Assessment of Genetic Diversity in Soybean (Glycine max (L.) Merrill) by Morphological, Biochemical and Molecular Markers
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Date
2016
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MPUAT, Udaipur
Abstract
Genetic divergence among 24 genotypes of Glycine max (L.) Merrill was assessed through morphological (10 phenotypic traits), biochemical (PER, EST and SOD isoenzyems) and molecular (RAPD, ISSR and Chloroplast) markers. Significant genetic variance was observed for plant height, days to maturity, days to 50% of flowering, number of branches per plant, number of pods/plant, number of seeds/pod, test weight (g), seed yield/plant (g), and harvest index (%) amongst the morphological characters chosen for study. Hierarchical Euclidean and Ward’s cluster analysis based on 10 morphological characteristics grouped all the 24 genotypes into sixteen and eight divergent clusters respectively.
A total of 15 alleles were generated by three isozymes viz., peroxidase, esterase and superoxide dismutase. Isozymes of SOD exhibited a maximum of seven activity zones followed by esterase and peroxidase showing six and two activity zones respectively. Isozymes of peroxidase and esterase showed 50 and 83.33 per cent of polymorphism with all alleles. Jaccard’s Similarity Coefficient values lay between 0.67 to 1.00 that divided all the genotypes into two main clusters including 22 genotypes in cluster I and two genotypes in cluster II. Genetic divergence and distance derived from isozyme analysis was found low due to the small number of polymorphic alleles.
RAPD analysis showed 100 per cent of polymorphism. A total of 164 amplified bands were obtained that ranged between 100 bp to 2500 bp, using 18 RAPD primers. All the 164 bands were polymorphic. ISSR analysis showed 97.25 per cent of polymorphism. A total of 182 amplified bands were obtained that ranged between 200 bp to 2500 bp using 19 ISSR primers. The 177 out of 182 bands were polymorphic. Three universal chloroplast primers were used viz., trnT, trnCD and PsbA-trnH amplified with single band. A total of 3 amplified bands were obtained of which all were monomorphic. The average PIC of RAPD and ISSR were found 0.295 and 0.344 respectively.
Jaccard’s similarity coefficient for RAPD and ISSR as well as their combined values lay between 0.12 to 0.70, 0.27 to 0.89 and 0.50 to 0.91, respectively. The similarity coefficients values were significant for all the markers. Dendogram based on genetic distance calculated by UPGMA method segregated the 24 soybean genotypes into three (RAPD), three (ISSR) and three (combined) major clusters, respectively.
Based upon protein content, morphological aspects, molecular data and performance of genotypes, cp-DNA sequence of 8 genotypes (JS-20-53, JS-20-79, NRC-98, NRC-107, RVS-2002-4, SL-955, PS-1539 and PS-1543) for each primer were also sequenced at Xceleris Pvt Ltd. Ahmadabad. The primer OPD-05 gave bright, constant and unique band of 400 bp in genotype NRC-98. This unique DNA band was excised from 1.2 percent agrose gel and DNA was purified by gel elution kit (Banglore Genei gel extraction kit) for further study.
Based on mean per se performance, genotypes NRC-107, Himso-1685, JS-20-79, PS-1539 and RVS-2002-4 showed superiority in yield attributes, hence could be further improved through pure line breeding. Therefore, the results of the present study could be gainfully utilized in soybean improvement programmes.
Description
Assessment of Genetic Diversity in Soybean (Glycine max (L.) Merrill) by Morphological, Biochemical and Molecular Markers
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Citation
Jain and Joshi, 2016