Molecular and serodiganosis of jatropha mosaic virus infecting Jatropha

Jatropha mosaic virus (JMV) infecting Jatropha curcas is one of the most destructive pathogen affecting the plant bringing down its yield. Serological and molecular detection is a fast means to detect the virus for its management. The present study thus aimed at standardizing indirect plate and Dot enzyme linked immunosorbent assay (ELISA) and Polymerase Chain Reaction (PCR) for the detection of the virus. Electron microscopically, the virus appeared as isometric particles of 25 X 24 nm in diameter typical of Gemini viruses infecting plants. Polyclonal antiserum was produced against the virus in rabbit. In Indirect plate ELISA, optimum OD was 1.062 and 0.672 at 405 nm with diseased plant sap and ultrapurified virus as the antigen, at a dilution of 1:100 and 1:10 respectively. Primary antibody dilution was 1:1000 and 1:2000 and that of secondary antibody 1:7000 and 1: 2000 for the two antigens respectively. In DOT-ELISA JMV was detected at a primary antibody dilution of 1:4000 and secondary antibody dilution of 1:2000. The DNA segment of the coat protein and replicase gene of JMV were PCR amplified using corresponding primers, resulting in the amplification of a 1400bp sized product.