Identification of tomato pathogens de novo

One of the most disease-prone crops is tomato (Solanum lycopersicum), which creates the need to identify the pathogens causing these diseases as accurately and completely as possible. One of the methods allowing to determine pathogens from scratch or their complex is sequencing of highly conserved DNA locus. The purpose of the work was to determine the species composition of pathogenic fungi and bacteria by this method. The study was conducted in 2023, in the Bakhchisaray district. The material was selected from tomato plants of Pink Heart variety (China) grown under protected soil conditions. During visual inspection, two types of lesions were detected on the plants, from which 15 isolates of bacteria and fungi were isolated into pure culture on LB medium, pea-carrot agar and potato-dextrose agar. From the grown cultures, samples with similar features were selected for isolation of total DNA, on which PCR with universal fungal and bacterial primers was performed. The amplicons obtained were sequenced. Nucleotide sequences homologous to Enterobacter ludwigii, E. kobei, E. cloaca and Pseudomonas veronii in bacteria were obtained from the first lesion type. The identity of the obtained homologues was 100% at 100% coverage. In fungi from the same type of lesions, the nucleotide sequences were homologous to Botrytis cinerea and the genus Penicillium, their homologues were, as in bacteria, 100% with 100% coverage. Sequences homologous to Cladosporium sp. and Penicillium chrysogenum were obtained from the second lesion type. The identity of the homologues was 100% with 100% coverage.

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