Expression of Xanthomonas oryzae pv. oryzae hrp Genes in XOM2, a Novel Synthetic Medium

Tsuge, Seiji; Furutani, Ayako; Fukunaka, Rie; Oku, Takashi; Tsuno, Kazunori; Ochiai, Hirokazu; Inoue, Yasuhiro; Kaku, Hisatoshi; Kubo, Yasuyuki

doi:10.1007/pl00013104

Cited by 104 publications

(141 citation statements)

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“…X. oryzae pv. oryzae strains were usually grown at 28°C in nutrient broth-yeast extract (NBY) medium (32) or in an hrp-inducing medium, XOM2 (31). Xanthomonas axonopodis pv.…”

Section: Methodsmentioning

confidence: 99%

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Evidence for HrpXo-Dependent Expression of Type II Secretory Proteins in Xanthomonas oryzae pv. oryzae

et al. 2004

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Xanthomonas oryzae pv. oryzae is a causal agent of bacterial leaf blight of rice. Recently, an efficient hrp-inducing medium, XOM2, was established for this bacterium. In this medium, more than 10 proteins were secreted from the wild-type strain of X. oryzae pv. oryzae. Many of these proteins disappeared or decreased in amount in culture on XOM2 when incubated with the strain that has a mutation in the hrp regulatory gene. Interestingly, the secretory protein profile of a mutant lacking a type III secretion system (TTSS), components of which are encoded by hrp genes, was similar to that of the wild-type strain except that a few proteins had disappeared. This finding suggests that many HrpXo-dependent secretory proteins are secreted via systems other than the TTSS. By isolating mutant strains lacking a type II secretion system, we examined this hypothesis. As expected, many of the HrpXo-dependent secretory proteins disappeared or decreased when the mutant was cultured in XOM2. By determining the N-terminal amino acid sequence, we identified one of the type II secretory proteins as a cysteine protease homolog, CysP2. Nucleotide sequence analysis revealed that cysP2 has an imperfect plant-inducible-promoter box, a consensus sequence which HrpXo regulons possess in the promoter region, and a deduced signal peptide sequence at the N terminus. By reverse transcription-PCR analysis and examination of the expression of CysP2 by using a plasmid harboring a cysP2::gus fusion gene, HrpXo-dependent expression of CysP2 was confirmed. Here, we reveal that the hrp regulatory gene hrpXo is also involved in the expression of not only hrp genes and type III secretory proteins but also some type II secretory proteins.In general, plant-pathogenic bacteria possess hypersensitive response and pathogenicity (hrp) genes, which are clustered in their chromosomes. hrp genes encode a type III secretion system (TTSS) that delivers virulence and avirulence factors from the bacteria to plant cells and are required for pathogenesis in host plants and for triggering a hypersensitive response in nonhost plants (1,38). Transcriptional regulation of hrp genes depends on environmental conditions. The expression of hrp genes is generally suppressed in complex media and induced in planta and under certain in vitro conditions (6,24,35,39).In xanthomonads, the hrp cluster comprises six hrp loci, hrpA to hrpF, which are all required for full pathogenicity, and their expression is regulated by two genes, hrpX and hrpG, which are located outside the hrp gene cluster region (7, 36). The HrpG protein belongs to the OmpR family of two-component regulatory systems and activates the expression of hrpA and hrpX (37). HrpX, an AraC-type transcriptional activator, has been reported to control the expression of the operons hrpB to hrpF, which contain the hrp genes encoding a component of TTSS (36). It has also been suggested that HrpX controls some effector proteins (5). Several genes that are regulated in a HrpX-dependent manner possess the consensus nucleotide ...

