very much useful in molecular
identi- fication of 3 important vector
mosquitoes belonging to the Cx.
vishnui subgroup, which are very
difficult to be separated in the adult
stage, as they have similar
morphological characters which
some times vary within species and Fig. 2. Gel picture showing differential PCR results
overlap between species in a identifying Culex triteaniorhynchus, Cx. vishnui &
proportion of specimens (16). They Cx. pseudovishnui with 536 bp, 246 bp & 344 bp
were also very much useful in respectively. Adapted from Dhananjeyan, et al. (8).
identifying the Aedes aegypti and
Ae. albopictus (8).

Two new species of mosquitoes namely Anopheles pseudosundaicus (17) and

Toxorhynchites tyagii (18) were discriminated from their closely related alleys, An.
sundaicus, An. subpictus and Tx. splendens, respectively by using the molecular
methods of species identification. The technique included the isolation of genomic
DNA and amplification of mitochondrial DNA, which was then used for the specific
amplification of Cytochrome C Oxidase subunit 1 gene. In both the cases, the
cladogram constructed based on the mitochondrial gene sequence, clearly placed the
new species precisely position in the tree, representing the phylogenetic status.
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Fig. 3. Gel picture showing ITS2 PCR differentiating Aedes aegypti (330 bp) and Ae. albopictus
(520 bp). Adapted from Dhananjeyan, et al., (8).

The molecular markers for species identification are very powerful when
combined with traditional taxonomical tools. The only drawback is data on
intra-specific and geographical variation is very limited. This makes it essential to
vouch and submit the specimen, assigned to the DNA and study the biology of the
species in query. And the use of non-destructive sources of genomic DNA enables a
researcher to comply with the needs.

4. References
1.

Sivaramakrishnan, K.G., K.A. Subramanian, M. Arunachalam, C.S. Kumar & S. Sundar (2011) Emerging
trends in molecular systematics and molecular phylogeny of mayflies (Insecta: Ephemeroptera). Journal
of Threatened Taxa 3(8): 19751980.
2. Reinert, J.E. (2000) New classification for the composite genus Aedes (Diptera: Culicidae: Aedini),
elevation of subgenus Ochlerotatus to generic rank, reclassification of the other subgenera, and notes on
certain subgenera and species. J. American Mosquito Control Association. 16(3):175-188.
3. Subbarao, S.K., Vasantha, K., Adak, T., Sharma, V.P. (1983) Anopheles culicifacies Complex: Evidence
for a New Sibling Species, Species C. Annals of the Entomological Society of America 76(6): 985-988.
4. Porter, C.H., Collins, F.H. (1991) Species-diagnostic differences in a ribosomal DNA internal transcribed
spacer from the sibling species Anopheles freeborni and Anopheles hermsi (Diptera: Culicidae). Am. J.
Trop. Med. Hyg. 45: 271-279.
5. Cornel, A.J., Porter, C.H., Collins, F.H. (1996) Polymerase chain reaction species diagnostic assay for
Anopheles quadrimaculatus cryptic species (Diptera: Culicidae) based on ribosomal DNA ITS2
sequences. J. Med. Entomol. 33: 109-116.
6. Goswami, G., Raghavendra, K., Nanda, N., Gakhar, SK., Subbarao, S.K. (2005) PCR-RFLP of
mitochondrial cytochrome oxidase subunit II and ITS2 of ribosomal DNA: markers for the identification
of members of the Anopheles culicifacies complex (Diptera: Culicidae). Acta Trop. 95: 92-99.
7. Toma, T., Miyagi, I., Crabtree, M.B. and Miller, B.R. (2000) Identification of Culex vishnui Subgroup
(Diptera: Culicidae) mosquitoes from the Ryuku archipelago, Japan: Development of a species diagnostic
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8.
9.
10.
11.
12.
13.
14.
15.
16.
17.

