American Journal of Plant Sciences, 2012, 3, 1241-1251
http://dx.doi.org/10.4236/ajps.2012.39150 Published Online September 2012 (http://www.SciRP.org/journal/ajps)
1241
Assessments of Biodiversity Based on Molecular Markers
and Morphological Traits among West-Bank, Palestine Fig
Genotypes (Ficus carica L.)
Rezq Basheer-Salimia1*, Murad Awad1, Joy Ward2
1Department of Plant Production and Protection, Faculty of Agriculture, Hebron University, West-Bank, Palestine; 2Department of
Ecology & Evolutionary Biology, University of Kansas, Lawrence, USA.
Received July 10th, 2012; revised August 6th, 2012; accepted August 17th, 2012
ABSTRACT
Both morphological characters and PCR-based RAPD approaches were used to determine the genetic diversity and re-
latedness among nine fig genotypes grown at the northern region of the West-Bank, Palestine. Although we tested 28
primers for the RAPD technique, only 9 produced reasonable amplification products. A total of 57 DNA loci were de-
tected in which 70.2% were polymorphic. DNA fragments presented a minimum of 3 and a maximum of 9 polymorphic
bands using primers OPT-10 and OPA-18, respectively. Primers exhibited collective resolving power values (Rp) of
18.826. The Mwazi genotype showed the highest genetic distances among all of the other genotypes. Morphologically,
considerable variations were found using 41 quantitative and qualitative traits. Adloni could be a very promising geno-
type for fresh consumption due to its very late maturation period, extended harvesting period, variable fruit size, and
easy skin peeling. In addition, 7 genotypes presented firm fruits, which are a very important criterion for exporting
purposes. Dendrogram constructed by UPGMA based on RAPD banding patterns appear somewhat contradictory to the
morphological descriptors particularly with Swadi and Biadi genotypes (closed genetically and distanced morphologi-
cally), which might be attributed to the phenotypic modifications caused by environmental differences across regions.
These preliminary results will make a fundamental contribution to further genetic improvement of fig crops for the re-
gion.
Keywords: Cluster Analysis; Ficus carica L.; Genetic Variability; Random Amplified Polymorphic DNA;
Morphological Descriptors
1. Introduction
Fig (Ficus carica L., Moraceae) is one of the oldest cul-
tivated fruit crops grown in the Mediterranean region.
Because of their nutritional, medicinal, and ornamental
values [1], figs have recently attracted a great deal of
attention for culinary purposes and therefore, are wide-
spread throughout the world. According to FAO statistics
[2], the world produces over one million metric tons of
figs yearly, of which 82% are produced in Mediterranean
countries. Middle East countries have been the most im-
portant center of fig production across several millennia
[3]. The discovery of carbonized figs in an early Neoli-
thic site in the Jordan Valley (between Jordan and Pale-
stine), dating back 11,400 - 11,200 years ago, suggests
that figs were first domesticated during the early Neoli-
thic Revolution preceding cereal domestication [4]. From
there, fig cultivation spread to neighboring western Asia
and other Middle-East regions, and subsequently across
the rest of the World [5].
In Palestine, fig trees are grown historically all over
the country and are mostly located on marginal lands, in
combination with other fruit trees (mainly olive and
grape), or are scattered at the periphery of orchards and
in home gardens. The long history of fig growth in Pale-
stine and the wide range of geographical and climatical
conditions under which it is grown, have combined to
produce a complex picture in which fig landraces and
genotypes are either misidentified or called by different
names in different areas. Additionally, fig names were
mainly given based on fruit skin color, internal color,
local geographic origin, maturity dates, or the name of
the orchard owner [3,6]. Therefore, it is crucial for discri-
mination between these landraces both for conservation
of plant genetic resources and for purposes of crop im-
provement [7,8].
Detection and analyzing of genetic variation could be
*Corresponding author.

Assessments of Biodiversity Based on Molecular Markers and Morphological Traits among West-Bank,
Palestine Fig Genotypes (Ficus carica L.)
1242
achieved either by morphological and/or DNA molecular
markers [9]. Here we make major advances in under-
standing the history and genetic variation of fig by com-
bining both morphological and DNA marker approaches
for fig trees located throughout the West-Bank, Palestine.
