Development of new PCR tests for diagnostics of the agent of bacterial leaf streak of wheat Xanthomonas translucens

In order to develop PCR tests that allow identifying the phytopathogen Xanthomonas translucens, which is significant for the export of Russian grain products, a search was made for a suitable PCR target. As a result of bioinformatic analysis of annotated proteins corresponding to the coding sequences of 10 genomic assemblies of the target bacterium – Xanthomonas translucens – and 161 genomic assemblies of 25 other species of bacteria of the genus Xanthomonas, downloaded from NCBI GenBank in March 2020, 6 genome regions specific for Xanthomonas translucens and suitable for use as a PCR target. The nucleotide sequences of these genes were used to design primers. For the discovered sequences, 12 pairs of primers were developed – 1F8/1R8, 1F10/1R10, 2F6/2R6, 3F3/3R3, 3F5/3R5, 3F9/3R9, 4F1/4R1, 4F3/4R3, 5F3/5R3, 5F6/5R6, 6F6/6R6 and 6F10/6R10, the species-specificity of which was tested in the course of the study with 35 strains of bacteria of the genus Xanthomonas, including Xanthomonas translucens. Analysis of the results of the PCR performed and the alignment of the obtained nucleotide sequences of the PCR products using the BLAST algorithm based on the NCBI showed that the primer pairs 1F8/1R8, 1F10/1R10, 4F1/4R1, 5F6/5R6, and 6F10/6R10 are suitable for their use in the diagnosis of the pathogen causing bacterial leaf streak of wheat, Xanthomonas translucens. New PCR tests can become part of the solution to the problem of establishing the phytosanitary state of consignments of Russian grain products, and will also make it possible to inspect the territories of the Russian Federation and inspect consignments of regulated products.

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