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European Journal of Plant Pathology

, Volume 104, Issue 9, pp 945–950 | Cite as

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

  • Pasquale Saldarelli
  • Adib Rowhani
  • Geoffrey Routh
  • Angelantonio Minafra
  • Michele Digiaro
Article

Abstract

RT-PCR with degenerate primers was used for the screening of the genome of some members of the Closterovirus, Vitivirus and Trichovirus genera. Two sets 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 allocation of grapevine viruses A, B, D and heracleum latent virus (HLV) in the genus Vitivirus, whereas, the detection of a HSP70 homologue in grapevine leafroll-associated viruses agrees with their assignment in the genus Closterovirus. The use of degenerate primers for the identification of grapevine viruses belonging to Vitivirus and Closterovirus genera is envisaged.

degenerate primers filamentous viruses RT-PCR phylogenetic analysis vitivirus trichovirus closterovirus grapevine 

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Copyright information

© Kluwer Academic Publishers 1998

Authors and Affiliations

  • Pasquale Saldarelli
    • 1
  • Adib Rowhani
    • 2
  • Geoffrey Routh
    • 2
  • Angelantonio Minafra
    • 1
  • Michele Digiaro
    • 3
  1. 1.Centro di Studio del CNR sui Virus e le Virosi delle Colture MediterraneeBariItaly
  2. 2.Foundation Plant Material ServiceUniversity of CaliforniaDavisUSA
  3. 3.CIHEAM-Istituto Agronomico MediterraneoBariItaly

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