Molecular marker analysis for bacterial blight resistance in cluster bean (Cyamopsis tetragonoloba (L.) Taub.)
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Date
2014
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CCSHAU
Abstract
Cluster bean (Cyamopsis tetragonoloba) is an important member of Leguminoseae family and
is a rich source of galctomannan content making it an industrial crop. In this crop, there are almost no
genomic resources or high quality co-dominant markers available for genetic analysis. At present there
is no report of SSR markers targeting bacterial leaf blight resistance available for this crop but available
EST database was used to develop EST-SSRs. Cluster bean EST sequences (16476) were retrieved and
used to identify EST-SSRs using SSRIT tool. A total of 858 SSRs were identified with maximum
proportion of trinucleotide repeats and least proportion of pentanucleotide repeats. CT/TC motif was
most abundant. Eighty sequences related to pathogen stress response were identified out of which 17
SSRs were used for in silico primer designing. Twenty seven genotypes were selected for molecular
marker analysis using these 11 EST-SSRs. Plants were also grown in net house for studying disease
incidence. Two wild speices, C. senegelensis (18.55 ± 1.86 %) and C. serrata (16.30 ± 0.93 %) showed
lowest disease incidence, while highest disease incidence was observed in case of Pusa Nav Bahar
(73.23 ± 0.92 %). All the 11 EST-SSR markers were polymorphic in molecular analysis, showing
different allele sizes in wild and cultivated genotypes. Diversity analysis suggested that two wild
species formed a different group at a similarity coefficient of 0.75 including some of the cultivated
genotypes. Four genotypes, C. senegelensis, HG 563, HG 75 and PNB, were selected for sequencing
and multiple sequence alignment against HES 1401. The alignment suggested the presence of SNPs
and In/ Dels in the sequences
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Keywords
Cluster bean, ESTs, EST-SSRs, bacterial leaf blight, C. senegelensis, C. serrata, PNB, disease incidence.