Morphological markers have been used for many years
for identification and characterization of genotypes. In
fig, several reports demonstrated the usefulness of these
markers in documenting variability among genotypes [6,
9-11]. However, morphological characters can often
yield ambiguous results due to high plasticity for many
traits, as well as phenotypic modifications caused by
environmental differences [12].
The limitations of phenotype-based genetic markers
led to the development of more general and now wide-
spread use of DNA-based markers [13], which proved to
be powerful tools to estimate genetic diversity of species,
as well as genotype identity. In fact, molecular markers
offer numerous advantages over conventional morpholo-
gical based approaches, since they are stable and detec-
table in all tissues regardless of growth, differentiation,
development, or defense status of the cell. In addition,
DNA markers are not confounded by the environment,
pleiotropic, and epistatic effects [13]. In figs, assessment
of genetic relatedness and diversity has been investigated
by using RFLP, AFLP, SSR, ISSR, and RAPD methods
[7,14-22]. Compared with other molecular techniques,
RAPD is based on random amplification of bases from
short primers. Interestingly, RAPD is a simple, fast,
efficient, and inexpensive method. Further, it does not
require prior knowledge of the sequences of the markers
and can produce abundant polymorphic fragments [22,
23]. Therefore, RAPD has become a powerful and accu-
rate tool for analyzing the genetic relatedness and diver-
sity in figs.
Combinations of morphological as well as molecular
markers prove to be essential since morphological mar-
kers continue to be a highly recommended first step for
the description and classification of any germplasm [24],
as well as useful tools for screening the accessions of any
collection [25]. The present study is the first attempt to
characterize and detect similarities among some fig geno-
types grown in the northern region of Palestine using
both the powerful combination of morphological and
molecular markers.
2. Materials and Methods
2.1. Molecular Analysis
2.1.1. G en etic Markers
This study was carried out during the growing season of
2011. A total of nine fig accessions including: Khorto-
mani, Enaki, Hmadi, Hmari, Khdari, Biadi, Mwazi, Swa-
di, and Adloni were surveyed throughout the northern
region of West-Bank, Palestine. The climate of the re-
gion is an atypical Mediterranean climate, with mild tem-
peratures (18˚C - 25˚C), rainy weather (580 - 800 mm/year)
in autumn and winter, and hot, dry summers. Generally,
all fig trees are cultivated under rain-fed conditions.
2.1.2. D NA Extracti on
Genomic DNA was extracted from fresh leaves of single
adult trees using the DNeasy Plant Mini Kit (Qiagen
Inc.).
2.1.3. RAPD Primers and PCR Reactions
A total of 28 RAPD primers (Sigma-Aldrich) were used
for the amplification of random DNA banding patterns
(Tables 1, 2). PCR reactions were repeated twice and
carried out in a 25 ml volume mixture containing: 5 µl of
total DNA (30 ng), 2 µl primer (5 µM), 2 µl dNTPs (200
mM) (Fermentas), 2.5 µl Taq buffer (10×), 2 µl magne-
sium chloride (25 mM) and 1.5 U of Taq DNA poly-
merase (Hy Labs). Consequently, DNA was amplified by
PCR on a Peltier Thermal Cycler-200 (MJ Research. Inc,
Watertown, MA) and the PCR program was: 1 cycle,
94˚C (3 min); 35 cycles, 94˚C (1 min), 35˚C (1 min), 72
(1; 30 min) 1 cycle, 72˚C (5 min), followed by storage at
4˚C.
Amplified products (25 µl) were mixed with 5 µl of
orange gel loading buffer and analyzed by electrophore-
sis in 2% agarose gels (Hy Labs) in 1× TAE buffer at 4
volt/cm for 4h as well as detected by staining with ethi-
dium bromide (Sigma). A 100 bp DNA ladder was used
as standard marker (Fermentas). Consequently, ampli-
cons were visualized with a UV transilluminator (Ima-
geMaster®VDS).
