International Journal of Bio-Technology

and Research (IJBTR)

ISSN(P): 2249-6858; ISSN(E): 2249-796X
Vol. 4, Issue 6, Dec 2014, 1-12
TJPRC Pvt. Ltd.

A MULTIDISCIPLINARY MOLECULAR MARKER APPROACHES TO ASSESS THE

GENETIC DIVERSITY IN EGYPTIAN DATE PALM
SAMI S. ADAWY & MOHAMED A. M. ATIA
Department of Genome Mapping, Agricultural Genetic Engineering Research Institute Ageri, Arc, Giza, Egypt

ABSTRACT
Date palm is one of the most successful fruit crops in tropical and sub-tropical habitats. In the present
investigation a multidisciplinary molecular marker approaches including three novel marker types (SCoT, CDDP and
ITAP) in addition to (SSR, AFLP and RAPD) were employed to assess the genetic diversity and genetic relationships
within and among different Egyptian cultivars. Males and females genotypes representing three or five Egyptian cultivars
were assayed using 14 AFLP, 48 SCoT, 14 SSR, 21 CDDP, 18 ITAP and 100 RAPD primers or primer combinations.
Amplification products yielded a total no. of bands 591, 484, 83, 192, 204 and 1084, with a percentage of polymorphism
37.4%, 41.3%, 59%, 31.7%, 34% and 24.9%, respectively. The average no. of bands/primer ranged from 5.9 for SSR to
42.2 for AFLP. The cluster analysis of the studied genotypes using these different marker systems revealed five
dendrograms exhibited unique topology with some similarities. The generated dendrograms from these marker systems
successfully clustered the studied genotypes based on cultivar or gender. The data scored from AFLP, SCoT, SSR, CDDP
and ITAP were combined and computed to generate more accurate relationships based on large and versatile genome
coverage. The dendrogram based on the combined data exhibited the closest relationships to those illustrated by the
AFLP-based dendrogram. On the other hand, the RAPD dendrogram exhibited also a unique topology with some
similarities comparing with other marker systems.
These results confirmed that different marker systems differ in the mechanism of detecting polymorphism,
genome coverage and the ease of application. Therefore, they could complement each other to draw more accurate
conclusions.

KEYWORDS: Date Palm, AFLP, SCot, SSR, CDDP, ITAP, RAPD, Molecular Markers
INTRODUCTION
The genus Phoenix, which includes the date palm (Phoenix dactylifera L) is the sole member of the tribe
Phoeniceae of the monocotyledonous family palmae (Bailey Hortatorium, 1976). The date palm (Phoenix dactylifera L)
is one of the excellent candidate crops in arid and semiarid regions of the world with high tolerance to environmental
stresses. In addition to its valuable fruit, the tree is cultivated for fuel, fiber and as shelter for ground crops. Egypt is the
leading date production country in the world. Egypt has very long history of date cultivation that dates back to 3200 BC.
The progress of any genetic preservation is dependent on understanding the amount and distribution of the genetic
variation present in the genetic pool (Jubrael et al, 2005). Due to developments in the field of molecular genetics, various
techniques have been emerged to analyze genetic variation in date palm.
DNA fingerprinting in plants is primarily used for identification of genetic diversity, protection of biodiversity or
germplasm conservation andidentifying markers associated with specific traits (Khanam et al, 2012). Molecular markers
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Sami S. Adawy & Mohamed A. M. Atia

