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, GA(3) 0.1 and subjected to chemotherapy with ribavirin (1-beta-ribofui-anosyl-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 PP V 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 V, 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.