2.1.4. D ata Analysi s of RAPD Mar k ers
For each primer, three independent researchers calculated
the total number of bands and the polymorphic bands in
order to avoid subject bias. The ability of the most in-
formative primers to differentiate between genotypes was
assessed by the estimation of their resolving power (Rp)
[26]. The Rp has been described to correlate strongly
with the ability to distinguish between genotypes ac-
cording to the following formula: Rp = ∑Ib, where Ib = 1
– (2 × |0.5 – p|) where p is the proportion of the nine geno-
types possessing the I band [27]. Banding profile data
were scored as present (1) or absent (0) for each sample.
Afterwards, RAPD bands were transformed into a binary
matrix. Next, a genetic distance matrix was estimated
(Table 3) based on Jaccard’s similarity coefficient using
the multilocus fingerprinting data sets containing missing
data (FAMD) software version 1.108 beta. Then a cluster
analysis was made using the un-weighted pair-group

Assessments of Biodiversity Based on Molecular Markers and Morphological Traits among West-Bank,
Palestine Fig Genotypes (Ficus carica L.)
1243
Table 1. List of the selected primers along with their sequences, number of banding patterns, resolving pow er, total and po-
lymorphic bands gener ate d that are resultant from all tested fig genotypes.
Resolving Power
(Rp)
Frequency
Number
of Patterns
Percentage
of
Polymorphic
Markers
Mono-
morphic
Bands
Poly-
Morphic
Bands
RAPD
Total Bands
Sequence 5'-3'Primer Code #
1.554
3/9
2/9
1/9
1
2
2
80 1 4 5 TCCCATGCTGZ-5 1.
3.166
3/9
1/9
1
6
88 1 7 8 GGGTGGGTAAZ-8 2.
1.999
2/9
1/9
1
7
60 2 3 5 CTCAGTCGCAZ-11 3.
2.443
2/9
1/9
2
7
63 3 5 8 CAATCGCCGTOPA-11 4.
2.722
2/9
1/9
1
7
63 3 5 8 CAGCACCCACOPA-13 5.
1.777
4/9
2/9
1/9
1
1
3
56 4 5 9 AGGTGACCGTOPA-18 6.
1.388
4/9
3/9
1/9
1
1
2
60 2 3 5 TCGGACGTGAOPH-02 7.
3.111 1/9 9 83 1 5 6 CTGACCAGCCOPH-19 8.
0.666
7/9
1/9
1
2
100 0 3 3 CCTTCGGAAGOPT-10 9.
2.092 4.44 6.33 Mean
18.826 70.2% 17 40 57 Total
Table 2. Summary of amplification presents gener ated by the random primers tested in this study.
Description Number/Frequency
Total number of primers screened with all the eleven fig genotypes19
Number of primers that produced polymorphic bands9
Total number of bands amplified by the primers that generated polymorphic bands57
Average number of bands per primer6.33
Total number of polymorphic bands40
Percentage of polymorphic bands70.2
Average number of polymorphic bands per primer4.44
Total number of primers that produced more than 75% polymorphic bands4
Total number of bands produced by these 4 primers23
Number of polymorphic bands produced by these 4 primers19
Percentage of polymorphic bands82.6
Average number of polymorphic bands per primer4.75
Average size of the fragments amplified190 bp - 1300 bp

Assessments of Biodiversity Based on Molecular Markers and Morphological Traits among West-Bank,
Palestine Fig Genotypes (Ficus carica L.)
1244
Table 3. Jaccard’s distance index generated for the 9 Palestinian fig genotypes based on RAP D mar kers.
Genotypes Swadi Biadi Hmadi Mwazi Khdari Adloni Enaki
Khorto
mani
Biadi 0.342
Hmadi 0.378 0.372
Mwazi 0.457 0.419 0.267
Khdari 0.333 0.368 0.286 0.444
Adloni 0.410 0.361 0.357 0.442 0.306
Enaki 0.368 0.405 0.357 0.477 0.306 0.432
Khorto
mani 0.419 0.452 0.256 0.302 0.325 0.400 0.270
Hmari 0.366 0.316 0.238 0.400 0.263 0.342 0.342 0.317
method with arithmetic averages (UPGMA) [28] and the
Tree view software (Win32, version 1.6.6).