based on AFLP (Amplified Fragment Length Polymorphism), RAPD (Random Amplified Polymorphic DNA) and SSR
(Simple Sequence Repeats) are powerful techniques, which can be used to identify and determine plant genomes or to
estimate the phylogenetic relationship among genotypes of date palm (Adawy et al. 2005 & 2014 and Elshibli and
Korpelainen, 2008).The molecular markers technologies continue to improve with development of simpler protocols with
greater reliability and lower cost. This makes them increasingly practical for routine applications to tropical or subtropical
species for which very limited research resources are available.
Recently, a molecular marker techniques termed (SCoT, CDDP and ITAP) were developed by Collard and
Mackill (2009a), Collard and Mackill (2009b) and Xiong et al. (2013), respectively. Start codon targeted (SCoT)
polymorphism; a simple and novel DNA marker technique, performed through PCR using single primer targeting the short
consensus conserved region flanking the ATG translation initiation codon in plant genes. This has been validated through
study on genetic diversity among rice varieties and marker segregation in rice backcross population (Collard and Mackill,
2009a). Also, Collard and Mackill (2009b) developed another novel method for generating plant DNA markers called
Conserved DNA-Derived Polymorphism (CDDP), based on data mining for short conserved amino acid sequences in
proteins and designing PCR primers based on the corresponding DNA sequence. This method uses single 15-to 19-mer
primers for PCR and an annealing temperature of50C. In addition, Xiong et al. (2013) developed a new molecular marker
technique called intron targeted amplified polymorphism (ITAP) based on the principles of sequence related amplified
polymorphism (SRAP), target region amplification polymorphism (TRAP) and conserved region amplification
polymorphism (CoRAP).
Therefore, the present investigation was conducted to define the genetic diversity among male and female
genotypes belonging to different Egyptian cultivars using AFLP, SSR and RAPD. In addition to introducing three novel
marker systems (SCoT, CDDP and ITAP) which applied for the first time on date palm for genotyping and fingerprinting
studies.

MATERIALS AND METHODS

Plant Material
The samples of superior date palm cultivars were collected from the two gander (male and female) as following:

For RAPD Analysis: Ten samples representing males and females of five Egyptian cultivars
(Bertamoda, Hayani, Samany, Siwi and Zaghloul) were collected to be subjected for molecular analysis.

For SSR, AFLP, SCoT, CDDP and ITAP analysis: Twelve samples representing males and females of three
Egyptian cultivars (Zaghloul, Hayani and Samany; two plants from each gender) were collected to be subjected
for molecular analysis. Two additional samples represent bulked males and bulked females were prepared.
High-quality genomic DNA was extracted from fresh leaves (100 mg) of all collected plants using a DNAeasy
Plant Mini Kit (QIAGEN, Santa Clarita, CA) and according to the manufacturers protocol.

RAPD Analysis
RAPD-PCR was carried out as described by Diab et al, (2013). A set of 100 random 10-mer primers were applied
against the ten date palm samples in order to screening the polymorphism between and among (based on gender) cultivars.

Impact Factor (JCC): 2.8872

Index Copernicus Value (ICV): 3.0

A Multidisciplinary Molecular Marker Approaches to Assess the Genetic Diversity in Egyptian Date Palm

Microsatellite Analysis
A total of 14 microsatellite date palm specific primer pairs were tested. These primers were developed by Elmeer
and Mattat (2012). PCR reaction was performed as described by Adawy et al, (2009).
AFLP Analysis
The AFLP analysis was performed using the AFLP Analysis System II (Invitrogen, USA) as described by Vos
et al. (1995). Initially, 24 AFLP primer combinations (PCs) were tested on male and female samples represent the three
date palm cultivars. Among these, the best fourteen PCs were selected on the basis of the number of bands, clarity of
pattern, and distribution on the gel. These were tested again on the twelve date palm samples (the males and females of
three Egyptian cultivars) in addition to the two bulks.
SCot Analysis
SCoT-PCR was performed as described by Adawy et al, (2014). A set of 23 18-mer primers were used against the
twelve date palm samples.
CDDP Analysis
A total of 21 CDDP primers were tested against the twelve date palm samples and two bulks. These primers were
originally developed by Collard and Mackill (2009b). PCR reaction was performed also as described by Collard and
Mackill (2009b).
ITAP Analysis
A total of 18 ITAP primer combinations were tested against the twelve date palm samples and two bulks. These
primer combinations were originally developed by Xiong et al.(2013). PCR reaction was performed as described by Xiong
et al. (2013).
Data Analysis
The generated/ amplified bands were scored visually. The bands were scored as present (1) or absent (0) to create
the binary data set. Polymorphism percentage was calculated by dividing number of amplified polymorphic bands by the
total number of amplified bands by the same primer or primer combination. To estimate the genetic similarity, Jaccards
coefficient (Jaccard, 1908) was used. A dendrogram was generated by cluster analysis using the un-weighted pair group
method of the arithmetic averages (UPGMA) for all different marker systems.

