Molecular Plant Breeding 2016, Vol.7, No.23, 1
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Figure 1 Amplification profile of lentil genotypes with ISSR markers
Note: a: UBC-888; b: UBC-891; c: UBC-886; d: UBC-815; e: UBC-889; f: UBC-858; Lane M: 100 bp DNA ladder; Lane 1-25: lentil
genotypes
Total 156 amplicons were generated by the 24 ISSR markers with an average of 6.5 bands per marker with the
amplicons size ranging between 160-3 000 bp. One hundred and forty eight fragments (94.87%) were found
polymorphic. Among the 24 polymorphic markers, 17 (UBC-810, UBC-812, UBC-814, UBC-815, UBC-817,
UBC-819, UBC-822, UBC-826, UBC-827, UBC-854, UBC-859, UBC-865, UBC-881, UBC-886, UBC-888,
UBC-889, UBC-891) showed 100% polymorphic bands (Table 2), among these markers most contained
dinucleotide repeats.The maximum numbers of amplicons were amplified by UBC-815(14) followed by
UBC-827(10), while UBC-824 and UBC-816 amplifiede minimum (2) number of amplicons. UBC-858 and
UBC-861 amplified three amplicons each. On the other hand, UBC-816 and UBC-824 exhibited minimum
polymorphism (50%).
1.1 DNA polymorphism and genetic relationship
ISSR markers analysis of lentil genotypes using Jaccard’s (1908) coefficient of genetic similarity are given in
Table 1. The similarity coefficients varied from 0.37 to 0.84, the average being 0.69. Genotypes LL147 and L4147
were found to have maximum similarity (84%) which was closely followed by L4147 and VL103 (82.9%). The
lowest similarity was found in between T36 and Precoz (37%) which was followed by IPL525 and T36 (40.9%).
A total of 11 unique bands were also obtained which is present only in a particular genotype. A 900 bp band was
obtained in genotype DPL 44 with UBC-808 marker. Similarly the marker UBC-815 amplified two unique bands
of 1 120 bp and 400 bp in case of Sehore 74-3 and IPL525. Table 2 includes the complete list of such genotype
specific bands.
1.2 Cluster analysis
The Jaccard’s (1908) similarity coefficients based on ISSR profiles were subjected UPGMA analysis and a
dendrogram of 25 lentil genotypes was constructed (Figure 2). It showed two main clusters. Cluster I contains
genotypes IPL525, PL234 and PL5 which are tolerant to rust and wilt disease, the similarity between IPL525 and
PL 234 is 68.10%. Cluster IIA contains genotypes DPL15, WBL58 and Sehore74-3. The similarity value between
WBL 58 and Sehore74-3 is 75.30%. These two genotypes were large seeded and had ash green colour foliage with
high pubescence. The cluster II B contains 4 sub-clusters; sub cluster IIB1 contains two small seeded varieties
PL406 and L-9-12. The similarity value between these genotypes is 77.7%. In Sub cluster II B2, NDL 1 and LH
84-8 genotypes were grouped together; the reason for this closeness may be due to a common parent L-9-12 in
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