2.2. Morphological and Pomological Analysis
2.2.1. P lant Mater ials and Descriptors
From the nine studied genotypes, random samples of 20
adult leaves and 20 mature fruits were collected from
three adult trees per genotype. 15 leaf morphological and
26 pomological traits (Tables 4, 5, respectively) were
determined according to the fig descriptors prepared by
IPGRI and CIHEAM [29], as well as Aljane and Ferchi-
chi [3], with some minor modifications that showed high
discrimination values.
2.2.2. Data Analysi s o f Mo rpholog i cal and
Pomological Traits
Relatedness as well as cluster analysis among genotypes
was established following the same method of RAPD
data analysis, in which each morphological descriptor
was scored in a dominant manner and transformed into
either a 1 (present) or 0 (absent).
3. Results
3.1. Molecular Results
3.1.1. Ge n etic Pol ymorphism and RAPD Patter n s
Examined primers revealed various banding patterns.
Among the 28 tested primers used for common fig
genotypes grown at the northern region West-Bank, Pal-
estine, only 9 primers produced reasonable amplification
products with high intensity and pattern stability (Table 1).
The remaining 19 primers exhibited ambiguous, light,
and non-clear complex amplification products, and there-
fore were excluded from our analysis. A total of 57 DNA
fragments (loci) separated by electrophoresis on agarose
gels, were detected (Table 1), ranging in size from 190
to 1300 bp. Of these fragments, 40 (70.2%) were poly-
merphic and 17 (29.8%) were monomorphic. Our results
also revealed an average of 6.33 loci per primer (Tables
1, 2). A minimum of three and a maximum of nine DNA
fragments were obtained using OPT-10 and OPA-18
primers, respectively (Figure 1). The maximum percent-
age of polymorphic markers was 100.0 (OPT-10).
3.1.2. Resolvin g P ower (Rp)
The tested primers exhibited a collective Rp value of
18.826, and varied from 0.666 for the (OPT-12) primer
to 3.166 for the (Z-8) with a mean of 2.092 (Table 2).
3.1.3. G e netic Di stances
The data matrix size analysed was 513 entries, 300 of
which were for present loci (1) and 213 for absent loci
(0). Accordingly, the Jaccard coefficient was calculated
and presented in (Table 3). The genetic distance matrix
showed an average distance range from 0.238 to 0.477
with a mean of 0.358. The maximum genetic distance
value of 0.477 was registered between Mwazi and Enaki
genotypes, whereas the lowest genetic distance of 0.238
(the highest similarities of 0.762) was exhibited between
the Hmadi and Hmari genotypes (Table 3).
3.1.4. D endrogram of Genetic Relationship
(Similarity Matrix and Cluster Analysis)
The average genetic relatedness among the genotypes is
illustrated in Figure 2. RAPD UPGMA dendrogram
analysis divided the genotypes studied into two main
clusters. The first (I) is made up of Biadi and Swadi
genotypes. The second cluster (II) is divided into two sub
clusters. The first sub cluster labelled (II.a) is made up of
(Khortomani and Enaki), as well as (Hmari and Hmadi)
related to Khdari. The second sub-cluster (II.b) is com-
posed of only genotype Adloni. However, Mwazi geno-
type is separated and identified as a distant genotype.
3.2. Morphological Descriptors
Fifteen quantitative and qualitative morphological traits

Assessments of Biodiversity Based on Molecular Markers and Morphological Traits among West-Bank,
Palestine Fig Genotypes (Ficus carica L.)
1245
Table 4. Morphological descriptors of some fig genotypes grown in the northern region of West-Bank, Palestine.
