WxcX is involved in bacterial attachment and virulence in Xanthomonas campestris pv. campestris

Abstract

Xanthomonas campestris pv. campestris (Xcc) is the causative agent of black rot in crucifers. Here, one EZ-Tn5 transposon mutant of Xcc, altered in bacterial attachment, was isolated. Further analysis revealed that the transposon was inserted in the wxcX gene (encodes a hypothetical protein) of the transposon mutant. Sequence analysis revealed that WxcX is highly conserved in Xanthomonas, but none has been characterized. In this study, it was indicated that mutation of wxcX resulted in enhanced bacterial attachment, reduced virulence on the host cabbage, and increased sensitivity to sodium dodecyl sulfate. The affected phenotypes of the wxcX mutant could be complemented to wild-type levels by the intact wxcX gene. Site-directed mutagenesis revealed that E408 and E411 are critical amino acid residues for WxcX function in bacterial attachment. Taken together, our results demonstrate the roles of wxcX in attachment, virulence, and tolerance to sodium dodecyl sulfate in Xanthomonas for the first time.

Functional characterization and proteomic analysis of lolA in Xanthomonas campestris pv. campestris

“…Previously, we subjected the X. campestris pv. campestris wild-type strain Xcc17 to transposon mutagenesis and subsequently screened approximately 1000 transposon mutants for bacterial attachment ability and biofilm formation [13, 29]. One mutant, H27, which exhibited reduced attachment ability, was characterized in this study.…”

“…oryzae strain KACC10331 [49]. Previously, we screened an EZ-Tn 5 library containing approximately 1000 Xcc17 mutants for their attachment ability and identified two novel biofilm-related genes ( prc and wxcX ) that have been further determined to play a role in pathogenicity [13, 29]. In this study, we extended our previous work to identify an attachment-reduced mutant (H27) that had a transposon inserted in the lolA gene and putatively encoded an outer membrane lipoprotein chaperone in Xcc17.…”

“…One transposon mutant, designated as H27, exhibiting reduced attachment was selected for further characterization. The site inserted by transposon in H27 was identified using the rescue cloning method using the TransforMax™ EC100D™ pir + electrocompetent E. coli (Epicentre), as previously described [29].…”

Functional characterization and transcriptional analysis of degQ of Xanthomonas campestris pathovar campestris

“…Transposon mutagenesis of Xcc17, screening of bacterial attachment, and transposon insertion site identification were conducted as previously mentioned [12,16]. The EZ-Tn5 TM < R6Kγori/KAN-2> Tnp transposome Kit used for transposon mutagenesis was supplied by Lucigen.…”

“…In our laboratory, a transposon insertion library of Xcc strain 17 (hereinafter, Xcc17) was previously constructed [12]. Transposon mutants have been used to screen for biofilmrelated genes by evaluating altered attachment ability compared with wild-type strains [12][13][14][15][16]. One mutant that had decreased attachment ability was identified, and this mutant was found to have a transposon inserted in the degQ gene.…”