Available online: www.notulaebotanicae.ro
Print ISSN 0255-965X; Electronic 1842-4309
Not Bot Horti Agrobo, 2015, 43(1):XXX-XXX. DOI:10.15835/nbha4319773
Morphological and Molecular Characterization and Relationships of Turkish
Local Eggplant Heirlooms
Hatice Filiz BOYACI*, Volkan TOPCU, Akin TEPE, Isilay Karasahin YILDIRIM,
Mehmet OTEN, Aytekin AKTAS
Bati Akdeniz Agricultural Res. Inst., Dept. of Vegetable Crops and Ornamentals, 07100 Antalya,
Turkey; filiz_boyaci@yahoo.com (*corresponding author)
Abstract
A total of 38 eggplant genotypes, of which 32 were heirloom accessions collected from different regions of Burdur province
five were different local genotypes from other provinces, and one was a cultivar, were used as reference in this study. The
phylogenetic relationships among these heirlooms were evaluated using 40 morphologic descriptors and five randomly
amplified polymorphic RAPD markers. The horizontal dendrograms were created by using UPGMA with both morphologic
and molecular data. Burdur heirloom accessions showed high genetic diversity based on morphological and molecular data.
The genetic similarity rates ranged from 0.29 to 0.91 according to the morphological data, and ranged from 0.84 to 0.98
according to the molecular data. Molecular data generated by RAPD method, compared to morphological data, were
insufficient to reveal genetic diversity. Therefore, in order to confirm genetic variations, studies based on other molecular
methods are necessary. The regional genetic populations include a wide eggplant genetic diversity which can be good source for
the breeding studies performed in the future.
Keywords: accession, breeding, diversity, genetic variation, phylogenetic relationship, RAPD, UPGMA
Introduction
Eggplant, which was called as the king of vegetables (Daunay
and Janick, 2007), is an indigenous plant of India (Weese and
Bohs, 2010). It has been well known since BC III and cultivated
for 1,500 years in Asia (Kashyap et al., 2003). It is cultivated as a
perennial in tropical areas, while it is cultivated as annual in
subtropical areas (Kowalska, 2008). Eggplant is placed in
Solanum genus and includes wide genotypic and phenotypic
variation (Fukuoka et al., 2010). Eggplant is thought to have
been developed from the wild ancestor, Solanum insanum has
small, round, green, thick-skinned and bitter taste fruits (Barchi
et al., 2010). Cultivation S. insanum of had been performed in
China, India and Thailand (Daunay et al., 2001). Large fruiting
eggplants were cultured in India in early time, and small fruiting
was cultured at IV century in China and at IX century in Africa
(Sekera et al., 2007). First cultivated eggplants were described as
high tall plants, with big spines on the calyx, small, bitter fruiting
and with high seed content in fruits (Swarup, 1995). Mutation,
natural pollination and hybridization, together with selection
gave rise to genetic diversity, as well as in decreasing of prickles
and bitterness at fruit, changing of fruit shape, size and color
(Frary et al., 2007). Genetic diversity accumulated and many
different heirlooms emerged in countries where it was cultivated
(Prohens et al., 2003). Entrance of the eggplant to Turkey was
Received: 18.12.2014. Accepted: 06.04.2015.
carried out by the silk-road. The genetic diversity accumulated in
producing areas and by trade of eggplant throughout centuries in
Anatolia (Janick, 2001). Eggplant cultivation was done in open
field until the second half of the 1970s in Turkey and then
cultivation under protected cultivation started. The eggplant
cultivation in greenhouse was begun with local varieties.
However, the growers preferred hybrid F1 varieties which have
the cylindrical and dark purple or black colored fruiting and
using of them has become commonly in a short period (Ekiz and
Boyaci, 2001). Also, the increase of use F1 hybrids in the open
field cultivation was observed in recent years. Steadily decrease
was seen in the cultivation of local varieties compared to the
hybrids (Cericola et al., 2013). The genetic diversity is low among
the genotypes which have dark purple-black fruits (MuñozFalcón et al., 2009). In recent years, one of the important
problems faced by eggplant breeding programs, as well as in
other species, is a narrowing of the genetic base. To create
variations, time-consuming and expensive methods are
needed, including mutation breeding, interspecific
hybridization and biotechnological approaches. The genetic
variation contained by heirlooms among is seen in the
previous studies (Demir et al., 2010; Muñoz-Falcón et al.,
2008, 2009; Prohens et al., 2003, 2008, 2011). Therefore,
collection and characterization of genetic resources is required
for the improvement of new varieties. In this respect,
Boyacı HF et al. / Not Bot Horti Agrobo, 2015, 43(1):X-X
X
molecular characterization is more reliable then
morphological characterization (Li et al., 2010).
Here we investigate the genetic relationships between
Turkish heirlooms using morphological and molecular data is
presented here.
Materials and methods
Morphological and molecular studies were conducted at Bati
Akdeniz Agricultural Research Institute, Antalya, Turkey in
2010-2013.
