MOLECULAR BIOTECHNOLOGY OF HOP CROWN GALL DIAGNOSTIC

Aim. The development of molecular biotechnology for crown gall diagnosis in hop. Methods. Isolation of pure culture cells, extraction of DNA, polymerase chain reaction (PCR), gel electrophoresis. Results. For the total DNA extraction from the tissue there were used the buffer, which recorded the presence in hop tissue such specific compounds as humulone, lupulon, xanthohumol and others. There were performed PCRs with primers for genes of virulence and pathogenicity of the Ti-plasmid and DNAs extracted from the pure culture of the crown gall causative Agrobacterium tumefaciens, healthy samples of hop, hop samples with visual crown gall symptoms and agrobacterium and hop DNA mixtures at the ratio of 1:1 1:5, 1:10. PCR products were obtained with the primers to virulence and pathogenic genes of Ti-plasmid and DNA extracted from a pure culture of A. tumefaciens, the hop tissue samples with visual symptoms of crown gall and the mixtures of DNA from hop. According to the results of PCRs with total DNA from healthy samples of hops and controls (PCR-mix without DNA) the amplification products were not observed. Infected by crown gall were considered those samples of hop in which the total DNA virulence and pathogenicity genes of Ti-plasmid were identified. Conclusion. The diagnosis of hops should include the detection of pathogenes and virulence genes ipt and virD2