18.

polymerase chain reaction assay based on sequence variation in ribosomal DNA spacers. J. Medical
Entomology 37(4): 554-558.
Dhananjeyan, K.J. Paramasivan, R., Tewari, S.C., Rajendran, R., Thenmozhi, V., Victor Jerald Leo, S.,
Venkatesh, A. and Tyagi, B.K. (2010) Molecular identification of mosquito vectors using genomic DNA
isolated from eggshells, larval and pupal exuvium. Tropical Biomedicine 27(1): 47-53.
Cook, S., Diallo, M., Sall, A.A., Cooper, A., Holmes, E.C. (2005) Mitochondrial markers for molecular
identification of Aedes mosquitoes (Diptera: Culicidae) involved in transmission of arboviral disease in
West Africa. J. Medical Entomology, 42(1): 19-28.
Bertholf, L.M. (1925) The moults of the honeybee. Journal of Economic Entomology 18: 380-384.
Gregory, P.G., Rinderer, T.E. (2004) Nondestructive sources of DNA used to genotype honey bee (Apis
mellifera) queens. Entomologia Experimentalis et Applicata 111: 173-177.
Su S., Stefan, A., Shaowu, Z., Sven, M., Shenglu, C., Honghu, D., Jrgen, T. (2007) Non-destructive
genotyping and genetic variation of fanning in a honey bee colony. J. Insect Physiology. 53(5): 411417.
Watts, P.C., Thompson, D.J., Daguet, C., Kemp, S.J. (2005) Exuviae as a reliable source of DNA for
population-genetic analysis of odonates. Odonatologica 34(2): 183-187.
Feinstein, J. (2004) DNA sequence from butterfly frass and exuviae. Conservative Genetics 5: 103-104.
Krosch, M.N. and Cranston, P.S. (2012) Non-destructive DNA extraction from Chironomidae, including
of fragile pupal exuviae, extends analysable collections and enhances vouchering. Chironomus. 25: 22-27.
Reuben, R., Tewari, S.C., Hiriyan, J. and Akiyama, J. (1994) Illustrated keys to species of Culex (Culex)
associated with Japanese encephalitis in Southeast Asia (Diptera: Culicidae). Mosquito Systematics 26:
7596.
Tyagi, B.K., Hiriyan, J., Tewari, S.C., Ayanar, K., Philip Samuel, P., Arunachalam, N., Paramasivan, R.,
Krishnamoorthy, R., Dhananjeyan, K.J., Victor Jerald Leo, S., and Rajendran, R. (2009) Description of a
new species, Anopheles pseudosundaicus (Diptera: Culicidae) from coastal Kerala, India. Zootaxa 2219:
4960.
Krishnamoorthy, R., Munirathinam, A., Dhananjeyan, K.J., Hiriyan, J., Mariappan, T., Philip Samuel, P.,
and Venkatesh, A. (2013) Description of a new species, Toxorhynchites (Toxorhynchites) tyagii (Diptera:
Culicidae), from Nilgiri hills, Western Ghats, South India. Zootaxa. 3701(4): 447-459.

Article History: Received 10th August, 2013; Revised 10th October, 2013; Accepted 1st January, 2014 and
Publsihed 30th October 2014.

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Insect Vectors

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Table Contents

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Page No.

Preface
Forward message
Contributors
Reviewers
Acknolwedgement

i
ii
iii
iv
v

Volume1
Section I: Insect Biochemical approaches

1. Introduction to Insect Molecular Biology.

Raman Chandrasekar, P.G., Brintha, Enoch Y.Park, Paolo Pelsoi, Fei Liu,
Marian Goldsmith, Anthony Ejiofor, B.R., Pittendrigh, Y.S., Han,
Fernando G. Noriega, Manickam Sugumaran, B.K., Tyagi, Zhong Zheng Gui,
Fang Zhu, Bharath Bhusan Patnaik, and P. Michailova

2.

Modulation of Botanicals on pests biochemistry.

57

Sahayaraj, K.

3.

Detoxication, stress and immune responses in insect antenna:

new insights from transcriptomics.

75

David Siaussat, Thomas Chertemps and Martine Maibeche

4.

Application of isotopically labeled compounds and tandem mass

spectrometry for studying metabolic pathways in mosquitoes.

99

Stacy Mazzalupo and PatriciaY.Scaraffia

5.

Field Response of Dendroctonus armandi Tsai & Li (Coleoptera:

Scolytinae) to Synthetic Semiochemicals in Shaanxi, China.

