Differentiation of Pseudomonas syringae pathovars originating from stone fruits

Abstract

SuMMARyDue to an overlapping host range, similar symptomatology and many common characteristics, Pseudomonas syringae pathovars originating from stone fruits can easily be misidentified. In order to select tests for rapid and efficient differentiation of P. s. pvs. syringae, morsprunorum and persicae, we studied the suitability and differentiating potential of some standard bacteriological and molecular methods. Differentiation of the strains was performed using LOPAT, GATTa and ice nucleation tests, nutrient sucrose broth growth and utilization of various carbon sources. PCR method enabled the detection of toxin-producing genes: syrB and syrD in P. s. pv. syringae, and cfl gene in P. s. pv. morsprunorum race 1. Syringomycin production by pv. syringae was confirmed in bioassay using Geotrichum candidum, Saccharomyces cerevisiae and Rhodotorula pilimanae as indicator organisms. Pathogenicity test on lemon and immature nectarine fruits, as well as on string bean pods, showed different intensity of reaction of the inoculated material which could separate pv. syringae from the other two pathovars. PCR-based repetitive sequences, Rep-PCR with REP, ERIC and BOX primers revealed different genetic profiles within P. syringae pathovars.

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

“…lachrymans -765R In this work, rep-PCR clearly indicated the differences between pathovars syringae and morsprunorum race 1. Thus, this method is useful in the identification of these pathogens globally, and in our country, as was previously reported by a number of authors (Vicente, Roberts, 2007;Gilbert et al, 2009;Ivanović et al, 2009;Bultreys, Kaluzna, 2010;Kaluzna et al, 2010;Gašić et al, 2012;Gavrilović et al, 2012;2013). The cumulative rep-PCR dendrogram revealed 58% differences between isolate groups I and II.…”

“…Those primer sets were previously used in a separate polymerase chain reaction (PCR) during P. s. pv. syringae testing (Natalini et al, 2006;Gašić et al, 2012;Ivanović et al, 2012). PCR amplification was carried out in a 30 µl reaction volume using GreenTaq Dream Master Mix (Thermo Scientific, Lithuania) with 1.2 µl of template DNA and 50 pmol of each primer (B1/B2 and SyD1/SyD2) (Metabion, International AG, Germany).…”

“…morsprunorum race 1 and produces an amplicon of 650 bp using primer set Primer1/2 (Metabion International AG, Germany). Amplification conditions adopted in this work followed the recommendations of Gašić et al (2012). PCR amplification was carried out in a 30 µl reaction volume using GreenTaq Dream Master Mix (Thermo Scientific, Lithuania) with 1.2 µl of template DNA and 50 pmol of primer1/primer2.…”

“…The authors indicated that this simultaneous detection method, including a large number of genes, is precise and rapid, and this assertion was confirmed in our investigation. Separate application of primer genes for syringomycin synthesis and secretion was reported by several authors Bultreys, Kaluzna, 2010;Gašić et al, 2012;Ivanović et al, 2012). According to Bultreys and Kaluzna (2010), some P. s. pv.…”

Bacterial halo blight of coffee crop: aggressiveness and genetic diversity of strains

“…juglandis (Scortichini et al 2001), P. syringae pv. syringae (natalini et al 2006) and studies of inter-and intraspecificity in strains of P. syringae (Vicente and Roberts 2007;Çepni and Gürel 2012;Gašić et al 2012).…”