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Background  The Gram-negative bacterium  Xanthomonas citri  subsp.  citri  ( Xac ) causes citrus canker, one of the most destructive diseases of citrus worldwide. In our previous work, a transposon mutant of  Xac  strain 306 with an insertion in the  XAC3110  locus was isolated in a screening that aimed at identifying genes related to biofilm formation. The  XAC3110  locus was named as  bdp24  for biofilm-defective phenotype and the mutant was observed to be affected in extracellular polysaccharide (EPS) and lipopolysaccharide (LPS) biosynthesis and cell motility. In this study, we further characterized the  bdp24  (XAC3110) gene (designated as  gpsX ) using genetic complementation assays and expanded the knowledge about the function of the  gpsX  gene in  Xac  pathogenesis by investigating the roles of  gpsX  in EPS and LPS production, cell motility, biofilm formation on host leaves, stress tolerance, growth  in planta , and host virulence of the citrus canker bacterium.    Results  The  gpsX  gene encodes a putative glycosyltransferase, which is highly conserved in the sequenced strains of  Xanthomonas . Mutation of  gpsX  resulted in a significant reduction of the amount of EPS and loss of two LPS bands visualized on sodium dodecylsulphate- polyacrylamide gels. Biofilm assays revealed that the  gpsX  mutation affected biofilm formation by  Xac  on abiotic and biotic surfaces. The  gpsX  mutant showed delayed bacterial growth and caused reduced development of disease symptoms in susceptible citrus leaves. The  gpsX  mutant was more sensitive than the wild-type strain to various stresses, including the H 2 O 2  oxidative stress. The mutant also showed attenuated ability in cell motility but not in flagellar formation. Quantitative reverse transcription-PCR assays indicated that mutation of  gpsX  did not affect the expression of virulence genes such as  pthA  in  Xac  strain 306. The affected phenotypes of the  gpsX  mutant could be complemented to wild-type levels by the intact  gpsX  gene.    Conclusions  Taken together, our data confirm that the  gpsX  gene is involved in EPS and LPS synthesis and biofilm formation in  Xac  and suggest that the  gpsX  gene contributes to the adaptation of  Xac  to the host microenvironments at early stage of infection and thus is required for full virulence on host plants.

bmc microbiology

The gpsX gene encoding a glycosyltransferase is important for polysaccharide production and required for full virulence in Xanthomonas citri subsp. citri