Molecular analysis of virus causing leaf curl disease in cotton (Gossypium hirsutum)
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Date
2013
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CCSHAU
Abstract
In the present study, leaf samples from cotton leaf curl diseased (CLCuD) plants were
collected from 115 diverse fields of four districts (Hisar, Fatehabad, Jind, Sirsa) of cotton growing belt
of Haryana. The principal objective was to carry out diversity analysis of virus causing the leaf curl
disease in cotton (Gossypium hirsutum) using molecular approaches. Genomic DNA from diseased
cotton leaves was isolated using modified CTAB method. Fifty quality DNA samples of concentration
>150ng/ul were selected for PCR amplification. Three pairs of cotton leaf curl virus (CLCuV) coat
protein gene specific primers were employed to carry virus specific amplification. PCR conditions were
standardized for optimum viral coat protein gene amplification. The amplicons were resolved using
agarose gel electrophoresis and a prominent band of size approx. 10 kb was found as CLCuV’s coat
protein gene specific amplicon. Out of 50, only 29 samples showed amplification which was rather
similar. Further to dissect out the differences in pathogenic viruses’s coat protein gene, the amplicons
were digested using three restriction enzymes (EcoR1, HinD III and Taq1α). Characteristically
differential restriction patterns were observed that revealed four variants of CLCuV in cotton growing
belt of Haryana.The maximum diversity was observed in district Fatehabad followed by Hisar, Jind and
Sirsa.
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Keywords
CLCuD, CLCuV, Diversity analysis, Gossypium hirsutum,.