In total 38 materials were used in the study of these. 32 were
local heirlooms collected from Burdur province during survey
studies. Materials origin, providing method and place/company
of origin are given in Table 1. Long purple commercial variety
(YRL 68) and heirlooms originating in other province (YRL 1,
YRL 3, YRL 6, YRL 57, and YRL 79) were used as reference
cultivars for classification of Burdur province heirlooms.
Five RAPD primers OPH-02, OPL-04, OPB-07, OPO10, OPL-16 (10 mer) reported as highly polymorphic in
previously studies (Demir et al., 2010; Nunome et al., 2001)
were selected to detect polymorphisms and identify genetic
relationship of the heirlooms.
locations of collected eggplants in Burdur province were
represented on the map (Fig. 1) and their geographic coordinates
were defined (Table 2).
Morphological observations
For each accession, the seeds were sown in seedling treys
containing peat moss. Seedlings in 4-5 leaves stages were
planted in glasshouse. Twenty plants were planted for each
genotype. Morphological observations were performed
according to 40 descriptors chosen among the International
Board for Plant Genetic Resources Institute (IBPGRI),
International Union for the Protection of New Varieties of
Survey
The survey studies were performed at nine different locations
in Burdur province in September both in 2010 and 2011. In Fig. 1. Geographic map of district and village in Burdur province of
total 32 materials were collected from different farms. The Turkey including collection places
Table 1. Origin, providing method and place/company of origin of the materials used in the study
No
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
Name of the materials
YRL 1
YRL 3
YRL 6
YRL 7
YRL 8
YRL 9
YRL 12
YRL 14
YRL 15
YRL 19
YRL 20
YRL 24
YRL 26
YRL 27
YRL 28
YRL 30
YRL 34
YRL 35
YRL 36
YRL 43
YRL 44
YRL 45
YRL 46
YRL 47
YRL 49
YRL 50
YRL 51
YRL 52
YRL 57
YRL 58
YRL 59
YRL 61
YRL 62
YRL 64
YRL 65
YRL 68
YRL 75
YRL 79
Origin
Antalya-Kumluca District
Antalya-Kumluca District
Antalya-Kumluca District
Burdur-Aglasun /Cine Village
Burdur-Aglasun /Cine Village
Burdur-Aglasun /Cine Village
Burdur/Askeriye Village
Burdur-Aglasun /Cine Village
Burdur-Aglasun /Cine Village
Burdur-Aglasun /Cine Village
Burdur-Aglasun /Cine Village
Burdur-Celtikci/Tekke Village
Burdur-Celtikci/Tekke Village
Burdur-Celtikci/Tekke Village
Burdur-Celtikci/Tekke Village
Burdur-Celtikci/Tekke Village
Burdur-Celtikci/Tekke Village
Burdur-Celtikci/Tekke Village
Burdur-Celtikci/Tekke Village
Burdur-Celtikci/Tekke Village
Burdur-Celtikci/Tekke Village
Burdur-Celtikci/Tekke Village
Burdur-Askeriye Village
Burdur-Askeriye Village
Burdur-Askeriye Village
Burdur-Askeriye Village
Burdur-Askeriye Village
Burdur-Askeriye Village
Mugla-Fethiye/Günesli Village
Burdur-Karamanlı/Manca Village
Burdur-Karamanlı/Manca Village
Burdur-Gölhisar/Sorkum Village
Burdur-Yesilova/Kayadibi Village
Burdur-Tefenni District
Burdur-Tefenni District
Long purple
Burdur City Centrum
Antalya-Akseki/Uzumdere Village
Providing method
Agricultural district offices
Agricultural district offices
Agricultural district offices
Survey in the field
Survey in the field
Survey in the field
Survey in the field
Survey in the field
Survey in the field
Survey in the field
Survey in the field
Survey in the field
Survey in the field
Survey in the field
Survey in the field
Survey in the field
Survey in the field
Survey in the field
Survey in the field
Survey in the field
Survey in the field
Survey in the field
Survey in the field
Survey in the field
Survey in the field
Survey in the field
Survey in the field
Survey in the field
Survey in the field
Survey in the field
Survey in the field
Survey in the field
Survey in the field
Survey in the field
Survey in the field
Purchased
Agricultural district offices
Survey in the field
Place/company of origin
Grower
Grower
Grower
Grower
Grower
Grower
Grower
Grower
Grower
Grower
Grower
Grower
Grower
Grower
Grower
Grower
Grower
Grower
Grower
Grower
Grower
Grower
Grower
Grower
Grower
Grower
Grower
Grower
Grower
Grower
Grower
Grower
Grower
Grower
Grower
Company
Grower
Grower
Boyacı HF et al. / Not Bot Horti Agrobo, 2015, 43(1):X-X
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Table 2. Geographical coordinates of local heirloom eggplant collected in Burdur
province of Turkey
Location
Aglasun /Cine Village
Geographical coordinates
37°35'0.07"N/30°38'1.34"E
Celtikci Tekke Village
37°34'36.86"N/30°26'40.11"E
Burdur/Askeriye Village
37°45'41.88"N/30°21'15.99"E
Karamanlı-Manca Village
37°18'18.74"N/29°53'19.89"E
Karamanlı-Centrum
37°22'2.35"N/29°49'19.71"E
Gölhisar-Sorkum Village
37° 9'54.49"N/29°34'39.38"E
Yesilova-Kayadibi Village
37°31'19.56"N/29°44'35.03"E
Tefenni-Centrum
37°18'27.39"N/29°46'40.57"E
Plants (UPOV) plant feature criteria and some of the criteria
for the breeders. Descriptors include plant, leaf, flower and
fruit traits observations and measurements (Table 3). Skin
color of the each eggplant specimens was measured with a
portable tristimulus reflectance colorimeter Minolta CR-400
Chroma Meter (Konica Minolta Sensing, Inc., Osaka, Japan),
and the parameters were expressed in CIE L*a*b* system,
where L* is lightness (brightness-darkness) ranged from 0 to
100 units, a* is light intensity in red (+) or green (–)
spectrum, b* is intensity in yellow (+) or blue (–) spectrum.
