Characterization of Clavibacter michiganensis subsp. michiganensis strains from recent outbreaks of bacterial wilt and canker in Serbia

Milijašević-Marčić, Svetlana; Gartemann, Karl-Heinz; Frohwitter, Jonas; Eichenlaub, Rudolf; Todorović, Biljana; Rekanović, Emil; Potočnik, Ivana

doi:10.1007/s10658-012-0046-x

Cited by 28 publications

(22 citation statements)

References 27 publications

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“…Moreover, gyrB gene was used in MLST schemes developed to type Cmm strains [7,33,37]. DnaA sequence was shown a good taxonomic marker to identify and classify plant pathogenic bacteria such as Clavibacter, Xanthomonas and Ralstonia [38].…”

Section: Methodsmentioning

confidence: 99%

“…A high homogeneity was also observed among strains isolated from 2002 to 2007 in Canary Islands suggesting a single introduction of the pathogen as a source of infection [6]. Primary infections in many countries were attributed to the introductions of contaminated tomato seeds and/or seedlings [7,10]. These findings indicate that seeds play an important role in long-distance spread of the pathogen.…”

Section: Introductionmentioning

confidence: 99%

“…A direct link between tomato cultivar, year or place of isolation and Cmm type mostly could not be recognized [6,8,9] except the outbreak in 2001 in Turkey where bacterial canker was detected only on one tomato cultivar ‘Target’ [11]. Interestingly, in Israel and Serbia Cmm strains showing the same haplotypes were repeatedly isolated from the same locations during several subsequent years [7,10]. Reoccurring outbreaks suggest that despite intensified efforts for eradication, reliable control of this disease remains an unattainable goal.…”

Section: Introductionmentioning

confidence: 99%

“…PFGE (pulsed-field gel electrophoresis of macro-restricted bacterial DNA), one of the oldest techniques used in epidemiology, is labor intensive and expensive but is still used as a gold standard in typing of some bacterial species [10,13]. PFGE was applied to study the diversity of Cmm strains from outbreaks in Serbia [7] and in Israel [10] where the results of PFGE showed similar resolution of those obtained by gene sequence analysis and rep-PCR, respectively. Also, AFLP, a high resolution molecular typing method was applied by De Leon and coworkers to study genetic diversity of Cmm strains from Canary Islands [6].…”

Section: Introductionmentioning

confidence: 99%

“…Highly reproducible multilocus sequence typing (MLST) was used to analyze Cmm population from Serbia. Cmm strains were divided into seven groups and the results were confirmed by PFGE analysis [7]. …”

Section: Introductionmentioning

confidence: 99%

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Multilocus Variable-Number-Tandem-Repeats Analysis (MLVA) distinguishes a clonal complex of Clavibacter michiganensis subsp. michiganensis strains isolated from recent outbreaks of bacterial wilt and canker in Belgium

Zaluga

Stragier

Vaerenbergh³

et al. 2013

BMC Microbiology

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BackgroundClavibacter michiganensis subsp. michiganensis (Cmm) causes bacterial wilt and canker in tomato. Cmm is present nearly in all European countries. During the last three years several local outbreaks were detected in Belgium. The lack of a convenient high-resolution strain-typing method has hampered the study of the routes of transmission of Cmm and epidemiology in tomato cultivation. In this study the genetic relatedness among a worldwide collection of Cmm strains and their relatives was approached by gyrB and dnaA gene sequencing. Further, we developed and applied a multilocus variable number of tandem repeats analysis (MLVA) scheme to discriminate among Cmm strains.ResultsA phylogenetic analysis of gyrB and dnaA gene sequences of 56 Cmm strains demonstrated that Belgian Cmm strains from recent outbreaks of 2010–2012 form a genetically uniform group within the Cmm clade, and Cmm is phylogenetically distinct from other Clavibacter subspecies and from non-pathogenic Clavibacter-like strains. MLVA conducted with eight minisatellite loci detected 25 haplotypes within Cmm. All strains from Belgian outbreaks, isolated between 2010 and 2012, together with two French strains from 2010 seem to form one monomorphic group. Regardless of the isolation year, location or tomato cultivar, Belgian strains from recent outbreaks belonged to the same haplotype. On the contrary, strains from diverse geographical locations or isolated over longer periods of time formed mostly singletons.ConclusionsWe hypothesise that the introduction might have originated from one lot of seeds or contaminated tomato seedlings that was the source of the outbreak in 2010 and that these Cmm strains persisted and induced infection in 2011 and 2012. Our results demonstrate that MLVA is a promising typing technique for a local surveillance and outbreaks investigation in epidemiological studies of Cmm.

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