Patil, F.S.Chavan, Sachin Gorakshnath2020-10-092020-10-092010-05-20T06067https://krishikosh.egranth.ac.in/handle/1/5810153065Cowpea (Vigna unguiculata L. Walp.) is an important leguminous food grain crop attacked by wide array of diseases of biological origin especially viruses which cause devastating effects and are a really constraint to increased yield of cowpea. Management of the viral diseases is based primarily on the development of cowpea resistant varieties. The present work is aimed at development of molecular diagnostic methods of some economically important seedbome viruses-of cowpea to provide a basis for the control of these viruses. Seed samples of cowpea were collected and tested for the presence of the blackeye cowpea mosaic strain of bean common mosaic virus, cowpea aphid-borne mosaic virus, cowpea mosaic virus, cucumber mosaic virus and bean yellow mosaic virus using growingon test, enzyme linked immunosorbent assay, tissue print immune assay, differential host test, and reverse transcription polymerase chain reaction. Sodium dodecyl sulphatepolyacrylamide gel electrophoresis showed that partially purified viruses contained coat protein polypeptide with an approximate molecular weight ITT 34 kDa. DAS ELISA kit specific for BlCMV showed positive results for BlCMV antigen. Total RNA extracted from infected tissues, using total RNA extraction kit was used for reverse transcription and subsequent amplification of viral sequences. Alignments of potyvirus and comovirus coat protein gene sequences allowed the selection of degenerate PCR primers in conserved coat protein region. BlCMV coat protein gene was successfully amplified by reverse transcriptionpolymerase chain reaction (RT-PCR) using degenerate primers. The amplified genome fragment 290 base pairs in size, amplified from the coat protein gene of virus, was analyzed by gel electrophoresis and visualized by Ethidium bromide staining.EnglishThesis