Molecular characterization of Pseudomonas syringae pvs. from different host plants by repetitive sequence-based PCR and multiplex-PCR

Abstract

Pseudomonas syringae pvs., isolated from different cultivars of sweet cherry grown in several locations in Serbia, were characterized and compared with strains collected earlier from sour and sweet cherry and oil pumpkin growing in this region, as well as with reference strains P. syringae pv. morsprunorum race 1 CFBP2119, P. s. pv. lachrymans 765R and P. s. pv. syringae H-1. By employing LOPAT and GATTa tests, isolates were identified as P. syringae pv. syringae and P. s. pv. morsprunorum race 1. Simultaneous detection of syringomycin synthesis (syrB) and syringomycin secretion (syrD) genes in multiplex-polymerase chain reaction (m-PCR) was used for P. syringae pv. syringae confirmation. All isolates detected as P. syringae pv. morsprunorum race 1 after biochemical characterization were positive for cfl gene amplification. Using a repetitive sequence-based PCR (rep-PCR) both syringae and morsprunorum race 1 pathovars were clustered separately, with 42% similarity. No significant differences between isolates in each pathovar were noted, although they were collected from different sweet cherry cultivars and varying localities. The most similar to the P. syringae pv. syringae isolates was strain T6 with 19% similarity, followed by strain Tk21 from oil pumpkin -25%. The isolates of both pathovars were detected on the same location (Selenca, Serbia) and the same cultivar ('Merchant'), albeit in two different years.

Evaluation of cherry cultivar susceptibility to bacterial canker and leaf spot disease

“…were cultivated, disease symptoms were detected only on V. Chkalov, Summit, B. Burlat and N. Star (which were rated as highly susceptible). Occurrence of bacterial spots on sweet cherries in a few locations in Vojvodina Province, Serbia, was reported earlier (Balaž et al, ; Iličić, ; Iličič, Balaž, et al, ). Bacterial spot can affect 15%–40% of sweet cherry leaves, and cvs.…”

“…Obtained isolates were tested for different biochemical characters: Gram reaction; presence of catalase; production of fluorescent pigment on King B medium; oxidative-fermentative metabolism of glucose; LOPAT and GATTa tests (Latorre & Jones, 1979;Lelliott & Stead, 1987;Lelliott et al, 1966). Molecular identification was based on the detection of genes for discriminative bacterial toxin production or synthesis, namely: coronatine production gene (cfl) using Primer1/2 primer set (Bereswill et al, 1994) and genes for syringomycin synthesis (syrB) and syringomycin secretion (syrD) by two sets of primers, B1/B2 and SyD1/SyD2 (Iličič, Balaž, et al, 2016). The reference strains KFB0120 (pv.…”

Journal of Plant Protection Research

“…Most phenotypic tests such as Gram staining, fluorescent pigmentation on KB medium and LOPAT were similar to strains previously reported (Mohammadi et al 2001;Natalini et al 2006;Ilicic et al 2016). In this study, strains isolated from barley could not produce levan polymers.…”

“…Syringomycin is an important bacterial phytotoxin with a number of syr genes involved in regulation (syrA, syrF, syrG and salA), biosynthesis (syrB1, syrB2, syrC and syrE) and secretion (syrD and syrP) (Bender et al 1999;Vaughn and Gross 2016;Adbellatif et al 2017). Pss strains were compared with each other on the basis of syrB, syrC and syrD genes (Natalini et al 2006;Gasic et al 2012;Ivanovic et al 2012;Ilicic et al 2016). In this study, we found a correlation between syringomycin production in vitro and amplification of syrB (752 bp) and syrD (446 bp) genes.…”

The presence of pathogenic bacteria Pseudomonas syringae in cereals in Lithuania

“…P. syringae pv. syringae can infect an extensive host range of monocotyledonous and dicotyledonous plants (Iličic et al, 2016;Choi et al, 2017). P. syringae pv.…”