# Descriptors Khort-
omani Enaki Hmari Hmadi KhdariBiadi Mwazi Swadi Adloni
1 Bud Break Mar,
15 - 30
Mar,
15 - 30
Mar,
1 - 15
Mar,
1 - 15
Mar,
1 - 15
Mar,
1 - 15
Mar,
15 - 30
Mar,
1 - 15
Mar,
15 - 30
2 Leaf Color Light green Green
dark green Dark greenDark green Dark greenDark greenLight green green
Green
dark green
Light
green
3 Leaf Shape Base cordate,
lobes spatulate
Base
cordate, lobes
spatulate
Base
cordate,
lobes
spatulate
Base
cordate, lobes
spatulate-base
calcarate, lobes
latate
Base
cordate,
lobes
spatulate
Base
cordate,
lobes
spatulate
Base
cordate,
lobes
spatulate
Base
calcarate
lobes
lineate
Base
cordate, lobes
spatulate
4 Lobes Number Five Five Five Five Five Five Five Five Five
5 Leaf Venation Apparent Apparent ApparentApparent Apparent Apparent Apparent Apparent Apparent
6 Apex Shape Tri
Anleobtuse
Tri
Angle-
obtuse
Obtuse
rounded
Triangle-
obtuse TriangleTriangleTriangle Triangle Obtuserounded
7 Counter Crenate
dentate
Crenate
dentate
Crenate
undulate
Crenate
undulate
Crenate
CrenateCrenate Crenate undulate Crenate
8 Leaf Roughness Fairly
rough
Fairly
rough
Fairly
rough
Fairly
rough
Fairly
rough
Fairly
rough
Fairly
rough Rough Fairly rough
9 Leaf Area (cm2) Medium Large Large Large Large Large Small Medium Small
10 Limb Length (mm) Short Medium Medium Long MediumLong Short Medium Short
11 Limb Width (mm) Medium Medium Medium Medium Large Large Small Medium Small
12 Lateral Sinus Depth
(mm) Medium Medium Small Medium MediumMediumMedium Long Medium
13 Petiole Length (mm) Medium Long Long Long MediumMediumMedium Medium Medium
14 Petiole Width (mm) Medium Medium Medium Medium Large Large Small Small Small
15 Beginning of
Defoliation Sep-01 Sep-15 Aug-01 Aug-01 Aug-01Aug-01 Aug-20 Jul-01 Sep-15
are shown in Table 4. Bud break was observed between
March 1-15 in five genotypes (Hmari, Hmadi, Khdari,
Biadi and Swadi), with the remainder occurring between
March 15-30.
Among all genotypes tested, leaf color ranged from
light green to dark green, leaf shape was always base
chordate with lobes spatulate (except for Hmadi and
Swadi), lobe number was five, leaf venation was appar-
ent, leaf apex shape was variable, leaf serration tended to
be crenate, leaf roughness was fairly rough except for the
Swadi (rough) genotype. The Biadi genotype presented
the greatest value for leaf area, leaf limb length and
width, whereas Adloni tended to have the smallest values
for each of these parameters. Lateral sinus depth was
small for Hmari and long for Swadi, with the other
genotypes being medium for this trait. Petiole length was
long for genotypes Enaki, Hmari, and Hmadi and it was
medium for the remaining genotypes. Additionally, peti-
ole width was large for Biadi and Khdari genotypes, me-
dium for (Khortomani, Enaki, Hmari, and Hmadi), and
small for (Mwazi, Swadi, and Adloni).Defoliation was
early (July 1) for the Swadi genotype, while it was latest
(Sept 15) for the Enaki and Adloni genotypes; others
were intermediate.
3.3. Pomological Descriptors
Twenty-six quantitative and qualitative pomological traits
are presented in Table 5 . In terms of fruit maturation, the
nine genotypes studied were categorized very early
(Hmari, Hmadi, Khdari, Biadi, and Swadi), early (Khor-
tomani and Mwazi), or mid-season (Enaki and Adloni).
However, no genotypes were present which would be
categorized as either late or very late. The same trend
was observed for full fruit maturity for each genotype,
with the exception of Adloni, where maturation extended
beyond October 1 (very late). For all tested genotypes,
the harvesting period observed ranged from medium
(Khortomani, Hmari, Hmadi, Khdari, Biadi, and Swadi),
to long (Enaki), to very long (Mwazi and Adloni). Fruit

Assessments of Biodiversity Based on Molecular Markers and Morphological Traits among West-Bank,
Palestine Fig Genotypes (Ficus carica L.)
1246
Table 5. Pomological descriptors of some fig genotypes grown in the northern region of West-Bank, Palestine.
