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20191
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Subject: Biotechnology and molecular Biology × Author: Chaudhary, Vrantika × Start date: 1996 ×
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    ThesisItemOpen Access
    Development of micro-RNA and EST-SSR markers for diversity analysis in clusterbean [Cyamopsis tetragonoloba (L.) Taub]
    (CCSHAU, Hisar, 2019-09) Chaudhary, Vrantika; Yadav, Neelam R
    Cluster bean [Cyamopsis tetragonoloba (L.) Taub] is an important industrial crop due to the presence of soluble dietary fibre, high protein content and high value gum. MicroRNA are tiny non-coding small RNAs master regulators that play important role in multiple biological processes by degrading targeted mRNAs or repressing mRNA translation. In silico computational analysis based on Cluster bean ESTs led to identification of 57 miRNA families along with 37 targets. The length of most of the mature miRNAs was found to be 21nt long and average MFE value of 25.4 (-kcal/mol). The gene ontology study revealed the genes encoding transcription factors or enzymes involved in the regulation of development, growth, metabolism, physiological processes and stress response. Identification of transcription factors and miRNA resulted into model prediction. Most abundant miRNA families predicted in this study were miR156, miR172 and miR2606. The miRNA related to gum metabolism were also identified i.e. miR5658, miR1533 and miR414. A total of 40 unigene SSRs were screened among hundred germplasm lines including selected cultivars. The molecular analysis divided the 100 cluster bean genotypes into two major clusters at similarity coefficient of 0.49. Genic SSRs related to galactomannan metabolism were also used for diversity analysis of low and high gum clusterbean genotypes. A new class of functional and regulatory markers i.e., miRNA-SSRs have been identified in Cluster bean with mononucleotides repeats in highest proportion (76%) followed by di- (10%), tri- (10%) and tetranucleotides (4%) in the pre-miRNA sequences. Cross transferability of Medicago truncatula miRNA SSRs showed 34% cross transferability among Cluster bean genotypes. A key enzyme in galactomannan metabolism, galactosyltransferase was used for structure prediction using in silico approaches. In silico molecular dynamics revealed its stability and structure was dominated by random coil and α-helix followed by β sheet region. Cluster bean galactosyltransferase sequence showed highest similarity with fenugreek and least similarity with Medicago truncatula