Chroma (C: √(a2+b2)) measures color saturation or intensity
and the hue angle (h ° = tan-1 (b*/a*) determines the red,
yellow, green, blue, purple, or intermediate colors between
adjacent pairs of these basic colors. The L*, a*, and b*, C and h
° values obtained from six samples of each eggplant accession.
Every record represents the average of three readings which
were taken from the equatorial region, spaced equidistantly.
DNA extractions and PCR analysis
DNA extractions from young leaves were performed
according to a modified Doyle and Doyle (1990) method by
using CTAB protocol (Mutlu et al., 2008). RAPD analysis was
performed according to Demir et al. (2010). The amplification
reactions were released containing 20 ng DNA, 0.5 unite Taq
polymerase, 2.5 L 10X buffer, 3.5 L 25 mM MgCl2, 2 L 2.5
mM dNTPs, 2 L RAPD primer for RAPD-PCR. DNA was
amplified in a thermal cycler. It was programmed for an initial 5
min denaturation step at 94 °C, followed by 35 cycles of a 30 s
denaturation step at 94 °C, 1 min annealing at 35 °C, 45 s
extension step at 72 °C, followed by a final 8 min extension step
at 72 °C.
Statistical analysis
Genetic similarity was analyzed by the UPGMA (Unweighted
pair-group method, arithmetic average) clustering procedure using
the software NTSYS (Numerical Taxonomy Multivariate
Table 3. Descriptors used for characterization and evaluation of eggplant accessions used in the study
Traits
Plant habit
Plant height
Stem thickness
Stem hairiness
Stem color
Shoot tip color
Length of internodes
Leaf color
Leaf size
Leaf hairiness
Presence of spine on petiole
Bud size
Bud hairiness
Presence of spine on bud
Flower color
Flower size
Calyx size
Fruit shape
Dominated fruit color
Range of dominated fruit color
Fruit stalk length
Presence of spine on fruit stalk
Fruit calyx prickles
Fruit brightness
Fruit end shape
Fruit curvature
Fruit end button size
Fruit length (cm)
Fruit diameter (cm)
Average fruit weight (g)
Presence of groove on fruit
Fruit flesh firmness
Fruit flesh color
Presence of hole in fruit
Degree of fruit curvature
Soluble solids
The length of fruit coated by calyx
Tendency to parthenocarpy
Presence of seed in fruit
Seed maturity
Description
Score range (1=Open, 3=Bushy, 5=Semi open)
Score range (1=Long, 3=Intermediate, 5=Short)
Score range (1=Thick, 3=Intermediate, 5=Thin)
Score range (1=Dense, 3=Intermediate, 5=Tenuous)
Score range (1=Grayish, 3=Green, 5=Green-purple, 7=Grayish-green-purple, 9=Grayish-green, 11= Grayish-purple, 13=Purple)
Score range (1=Grayish, 3=Green, 5=Green-purple, 7=Grayish-green-purple, 9=Grayish-green, 11=Grayish-purple, 13=Purple)
Score range (1=Long, 3=Intermediate, 5=Short)
Score range (1=Light green, 3=Green, 5= Dark green)
Score range (1=Large, 3=Intermediate, 5=Small)
Score range (1=Dense, 3=Intermediate, 5=Tenuous)
Score range (1=Many, 3=Intermediate, 5=Few, 7=Absent)
Score range (1=Large, 3=Intermediate, 5=Small)
Score range (1=Dense, 3=Intermediate, 5=Tenuous, 7=Absent)
Score range (1=Many, 3=Intermediate, 5=Few, 7=Absent)
Score range (1=Light purple, 3=Purple, 5= Dark purple)
Score range (1=Large, 3=Intermediate, 5=Small)
Score range (1=Large, 3=Intermediate, 5=Small)
Score range (1=Long, 3=Intermediate, 5=Short, 7=Ovoid, 9=Pear shaped)
Score range (1=White, 3= Green, 5=Purple, 7=Black)
Score range (1=Regular, 3=Mottled, 5=Stripe, 7=Mealy)
Score range (1=Long, 3=Intermediate, 5=Short)
Score range (1=Few, 3=Intermediate, 5=Many, 7=Absent)
Score range (1=Few (1-5 pricks), 3=Mid (6-20 pricks) , 5=Many (more than 20)
Score range (1=Bright, 3=Matt)
Score range (1=Flat, 3= Pointed, 5=Round)
Score range (1=Present, 3=Absent)
Score range (1=Large, 3= Intermediate, 5=Small)
The average measurement of ten fruits
The average measurement of ten fruits
The average measurement of ten fruits
Score range (1=Present, 3=Absent)
Score range (1=Tightly, 3= Floppy, 5=Spongy)
Score range (1=Greenish, 3=Greenish-cream, 5=White, 7=White- cream, 9=Greenish-white, 11=Cream)
Score range (1=Present, 3=Absent)
Score range (1=Slight, 3= Mid, 5=Much)
The average measurement of five fruits juice samples
Score range (1=Less than 20%, 3= between 20-70%, 5=More than 70%)
Score range (1=Present, 3=Absent)
Score range (1=Few, 3=Intermediate, 5=Many)
Score range (1=Immature, 3=Mature)
Boyacı HF et al. / Not Bot Horti Agrobo, 2015, 43(1):X-X
X
Analysis System) pc 2.2 version (Rohlf, 1998), using
morphological and molecular data. For morphological data, each
genotype was characterized with description number (Table 3).
