Serological and molecular detection of an isolate of Cucumber Mosaic Virus (CMV) infecting cucumber (Cucumis sativus) and cloning of its coat protein gene

Cucumber mosaic virus (CMV) is a widely prevalent plant virus infecting important vegetable, plantation and flower crops. Methods for early detection of viruses in plants and vectors transmitting them play a critical role in plant virus disease management. Direct plate and Dot- Enzyme Linked Immunosorbent Assay (ELISA) was standardized for detection of CMV. Optimum OD of 1.249 (1.9 ng/μl) and 1.242 (1.52 ng/μl) was observed in 1:20 and 1:50 dilution of crude and ultrapurified antigen respectively, at a dilution of 1:1000 of both primary and secondary antibody. Polymerase Chain Reaction (PCR) using CMV coat protein (CMV CP) gene specific primers amplified a 657 base pair (bp) fragment, which was then  cloned in pTZ57R/T cloning vector and positive clones were identified by band shift assay and colony PCR. This will aid in developing field diagnostic kits for detection of CMV in different crops and also in developing transgenics with the CP gene.