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Archives of Virology

, Volume 144, Issue 6, pp 1071–1079 | Cite as

Synthesis of infectious transcripts of olive latent virus 1: genes required for RNA replication and virus movement

  • V. Pantaleo
  • F. Grieco
  • M. A. Castellano
  • G. P. Martelli
Article

Summary

Genomic RNA of olive latent virus 1 (OLV-1) contains five open reading frames (ORFs) encoding proteins of 23, 82, 8, 6 and 30 (CP) kDa. A full-length cDNA copy of OLV-1 RNA was prepared and cloned in a low-copy-number vector (pMUC-19) downstream of or T7 RNA polymerase promoter. Transcripts derived from this template, denoted pMUC-OLV, were highly infectious when inoculated in local and systemic hosts and infected tissues contained virus-like particles. Genes required for replication and virus movement were mapped by site-directed and deletion mutogenesis of the pMUC-OLV. ORF1 and ORF2 mutants were not viable, suggesting that replication requires the 23 and 82 kDa proteins. The 8 and 6 kDa polypeptides were involved in cell-to-cell movement, since their absence did not interfere with RNA replication but prevented systemic infection of inoculated plants. Mutant clones in R and S domains of the CP gene could replicate, but they did not systemically infect Nicotiana benthamiana, indicating that the CP gene is required for OLV-1 long-distance translocation. Mutant clones with large deletions in the CP gene were not viable, probably due to loss of 3′-proximal sequences required for RNA replication.

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Copyright information

© Springer-Verlag 1999

Authors and Affiliations

  • V. Pantaleo
    • 1
  • F. Grieco
    • 1
  • M. A. Castellano
    • 1
  • G. P. Martelli
    • 1
  1. 1.Dipartimento di Protezione delle Piante, Università degli Studi and Centro di Studio del CNR sui Virus e le Virosi delle Colture Mediterranee, University of Bari, ItalyItaly

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