127

Shou-An Xie, Shu-Jie L.V., Hui-Chen, Raman Chandrasekar

xvii

Section II: Insect Growth

6. Insect Cuticular SclerotizationHardening Mechanisms and Enzymes.

149

Manickam Sugumaran

7. New Approaches to Study Juvenile Hormone Biosynthesis in Insects.

185

Crisalejandra Rivera-Perez, Marcela Nouzova and Fernando G. Noriega

8. The regulatory biosynthetic pathway of juvenile hormone.

217

Zhentao Sheng and Raman Chandrasekar

Section III:

Insect Immunity

9. The innate immune network in a hemimetabolous insect, the brown

planthopper, Nilaparvata lugens.

233

Yanyuan Bao, Raman Chandrasekar, Chuan-Xi Zhang

10. Immune Pathways in Anopheles gambiae.

253

Maria L. Simes and Raman Chandrasekar

11. Key biochemical markers in silkworms challenged with immuno-

271

elicitors and their association in genetic resistance for survival.

Somasundaram, P., Chandraskear, R., Kumar,K.A., and Manjula, A.

Section IV:

Insect Molecular Genetics

12. The recent progress of the W and Z chromosome studies of the

291

silkworm, Bombyx mori

Hiroaki Abe, Tsuguru Fujii and Raman Chandrasekar

13. Molecular characterization and DNA barcoding for identification of

317

agriculturally important insects.

Rakshit Ojha, Jalali, S.K., and Venkatesan, T.

14. Polytene chromosomes and their significance for Taxonomy,

331

Speciation and Genotoxicology

Paraskeva V. Michailova

15. Insect exuvium extracted DNA marker: a good complementary

molecular taxonomic characteristics with special reference
to mosquitoes.

355

Dhanenjeyan, K. J., Paramasivam, R., Thanmozhi, V., Chandrasekar,R., and Tyagi, B.K.
Index

363

xviii

Volume2
Section V:

Molecular Biology of Insect Pheromones

16. Understanding the functions of sex-peptide receptors?

373

Orly Hanin, Ada Rafaeli

17. Current views on the function and evolution of olfactory receptors

385

in Lepidoptera.

Arthur de Fouchier, Nicolas Montagn, Olivier Mirabeau, Emmanuelle Jacquin-Joly

18. Molecular architecture, phylogeny and biogeography of pheromone

409

biosynthesis and reception genes / proteins in Lepidoptera.

Jian-Cheng Chang, P. Malini, R. Srinivasan

Section VI:

Insect Molecular Biology

19. Application of Nanoparticles in sustainable Agriculture :

429

Its Current Status.

Atanu Bhattacharyya , Raman Chandrasekar, Asit Kumar Chandra,

Timothy T. Epidi and Prakasham, R.S.

20. Mosquito Ribonucleotide Reductase: A Site for Control.

449

Daphne Q.-D. Pham, Victor H. Perez, Lissette Velasquez, Dharty Bhakta,

Erica L. Berzin, Guoli Zhou, and Joy. J. Winzerling.

21. Green protocol for synthesis of metal nanoparticles

to control insect pests.

473

Murugan, K., Chandrasekar, R., Panneerselvam, C., Naresh Kumar, A.,

Madhiyazhagan, P., Mahesh Kumar, P., Jiang-Shiou Hwang, Jiang Wei

22. Aquaporins in Blood-Feeding Arthropods.

497

Lisa L. Drake, Hitoshi Tsujimoto, Immo A. Hansen

23. Mimetic analogs of three insect neuropeptide classes

509

for pest management.

Ronald J. Nachman

xix

Section VII:

Insect Pest Management through

Biochemical and Molecular approaches

24. Induced resistance in plants against insect pests and

533

counter-adaptation by insect pests.

Abdul Rashid War and Hari C Sharma

25. Insect Chemical communication - an important component of

549

novel approaches to insect pest management.

Usha Rani, P.

26. Mosquito control using biological larvicides: Current Scenario.

575

Subbiah Poopathi, C. Mani and R. Chandrasekar

27. Application of RNAi toward insecticide resistance management.

595

Fang Zhu, Yingjun Cui, Douglas B. Walsh, Laura C. Lavine

Section VIII:

Insect Bioinformatics

28. Entomo-informatics: A prelude to the concepts in Bioinformatics.

621

Habeeb, S.K.M. and Raman Chandrasekar

29. Molecular expression and structure-function relationships of

633

apolipophorin III in insects with special reference to innate immunity.