RESULTS AND DISCUSSIONS

RAPD Analysis
Ten genotypes representing the males and females of five date palm Egyptian cultivars were amplified using 100
RAPD oligonucleotide. Amplification products of 10 genotypes with these 100 RAPD primers yielded a total of 1084
scorable bands, out of which 270 (24.91%) were found to be polymorphic with an average 10.8 band/primer (Table 1).
A dendrogram (Figure 1) based on UPGMA analysis comprised two main clusters, the first cluster (The major)
was subsequently divided into two subclusters; the first subcluster comprised three female genotypes and one male
genotype, while, the second subcluster including mainly male genotypes(four out of five males). Meanwhile, the second
cluster involved only two female genotypes. The highest genetic similarity (96.0%) was observed between Hayani female
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and Samany female. While the lowest genetic similarity (89.0%) was detected between Bertamoda male and Siwi female
(Table 2).
In this context, Srivashtav et al. (2013) defined the genetic diversity among 8 female and male genotypes grown
in the Kutch region using 13 RAPD and 18 ISSR primers. They reported that RAPD assay was more efficient than ISSR
assay with regard to polymorphic detection; since, RAPD markers detected 39.77% as compared to 23.07% of ISSR
markers. Moreover, the cluster analysis by UPGMA showed that the dendrogram obtained by RAPD and combined
(RAPD + ISSR) were similar. The dendrogram came into two clusters; Cluster A consisted of early maturing, Ghanshyam
and Late maturing female genotypes with 0.81 to 0.88 Jaccards similarity range. Cluster B consisted of seasonal female,
Male-1, Male-2, Male-3 and Male-4 genotypes with 0.82 to 0.91 similarity range. Meanwhile, Hussein et al. (2005)
assessed the genetic relationships among 14 date palm accessions representing six Egyptian date palm cultivars using 27
RAPD and 10 ISSR primers. They demonstrated that the level of polymorphism revealed by RAPD and ISSR was 25.2%
and 28.6%, respectively. They interpreted these low levels of polymorphism due to the narrow genetic background of these
accessions. They also reported that RAPD and ISSR based dendrograms were clustered the accessions belonging to each of
the 3 cultivars Fraihy, Siwi and Gandila in separate groups. However, the reshuffling in the position of some of the
accessions belonging to the other cultivars in the different dendrograms revealed that they share common genetic
background. Moreover, Al-Khalifah and Askari (2003) analyzed the genetic diversity among 13 different commercial
cultivars of date palm originating from various plantation areas of the Kingdom of Saudi Arabia using 144 RAPD primers.
The cluster analysis by UPGMA showed two main clusters. Cluster A consisted of five cultivars (Shehel, Om-Kobar,
Ajwa, Om-Hammam and Bareem), cluster B consisted of seven cultivars (Rabeeha, Shishi, Nabtet Saif, Sugai, Sukkary
Asfar, Sukkary Hamra and Nabtet Sultan). On the other hand, Adawy et al. (2002) employed 15 RAPD and 7 ISSR
primers to assess the polymorphism among four Egyptiandate palm cultivars (Samany, Zaghloul, Siwi and Hayany) and to
identify additional unique markers characterizing each cultivar. They reported that all tested primers exhibited intravarietal
polymorphism.
Table 1: Marker Types, No. of Primers, Total Bands, Polymorphic Bands,
Percentage of Polymorphism and Average for Each Marker Type
Marker Type
RAPD
SSR
CDDP
ITAP
SCoT
AFLP

