“MOLECULAR CHARACTERIZATION OF BLACKGRAM [Vigna mungo (L.) Hepper] GENOTYPES FOR SALINE WATER TOLERANCE”
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Date
2018-07
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JAU,JUNAGADH
Abstract
Blackgram [Vigna mungo (L.) Hepper], Family: Fabaceae and sub family: faboideae is the most widely grown type of pulse. To alleviate protein energy malnutrition, a minimum of 50g pulses/capita/day should be available in addition to other sources of protein such as cereals, milk, meat and eggs. The present experiment on “MOLECULAR CHARACTERIZATION OF BLACKGRAM [Vigna mungo (L.) Hepper] GENOTYPES FOR SALINE WATER TOLERANCE” was conducted at Department of Biotechnology, Junagadh Agricultural University, Junagadh with objectives to analyze molecular diversity of different blackgram genotypes by PCR based molecular markers to find out the phylogenetic relationship among different blackgram genotypes tolerance to salinity.
On the basis of physiological parameters, the blackgram genotypes were discriminated into tolerant, moderate and sensitive to salinity stress. Out of 22 genotypes, five genotypes viz., SKNU-03-03, SKNU-07-06, SKNU-06-03, SKNU-07-01 and IC-214520 were found to be tolerant to salinity. Thirteen genotypes were found to be moderately tolerant and four genotype viz., GJU-1506, JAWAHAR URD-3, JAWAHAR URD-2 and GJU-1509 are sensitive to salinity.
Germination per cent decreased by 67.77 %, root length reduced by 64.11 %, shoot length reduced by 67.91 % while seedling length decreased by 66.44 % in T4 treatment as compared to the T1 (control) treatment among all the blackgram genotypes. Seedling dry weight reduced by 1.40 fold in T4 treatment as compared to the T1 (control) treatment. Looking to the vigor index, seedling vigor index-I (length basis) and seedling
vigor index-II (Dry weight basis) decreased by 2.21 and 2.03 folds, respectively in T4 ECe dSm-1.
For molecular characterization, 11 RAPD, 10 ISSR and 10 SSR primers were used. Amplification of genomic DNA of 20 blackgram genotypes, using RAPD analysis, yielded 80 polymorphic fragments with an average of 99.99 % polymorphism. Number of amplified fragments with RAPD primers ranged from 3 to 15 bands and varied in size from 102 to 3344 bp. A dendrogram based on UPGMA analysis grouped the 20 blackgram genotypes into two main clusters named cluster-A and cluster-B with 10 % similarity, with Jaccard’s similarity coefficient ranging from 0.047 to 0.857. The 10 ISSR primers produced 44 bands across 20 blackgram genotypes, which were polymorphic. The size of amplified bands varied from 61 to 2267 bp. A dendrogram based on UPGMA analysis of 20 blackgram genotypes generated for ISSR data formed two main clusters, named cluster-A and cluster-B with 35 % similarity and Jaccard’s similarity coefficient ranged from 0.240 to 0.741. The 10 SSR primers produced 14 bands across 20 blackgram genotypes, which were polymorphic. A dendrogram based on UPGMA analysis of 20 blackgram genotypes generated by SSR molecular marker data formed two main clusters, named cluster-A and cluster-B with 40 % similarity and Jaccard’s similarity coefficient ranged from 0.100 to 1.00.
The pooled study of all markers revealed a dendrogram consisted of two clusters. Cluster-I consisted of 5 blackgram genotypes, while cluster-II consisted of 17 genotypes in different cluster. From the dendrogram depicted genotype JAWAHAR URD-3 found to be most diverse from the other genotypes.
The molecular markers (RAPD, ISSR and SSR) revealed phylogenetic relation of 20 blackgram genotypes and showed the different dendrogram pattern for different genotypes taken for study. RAPD abled to discriminate most diverse genotype SKNU-07-01 into cluster-B separately. The ISSR revealed one genotype JAWAHAR URD-3 in cluster-B found to be most diversified genotype across the 20 blackgram genotypes. SSR depicted one diverse genotypes JAWAHAR URD-3 into one cluster-B which were able to be showed diverse position in cluster.
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BIOTECHNOLOGY