Bharat Bhusan Patnaik, Raman Chandrasekar, Yeon Soo Han

30. Computer-aided pesticide design: A short view

685

Jitrayut Jitonnom

Index

709

xx

ISBN No. 978-1-63315-205-2 (USA)

First Edition: Volume 1, 2 October 2014

Total No. Pages: 398 + 372 = 770

Edited by Raman Chandrasekar

B.K. Tyagi
Zhong Zheng Gui
Gerald R. Reeck
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Volume 1 & 2, October 2014

Short Views on Insect Biochemistry

and Molecular Biology

PREFACE
Entomology as a science of inter-depended branches like biochemistry, molecular entomology, insect
biotechnology; has made rapid progress in its attributes in the light of modern discoveries. This also
implies that there is an urgent need to manage the available resources scientifically for the good of man.
In the past five decades, entomology in the world/country has taken giant steps ahead. Continued
research has evolved better pest management through molecular approaches. The aim of the Short
Views on Insect Biochemistry and Molecular Biology book is to integrate perspectives across
biochemistry and molecular biology, physiology, immunology, molecular evolution, genetics,
developmental biology and reproduction of insects. This century is proclaimed as the Era of
Biotechnology and its consists of all types of Mol-Bio applications, which is an essential component for
a through understanding of the Insect Biology. This volume 1 & 2 (8 section with 30 chapters)
establishes a thorough understanding of physiological and biochemical functions of proteins, genes in
insects life processes; the topics dealt with in the individual chapters include chemistry of the insect
cuticle, hormone and growth regulators; biochemical defenses of insects; the biochemistry of the toxic
and detoxification action; modern molecular genetics and evolution; inter- and intra-specific chemical
communication and behavior; insect pheromone and molecular architecture, phylogeny and chemical
control of insect by using insect pheromones biotechnology; insect modern biology and novel plant
chemical and microbial insecticides for insect control, followed by a discussion of the various
mechanisms of resistance (both behavioral and physiological) and resistance management; modern insect
pest management through biochemical and molecular approaches; Mimetic analogs of insect
neuropeptide for pest management; entomo-informatics and computer-aided pesticide designing. In short
this book provides comprehensive reviews of recent research from various geographic areas around the
world and contributing authors area recognized experts (leading entomologist/scientist) in their
respective filed of molecular entomology. We will miss this collaboration now it has ended, but will feel
rewarded if this book is appreciated by our team/colleagues and remarkable mile stone in entomology
field.
This book emphasizes upon the need for and relevance of studying molecular aspects of entomology in
Universities, Agricultural Universities and other centers of molecular research. To encompass this
knowledge and, particularly disseminate it to the scientific community free of cost, was the major
inspiring force behind the launch of Short Views on Insect Biochemistry and Molecular Biology.

Editors

Raman Chandrasekar
Brij Kishore Tyagi

ii

iii

iv

vi

ShortViewson

InsectBiochemistryand
MolecularBiology
Editedby
Raman Chandrasekar, Ph.D.,
Kansas State University, USA.
B.K.Tyagi, Ph.D.,
Centre for Research in Medical Entomology (ICMR), India.
Zhong Zheng Gui, Ph.D.,
Jiangsu University of Science and Technology,
Sericultural Research Institute, Chinese Academy of
Agricultural Sciences, China.
Gerald R. Reeck, Ph.D.,
Kansas State University, USA.

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Contributing Authors

Dr. B.K.Tyagi

Prof.Fernando G. Noriega

Centre for Research in Medical Entomology,

4Sarojini Street, Chinna Chokkikulam,
Madurai 625002 (TN), India

Department of Biological Sciences

HLS 227, Florida International University
11200 SW 8th St, Miami, FL 33199, USA.

Prof. Gui Zhongzheng

Dr. Zhentao Sheng

Sericultural Research Institute,

Chinese Academy of Agricultural
Sciences, Zhenjiang, 212018,
Jiangsu, P. R. China.

Chicogo University, Chicogo, USA.

Prof. K. Sahayaraj

Prof.Yanyuan Bao
Institute of Insect Science,
Zhejiang University, China.

