The use of in vitro chemotherapy in grapevine virus elimination
The aim of the work was to eliminate single infection with grapevine fanleaf virus (GFLV) and grapevine leafroll-associate virus 1 (GLRaV-1) in two Vitis vinifera L. genotypes, 'Fetească neagră 7Od' and 'Frâncuşă 15Od', respectively, by chemotherapy with ribavirin and oseltamivir used concomitantly, in controlled conditions of in vitro culture. Apices and axillary buds collected from infected plants have been cultured on solidified MS basic medium supplemented with 1 mg L-1 BAP and 0.5 mg L-1 β-IAA. The viricide mixtures have been used in three variants of concentration during three consecutive subcultures, as follows: V1 = 40 mg L-1 ribavirin + 40 mg L-1 oseltamivir; V2 = 20 mg L-1 ribavirin + 40 mg L-1 oseltamivir; V3 = 20 mg L-1 ribavirin + 80 mg L-1 oseltamivir. The microshoots originated from stabilized infected cultures have been cut in single node fragments and subcultured on media containing viricides. In order to establish the efficiency of the method, three month-old acclimated plants have been analyzed by ELISA. The combination ribavirin-oseltamivir did not induce the mortality of explants, the significance of the differences of multiplication rates being influenced by the regenerative potential of grapevine genotype and virus infection. The virus elimination in grapevine by combining the two chemotherapeutics did not lead to the increased sanitation rate as expected. The highest percentage of GLRaV-1-free plants, 13.3%, was registered on V1 after two subculture treatments. In the case of GFLV-elimination, the best result, 6.8% healthy plants, has been obtained on V2 after one subculture.
Guţă, I.-C., Buciumeanu, E.-C., Topală, C.M. and Tătaru, L.D. 2017. The use of in vitro chemotherapy in grapevine virus elimination. Acta Hort. (ISHS) 1155:425-430
oseltamivir tissue culture, phytotoxicity, ribavirin, sanitation, Vitis