Assessment of genetic variation in lucerne (Medicago sativa L.) using protease inhibitor activities and RAPD markers.
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Date
2011-09
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Abstract
The aim of the present study is to identify and characterize lucerne lines resistance to weevil infestation. After
three years of field screening for resistance to weevil infestation, 13 lines of lucerne were selected to assess the
genotypic variations for lucerne weevil (Hypera postica Gyll.) at biochemical and molecular levels. Total
phenols varied from 0.15 to 0.91 mg g-1 (DM) in these genotypes. The highest trypsin (11.11 unit mg-1 protein) and
chymotrypsin (93.0 unit mg-1 protein) inhibitors activities were recorded in G-1-02 and B-4-03 lines respectively,
whereas highest a-amylases inhibitor activity (14.2 unit mg-1 protein) in C-6-01. Zymogram patterns for trypsin
inhibitor activity showed quantitative variations among the lines. In total 262 DNA fragments were generated
when 45 deca-mer random primers were employed. Genetic variation in terms of genetic distance ranged from
0.65 to 0.85. Sequential Agglomerative Hierarchical and Nested (SAHN) clustering using the Un-weighted Pair
Group Method with Arithmetic mean (UPGMA) algorithm yielded two clusters (cluster I and II) which converged
at 72% similarity level. Cluster I contained most of the lines having low level of weevil infestation. High
bootstrap values (>40) indicated the significance of nodes embodied in these two clusters. However, SDSPAGE
analysis of the leaf proteins of these 13 lines showed no major variations except minor difference in the
protein bands of molecular weights between 14 to 20 kD.
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Keywords
Genotypic variation, Heat stable proteins, Insect resistance, Lucerne, Proteinase inhibitor, RAPD, SDS-PAGE
Citation
Chandra Amaresh, Pandey Krishna Chandra. Assessment of genetic variation in lucerne (Medicago sativa L.) using protease inhibitor activities and RAPD markers. Journal of Environmental Biology. 2011 Sept; 32(5): 559-565.