Dept. of Advanced Zoology and Biotechnology,

St. Xavier's College
Palayamkottai 627 002, Tamil Nadu, India.

Prof. Chuan-Xi Zhang,

Prof. David Siaussat

Dr. Maria L. Simes

Universit Pierre et Marie Curie (Paris 6/UPMC),

UMR 1272A Physiologie de l'Insecte:
Signalisation et Communication (PISC),
7 Quai Saint Bernard, Batiment A - 4me tage bureau 410, 75252 Paris Cedex 05, France.

Prof. PatriciaY.Scaraffia
Department of Tropical Medicine,
Tulane University, New Orleans,
LA 70112, USA.

Prof. Shou-An Xie

Institute of Insect Science,

Zhejiang University, China.

UEI Parasitologia Mdica,

Centro de Malria e Outras Doenas Tropicais,
Instituto de Higiene e Medicina Tropical,
Rua da Junqueira 96, 1300 Lisboa,
Portugal.

Dr. P. Somasundaram
Central Sericultural Germplasm Resources Centre,
P.B.No.44, Thally Road,
Hosur-635109,
Tamilnadu, India.

College of Forestry,
Northwest A & F University
Yangling, Shaanxi 712100, China

Dr. Hiroaki Abe

Dr. Raman Chandrasekar

Dr. S.K. Jalali

Department of Biochemistry and Molecular

Biophysics, Kanas State University,
Manhattan, 66506, KS, USA.

Prof. Gerald R. Reeck

Department of Biochem. and Molecular
Biophyscis, Kansas State University, KS, USA.

Prof. Manickam Sugumaran

Department of Biology
University of Massachusetts Boston
100 Morrissey Blvd,
Boston, MA 02125, USA.

Tokyo University of Agriculture and Technology,

Japan.

National Bureau of Agriculturally Important

Insects, ICAR, India.

Prof. Paraskeva V. Michailova

Institute of Biodiversity and
Ecosystem Research,
1 Tzar Osvoboditel boulv
Bulgarian Academy of Sciences
Sofia 1000, Bulgaria.

Prof. Ada Rafaeli

Associate Director for Academic Affairs &
International Cooperation
Agricultural Research Organization,
The Volcani Center, P. O. Box 6,
Bet Dagan 50250, Iseral.

ix

Prof. Emmanuelle Jacquin-Joly

Dr. Fei Liu

UMR PISC Physiologie de l'insecte

INRA, Route de Saint-Cyr
78026 Versailles cedex, France..

Department of Biological Science & Technol.,

Shaanxi Xueqian Normal University,
Shaanxi, China.

Dr. R. Srinivasan

Prof. Marian Goldsmith

Entomologist and Head of Entomology Group

AVRDC-The World Vegetable Center
60 Yi Ming Liao, Shanhua
Tainan 74151, Taiwan.

Biological Sciences Department,

University of Rhode Island,
Kingston, RI 02881, USA

Prof. Atanu Bhattacharyya

Prof. Anthony Ejiofor

Vidyasagar College for Women,

Post Graduate Department of Environmental
Science,
University of Kolkata, India.

Department of Biological Sciences,

College of Agriculture, Human & Natural
Sciences, Tennessee State University,
3500 John A Merritt Blvd., Nashville,
Tennessee 37209, USA.

Prof. Daphne Q.-D. Pham

Dr. Bharath Bhusan Patnaik

Dept of Biological Sciences,

University of Wisconsin-Parkside,
900 Wood Road, Kensoha,
WI 53144, USA.

School of Biotechnology,
Trident Academy of Creative Technology
(TACT), Bhubaneswar 751013 Odisha, India.

Prof. Jitrayut Jitonnom

Prof. B.R. Pittendrigh

School of Science
University of Phayao, Thailand.

Department of Entomology,
University of Illinois, Urbana-Champaign, IL,
61801, USA
.

Prof. K. Murugan

Dr. Subbiah Poopathi

Department of Zoology, School of Life Sciences,

Bharathiar University,
Coimbatore - 641 046, India.

Prof. Immo A. Hansen

Department of Biology,
New Mexico State University,
Las Cruces, NM, USA.

Dr. Ronald J. Nachman

USDA-ARS,
Food Animal Protection Research Laboratory,
USA.

