Development of Methodologies for Rapid Detection of Grapevine Viruses
We have produced specific polyclonal antibodies to GLRaV-1 and GVA Evaluated the antiserum produced to GVA, GVB, GLRaV-1, GLRaV-4, and GFKV. We have produced specific polyclonal antibodies in goat for use in I-ELISA for GLRaV-1 and GLRaV-4. These antibodies have given us good reaction and low non-specific background. We have investigated the existence of different strains of GRSPaV. So far we have found three different strains with about 80-85%homology among their coat protein gene sequence. We have developed strain-specific and universal primers for the detection of GRSPaV in RT-PCR technique. We have found that by excising meristem tip of 0.5 mm or less from RSP-infected vines and grew in tissue culture media would eliminate GRSPaV with high efficiency. We have cloned and sequenced pieces of the genome of a new virus causing leafroll disease in grapevine (different from GLRaV-1 to -5) and its characterization is in progress. We have developed a sample preparation protocol for grapevine tissue for RT-PCR methodology.