Molecular characterization of Pseudomonas syringae isolates from fruit trees and raspberry in Serbia

Ivanović, Žarko; Stanković, Slaviša; Živković, Svetlana; Gavrilović, Veljko; Kojić, Milan; Fira, Djordje

doi:10.1007/s10658-012-9978-4

Cited by 13 publications

(12 citation statements)

References 27 publications

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“…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).…”

Section: Methodsmentioning

confidence: 99%

“…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.…”

Section: Molecular Characterization Of Pseudomonas Syringae Pvs Frommentioning

confidence: 96%

“…syringae strains, with the findings revealing high genetic heterogeneity between P. syringae pv. syringae strains originating from different and the same hosts (Natalini et al, 2006;Gilbert et al, 2009;Kaluzna et al, 2010;Ivanović et al, 2012). Repetitive element sequence-based PCR (rep-PCR) is based on three main sets of repetitive DNA elements: the repetitive extragenic palindromic (REP) elements (REP-PCR), the enterobacterial repetitive intergenic consensus (ERIC) sequences (ERIC-PCR) and BOX elements (BOX-PCR), and this method is successful in typing a variety of bacteria, as well as P. syringae (Vicente, Roberts, 2007;Marques et al, 2008;Kaluzna et al, 2010).…”

Section: Molecular Characterization Of Pseudomonas Syringae Pvs Frommentioning

confidence: 99%

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

Iličić

Balaž

Stojšin

et al. 2016

Zemdirbyste-Agriculture

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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.

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Section: Methodsmentioning

confidence: 99%

Section: Molecular Characterization Of Pseudomonas Syringae Pvs Frommentioning

confidence: 96%

Section: Molecular Characterization Of Pseudomonas Syringae Pvs Frommentioning

confidence: 99%

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

Iličić

Balaž

Stojšin

et al. 2016

Zemdirbyste-Agriculture

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“…Two sets of primers were simultaneously used in PCR: B1/B2 specific to syr B gene, and SyD1/SyD2, specific to syr D gene. Those primer sets were previously used in PCR during Pss testing (Gasic et al., ; Ivanovic et al., ; Natalini, Rossi, Baronovi, & Scortchini, ). PCR cycling was performed in a total volume of 25 μl containing 2.5 mM MgCl2, 0.2 mM dNTPs, 0.2 μM of each primer (B1/B2 and SyD1/SyD2), 2.5 μl of 10× buffer (100 mM Tris‐HCl, 500 mM KCl, pH 8.4), 1.25 U Taq DNA polymerase (CinnaGen) and 3 μl of template DNA.…”

Section: Methodsmentioning

confidence: 99%

Characterization and genetic diversity of Pseudomonas syringae isolates from stone fruits in north‐western Iran

Nosratnezhad

Rouhrazi

Khezrinezhad

2018

Journal of Phytopathology

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From 33 Iranian fluorescent Pseudomonas isolates originating from symptomatic tissues of peach (Prunus persica), plum (Prunus domestica), sweet (Prunus avium) and sour cherry (Prunus cerasus), 27 were identified as Pseudomonas syringae using LOPAT tests. Further characterization of those isolates by GATTa and L‐lactate utilization tests and the detection of syringomycin and coronatine and yersiniabactin coding genes showed that five of them belonged to race 1 and four to race 2 of P. syringae pv. morsprunorum (Psm) and eighteen other isolates were identified as P. syringae pv. syringae (Pss). Based on the analysis of the fingerprint patterns generated by REP, ERIC and BOX‐PCR, the strains were differentiated into three main groups at the 67% similarity level. Strains of the groups 1, 2 and 3 belong to Psm race 1, Psm race 2 and Pss, respectively. Rep‐PCR analysis showed high intra‐pathovar variation within the Pss isolates, which grouped into four distinct clusters. Using the REP primers, the percentage of polymorphic loci was 74.61%, whereas with BOX and ERIC primers, it was 60.5 and 55.21%, respectively. Finally, this study is the first report of the isolation of P. syringae pv. morsprunorum race 1 and 2 strains from stone fruit trees in Iran.

