Cloning and Characterization of Coat Protein Gene from CVMV

Chilli leaf curl is the most destructive syndrome causing substantial economic losses in chilli production. Severe leaf curling, leaf distortion, dark green mottle, vein banding, reduced leaf size and stunted growth are typical symptoms of the viral infection. For a reliable detection specific primer pairs were designed, after PCR amplification using gene specific primers of CVMV, 531 bp DNA fragment was amplified in all diseased samples of chilli and no amplification was obtained in healthy samples. PCR amplicon of CVMV-CP gene was cloned into pTZ57R/T cloning vector and transformed into E. coli DH5a. Transformed white colonies were picked after blue-white colony assay and confirmed by colony PCR and restriction analysis. The sequence results clearly revealed that the coat protein gene showing 95% identity with the reported CVMV-CP gene sequence. Thus, the investigation focused on molecular characterization of the virus convincingly revealed potyvirus association with leaf curl disease in chilli.