For molecular analysis data, each genotype was identified for each
primer based on the presence (1) and absence (0) of bands. In
order to show the variations STANDARDIZATION module
was used. Correlation matrix adapted to SIMINT module was
used to determine of correlation coefficient. The dendrograms
were drawn using the clustering method UPGMA via the
SHAN module. The cophenetic correlation coefficient was
calculated with Mantel method to evaluate the efficiency of
clustering.
Results
A high morphological diversity was observed among
eggplant genotypes characterized by quantitative and qualitative
descriptors. Some of the phenotypic observation data related
with important variable traits (calyx prickles, fruit size, weight,
shape and curvature, skin color, groove etc.) are reported in Table
4. The fruits calyx of all genotypes had prickles. However, nearly
50% of them had lower numbers of prickles. Most of the
genotypes had between 20-70% of their fruit length covered by
the calyx. The degree of fruit curvature in 50% of the genotypes
was slight. Nearly 30% percent of the genotypes had fruits with
grooved appearance. Dominant fruit color was mostly purple
and distribution of this color was 50% regular, 22% mealy, 18%
mottled, and 2% striped according to the genotypes. Fruit flesh
color was generally white and hole in the fruit was mostly absent.
The lowest total soluble solid content (brix value) was 2.8. The
highest brix value (6.1) was determined in YRL 15 which was
collected from Cine village The average diameter of the fruits
ranged from 32.67 to 73.22 mm. Fruit length and fruit weight
average ranged between 10.9 and 23.3 cm, 0.100 and 0.235 kg,
respectively.
Skin color characteristics of the eggplant heirlooms are
shown in Table 5. Fruit color varied greatly among different
genotypes. Regarding the skin Lightness (L*), higher values
were obtained from genotypes which were mostly, cream,
yellowish, orange-yellowish or goldenred in color (L* > 70).
When L* values were ranged from 48 to 58 together with h°
values ranged from 80 to 112, it is observed that these
genotypes’ skin color were generally greenish-yellow, chartreuse
or green in color. Regarding the b* values, very low values near
or below zero mostly indicated that the skin color was cyan (if h
° > 285); blue (if h ° > 240); purple (if h ° > 285); magenta (if h °
> 350). It is observed from the research, that fruits which had
lower h ° values (1.05 < h ° < 12.60) were found to be red in
color. Chroma values were varied from 3.82 to 24.26.
Totally 40 basic morphological descriptors were used for to
determine the phylogenetic relationships among the Burdur
local heirlooms. The Eigen value was 84. A 2-way Mantel test
(Mantel, 1967) method was performed. Approximate Mantel
t- statistic test were t = 10.0925, p = 1.0000. The matrix
correlation (r) was 0.72. The similarity rates according to the
coefficient similarity of genotypes ranged between 0.29 and
0.91.
Two major groups were revealed using the dendrogram
generated by the UPGMA method using morphological data
(Fig. 2). First group (Group A) was consisted of YRL 75, YRL
65, YRL 61 and YRL 59. These genotypes showed low genetic
similarity with reference genotypes and cultivar (Long purple).
Second group was divided into three subgroups. Group B
consist of YRL 19 which showed high genetic similarity with
reference cultivar YRL 68 (Long purple). The highest genetic
similarity was observed in Group D between genotypes YRL
47 and YRL 51. The fruits of eggplant Burdur heirlooms
belonging to the Group C are shown in Fig. 3.
