Анализ распространения фитоплазм почернения древесины и золотистого пожелтения на виноградниках Краснодарского края

Abstract

The purpose of research is to study the spread of phytoplasma infection in the vineyards of the Krasnodar Region using the PCR method. The object of the study is grape plants of various varieties with external symptoms of phytoplasma infection. The material for total DNA extraction was symptomatic leaves (their veins and petioles) collected from plantings of Shiraz, Chardonnay, Cabernet Sauvignon, Saperavi, Mizar, Moldova, Augustine, Pinot Blanc, Riesling, and Aligote varieties. DNA extraction was carried out using a buffer based on the cationic detergent CTAB (cetyltrimethylammonium bromide), according to the protocol recommended by EPPO (European plant protection organization) for the isolation of phytoplasmas from grape plant material. Sampling was carried out in industrial vineyards of the Krasnodar Region in 9 different geographical locations of the Novorossiysk, Temryuk and Anapa Regions. DNA was isolated from symptomatic leaves according to the protocol recommended by EPPO (European plant protection organization). Determination of the presence of phytoplasma infection was carried out by real-time PCR on a Quant Studio 5 amplifier using a commercial kit designed to detect phytoplasmas – Candidatus Phytoplasma solani and Candidatus Phytoplasma vitis (Synthol) and a test system recommended by EPPO. Using a kit produced by Synthol, 239 samples were analyzed, including 12 rechecked with markers recommended by EPPO. As a result of the analysis, it was found that DNA of Candidatus Phytoplasma solani was present in 113 samples studied. DNA from the phytoplasma Candidatus Phytoplasma vitis, which causes golden yellowing of leaves, was not detected in any of the analyzed samples.