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
P. Saldarelli et al., 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, EUR J PL P, 104(9), 1998, pp. 945-950
RT-PCR with degenerate primers was used for the screening of the genome of
some members of the Closterovirus, Vitivirus and Trichovirus genera. Two se
ts of primers, targeted to conserved sequences of the heat shock protein 70
homologue of closteroviruses or to the RNA dependent RNA polymerase genes
of tricho- and vitiviruses, amplified the expected fragments from total RNA
extracts or double-stranded RNAs of infected plants. Amplified cDNAs were
cloned, sequenced and phylogenetically analyzed. Results support the alloca
tion of grapevine viruses A, B, D and heracleum latent virus (HLV) in the g
enus Vitivirus, whereas, the detection of a HSP70 homologue in grapevine le
afroll-associated viruses agrees with their assignment in the genus Closter
ovirus. The use of degenerate primers for the identification of grapevine v
iruses belonging to Vitivirus and Closterovirus genera is envisaged.