# Genotypes Khort-
omani Enaki Hmari Hmadi Khdari Biadi Mwazi Swadi Adloni
16 Beginning of
Maturation Early Mid season Very earlyVery earlyVery earlyVery earlyEarly Very earlyMid season
17 Full Maturity Early Mid-season Very earlyVery earlyVery earlyVery earlyEarly Very earlyVery late
18 Harvest Period Medium Long Medium Medium MediumMedium Very long Medium Very long
19 External Color Green-
purple
Green-
purple
Green-
purple
Green-
yellow
Green-
yellow
Green-
yellow
Green-
yellow
Black-
purple
Green-
yellow
20 Skin Cracks Cracked
skin
Scarce
longitudinal
cracks
Cracked
skin
Cracked
skin
Cracked
skin
Scarce
longitudinal
cracks
Cracked
skin
Scarce
longitudinal
cracks
Cracked
skin
21 Fruit Shape Pyriform Pyriform Ovoid Ovoid Ovoid PyriformPyriform Pyriform Pyriform
22 Fruit Symmetry No No Yes Yes Yes Yes Yes No Yes
23 Size Uniformity Uniform Variable Variable UniformVariable Variable Variable Variable Variable
24 Fruit Weight (g) Medium Large Medium Large MediumMedium Medium Medium Small
25 Fruit Firmness Soft Soft Soft Medium Firm Firm Medium Soft Soft
26 Fruit Length
(mm) Medium Long Short Medium Short Medium Medium Medium Medium
27 Fruit Width (mm) Medium Medium Medium Medium MediumMedium Medium Medium Small
28 Neck Length- mm Medium Medium Short Short Short Medium Short Short Medium
29 Neck Width (mm) Medium Medium Medium Small MediumLarge Large Medium Medium
30 Stalk Width (mm) Large Large Large Large Large Large Medium Large Small
31 Stalk Length (mm) Medium Medium Medium Long MediumLong Short Long Long
32 Ostiole Type Semi-openOpen Semi-openSemi-openClosed Semi-open Closed Semi-openClosed
33 Ostiole Drop Present Present Absent Absent Absent Absent Absent Absent Absent
34 Drop Color Transparent Transparent
35 Ostiole Width-mm Very largeVery large Very largeVery largeVery largeVery largeLarge Very largeLarge
36 Skin Peeling Medium Medium Medium Medium MediumEasy Easy Medium Easy
37 Internal Color Amber Amber Red White-redWhite-rosyWhite-redRosy-red White-redDark red
38 Flesh Thickness mm Medium Large Medium Medium Small Medium Large Medium Medium
39 Pulp Texture Fine Fine Medium Medium Medium Medium Fine Medium Medium
40 Pulp Flavor Aromatic Strong Strong Little flavorLittle flavorLittle flavor Strong Little flavorLittle flavor
41 TSS [%] High High High Very HighHigh High High High Low
Figure 1. Example of RAPD banding patterns generated in
Palestinian fig genotypes using OPA-19 primer. L: 1 Kb
ladder. 1: Swadi, 2: Biadi, 3: Adloni, 4: Enaki, 5: Khorto-
mani, 6: Khdari, 7: Mwazi, 8: Hmadi, and 9: Hmari.
external color for all genotypes was either green-purple
or green-yellow, except for the Swadi genotype, which
was black-purple. Regarding skin cracks, none of the
genotypes in our study were categorized as minute. In
our nine genotypes, the frequency of fruit shape observed
was six (pyriform) and three (ovoid); none of our geno-
types were bell-shaped. In addition, six genotypes de-
monstrated symmetrical fruits and two-presented unifor-
mity of size.
The largest fruit weight observed (55.25 g) was ob-
tained with Enaki and the smallest (17.64 g) was ob-
tained with Adloni; other varieties were intermediate.
Similar trends were observed with fruit length and fruit

Assessments of Biodiversity Based on Molecular Markers and Morphological Traits among West-Bank,
Palestine Fig Genotypes (Ficus carica L.)
1247
Figure 2. Dendrogram of 9 local Palestinian fig genotypes constructed by UPGM A base d on RAPD banding patterns.
width. Genotypes Khdari and Biadi presented firm fruits,
whereas soft fruits were observed for genotypes Khorto-
mani, Enaki, Hmari, Swadi, and Adloni. Fruit neck
length, neck width, stalk width, and stalk length varied
among genotypes in our study.