No. of
Primers

Total
Bands

Polymorphic
Bands

% of
Polymorphism

Average

100
14
21
18
48
14

1084
83
192
204
484
591

270
49
61
70
200
221

24.9
59.0
31.7
34.0
41.3
37.4

10.8
5.9
9.1
11.3
10.1
42.2

Table 2: Genetic Distances Values among the 10 Date Palm Genotypes Consisting of 1-5 are Males
(Bertamoda, Hayani, Samany, Siwi and Zaghloul) and 6-10 are Females (Bertamoda,
Hayani, Samany, Siwi and Zaghloul) as Revealed by RAPD Data

Impact Factor (JCC): 2.8872

Index Copernicus Value (ICV): 3.0

A Multidisciplinary Molecular Marker Approaches to Assess the Genetic Diversity in Egyptian Date Palm

Figure 1: Dendrogram Depicting the Genetic Relationship among 10 Date Palm Genotypes
Consisting of 1-5 are Males (Bertamoda, Hayani, Samany, Siwi and Zaghloul) and 6-10 are Females
(Bertamoda, Hayani, Samany, Siwi and Zaghloul) Based on RAPD Data Microsatellite Analysis
Twelve date palm genotypes representing the males and females of three Egyptian cultivars were amplified using
14 SSR primer pairs. Amplification products yielded a total no. of 83 scorable bands, out of them 49 (59%) were found to
be polymorphic with an average 5.9 band/primer (Table 1). A dendrogram based on UPGMA analysis comprised three
main clusters, the first cluster including four male genotypes (two Zaghloul, one Hayani and one Samany), while, the
second cluster comprised three Hayani genotypes (two females and one male) in addition to one Zaghloul female. Finally,
the third cluster including three Samany genotypes (two females and one male)(Figure 2). The highest genetic similarity
was (99.0%) observed between Hayani females. While, the lowest genetic similarity (70.0%) was detected between
Zaghloulmale and Hayani females (Table 2).
In this context, Khierallah et al. (2011a) characterized 30 Iraqi date palm cultivars (representing 24 female and six
male cultivars) using 22 microsatellite SSR markers. They revealed that 188 alleles were detected at the 22 loci.
The genetic distance among cultivars varied from 0.171 to 0.938 indicating diverse relationships. Moreover, Elmeer et al.
(2011) used recently 23 developed new microsatellite markers to assess the genetic diversity among 11 cultivars chosen
randomly from different locations in Qatar. These primers generated a total of 77 alleles with a mean of 7.7 alleles per
locus. The average of gene diversity was 0.80 ranging from 0.6 to 0.9. Furthermore, Adawy et al. (2009) estimated the
genetic relatedness and generate unique fingerprint characterizing each genotype among 9 Egyptian date palm cultivars
using 25 SSR primers. These primers generated a total of 99 alleles with an average of 3.96 alleles with an average level of
polymorphism was 50.1%. They reported that the obtained dendrogram was divided into two main clusters, one cluster
included dry cultivars. While, the second cluster included two subclusters, the first subcluster included semi dry cultivars
and the second subcluster included soft cultivars. In addition, Elshibli and Korpelainen (2008) investigated the genetic
diversity of date palms in Sudan and to reveal the genetic relationships among 37 date palm cultivars using microsatellite
markers. They used 16 microsatellite markers developed for Phoenix dactylifera by Billotte et al. (2004). They detected a
total of 343 alleles at the 16 loci. The number of alleles per marker ranged from 14 to 44 with an average of 21.4 per locus.
They reported that the genetic groups of the Sudan cultivars and/or males do not follow a clear geographic pattern.
Also, Billotte et al. (2004) published a new SSR resources for P. dactylifera, to be used for cultivar identification, pedigree
analysis, germplasm diversity as well as genetic mapping studies. They designed a set of 16 PCR primer pairs for
microsatellite amplification based on appropriate consensus sequences. They performed PCR amplification using these
developed primers on P. dactylifera from various European, African and Middle East origins. They found that all 16 SSR
primer pairs amplified successfully in P. dactylifera with a total no. of alleles 166 with an average total number of alleles
