Type-II secretion pathway structural gene xpsE, xylanase- and cellulase secretion and virulence in Xanthomonas oryzae pv. oryzae

Abstract

Rice bacterial blight caused by Xanthomonas oryzae pv. oryzae is one of the most important diseases in rice-growing areas worldwide. Four virulence-deficient mutants were identified from a transposon mutagenesis library of X. oryzae pv. oryzae. Sequence-analysis revealed that the transposon of the four mutants inserted at different sites in the same ORF, which is homologous to the xpsE gene encoding a component of the type-II secretion system in many bacterial pathogens. Extracellular enzymes, such as xylanase and cellulase, were not secreted to the extracellular space in the mutants. Analysis of the protein profile of the extracellular, periplasmic and intracellular fractions indicated that at least two secreted proteins accumulated in the periplasmic space in the mutants. After genetic complementation of these mutants with a functional xpsE gene, the xpsE gene could express normally and the pathogenicity of the mutants and their secretion of extracellular enzymes were restored. Western blot analysis with an anticellulase antiserum also showed that cellulase was secreted normally in the complemented strains. The results show that the type-II secretion pathway structural gene xpsE is required for xylanase and cellulase secretion and full virulence in X. oryzae pv. oryzae.