Dr. Hari C Sharma

International Crops Research Institute for the
Semi-Arid Tropics (ICRISAT), Patancheru502324,
Andhra Pradesh, India.

Prof. Paolo Pelsoi

State Key Laboratory for Biology Plant Diseases
and Insect Pests, Institute of Plant Protection,
Chinease Academy of Agricultural Sciences,
Bejing, China.

Unit of Microbiology and Immunology,

Vector Control Research Centre
(Indian Council of Medical Research),
Medical complex, Indira Nagar,
Puducherry 60 5006, India.

Dr. P.Usha Rani

Biology and Biotechnology Division
Indian Institute of Chemical Technology
(CSIR)Taranaka,
Hyderabad - 500 007 (AP), India.

Dr. Fang Zhu

Irrigated Agriculture Research and Extension
Center, Dept.of Entomology,
Washington State University,
Prosser, WA, USA.

Prof. S.K.M. Habeeb

Department of Bioinformatics,
Faculty of Engineering & Technology,
SRM University, Kattankulathur,
Chennai 603203, Tamilnadu, India.

Prof. Yeon Soo Han

Division of Plant Biotechnology,
College of Agriculture & Life Science,
Chonnam National University,
Gwangju 500-757, South Korea

SION
MIS

TERNA
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N AL B OO

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Reviewer & External supportive members

Prof. Michael Riehle, Department of Entomology, University of Arizona, USA.

Dr. Dawn L.Geiser, College of Agriculture and Life Sciences, University of Arizona, USA.
Prof. Young Jung Kwon, School of Applied Biosci., Kyungpook National University, South Korea.
Dr. Kaliappandar Nellaiappan, CuriRx Inc. USA.
Prof. Patricia Y. Scaraffia, Department of Tropical Medicine, Tulane University, USA.
Prof. Richard Newcomb, Plant & Food Research, University of Auckland, New Zealand.
Dr. S. Krishnaswamy, School of Biotechnology, Madurai Kamaraj University, South India.
Dr. Mary-Anne Hartley, University of Lausanne, Switzerland.
Dr. Igor F. Zhimulev, Institute of Molecular and Cellular Biology, Novosibirsk, Russia.
Dr. S. Subramanin, Indian Agricultural Research Institute. India.
Prof. Gustavo F. Martins, Departament de Biologia Geral, Universidade Federal de Vicosa, Brazil.
Prof. Helena Janols, Infektionsklinien, Skanes Universitetsisjukhus, Sweden.
Prof. Donald R.Barnard, USDA, Agricultural Research Service, CMAVE, USA.
Dr. Keith White, Faculty of Life Science, University of Manchester, UK.
Prof. Marten J.Edwards, Biology Department, Muhlenberg College, USA.
Prof. E. Warchalowska-Sliwa, Polish Academy of Sciences, Poland.
Dr. K. Balakrishnan, Department of Immunology, Madurai Kamaraj University, India.
Dr. J.Joe Hull, USAD-ARS, Arid Land Agricultural Research Centre, USA.
Dr. Neil Audsley, The Food & Environment Research Agency, UK.
Dr. Raman Chandrasekar, Kansas State University, USA.
Dr. B.K. Tyagi, Centre for Research in Medical Entomology (ICMR), Madurai, TN, India.
Prof. Zhongzheng Gui, Sericulture Research Institute, Chinese Academy of Agricultural Sci., China.
Dr. Fang Zhu, Irrigated Agril. Research and Extension Center, Washington State University, USA.
Prof. K. Murugan, Department of Zoology, Bharathiar University, Coimbatore, India.
Dr. Xiao-Wei Wang, Institute of Insect Science, Zhejiang University, China.
Dr. Haijun Xu, Institute of Insect Science, Zhejiang University, China.
Dr. Alisha Anderson, CSIRO Ecosystem Sciences, Australia.
Prof. Eric D.Dodds, Department of Chemistry, University of Nebraska-Lincoln, USA.
Prof. P. Mosae Selvakumar, Department of Chemistry, Karnaya University, Coimbatore, India.
Prof. A.K.Dikshit, Indian Agriculture Research Institute, New Delhi.
Prof. K.R.S. Sambasiva Rao, Dept. of Biotech. & Zoology, Acharya Nagarjuna University, India
Dr. R. Rangeshwaran, National Bureau of Agriculturally Important Insects, Banglore, India.
Dr. V. Selvanarayanan, Faculty of Agriculture, Annamalai University, Tamil Nadu, India.
Prof. Fernando G. Noriega, Florida International University, Miami, USA.
Prof. Ada Rafaeli, Department of Food Quality and Safety, A.R.O., Israel.
Prof. Daphne Q.-D. Pham, Dept. of Biological Sciences, University of Wisconsin-Parkside, USA.
Prof. Emmanuelle Jacquin-Joly, INRA, UMR 1272 Physiologie de lInsecte, Versailles, France.
Prof. Manickam Sugumaran, University of Massachusetts Boston, USA.
Prof. Nannan Liu, Auburn University, USA.
Prof. Michihiro Kobyashi, Nagoya University, Japan.
Prof. Enoch Y.Park, Innovative Joint Research Center, Shizuoka University, Japan.
Prof. Luiz Paulo Moura ANDRIOLI, Universidade de So Paulo, SP - Brazil
Prof. SHIMADA Toru, The University of Tokyo, Japan.
Prof. Erjun Ling, Institute of Plant Physiology and Ecology, China.

