To receive more information about evolutionary relatedness of strains from Belgium and France the MLVA data was analyzed taking into account the number of repeat differences (Additional file 1: Figure S1). Interestingly, Belgian strain PD 5737 and French strain PD 5749 clustered closer to ES2686.1 and CL01TF02 strains isolated in Spain during bacterial

canker outbreak in 2002–2003. Moreover, these four strains showed to have a more similar MLVA haplotype to the group of strains from recent Belgian Ferrostatin-1 datasheet outbreaks 2010–2012. Figure 3 Minimum spanning tree of 56 Cmm strains based on eight VNTR loci. Each circle represents an MLVA type with a size corresponding to the number of strains that share an identical MLVA type. MLVA PF 01367338 types connected by a thick solid line differ from one another by one VNTR locus, while MLVA types connected by a thin solid line differ by two VNTR loci. MLVA types that differ from each other by three, four or more VNTR loci are connected by dashed and dotted lines. MLVA types were distinguished to define clonal complexes and to group in zones MLVA types that differ from one another by at most two locus variants. Letters visible on each circle are corresponding to strains described in Table 1. CC-Clonal complex. Discussion and conclusion Over the last few decades, bacterial canker has been frequently detected in tomato production areas,

leading to substantial financial and economical losses. Only during the last three years several local outbreaks of Cmm were reported in Belgium. In some cases, reoccurring infections were detected in the primarily contaminated farms, suggesting a persistence of an initial infection source. Despite a quite frequent detection of tomato canker and wilting over in Belgian tomato production areas there is little known about the genetic diversity of Cmm strains which hinders the correct conclusions about the probable sources of epidemics and transmission routes of Cmm. This study is the first MLVA approach developed for efficient genotyping of Cmm strains. To date typing

of Cmm strains was performed by RAPD-PCR [6], BOX-PCR [8, 48], AFLP [6], PFGE [10] and MLST [7]. Despite the fact that some of these methods were found to have a good resolution most of them have limitations such as a poor interlaboratory portability or limited this website exchangeability of results that were generated on a specific machine or compared to an in-house database. Nowadays, fully sequenced genomes give a unique opportunity for a development of more robust and accurate typing methods such as MLVA. Its advantages, such as, high reproducibility, exchangeability of results and the possibility to add loci greatly facilitates epidemiological studies of economically important pathogens such as Cmm. In this work, Clav-VNTR5 showed to be the most polymorphic loci with five different alleles and the highest HGDI of 0.664.