Node support: ML bootstrap/MrBayes posterior probability, values <70 or 0.70 are not shown. Using the same primer set as for wVulC ( Additional file 1: Table S1), the taxonomic distribution of pk1 and pk2 genes

was extended by PCR to seven Wolbachia strains that induce either CI or feminization in isopods. All these strains of selleck kinase inhibitor isopods are known to belong to the B-supergroup of Wolbachia whatever the phylogenetic marker used [2]. They do not form separate monophyletic clades according to the buy Necrostatin-1 phenotype they induce in their hosts based on the wsp gene ( Additional file 1: Figure S2). We also investigated the copy number variation by Southern blot analyses of EcoRI or BamHI digested DNA using pk1a pk1b and pk2b1 probes which, according to sequence identities, preferentially hybridized on pk1a pk1b and pk2b types, respectively (Table 2 & Additional file 1: Figure S1). In congruence with amplification and sequencing data, the pk1a and pk1b probes revealed two to six copies of the pk1 gene in the studied strains

(Table 2). By direct sequencing of the PCR products, we found VX-680 price that the pk1a gene of Wolbachia strains of C. convexus P. pruinosus A. vulgare (wVulM) and A. nasatum harboured 1, 1, 2 and 3 EcoRI sites, respectively, explaining the discrepancy between the number of bands observed by Southern blots, and the number of different sequences obtained (Table 2 & Additional file 1: Figure S1). Similarly, two pk1b alleles of the Wolbachia strain of A. nasatum contained one BamHI restriction site. Each of the two more intense Southern Florfenicol Blot signals ( Additional file 1: Figure S1) revealed the presence of two identical copies wVulC pk1b alleles, as confirmed by the analysis of contigs. Furthermore, Southern blots using a pk2b1

probe in combination with sequencing data revealed three copies of the pk2 gene in all strains tested except one (Table 2 & Additional file 1: Figure S1). In the Wolbachia strain of P. pruinosus, sequences of PCR products revealed two identical pk2 alleles, each containing one BamHI restriction site explaining the five signals obtained by Southern blotting (Table 2 & Additional file 1: Figure S1). Moreover, no signal was obtained from digested and undigested DNA of Wolbachia-free ovaries of isopod (non-infected population from Nice, France), which confirmed the Wolbachia origin of the pk1 and pk2 genes.