Investigation of genetic variation of lentil lines using random amplified polymorphic DNA (RAPD) and intron-exon splice junctions (ISJ) analysis

ABSTRACT: In order to investigation of genetic variation among lentil lines and genetic relationship these to Cistan local cultivar, DNA of 20 lentil lines and Cistan local cultivar were analysed using random amplified polymorphic DNA (RAPD) and intron-exon splice junctions (ISJ) markers. PIC, MI, percentage of polymorphic bands (P), Nei , s gene diversity (H), Shannon, s information index (I) were calculated for both random, semi random and combined of random and semi random primers. For both matrices, genetic similarity (GSRAPD and GSISJ) were calculated according to the Jaccard similarity coefficient. Correlation between two similarity matrices obtained with two markers types was estimated. The similarity matrix per marker was subjected to cluster analysis by the unweighted pair-group method (UPGMA). The of RAPD was as as ISJ, but MI index of ISJ (average 6.095) was much higher than RAPD (average 4.49). Mean of genetic similarity of RAPD marker was higher than ISJ marker, and combined of RAPD and ISJ markers. Thus in order to estimate of genetic diversity in lentil lines, ISJ marker was better than RAPD marker and combined of RAPD and ISJ markers was also better than RAPD marker. For RAPDs the similarity values ranged from 0.166 between TN2026 line and TN2464 line, to 0.697 between TN1086 line and TN1087 line. For ISJ, the values ranged from 0.052 between TN1084 line and TN2462 line, to 0.312 between TN2457 line and KC210031 line, and for RAPD +ISJ the values ranged from 0.147 between TN2420 line and TN2026 line, to 0.471 between TN1087 line and TN1086 line. In the ISJ markers the UPGMA cluster diagram have showed 3 major clusters: one cluster included the TN2026 line, the second cluster were contained TN2128, TN2022, TN2461, TN2464, TN1885, TN2463 and the third cluster were contained rest of lentil lines. In RAPD and combined of RAPD and ISJ markers, the UPGMA cluster diagram have showed 2 major clusters. Cistan local cultivar had higher genetic distance than TN2026 line in each three methods of grouping. Therefore in order to favourable genes transfer from TN2026 line to Cistan local cultivar should be considerable genetic distance.