Genetic diversity of pseudomonas syringae pv. Syringae isolated from sweet cherry in southern and northern regions in Serbia

Abstract

Bacterial canker and leaf spot caused by plant pathogenic bacterium Pseudomonas is among the most destructive cherry diseases worldwide. Nowadays in Serbia, sweet cherry production significantly increased and the new plantations, mainly grown from imported planting material are being raised every year. During spring, 2018 and 2019, occurrence of bacterial canker and leaf spot symptoms was observed on a newly planted sweet cherry plantations in two localities, Zitorada (Southern region) and Karavukovo (Northern region-Vojvodina). Typical P. syringae colonies were isolated on Nutrient Sucrose Agar supplemented with 5% sucrose (NSA). A total of fifteen isolates were selected and identified. Results of the LOPAT test (+---+) determined them to belong to fluorescent Pseudomonas Group Ia, while results of G(+)A(+)T(-)Ta(-) tests indicate presence of Pseudomonas syringae pv. syringae. Pathogenicity was confirmed on immature sweet and sour cherry fruitlets by forming of black, sunken lesions for all tested isolates. Genes syrB and syrD were successfully detected in all tested isolates. DNA sequencing using gapA, gltA, gyrB and rpoD housekeeping genes determined tested isolates to belong to P. s. pv. syringae using the National Center for Biotechnology Information (NCBI) nucleotide BLAST. The Serbian isolates shared 99.47% to 100% (Zitorada) and 99.38% to 100% (Karavukovo) identity with bacterium P. s. pv. syringae. Phylogenetic analysis grouped isolates from Zitorada in one tree cluster, separate from the Karavukovo isolates,indicating presence of two genetically diverse groups of causal pathogen P. s. pv. syringae, obtained from two geographically distinct localities in Serbia. Phylogeographic analysis grouped isolates from Zitorada in multilocus haplotype coded as REz and isolates originated from Karavukovo in multilocus haplotype coded as REk. Considering that during last few years P. syringae continuously occurs mainly in young sweet cherry plantations, where imported material is used for raising, health status check is recommended to be included as obligatory measure when nursery material is used from import.