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“…Ova istraživanja uključuju inokulacione testove, biohemijske karakteristike (GATT), korišćenje REP-PCR i BOX prajmera, kao i onih za detekciju gena za produkciju siringomicina i koronatina (Bultreys and Kaluzna, 2010;Ivanović, 2011;Ivanović et al, 2012).…”

Section: Rezimeunclassified

Pseudomonas syringae: Causal agent of sweet cherry fruit necrosis

Gavrilović¹,

Stošić²,

Stevanović³

2014

Plant protection

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U radu su prikazani rezultati proučavanja sojeva bakterija izolovanih iz nekrotičnih plodova trešnje sorte Samit (područje Šapca). Nekroza zahvata u proseku oko 1/3 ploda koji potpuno gubi tržišnu vrednost. Bolešću biva zahvaćeno oko 30% plodova. Primenom standardnih bakterioloških metoda (izolacijom na hranljivim podlogama), proverom patogenosti i proučavanjem bakterioloških karakteristika, zaključeno je da pomenute simptome prouzrokuje fitopatogena bakterija Pseudomonas syringae, široko rasprostranjen patogen naročito koštičavih voćaka.Ključne reči: trešnja, nekroza ploda, Pseudomonas syringae, patogenost, bakteriološke odlike REZIME Simptomi nekroze plodova trešnje (sorta Samit), zapaženi su posle perioda obilnih padavina tokom 2014. godine. Procenat obolelih plodova je bio oko 30%. Simptomi se ispoljavaju u vidu mrkih nekrotičnih, naboranih pega, koje zahvataju 1/3 ploda. (Slika 1). Slične simptome na plodovima trešnje može prouzrokovati i fitopatogena gljiva Monilinia spp., ali je zapaženo odsustvo fruktifikacije na obololeim plodovima, što ukazuje da bi prouzrokovač bolesti mogao biti i druge prirode. S obzirom da i fitopatogene bakterije roda Pseudomonas mogu prouzrokovati slične simptome (Sutton and Jones, 1996), cilj rada je bio da se utvrdi prouzrokovač ove bolesti, koji na osnovu zapažanja u zasadu trešnje može naneti velike štete.Oboleli plodovi trešnje prikupljani su polovinom maja, a izolovanje patogena je izvršeno na podlozi King B i hranljivoj podlozi (hranljivi agar, Torlak) obogaćenoj sa 5% sahroze (NAS). Patogenost dobijenih sojeva je proverena inokulacijom plodova trešnje i inokulacijom listova duvana (HR), (Lelliot and Stead, 1987; Aresenijević, 1997). Plodovi trešn-je su inokulisani injektiranjem suspenzije bakterije (10 7 cfu), medicinskim špricem neposredno ispod epidermisa ploda. Inokulisani su plodovi sorte Samit, na kojoj su i primećeni simptomi bolesti i sorte Sambarst koja sazrava polovinom juna i čiji su plodovi u vreme inokulacije bili potpuno zeleni. Inoklisano je po tri ploda svakim sojem. Plodovi su posle inokulacije odlagani u plastiče kutije obložene vlažnim filter papirom, radi održavanja optimalne vlažnosti neophodne za razvoj patogena.Od bakterioloških odlika proučene su: reakcija po Gramu, O/F test, aktivnost oksidaze, arginin-dehidrolaze i aktivnosti pektolitičkih enzima inokulisanim kriškama krompira (LOPAT test), ( Lelliott et al., 1966; Arsenijević, 1997). Kao kontrolni soj pri ovim ispitivanjima korišćen je P.syringae -CFBP 11, poreklom iz Nacionalne kolekcije fitopatogenih bakterija u Francuskoj.Posle tri dana razvoja na King B podlozi se pojavljuju bakterijske kolonije bele boje, ravnih ivica, blago ispupčene, koje stvaraju fluorescentni pigment. Na podlozi obogaćenoj saharozom (NAS) nakon tri dana patogen formira izrazito ispupčene kolonije, prečnika oko 3 mm, sluzaste, (levan tipa).Nekroza mrke boje i naborane površine, pojavljuje se na inokulisanim plodovima posle 3 dana i po izgledu odgovara nekrozi primećenim pri prirodnim infekcijama. Identični simptomi su za...

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Molecular characterization of Pseudomonas syringae isolates from fruit trees and raspberry in Serbia

Cited by 13 publications

References 27 publications

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

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

Characterization and genetic diversity of Pseudomonas syringae isolates from stone fruits in north‐western Iran

Pseudomonas syringae: Causal agent of sweet cherry fruit necrosis

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