Table 4. Some of the phenotypic descriptors related important variable traits of eggplant
Fruit
calyx
prickles
few
few
mid
mid
mid
few
few
mid
mid
few
mid
few
few
mid
few
mid
mid
mid
mid
mid
mid
mid
mid
few
Fruit length
covered by the
calyx
between 20-70%
between 20-70%
between 20-70%
less than 20%
less than 20%
between 20-70%
less than 20%
between 20-70%
between 20-70%
between 20-70%
between 20-70%
less than 20%
between 20-70%
between 20-70%
less than 20%
less than 20%
between 20-70%
between 20-70%
between 20-70%
between 20-70%
between 20-70%
between 20-70%
less than 20%
less than 20%
YRL 49
few
YRL 50
YRL 51
YRL 52
YRL 57
YRL 58
YRL 59
YRL 61
YRL 62
YRL 64
YRL 65
YRL 68
YRL 75
YRL 79
mid
few
few
few
few
mid
mid
few
few
mid
mid
mid
few
Genotype
YRL 1
YRL 3
YRL 6
YRL 7
YRL 8
YRL 9
YRL 12
YRL 14
YRL 15
YRL 19
YRL 20
YRL 24
YRL 26
YRL 27
YRL 28
YRL 30
YRL 34
YRL 35
YRL 36
YRL 43
YRL 44
YRL 45
YRL 46
YRL 47
Distribution of
dominant
color
mottled
striped
mottled
regular
regular
regular
regular
regular
regular
regular
regular
regular
mealy
mealy
regular
mealy
mealy
mealy
mealy
mottled
mealy
mealy
regular
regular
Fruit
flesh
firmness
floppy
tightly
floppy
floppy
tightly
tightly
spongy
tightly
tightly
tightly
spongy
spongy
floppy
floppy
floppy
spongy
spongy
spongy
floppy
spongy
spongy
tightly
floppy
floppy
Degree of
fruit curvature
Fruit
grooved
Dominated
fruit color
slight
slight
slight
midslight
slight
slight
none
slight
slight
slight
slight
midslight
midmidslight
midmidnone
slight
slight
midmid-
absent
present
absent
absent
present
absent
absent
absent
present
absent
absent
absent
absent
absent
absent
absent
present
absent
absent
present
absent
present
present
present
green
purple
green
black
purple
black
purple
black
black
black
black
purple
green
purple
black
purple
purple
purple
purple
purple
purple
purple
purple
purple
less than 20%
mid-
absent
purple
regular
tightly
less than 20%
less than 20%
less than 20%
between 20-70%
less than 20%
between 20-70%
between 20-70%
between 20-70%
less than 20%
less than 20%
between 20-70%
between 20-70%
between 20-70%
midmidmidmidmidmidnone
slight
slight
slight
slight
none
slight
absent
present
absent
absent
absent
absent
present
absent
absent
present
present
present
present
purple
purple
purple
green
purple
purple
purple
black
purple
purple
black
black
green
regular
regular
regular
mottled
regular
mottled
regular
regular
regular
mottled
regular
regular
mottled
tightly
floppy
tightly
tightly
tightly
floppy
tightly
tightly
floppy
tightly
tightly
tightly
floppy
Hole
in the
fruit
absent
present
absent
absent
present
absent
present
present
present
absent
present
present
present
present
present
present
present
present
present
present
present
present
absent
absent
Fruit
diameter
(mm)
34.80
49.79
36.60
40.33
46.86
45.38
40.98
43.60
35.07
47.32
42.17
38.96
45.73
58.13
53.13
47.43
54.11
53.66
52.05
57.87
51.29
50.50
42.79
35.05
Fruit
lengtht
(cm)
19.8
14.2
23.3
18.7
15.7
18.5
20.5
17.2
18.8
17.5
20.6
18.3
16.7
18.0
16.3
14.5
22.1
17.5
14.8
15.1
15.2
14.6
16.4
18.0
Fruit
weight
(kg)
0.116
0.140
0.140
0.133
0.133
0.138
0.125
0.133
0.110
0.146
0.160
0.100
0.143
0.213
0.177
0.133
0.232
0.207
0.163
0.187
0.168
0.173
0.125
0.127
Fruit flesh
color
Seed
content
Seed
maturity
greenish-cream
white
greenish-cream
cream
greenish-cream
greenish-cream
white
greenish-cream
greenish-cream
greenish-cream
greenish-cream
greenish-cream
white
white
white
white
white
white
white
white
white
white
white
white
intermediate
many
intermediate
intermediate
