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2023, № 106

UDC: 634.8:632.3:632.9:579.64
GSNTI: 68.35.55:68.37.31:34.27.15

PCR diagnosis of crown gall infection in vineyards of the Krasnodar Territory

Crown gall is of economic importance for the domestic viticulture industry. The causative agents of the disease are pathogenic agrobacteria Allorhizobium vitis, Agrobacterium tumefaciens and Rhizobium rhizogenes. The disease has a latent period of infection, and all organs of the infected plant, in particular the vine, are carriers of the pathogen. Reliable identification of grape crown gall infection by molecular genetic methods in planting material and uterine plantations is an urgent issue in the grape nursery industry. Purpose of the study: using DNA markers to determine the species composition of causative agents of crown gall in the vineyards of the Krasnodar Territory, to propose an effective protocol for PCR diagnostics of latent infection. A total of 351 DNA samples were analyzed. The study was conducted from 2015 to 2023. DNA samples were isolated by a modified CTAB method. Crown gall was diagnosed by PCR using the UF/B1R/B2R/AVR2, virD2A/virD2C, and PGF/PGR/VIRFF1/VIRFR2/VIRD2S4F716/VIRD2S4R1036 test systems, followed by horizontal electrophoretic separation of the amplified fragments in agarose gel. When analyzing DNA samples from the affected plant tissue, it was determined that the main causative agent of crown gall of grapes in the vineyards of the Krasnodar Territory is agrobacteria All. vitis (90,5%), A. tumefaciens in 9,5% of samples, and R. rhizogenes (46,6 %) in mixed infection. Evaluation of asymptomatic material showed that for effective diagnosis of the causative agent of crown gall of grapes in the latent stage, all 3 test systems should be used in the following sequence: UF/B1R/B2R/AvR, PGF/PGR/VIRFF1/VIRFR2/VIRD2S4F716/VIRD2S4R1036, virD2A/virD2C. Analysis of plants without signs of crown gall, but potentially infected, revealed the presence of All. vitis (27%), A. tumefaciens (20%), as well as specimens with mixed infection - All. vitis + A. tumefaciens (14%) and All. vitis + A. tumefaciens + R. rhizogenes (5%) and free from agrobacterial infection (30%), in 4 % of the samples the pathogen type was not determined.
Keywords: Crown gall, grapevine, Allorhizobium vitis, Agrobacterium tumefaciens, Rhizobium rhizogenes, DNA markers.
DOI: 10.21515/1999-1703-106-273-277

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Authors:

1. Makarkina Marina Victorovna, Junior Research Associate, Federal State Budget Scientific Institution "North Caucasian Federal Scientific Center of Horticulture, Viticulture, Wine-making".
2. Ilnitskaya Elena Tarasovna, Cand. Biol. Sci., Head of Laboratory, Federal State Budget Scientific Institution "North Caucasian Federal Scientific Center of Horticulture, Viticulture, Wine-making".