BIOCHEMICAL AND MOLECULAR CHARACTERIZATION OF INDIAN MUSTARD [Brassica juncea (L.) CZERN & COSS] GENOTYPES
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Date
2014
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AAU, Anand
Abstract
Brassica is an agriculturally important genus containing species with highly diverse morphology and wide ranging utility. Brassica juncea L. is an important oil seed crop which belongs to family Brassicaceae. Brassicajuncea is one of the major sources of oil in the subcontinent for centuries. To improve quality and quantity of Brassica spp., the presence of sufficient genetic diversity is very important for quality, quantity and utilization of oil in human nutrition. Improvements in molecular marker technology offer a potential tool for the efficiency and affordability of variety testing. In the present study, quality of seed was tested using different qualitative tests. Biochemical markers namely isozymes (Peroxidase and Esterase), SDS-PAGE protein and molecular markers viz., SSR, ISSR were used to study the polymorphism amongst different cultivars of Indian mustard. The moisture percentage of all cultivars was found in the ranged of 3.93-10.02%. Oil content in genotype SKM 9033 showed highest oil and RAYAD- 9602 showed lowest oil. Protein content and true protein in mustard cuhivars ranged from 27.90-33.69% and 10.75-20.01%, respectively. Sinigrin content in mustard cultivars found in the range of 2.49-46.80 nmole/g. Genotype Bio-Q-44-279 had the highest sinigrin concentration 46.80 nmole/g and SKM-9033 had the lowest sinigrin concentration 2.49 ^mole/g. Isozyme pattern of peroxidase and esterase generated significant correlation amongst the clusters. Electrophoresis of seed protein showed a total 28 bands in mustard genotypes and the intensity of the bands varied among all the genotypes. Simple Sequence Repeats amplification of mustard genomic DNA using 8 primers generated 252 scorable bands with average of 31 bands per primer. Percent polymorphism ranged from 33.33% (BRMS-08) to 100%. Polymorphic information content (PIC) ranged from 0.37 (BRMS-06) to 0.83 (BRMS-07). The similarity coefficient of all mustard cuhivars ranged between 0.036 to 0.947. This result indicated existence of genetic variation occurred among different genotypes. Clustering pattern of dendrogram generated by using the pooled SSR data formed two major clusters viz. having similarity coefficient of 0.39 to 0.94. It indicated genetic variability among the different genotypes. Cophenetic correlation was found r = 0.89, reflecting very good fit of the dendrogram. Both Simple Sequence Repeat (SSR) and Inter Simple Sequence Repeat (ISSR) markers were efficient to assess genetic diversity within species of mustard. Dendrogram obtained from both markers showed two major clusters A and B. Cophenetic correlation were r = 0.89 for SSR and for ISSR r = 0.85, reflecting very good fit for dendrograms. SSR and ISSR showed average polymorphism of 82.91% and 96.02%, respectively. It showed that ISSR was more polymorphic than that of SSR. Cophenetic correlation of both SSR and ISSR similarity matrices were r = 0.80, suggested suitable fit for cluster analysis. From these results it could be concluded that clusters produced by SSR and ISSR were conserved and were highly correlated with each other. The resuUs showed that, ISSR was more informative than SSR in assessing genetic diversity of mustard genotypes.
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