intermediate
intermediate
intermediate
few
few
intermediate
many
few
intermediate
intermediate
intermediate
intermediate
few
few
intermediate
intermediate
intermediate
few
intermediate
intermediate
mature
mature
immature
mature
mature
mature
mature
mature
immature
mature
immature
immature
mature
mature
mature
immature
mature
mature
mature
immature
mature
mature
mature
immature
white
intermediate
immature
present
3.8
44.98
19.7
0.153
white
white
white
greenish-cream
white
white
white
greenish-cream
white
white
greenish-cream
greenish-cream
greenish-cream
intermediate
intermediate
intermediate
intermediate
few
intermediate
intermediate
few
intermediate
intermediate
medium
intermediate
intermediate
mature
mature
mature
immature
mature
mature
mature
immature
mature
mature
immature
mature
immature
present
absent
present
present
present
present
present
present
present
present
present
present
present
3.1
3.1
2.9
5.1
4.2
4.7
4.1
4.9
3.8
3.8
3.1
4.2
4.2
43.15
44.82
42.73
32.67
47.03
51.40
64.76
46.09
51.86
73.22
50.76
65.97
47.71
19.2
17.4
17.5
19.1
18.5
13.0
10.9
16.3
22.9
11.6
14.2
11.9
13.6
0.143
0.147
0.133
0.100
0.148
0.155
0.183
0.143
0.228
0.235
0.150
0.137
0.122
Brix
5.2
4.2
4.2
5.1
4.7
3.8
2.8
4.3
6.1
4.1
5.2
5.2
3.2
4.2
3.4
3.1
3.1
3.0
3.2
3.2
3.4
3.9
4.1
4.1
Boyacı HF et al. / Not Bot Horti Agrobo, 2015, 43(1):X-X
X
Table 5. Skin color characteristics of eggplant
Genotype
YRL 1
YRL 3
YRL 6
YRL 7
YRL 8
YRL 9
YRL 12
YRL 14
YRL 15
YRL 19
YRL 20
YRL 24
YRL 26
YRL 27
YRL 28
YRL 30
YRL 34
YRL 35
YRL 36
YRL 43
YRL 44
YRL 45
YRL 46
YRL 47
YRL 49
YRL 50
YRL 51
YRL 52
YRL 57
YRL 58
YRL 59
YRL 61
YRL 62
YRL 64
YRL 65
YRL 68
YRL 75
YRL 79
X
L*
48.33 ± 6.38
73.36 ± 4.06
51.54 ± 8.50
26.21 ± 0.66
31.83 ± 3.32
29.28 ± 3.64
31.71 ± 3.20
25.83 ± 0.99
25.23 ± 0.22
25.48 ± 0.51
25.82 ± 0.92
29.02 ± 3.24
55.22 ± 5.86
68.54 ± 5.60
29.89 ± 2.86
66.80 ± 2.88
77.73 ± 3.23
75.07 ± 2.53
72.34 ± 4.46
56.55 ± 9.09
72.50 ± 8.32
76.29 ± 4.16
37.41 ± 5.48
32.80 ± 4.44
32.04 ± 3.54
36.77 ± 6.11
37.55 ± 4.73
35.51 ± 2.61
44.44 ± 8.19
37.80 ± 5.11
56.43 ± 6.66
51.83 ± 4.36
31.03 ± 5.72
37.51 ± 4.79
73.19 ± 4.48
35.81 ± 5.99
25.39 ± 0.72
57.34 ± 3.69
a*
-0.67 ± 4.80
9.05 ± 2.52
-6.18 ± 5.43
6.03 ± 1.76
11.44 ± 2.19
10.22 ± 3.40
21.64 ± 2.52
4.97 ± 0.79
4.07 ± 1.31
3.81 ± 0.78
4.84 ± 1.15
17.85 ± 4.63
-7.43 ± 2.45
8.83 ± 2.37
9.70 ± 0.84
11.23 ± 1.66
4.64 ± 1.69
6.01 ± 1.29
8.41 ± 2.74
19.35 ± 5.14
7.51 ± 4.42
5.00 ± 2.26
22.90 ± 1.68
20.59 ± 3.19
23.02 ± 2.99
22.17 ± 2.18
21.85 ± 3.29
23.76 ± 1.51
4.04 ± 6.62
23.66 ± 2.46
18.22 ± 4.99
14.86 ± 2.14
10.50 ± 2.59
24.23 ± 0.59
8.17 ± 2.89
9.95 ± 2.41
6.03 ± 1.01
0.12 ± 2.26
C
19.04 ± 3.88
11.27 ± 1.52
23.75 ± 6.71
6.05 ± 1.78
11.78 ± 2.31
10.38 ± 3.48
21.67 ± 2.54
5.03 ± 0.93
4.08 ± 1.31
3.82 ± 0.79
4.85 ± 1.15
17.86 ± 4.65
19.10 ± 4.20
10.88 ± 1.00
9.93 ± 0.72
12.99 ± 1.04
10.98 ± 0.16
10.38 ± 0.33
11.30 ± 1.24
19.81 ± 4.51
11.58 ± 2.54
12.40 ± 1.05
22.94 ± 1.68
20.64 ± 3.23
23.04 ± 3.00
22.20 ± 2.15
21.88 ± 3.30
23.81 ± 1.49
13.35 ± 1.43
23.72 ± 2.43
18.55 ± 4.72
15.45 ± 1.56
10.96 ± 2.90
24.26 ± 0.61
11.66 ± 1.61
12.01 ± 3.63
6.03 ± 1.01
19.64 ± 2.08
b*
18.38 ± 4.04
6.31 ± 1.09
22.47 ± 6.03
0.36 ± 0.51
2.69 ± 1.07
1.25 ± 1.45
-0.98 ± 0.75
0.36 ± 0.88
0.002 ± 0.26
-0.03 ± 0.22
0.003 ± 0.28
-0.42 ± 0.54
17.56 ± 3.60
5.71 ± 1.76
1.57 ± 1.36
6.30 ± 1.17
9.76 ± 1.00
8.28 ± 1.20
6.96 ± 1.55
2.09 ± 2.76
7.49 ± 2.90
10.97 ± 2.10
-0.49 ± 1.07
-0.50 ± 1.36
-0.51 ± 0.47
-0.07 ± 1.29
-0.89 ± 0.93
-1.37 ± 0.48
9.70 ± 5.12
-1.61 ± 0.66
1.86 ± 2.45
