Slyvų devirusavimas in vitro sistemoje

Abstract

Plum virus elimination experiment was performed on June of 2015 in orchad plant genetics and biotechnology laboratory of Lithuanian Research Centre for Agriculture and Forestry (LRCAF). Three plum species: Magna glauca, Mlejevčenka and Čačanska Včesna microshoots, which were infected by ACLSV and PNRSV viruses, were affected by chemotherapy with ribavirin with the aim to eliminate the viruses from microshoots. Microshoots of 5 variants with different ribavirin concentrations (10, 20, 30, 40, 50 mg l-1) and one control variant without ribavirin, 7 days were grown on MS agar medium (Murashige and Skoog, 1962). After 7 days of chemotherapy microshoots were replaced on the agar medium without ribavirin. Experiment was performed in 5 replications for every plum species. Propagation coefficient of the microshoots after chemotherapy was assessed after 30 and 60 days. Propagated microshoots after chemotherapy were tested for viruses by PGR method after 60 days. RNA purified from homogenized leaves using GeneJET Plant RNA Purification mini Kit (Thermo Scientific) according to producers protocol. Purified RNA was used for complementary DNA (cDNA) synthesis. cDNA was synthesized using RevertAid First Strand cDNA Synthesis Kit (Thermo Scientific) according to producers protocol. Specific virus protein shell oligonucleotide primers were used for detection of ACLSV and PNRSV viruses.