Use of degenerate primers in a RT-PCR assay for the identification and analysis of some filamentous viruses, with special reference to clostero- and vitiviruses of the grapevine.

摘要: RT-PCR with degenerate primers was used for the screening of genome some members Closterovirus, Vitivirus and Trichovirus genera. Two sets primers, targeted to conserved sequences heat shock protein 70 homologue closteroviruses or RNA dependent polymerase genes tricho- vitiviruses, amplified expected fragments from total extracts double-stranded RNAs infected plants. Amplified cDNAs were cloned, sequenced phylogenetically analyzed. Results support allocation grapevine viruses A, B, D heracleum latent virus (HLV) in genus Vitivirus, whereas, detection a HSP70 leafroll-associated agrees their assignment Closterovirus. The use identification belonging Closterovirus genera is envisaged.