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Acknowledgements
Writing and publishing a book requires the assistance of individuals who are
creative, talented, and hard-working. All of these qualities were present in the
individuals assembled to produce this book volume. I would like to express my
heartfelt gratitude to my former teacher Prof. Seo Sook Jae, (GSNU, South Korea),
Prof. Subba Reddy Palli (University of Kentucky, USA), and other external mentors
Prof. Marian R. Goldsmith (University of Rhode Island, USA), Prof. Enoch Y. Park
(Shizuoka University, Japan), Prof. M. Kobayashi (Nagoya University, Japan), Prof.
CHU Jang Hann (National University of Singapore, Singapore), Prof. Thomas W.
Sappington (USDA-ARS, USA), Prof. Fernando G. Noriega (Florida International
University, USA), Dr. Srinivasan Ramasamy, AVRDC, The World Vegetable
Center, Taiwan), Dr. H.C. Sharam (ICRISAT, India), who inspiration and
supported me at many ways for the commencement of this International Book
Mission Program. The book mission program was initiated on May 2010,
completed on March 2014 and published on October 2014. I have no words to
express my feeling for all those who provided valuable contributions from USA,
South Korea, Japan, China, India, Thailand, Taiwan, Bulgaria, France, Iseral, and
Portugal (Contributors name list, see page no. v) and made the completion of this
book possible. We express our appreciation to the following people (Reviewer
name list, see page no. vii) who reviewed various part of the manuscript as it was
being developed and improved quality of each chapter. I thank the ICMR, New
Delhi, and Chinese Academy of Agricultural, China, and Kansas State University for
support from several aspects. Many others (scientists and publishers) have also
allowed us to use their materials in the various chapters, their color image have then
been converted to gray color/BW. Iam especially indebted to International Book
Mission Organization, Academic Publishing Services for the production of book. I
thank my Co-Editors for their continuous vigilance over the book project and for
always giving advance notice of the editing and proofreading schedules. I thank also
my Brintha, P.G., (my wife), who in all possible way, encouragement helped
transform our original efforts into an acceptable final form. I apologize to those
whose work could not be cited owing to space considerations limitation. Further, I
wish to recognize the moral support extended by colleagues and friends. I hope that
this volume will inspire interest on the diverse aspects of insect biochemistry and
molecular biology in aspiring and established scientists.
Raman Chandrasekar

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A Note from the Publisher

Dear Readers,
This edition represents the first number of the Short Views on Insect
Biochemistry and Molecular Biology book series published by International
Book Mission. It serves to show the public how important entomology field in
expanding basic knowledge or in the development of new technologies nowadays,
in virtually all fields of knowledge. We called for piece of work falling into two
volumes (Basic and Advance aspects).
Far from being complete, the 30 chapters clearly structured and simply explained
experts contributions may provide an overview about current and prominent
advances in insect biochemistry and molecular biology which will help students and
researchers to broaden their knowledge and to gain an understanding of both the
challenges and the opportunities behind each approach.
We look forward to receiving new proposals for the new edition 2015 - 2017.
International Book Mission
Academic Publisher
Manager

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