per locus of 14 and no more than two displayed bands (nuclear SSR loci) except for some erratic amplification for the
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locus mPdCIR044.
AFLP Analysis
The twelve date palm genotypes were amplified using 14 AFLP primer combinations. Amplification products
yielded a total no. of 591 scorable bands, out of them 221 (37.4%) were found to be polymorphic with an average 42.2
band/primer (Table 1). A dendrogram comprised three main clusters, the major cluster (including 8 out of the 12
genotypes) divided into two subclusters; the first subcluster including three genotypes (two Hayani females and one
Hayani male), while, the second subcluster comprised five genotypes (three Zaghloulgenotypes and two Samany
genotypes). While, the second cluster collected three males genotypes representing the three cultivars. Meanwhile, the third
cluster including only one Samany female genotype (Figure 2). The highest genetic similarity was (96.0%) detected
between Hayani females. While the lowest similarity (89.0%) was detected between Zaghloul male and Hayani male
(Table 2).
In this manner, Khierallah et al. (2011b) used six AFLP primer combinations to identify the genotypes and
genetic characterization of 18 varieties (11 females and 7 males) of Iraq date palm. A total of 252 easily scorable bands
were generated. Among them 83 polymorphic AFLP fragments were detected with an average of 13.8 polymorphic
fragments/primer combination. Genetic distance ranged from 0.07 to 0.75. They found that UPGMA analysis ordered date
palm varieties into two main clusters independently of their origin and sex. While, El-Assar et al. (2005) used four
fluorescent-labeled AFLP primer sets to study the genetic variation of date accessions from Egypt (forty-seven accessions
from Egypt and two accessions from USA). A total of 350 bands were scored and 233 (66.6%) were polymorphic.
They reported that twenty-seven Egyptian accessions and Medjool and Deglet Noor accessions from California were
classified into the major cluster. They concluded that six groups of accessions of which had the same names, revealed
similar but not identical AFLP profiles. They suggested that these accessions might be derive from seedlings rather than
through clonal offshoot propagation. Meanwhile, Adawy et al. (2005) estimated the level of variability and assessed the
genetic relationships among six date palm cultivars (Fourteen accessions) using 16 AFLP primer combinations. AFLP
analysis generated a total of 657 amplicons representing a level of polymorphism of 45.8%. The level of polymorphism
ranged from 17.6% to 80.8%. They reported that the dendrogram comprised two main clusters, one containing Siwi and
Fraihy while the second cluster revealed two subclusters; Malkaby was separated in one subcluster while all the other
cultivars formed the second subcluster. Besides that, El- Khishin et al. (2003) estimated the level of polymorphism among
five Egyptian cultivars and determined the genetic relationships among these genotypes using AFLP analysis. A total of
433 amplification products were generated from six primer combinations with a mean of 72.17 amplicons.
The dendrogram comprised two major clusters; Siwi and Hayani being the most genetically similar cultivars, and in the
second cluster Amhat and Samany being next, while Zaghloul was the most distinct cultivar.
SCot Analysis
The SCoT marker technique was employed in the present study for several reasons. Firstly, it is a type of targeted
molecular marker technique characterized by simplicity and reproducibility. Its PCR products are resolved by performing
agarose gel electrophoresis. Compared to arbitrary markers such as RAPD, SCoT markers are highly reproducible due to
the use of longer primers. In general, recent few studies were employed SCoT-PCR to evaluate the genetic relationships in
rice (Collard and Mackill, 2009a), mango (Luo et al, 2010), peanut (Xiong et al, 2011) and Chinese sugarcane (Que et al,
2014). Particularly, only one study was conducted on date palm (Adawy et al, 2014). Therefore, the current study
Impact Factor (JCC): 2.8872

