Grapevine vitivirus A eradication in Vitis vinifera explants by antiviral drugs and thermotherapy

J Virol Methods. 2007 Dec;146(1-2):129-35. doi: 10.1016/j.jviromet.2007.06.008. Epub 2007 Jul 23.

Abstract

Grapevine shoot cultures infected by Grapevine vitivirus A (GVA) were grown on Quorin-Lepoivre basic medium and submitted to in vitro chemotherapy and thermotherapy sanitation techniques. Ribavirin (Rb) at 20gml(-1), dihydroxypropyladenine (DHPA) at 60gml(-1) and their combination (RbDH) were added to the proliferating medium for three subsequent subcultures of 30 days each. Phytotoxicity was observed on drug-treated plantlets, which displayed a high percentage of mortality for each drug at doses higher than those aforementioned. Sequential ELISA were performed at the end of each subculture and ELISA-negative explants were submitted to RT-PCR. ELISA showed no antiviral activity following DHPA administration. Rb and RbDH treatment produced ELISA-negative explants which were assayed by RT-PCR and nested PCR. Biomolecular results showed no virus eradication in Rb treated explants but RbDH administration generated a percentage (40.0%) of GVA-free plantlets that permitted restoration of a new healthy generation of explants. Sixty percent (60%) of GVA eradication as confirmed by RT-PCR was obtained by in vitro thermotherapy at 36 degrees C for 57 days.

MeSH terms

  • Adenine / analogs & derivatives*
  • Adenine / pharmacology
  • Antiviral Agents / pharmacology*
  • Flexiviridae / drug effects*
  • Flexiviridae / isolation & purification
  • Hot Temperature*
  • Plant Diseases / virology
  • Ribavirin / pharmacology*
  • Vitis / virology*

Substances

  • Antiviral Agents
  • Ribavirin
  • 9-(2,3-dihydroxypropyl)adenine
  • Adenine