0.41 ± 3.95
2.53 ± 2.32
0.02 ± 1.29
7.90 ± 0.91
6.17 ± 3.81
0.12 ± 0.14
19.50 ± 2.14
h°
88.77 ± 15.99X
36.39 ± 11.67
101.46 ± 13.81
62.44 ± 131.52
12.60 ± 4.48
5.85 ± 6.76
357.56 ± 1.74
182.80 ± 175.41
299.07 ± 131.60
238.99 ± 167.40
299.43 ± 131.95
243.94 ± 146.42
112.31 ± 3.60
34.29 ± 14.89
9.29 ± 8.33
29.80 ± 7.71
64.53 ± 10.30
53.91 ± 9.74
41.16 ± 14.66
68.93 ± 128.77
48.10 ± 21.20
64.49 ± 13.53
298.96 ± 131.62
357.03 ± 2.04
240.48 ± 166.70
179.90 ± 177.00
285.88 ± 142.39
356.67 ± 1.24
63.87 ± 38.34
356.05 ± 1.90
80.11 ± 137.94
242.80 ± 156.44
12.19 ± 9.68
285.99 ± 142.37
45.71 ± 12.66
27.50 ± 15.13
1.05 ± 1.04
88.98 ± 6.91
color
chartreuse
yellowish-orange
chartreuse
yellowish
redish
redish
magenta
cyan
purple
blue
purple
blue
greenish
orange
red
orange
cream-yellowish
yellow
orange-yellow
yellow
goldenrod
yellow
purple
magenta
blue
cyan
purple
magenta
yellowish
magenta
greenish-yellow
blue
redish
purple
orange-yellow
dark-orange
redish
chartreuse
: Means represent three 10-fruit samples ± SD.
Fig. 2. UPGMA dendrogram showing phylogenetic relationships of
local Burdur eggplant heirlooms together with reference cultivars
using morphological data
Fig. 3. The fruits of Burdur eggplant heirlooms belonging to the Group C
A total of 65 amplified RAPD bands were generated.
Twenty nine bands were polymorphic and the mean percentage
of polymorphism was 44.61%. OPO-10 primer produced the
maximum numbers of bands (18). Although the OPB-07
primer produced the minimal number of bands (8), it revealed a
100% polymorphism (Table 6). The OPH-2, OPB-07, OPO10 and OPL-16 primers’ PCR products and their band patterns
are shown in Fig. 4a-d.
percent polymorphism of each RAPD primer used
Table 6. Primer code, sequence, GC content, number of polymorphic bands and
Primer
code
OPH-02
OPL-04
OPB-07
OPO-10
OPL-16
TOTAL
Nucleotide sequence
5’-TCG GAC GTG A-3’
5’-GAC TGC ACA C-3’
5’-GGT GAC GCA G-3’
5’-TCA GAG CGC C-3’
5’-AGG TTG CAG G-3’
No. of
amplified
bands
14
16
8
18
9
65
No. of
polymorphic
bands
7
4
8
5
5
29
Polymorphic
bands (%)
50
25
100
27.77
55.55
44.61
Boyacı HF et al. / Not Bot Horti Agrobo, 2015, 43(1):X-X
X
The phylogenetic relationships among 38 genotypes were
evaluated using 29 polymorphic loci of the genomic DNA
generated using randomly amplified polymorphic DNA
(RAPD) technique. The approximate Mantel t-test statistic were
t = 8.9263, p = 1.0000. The matrix correlation (r) was found to
be 0.64. The similarity rates according to the coefficient similarity
of genotypes ranged between 0.84 and 0.98.
Using molecular data two major groups emerged in the
dendrogram generated with UPGMA method (Fig. 5). First
group branched into two subgroups and were showed in
brackets as Group A and Group B. Group A was consisted of
YRL 15 and YRL 27. Group E included most of the genotypes
had high similarities with reference cultivar. The highest genetic
similarity was observed in Group D with 98% percentage
similarity index between genotypes YRL 6 and YRL 44. YRL 1
and YRL 27 were detected as the most distantly genotypes from
each other in the cluster. Fruits of Burdur eggplant heirlooms
YRL 6 and YRL 44, which showed highest genetic similarity
with 98% percentage are shown in Fig. 6. Also, the fruits of
Fig. 7. Fruits of eggplant Burdur heirlooms with reference cultivar (YRL
68) situated in GROUP E generated by UPGMA using molecular data
Burdur heirlooms which grouped together in cluster E in the
UPGMA dendrogram using molecular data and reference
culture are shown in Fig. 7.