Index Copernicus Value (ICV): 3.0

A Multidisciplinary Molecular Marker Approaches to Assess the Genetic Diversity in Egyptian Date Palm

represents the first case focusing on the uses of SCoT assay in evaluation of the genetic diversity and genetic relationships
on date palm.
In the present study, 48 SCoT primers were used to amplify the twelve date palm genotypes, which yielded a total
scorable bands 484, out of them 200 (41.3%) were found to be polymorphic with an average 10.1 band/primer (Table 1).
A dendrogram based on UPGMA analysis comprised two main clusters, the major cluster divided into two subclusters; the
first subcluster comprised five female genotypes and only one male genotype. While, the second subclusters involved four
male genotypesand only one female genotype. Meanwhile, the second cluster comprised only one Samany male genotype
(Figure 2). The highest genetic similarity (95.0%) was observed between Hayani females. While, the lowest genetic
similarity (85.8%) was detected between Samany male and Zaghloul female (Table 2).
In this context, Adawy et al. (2014) used twenty-three SCoT primers against twelve date palm male and female
samples in order to identify any sex-specific markers. They successfully identified two differential amplicons/bands can
distinguish between males and females in Egyptian cultivars.
CDDP Analysis
Collard and Mackill (2009b) exploited conserved DNA regions within a selection of well characterized plant
genes primarily involved in response to abiotic and biotic stresses or plant development to generate DNA markers. It was
reasoned that these short conserved gene sequences should be present at multiple sites within plant genomes and, therefore,
provide multiple primer binding sites. We refer to this method as the conserved DNA-derived polymorphism (CDDP)
method. Compared to the RAPD method, this method uses longer primers and higher annealing temperatures that should
improve reproducibility. Furthermore, this method focuses on gene regions which may have advantages over random
markers for applications in QTL mapping (Andersen and Lubberstedt, 2003). To the best of our knowledge, no report has
focused on the applications of CDDP technique on date palm to date.
In the present investigation, a total of 21 CDDP primers were tested against the twelve date palm samples and two
bulks. These yielded a total scorable bands 192, out of them 61 (31.7%) were found to be polymorphic with an average 9.1
band/primer (Table 1). A dendrogram comprised two clusters, the major cluster divided into two subclusters; the first
subcluster comprised eight genotypes (five female genotypes and three male genotypes). While, the second subclusters
involved two male genotypes. Meanwhile, the second cluster comprised only two Samany genotypes (male and female)
(Figure 2). According to the cluster analysis, the CDDP successfully clustered males and females belonging to the same
cultivar in the same group. The highest genetic similarity (98.0%) was observed between Hayani females. While, the
lowest genetic similarity (85.8%) was detected between Samany male and Zaghloul female (Table 2). In this context,
Collard and Mackill (2009b) developed and applied twelve CDDP primers in order to fingerprint ten important rice
varieties and breeding lines (comprising eight indica and two japonica lines). A total of 54 polymorphic markers were
generated. Clustering of the rice genotypes was generally consistent with known taxonomic and pedigree information and
the results from Collard and Mackill (2009a).
ITAP Analysis
Xiong et al. (2013) originally developed PCR primers based on the 3 widely distributed consensus intron-exon
splice junction sequences which can be utilized in combination with the other arbitrary primers retrieved from SRAP
technique. They technique called intron targeted amplified polymorphism (ITAP) which described and used for
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fingerprinting in banana, longan, and cultivated peanut.