Among all tested genotypes, three (Khdari, Mwazi,
and Adloni) presented closed ostiole type, five (Khorto-
mani, Hmari, Hmadi, Biadi, and Swadi) presented semi-
open, and one (Enaki) had open ostiole. Furthermore,
transparent ostiole dew (drop) was observed in Khorto-
mani and Enaki genotypes. Additionally, most of the
genotypes exhibited very large ostiole width. Three ge-
notypes (Biadi, Mwazi, and Adloni) presented easy skin
peeling and the remainders were medium for this trait.
Internal fruit color was highly variable in this study,
ranging from amber to dark red. Flesh thickness was
large for Enaki and Mwazi genotypes, and small for
Khdari genotypes; other genotypes were medium.
Pulp texture in all genotypes was either fine or me-
dium, whereas the strongest pulp aromatic flavor was
observed in Enaki, Hmari, and Mwazi genotypes. Total
soluble solid (TSS) was either high or very high in all
genotypes, with the exception of Adloni, which had low
TSS.
3.4. Dendrogram of Morphological and
Pomological Relationship (Similarity Matrix
and Cluster Analysis)
Genetic distances ranged from 0.374 to 0.654 (Table 6).
“Hmari and Khdari” were the most closely related geno-
types, followed by Hmari and Hmadi. In contrary, Adloni
and Enaki were the most distantly related ones. UPGMA
dendrogram clustered the genotypes into two main clus-
ters (Figure 3). The smallest cluster, I, was composed of
Adloni and Mwazi genotypes. The largest cluster, II,
consisted of two sub-clusters, namely IIa, and IIb. Sub
cluster (II.a) was composed of the Khortomani and Enaki
genotypes, whereas the major sub cluster (II.b) included
highly related Khdari and Biadi as well as the two other
related genotypes Hmari and Hmadi, all of which derived
from the Swadi genotype.
4. Discussion
Fig (Ficus carica, Moraceae) is the only species among
700 species belonging to the Ficus genus that bear edible
fruits with significant commercial value. During the last
20 years, many Mediterranean countries have investi-
gated the genetic diversity of this crop; however, up to
now, therereally has not been substantial progress made
on this issue. This study is the first attempt to charac-
terize the figs grown in Palestine using a combination of
the PCR-based RAPD technique and morphological
characterization.
At the molecular level, our results (Tables 1, 2) and
comparable studies in the literature presented high poly-
morphism ratio (70.2% in 9 RAPD primers) among fig
genotypes grown in the Mediterranean countries which
commonly ranged between 39% - 81% within the same
marker (39% in 12 RAPD primers [14]; 67% in 7 RAPD
primers [19]; 72% in 6 RAPD primers [10]; 70% in 13
RAPD primers [30]; 77% in 6 RAPD primers [7]; 81% in
7 RAPD primers [21]). Indeed, the high polymorphism
that was observed here is indicative of more divergent
genotypes, and consequently a potential for success in
future selection programs [31]. Additionally, the average
of 6.33 amplicons (loci) per primer presented in this
study (Table 2) was sufficient to produce useful fin-ger-
prints for genotype and clone discrimination [14,17].

Assessments of Biodiversity Based on Molecular Markers and Morphological Traits among West-Bank,
Palestine Fig Genotypes (Ficus carica L.)
1248
Table 6. Jaccard’s distance index generated for the 9 palestinian fig genotypes based on pomological & morphological de-
scriptors & RAPD markers.
Genotypes Swadi Biadi Hmadi Mwazi Khdari Adloni Enaki Khortomani
Biadi 0.441
Hmadi 0.509 0.439
Mwazi 0.625 0.581 0.566
Khdari 0.500 0.386 0.434 0.589
Adloni 0.632 0.542 0.596 0.510 0.580
Enaki 0.567 0.602 0.586 0.646 0.569 0.654
Khortomani 0.547 0.581 0.514 0.537 0.562 0.594 0.406
Hmari 0.490 0.479 0.378 0.628 0.374 0.610 0.515 0.510
Figure 3. Dendrogram of 9 local Palestinian fig genotypes constructed by UPGMA based on morphologica l and pomological
characters.
