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In Vitro Production of Plum Pox Virus - Free Plums by Chemotherapy with Ribavirin

ABSTRACT Plum pox virus (PPV)-free plants of Prunus domestica L. cv Cacanska Lepotica were obtained from PPV-infected shoots by chemotherapy in vitro. The infected shoots were multiplied on MS medium supplemented with (in mg l−1): BAP 1.0, IBA 0.1, GA3 0.1 and subjected to chemotherapy with ribavirin (1-β-ribofuranosyl-1,2,4-triazole-3-carboxamide) at the concentrations of 10, 20, 40, 60, 80, and 100 mg l−1 during a six-week period. After chemotherapy, regenerated and rooted plants were grown for two years under the screen house conditions. The plants were re-tested for the presence of PPV by DAS-ELISA before and immediately after the treatments, and two years after, by both DAS-ELISA and IC-RT-PCR. Chemotherapy with ribavirin at concentrations of 40 and 60 mg l−1 was found to be effective in raised PPV-free plants, i.e. 15.38% and 16.66% respectively, as proved by IC-RT-PCR. At lower concentrations, 10 and 20 mg l−1, ribavirin was entirely ineffective, and accordingly, all obtained plants were infected. However, at higher concentrations, 80 and 100 mg l−1, ribavirin exhibited distinctive signs of phytotoxicity and shoots failed to endure the treatment.