In the present study, 18 ITAP primer combinations applied on 12 date palm genotypes yielded a total scorable
bands 204, out of them 70 (34%) were found to be polymorphic with an average 11.3 band/primer (Table 1).
A dendrogram comprised two clusters, the major cluster involved all genotypes except only one Zaghloul male. The major
cluster divided into two subclusters; the first subcluster comprised all Hayani genotypes except one. While, the second
subclusters collected all Samany genotypes and all Zaghloul genotypes except one Zaghloul male (Figure 2). The highest
genetic similarity (96.0%) was observed between Samany females. While, the lowest genetic similarity (89.6%) was
detected between Zaghloul male and Hayani male (Table 2). In this context, Xiong et al. (2013) applied 5, 6 and 34 ITAP
primer combinations on three plant species including banana, longan and cultivated peanut, respectively. These generated a
total bands 40, 53 and 117 bands. Of these bands, 80%, 79.25% and 45.30% were polymorphic. They reported that cluster
analysis of ITAP markers was largely consistent with previous studies.
Combined Data
The cluster analysis of the 12 studied genotypes using these different marker systems revealed five dendrograms
exhibiting unique topology with some similarities. The data scored from SSR, AFLP, SCoT, CDDP and ITAP were
combined and computed to generate more accurate relationships based on large and versatile genome coverage.
A dendrogram based on UPGMA analysis comprised two main clusters, the major cluster divided into two
subclusters; the first subcluster comprised three Hayani genotypes. While, the second subclusters involved three Zaghloul
genotypes and three Samany genotypes (males and females). Meanwhile, the second cluster comprised three male
genotypes from Samany, Hayani and Zaghloul (Figure 2). The highest genetic similarity (96.0%) was observed between
Hayani females. While, the lowest genetic similarity (88.7%) was detected between Zaghloul male and Hayani females
(Table 2).The interpretation of this results was in a good agreement with Hussein et al, (2002)results, which reported that
the reshuffling in the position of some of the genotypes within the different dendrograms could be attributed to the narrow
genetic background of these date palm cultivars, the dioecious nature of date palm or the occurrence of gene flow among
these genotypes. In addition, incomplete similarity of the dendrograms could be also attributed to the nature of
evolutionary mechanisms underlying the variation measured by the different types of markers (Hussein et al, 2002).In this
context, Adawy et al. (2005) compared the efficiency of AFLP markers in date palm with RAPD and ISSR markers.
They used 16 AFLP primer combinations, 27 RAPD and 10 ISSR primers to estimate the level of variability and genetic
relationships among six date palm cultivars (Fourteen accessions). They reported that the dendrogram based on the
combined data from the different types of markers (RAPD, ISSR and AFLP) was closest to the AFLP-based dendrogram.

Impact Factor (JCC): 2.8872

Index Copernicus Value (ICV): 3.0

A Multidisciplinary Molecular Marker Approaches to Assess the Genetic Diversity in Egyptian Date Palm

Figure 2: Dendrogram Depicting the Genetic Relationship among 12 Date Palm Genotypes
Consisting of 1-6 are Males (1, 2: Zaghloul, 3, 4: Hayani and 5,6: Samany) and 7-12 are Females (7, 8: Zaghloul, 9,
10: Hayani and 11, 12: Samany) Based on AFLP, CDDP, ITAP, SCot, SSR and Combined Data
Table 3: Genetic Distances Values among the 12 Date Palm Genotypes Consisting of 1-6 are Males
(1, 2: Zaghloul, 3, 4: Hayani and 5, 6: Samany) and 7-12 are Females (7, 8: Zaghloul, 9, 10: Hayani and 11, 12:
Samany) as Revealed By AFLP, CDDP, ITAP, Scot, SSR and Combined Data

CONCLUSIONS
The present study successfully developed a multidisciplinary molecular marker approaches including three novel
marker types (SCoT, CDDP and ITAP) in addition to (SSR, AFLP and RAPD) to define the genetic diversity among male
and female genotypes belonging to different Egyptian date palm cultivars. The results demonstrated that different marker
systems differ in the mechanism of detecting polymorphism, genome coverage and the ease of application. Therefore, they
could complement each other to draw more accurate conclusions.

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10

Sami S. Adawy & Mohamed A. M. Atia

ACKNOWLEDGEMENTS
This research project was carried out in Agricultural Genetic Engineering Research Institute and funded by the
Science and Technological Development Fund (STDF) under the program of U.S.- Egypt Joint Board on Scientific and
Technological Cooperation, Contract No. 4607 titled Germplasm conservation, genetic characterization, next-generation
sequencing, molecular markers and bioinformatics analysis to distinguish the genders of date palm trees.

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