Discussions
Fig. 4. PCR products of Burdur eggplant heirlooms generated using
RAPD primers a:OPB-07, b:OPH-02, c:OPO-016, d:OPL-04
Fig. 5. UPGMA dendrogram showing phylogenetic relationships of
local Burdur eggplant heirlooms together with reference cultivars using
molecular data
Fig. 6. Fruits of Burdur eggplant heirlooms YRL 6 and YRL 44, which
showed highest genetic similarity with 98% percentage
Eggplant has a wide genetic diversity in the regions where
it is cultivated, although they are not native to the region
(Muñoz-Falcón et al., 2008). In spite of the fact that Turkey
is not the place origin of eggplant, wide genetic diversity has
been reported in Turkey (Demir et al., 2010; Tümbilen et al.,
2011a, 2011b). It is clearly evident in this study data that
Burdur, which is a small geographical region, had a rich
genetic diversity. Also, a wide genetic variability was
determined in both Spain and Jordan local genotypes
(Prohens et al., 2003). Local genotypes can contribute to
enhancing the gene pool used in breeding studies and to help
increase heterosis (Muñoz-Falcón et al., 2009). In recent
years, some factors like cultivation of commercial varieties
instead of heirlooms, construction of buildings on agricultural
land, and innovation in cultivation methods have led to
erosion of plant genetic resources (Cericola et al., 2013).
Therefore, there is a need collecting and identification of local
heirlooms before they disappear (Muñoz-Falcón et al., 2008).
Some characters that contributed to genetic diversity were
as flowering dates, the number of seeds per fruits, fruit
features, and the growth pattern of plants. These features are
controlled by several genes in eggplant (Frary and Doğanlar,
2003). Solanum melongena accessions could characterize
these descriptors like bigger and flabby fruits, less
flowers/inflorescence, few fruits/plant and higher acidity etc.
compared to the wild relatives (Polignano et al., 2010).
Consistent with previously works, a higher diversity for most
morphological descriptors was recorded in the collection of
Burdur local heirlooms identified in this study. Fruit color
can be cream, green, red, reddish-purple, dark purple or black,
and some varieties produce fruit which is where the genetic
variation necessary for future varietal improvement and for
addressing future breeding challenges will be found.
Molecular markers linked with agronomic traits are useful
tools for marker assisted selection and mapping candidate
genes studies in breeding programs (Nunome et al., 2009;
Boyacı HF et al. / Not Bot Horti Agrobo, 2015, 43(1):X-X
X
Wang et al., 2010). Some of the RAPD markers used were
determined to have relationship with coloring of stem, calyx
and fruiting in eggplant (Frary et al., 2003). This provides a
great advantage for identifying features affecting by ecological
conditions (Biswas et al., 2009; Nunome et al., 2001).
Relationships among eggplant materials have been studied by
molecular studies for use by eggplant breeders (Furini and
Wunder, 2004). As mentioned above the RAPD markers
revealed as a potential useful tool for determination of genetic
diversity. It was found that RAPD analysis in eggplant with
four primers had been adequate to identify of genetic diversity
(Biswas et al., 2009). Also, RAPD markers were more
effective than ISSR for revealing genetic diversification as
reported by Ali et al. (2011). Tiwari et al. (2009) previously
reported that even two of the 29 RAPD primers were found
to be sufficient for identification of local cultivars. All RAPD
markers used in the study have produced polymorphic bands
as expected. However, if fruit criteria are taken into
consideration for distinguishing in the dendrogram and
grouping, it is not enough for the breeders. More informative
DNA markers can be used to provide better progress in
eggplant breeding studies (Stàgel et al., 2008). Simple
Sequence Repeats (SSRs) methods were found more
successful for distinguishing closely related eggplant cultivars
(Hurtado et al., 2012; Muñoz-Falcón et al., 2009; Prohens et
al., 2008). The approach of using SSR markers instead of
RAPD markers can provide better results in the phylogenetic
relationships studies. Muñoz-Falcón et al. (2009) reported
that if the morphological and molecular data are considered
together, they can be provide sufficient and useful
information for the breeders. Similarly to the findings of
other studies, we suggest that the molecular evidences need to
be supplemented by morphological data to validate the
phylogenetic relationships among the genotypes. It is very
important to note that the genetic variations can’t be detected
by only showing a dendrograms generated by molecular data.
Conclusions
The local populations are of great importance for the
breeders so that they adapted well to their cultivated areas.
There is a need collecting and identification of these
heirlooms before integrated to the breeding programs. The
aim of the study was to investigate the diversity among
heirlooms cultivated in Burdur province. A high genetic
diversity was determined widely among them. These
materials can be of a potential value for the breeders.
Acknowledgements
This research was financially supported by grants from
the General Directorate of Agricultural Research and
Policy, Republic of Turkey Ministry of Food, Agriculture
and Livestock under BBSS-10-12 project number.
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