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Role of DetR in defence is critical for virulence of Xanthomonas oryzae pv. oryzae.

ABSTRACT: Like other bacteria, Xanthomonas oryzae pv. oryzae (Xoo), the causal agent of bacterial leaf blight disease in rice, possesses intracellular signalling systems, known as two-component regulatory systems (TCSs), which regulate pathogenesis and biological processes. Completion of the genome sequences of three Xoo strains has facilitated the functional study of genes, including those of TCSs, but the biological functions of most Xoo?TCSs have not yet been uncovered. To identify TCSs involved in Xoo pathogenesis, we generated knockout strains lacking response regulators (RRs, a cytoplasmic signalling component of the TCS) and examined the virulence of the RR knockout strains. This study presents a knockout strain (detR(-) ) lacking the PXO_04659 gene which shows dramatically reduced virulence relative to the wild-type. Our studies to elucidate detR function in Xoo pathogenesis revealed a reduction in extracellular polysaccharide (EPS), intolerance to reactive oxygen species (ROS) and deregulation of iron homeostasis in the detR(-) strain. Moreover, gene expression of regulatory factors, including other RRs and transcription factors (TFs), was altered in the absence of DetR protein, as determined by reverse transcription-polymerase chain reaction (RT-PCR) and/or real-time quantitative RT-PCR analyses. All evidence leads to the conclusion that DetR is essential for Xoo virulence through the regulation of the Xoo defence system including EPS synthesis, ROS detoxification and iron homeostasis, solely or cooperatively with other regulatory factors.

SUBMITTER: Nguyen MP

PROVIDER: S-EPMC6638461 | biostudies-literature | 2016 May

REPOSITORIES: biostudies-literature

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Role of DetR in defence is critical for virulence of Xanthomonas oryzae pv. oryzae.

Nguyen Minh-Phuong MP Park Jongchan J Cho Man-Ho MH Lee Sang-Won SW

Molecular plant pathology 20151019 4

Like other bacteria, Xanthomonas oryzae pv. oryzae (Xoo), the causal agent of bacterial leaf blight disease in rice, possesses intracellular signalling systems, known as two-component regulatory systems (TCSs), which regulate pathogenesis and biological processes. Completion of the genome sequences of three Xoo strains has facilitated the functional study of genes, including those of TCSs, but the biological functions of most Xoo TCSs have not yet been uncovered. To identify TCSs involved in Xoo ...[more]

PMID: 26315668

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Project description:In Xanthomonas oryzae pv. oryzae, the bacterial blight pathogen of rice, there are over 20 genes encoding GGDEF, EAL, and HD-GYP domains, which are potentially involved in the metabolism of second messenger c-di-GMP. In this study, we focused on the characterization of an EAL domain protein, EdpX1. Deletion of the edpX1 gene resulted in a 2-fold increase in the intracellular c-di-GMP levels, which were restored to the wild-type levels in the complemented ΔedpX1(pB-edpX1) strain, demonstrating that EdpX1 is an active phosphodiesterase (PDE) in X. oryzae pv. oryzae. In addition, colorimetric assays further confirmed the PDE activity of EdpX1 by showing that the E153A mutation at the EAL motif strongly reduced its activity. Virulence assays on the leaves of susceptible rice showed that the ΔedpX1 mutant was severely impaired in causing disease symptoms. In trans expression of wild-type edpX1, but not edpX1 E153A, was able to complement the weakened virulence phenotype. These results indicated that an active EAL domain is required for EdpX1 to regulate the virulence of X. oryzae pv. oryzae. We then demonstrated that the ΔedpX1 mutant was defective in secreting exopolysaccharide (EPS) and forming biofilms. The expression of edpX1 in the ΔedpX1 mutant, but not edpX1 E153A, restored the defective phenotypes to near-wild-type levels. In addition, we observed that EdpX1-green fluorescent protein (EdpX1-GFP) exhibited multiple subcellular localization foci, and this pattern was dependent on its transmembrane (TM) region, which did not seem to directly contribute to the regulatory function of EdpX1. Thus, we concluded that EdpX1 exhibits PDE activity to control c-di-GMP levels, and its EAL domain is necessary and sufficient for its regulation of virulence in X. oryzae pv. oryzae.IMPORTANCE Bacteria utilize c-di-GMP as a second messenger to regulate various biological functions. The synthesis and degradation of c-di-GMP are catalyzed by GGDEF domains and an EAL or HD-GYP domain, respectively. Multiple genes encoding these domains are often found in one bacterial strain. For example, in the genome of X. oryzae pv. oryzae PXO99A, 26 genes encoding proteins containing these domains were identified. Therefore, to fully appreciate the complexity and specificity of c-di-GMP signaling in X. oryzae pv. oryzae, the enzymatic activities and regulatory functions of each GGDEF, EAL, and HD-GYP domain protein need to be elucidated. In this study, we showed that the EAL domain protein EdpX1 is a major PDE to regulate diverse virulence phenotypes through the c-di-GMP signaling pathway.

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Role of DetR in defence is critical for virulence of Xanthomonas oryzae pv. oryzae.

Publications

Role of DetR in defence is critical for virulence of Xanthomonas oryzae pv. oryzae.

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