Molecular Plant Breeding 2016, Vol.7, No.23, 1
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genetic linkage map in lentil. Several reports from our lab have established transferability of SSR markers from
one legume genera/species to other (Datta et al., 2010a, 2010b; 2011; 2012; 2013a, 2013b; 2015), and observed
high level of sequence conservation of microsatellite markers in legumes.
ISSR (Inter Simple Sequence Repeat) markers are inexpensive and readily adaptable technique for routine
germplasm fingerprinting and evaluation of genetic relationship between accessions or genotypes (Sardana et al.,
1998; Dixit et al., 2004, Edossa et al., 2007) and construction of genetic linkage maps (Abo-Elwafa et al., 1995).
This technique has been used to assess genetic diversity in germplasm collection (Gilbert et al., 1999; Salimath et
al., 1995), to identify cultivars (Prevost et al., 1999). The present investigation was undertaken to assess the
diversity and genetic relatedness in Indian and exotic lentil genotypes with the objectives, to study the
polymorphism and genetic relationship among and to identify genotype specific markers.
1 Results and Analysis
A total of 100 ISSR markers were used to test their ability in detection of DNA polymorphism. Only 24 markers
produced reproducible amplification in all the selected genotype and therefore only these were considered for
polymorphism assay (Table 1). The ISSR profiles generated with markers UBC-888, UBC-891, UBC-886,
UBC-815, UBC-889 and UBC-858 respectively showed (Figure 1).
Table 1 Characteristics and details of 24 ISSR markers amplified in 25 lentil genotypes
Primers
Sequence (5'-3')
Tm(
℃
)
ES (bp)
NA NP
%P
PIC (H
AV
)
P
MI
UA
No G(s)
UBC 808 AGAGAGAGAGAGAGAGC
51.3
160-1000
6
5
83.33
0.46 0.092
0.49 1 DPL44(900)
UBC-815 CTCTCTCTCTCTCTCTG
51.3
280-3000
14
14
100.00 0.79 0.056
0.36 2 Sehore74-3(1120), IPL525 (400)
UBC-824 TCTCTCTCTCTCTCTCG
51.3
1100-1200 2
1
50.00
0.88 0.440
1.43 0 -
UBC-826 ACACACACACACACACACC 51.3
820-2500
8
8
100.00 0.84 0.105
0.68 0 -
UBC-827 ACACACACACACACACG
51.3
400-200
10
10
100.00 0.79 0.079
0.51 0 -
UBC-847 CACACACACACACACARC
52.1
520-1600
7
6
85.77
0.54 0.074
0.41 1 IPL525 (500)
UBC-854 TCTCTCTCTCTCTCRG
52.1
900-3000
5
5
100.00 0.72 0.144
0.93 1 ILL8114 (3000)
UBC-859 TGTGTGTGTGTGTGTGRC
52.1
300-1400
7
7
100.00 0.52 0.074
0.48 0 -
UBC-822 TCTCTCTCTCTCTCTCA
48.9
650-3000
8
8
100.00 0.71 0.088
0.57 1 DPL15 (3000)
UBC-810 GAGAGAGAGAGAGAGAT
48.9
300-1600
8
8
100.00 0.46 0.057
0.37 1 IPL525 (300)
UBC-812 GAGAGAGAGAGAGAGAA
48.9
370-1200
8
8
100.00 0.29 0.036
0.23 1 DPL44 (1000)
UBC-814 CTCTCTCTCTCTCTCTA
48.9
500-2000
7
7
100.00 0.66 0.094
0.61 1 P2016 (500)
UBC-816 CACACACACACACACAT
48.9
600-1000
2
1
50.00
0.30 0.300
0.97 0 -
UBC-817 CACACACACACACACAA
48.9
480-2200
6
6
100.00 0.63 0.105
0.68 0 -
UBC-819 GTGTGTGTGTGTGTGTA
48.9
480-2500
6
6
100.00 0.71 0.118
0.77 0 -
UBC-834 AGAGAGAGAGAGAGAGYT 48.9
900-3000
5
3
60.00
0.17 0.056
0.22 1 DPL44 (3000)
UBC-858 TGTGTGTGTGTGTGTGRT
49.8
710-1160
3
2
66.66
0.98 0.490
2.12 0 -
UBC-886 VDVDTCTCTCTCT
46.5
300-1450
9
9
100.00 0.75 0.083
0.53 0 -
UBC-888 VDVCACACACACACACA
46.5
310-2000
10
10
100.00 0.57 0.057
0.37 0 -
UBC-889 DVDACACACACACACAC
46.5
400-1600
6
6
100.00 0.59 0.098
0.63 0 -
UBC-891 HVHTGTGTGTGTGTGTG
46.5
400-1200
5
5
100.00 0.43 0.086
0.55 0 -
UBC-861 ACCACCACCACCACCACC
58.9
860-1160
3
2
66.66
0.16 0.080
0.34 0 -
UBC-881 GGGTGGGGTGGGGTG
57.7
850-3000
6
6
100.00 0.45 0.075
0.48 1 VL103 (1570)
UBC-865 CCGCCGCCGCCGCCGCCG
72.6
960-2000
5
5
100.00 0.50 0.100
0.65 0 -
Total
156 148 --------
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Note: Tm: Temprature of melting; ES: Estimated size of amplicons in bp; NA: Total number of amplicons; NP: Number of
polymorphic amplicons; % P: Percentage polymorphism; PIC: Polymorphism information content; (H
AV
)
P
: Average polymorphic heterozygosity;
MI: Marker index; UA: Unique amplicons; G(s): Genotype and size of unique amplicon