Therefore, we may confidently assume that the RAPD
technique can solve one of the major problems associated
with varietal identification in Palestinian figs.
Resolving power of the collective examined primers
showed relatively high values of 18.826 in which primers
Z-8 and OPH-19 seemed to be the most useful RAPD
primers to assess the genetic diversity since they revealed
relatively high collective Rp rates of 3.166 and 3.111,
respectively. A similar result was also reported for other
Mediterranean figs by Salhi-Hannachi [10] with Rp value
of 21.771 in 6 RAPD primers.
Among all tested genotypes, the Mwazi genotype
tends to show the highest genetic distance values from
the others. The lowest genetic distance of 0.238 between
Hmadi and Hmari genotypes (Table 3), suggests there
are very closely related, and possibly might be the same
genotype, but with different names. For the remaining
genotypes, the genetic distance matrix showed a high
level of divergence at the DNA level. Similar result was
reported by Baraket [32].
Collectively, the high polymorphism ratio, relatively
high Rp values, and good genetic distances presented in
our study might suggest high genetic diversity in Pales-
tinian fig population at the DNA level.
At the morphological and pomological levels, more
than forty-one informative and economical traits (both
quantitative and qualitative) were conducted in this study,
which was much greater than most regional studies per-
formed on fig genotypes in the past: 11 traits [33]; 16
trait [19]; 22 traits [34]; 26 traits [35]; and 39 traits [11].
Except for the leaf venation parameter that was found to
be similar for all tested genotypes, considerable variation

Assessments of Biodiversity Based on Molecular Markers and Morphological Traits among West-Bank,
Palestine Fig Genotypes (Ficus carica L.)
1249
among the genotypes was exhibited for the remaining
fourteen leaf traits (Ta ble 4), which may be highly effec-
tive in differentiating among fig genotypes [9,36].
A great diversity of fruit harvesting period, fruit size,
fruit weight, fruit uniformity, and fruit external color ex-
hibited in our examined genotypes (Table 5) could be
potentially incorporated to both local and regional breed-
ing programs. Such a high degree of morphological vari-
ation and fruit characteristics could be utilized to interest
farmers in diversifying fig genotypes [37], increasing fig
production [34], and stimulating attraction of consumers
for fresh fruit consumption. Additionally, the majority of
our genotypes presented firm fruits, short fruit neck, and
acceptable TSS (Tab le 5 ), which are very important qua-
lity criterion particularly for exporting purposes.
Dendrogram constructed by UPGMA based on RAPD
banding patterns (Figure 2) appear contradictory to the
morphological descriptors in some cases (Figure 3). This
was particularly the case for the Swadi and Biadi geno-
types, which were found to be close to each other ge-
netically, but were morphologically distanced. However,
for the remaining genotypes they were often similar. This
difference could be attributed to the phenotypic modify-
cations caused by the harshest prevailing weather condi-
tions in the Mediterranean basin where fig can grow [7,
12], particularly since plasticity in morphology is present
in fig [9]. However, RAPD is not confounded by the en-
vironment, pleiotropic and epistatic effects [13], and has
been found to be a useful and powerful tool to estimate
genetic diversity of species and genotype identity.
It is also important to point out through morphological,
pomological and genetic comparisons that the Mwazi
genotype was separated and identified as a distant geno-
type (Table 6) indicating that its genome may differ from
the other common fig genotypes [7].
5. Conclusion
Morphological and pomological results will be very use-
ful in characterizing each considered genotype and to
create the first reference and catalogue of the local Pales-
tinian fig genotypes. RAPD is a very useful technique in
characterizing Palestinian fig genotypes. Here we com-
bined morphological and genetic characters to further
refine this Palestinian fig database for use by both re-
searchers and farmers.
6. Acknowledgements
We thank the U.S. National Academy of Sciences and
the Kuwait Institute for Scientific Research for hosting
the Arab-American Frontiers Program that helped make
this collaboration among the authors possible. We also
thank both of these agencies especially to Kathrin Hum-
phrey and Dalal Najib for the seed grants to help support
preparation of the manuscript.
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