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“…X. oryzae pv. oryzae strains were usually grown at 28°C in nutrient broth-yeast extract (NBY) medium (32) or in an hrp-inducing medium, XOM2 (31). Xanthomonas axonopodis pv.…”

Section: Methodsmentioning

confidence: 99%

“…oryzae is a causal agent of bacterial leaf blight of rice (28). Recently, an efficient hrp-inducing medium, XOM2, was established for the bacterium (31). Using this medium, we have identified Hpa1 as one of the HrpXoregulated type III secretory proteins in X. oryzae pv.…”

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confidence: 99%

Evidence for HrpXo-Dependent Expression of Type II Secretory Proteins in Xanthomonas oryzae pv. oryzae

et al. 2004

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“…In many gram-negative phytopathogenic bacteria, hrp (hypersensitive response and pathogenicity) genes are required for pathogenicity on host plants and for the HR on nonhost plants (3,4,5,8,64) and are repressed in nutrient-rich media but induced in nutrient-deficient media and inside the host (9,46,54,58,60,66). In xanthomonads, hrp genes are regulated by HrpG and HrpX (39,54,61,62).…”

Section: Resultsmentioning

confidence: 99%

hpaR, a PutativemarRFamily Transcriptional Regulator, Is Positively Controlled by HrpG and HrpX and Involved in the Pathogenesis, Hypersensitive Response, and Extracellular Protease Production ofXanthomonas campestrisPathovar campestris

Wei

Tang

et al. 2007

J Bacteriol

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The MarR family of transcriptional regulators of bacteria are involved in the regulation of many cellular processes, including pathogenesis. In this work, we have demonstrated genetically that hpaR (hpa, hrp associated), which encodes a putative MarR family regulator, is involved in the hypersensitive response (HR), pathogenicity, and extracellular protease production of the phytopathogenic bacterium Xanthomonas campestris pathovar campestris. A mutation in hpaR resulted in complete loss of virulence in the host plant cabbage, a delayed and weakened HR in the nonhost plant pepper ECW-10R, and an increase in extracellular protease production. Detection of the ␤-glucuronidase activity of a plasmid-driven hpaR promoter-gusA reporter revealed that the expression of hpaR is positively controlled by HrpG and HrpX and is suppressed in rich medium while being strongly induced in minimal and hrp-inducing media and inside the host. These findings indicate that hpaR belongs to the hrpG and hrpX regulon and that HrpX regulates the extracellular protease production via hpaR in X. campestris pv. campestris.

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“…Wild-type strain Xoo KACC10859 was obtained from the Korean Agricultural Culture Collection (KACC) at the National Institute of Agricultural Biotechnology, Suwon, Korea. Xoo strains were cultured at 28°C on peptone sucrose agar (PSA: peptone, 10 g/L; sucrose, 10 g/l; and agar, 15 g/L) or XOM2 medium [27]. E. coli was grown in Luria-Bertani (LB) broth (Difco, Detroit, MI, USA) at 37°C for 18 h. Antibiotics were added at the following final concentrations for E. coli and Xoo, respectively: ampicillin, 80 μg/mL and 50 μg/mL; gentamycin, 50 μg/mL and 20 μg/mL; and kanamycin, 50 μg/mL and 20 μg/mL.…”

Section: Bacterial Strains Plasmids and Culture Conditionsmentioning

confidence: 99%

Expressional Regulation of the Virulence Gene eglXoA Encoding Endoglucanase, Dependent on HrpXo and Cyclic AMP Receptor-Like Protein (Clp) in Xanthomonas oryzae pv. oryza

Temuujin¹,

Hw²

2017

J Plant Pathol Microbiol

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Expression of Xanthomonas oryzae pv. oryzae hrp Genes in XOM2, a Novel Synthetic Medium

Cited by 104 publications

References 24 publications

Evidence for HrpXo-Dependent Expression of Type II Secretory Proteins in Xanthomonas oryzae pv. oryzae

Evidence for HrpXo-Dependent Expression of Type II Secretory Proteins in Xanthomonas oryzae pv. oryzae

hpaR, a PutativemarRFamily Transcriptional Regulator, Is Positively Controlled by HrpG and HrpX and Involved in the Pathogenesis, Hypersensitive Response, and Extracellular Protease Production ofXanthomonas campestrisPathovar campestris

Expressional Regulation of the Virulence Gene eglXoA Encoding Endoglucanase, Dependent on HrpXo and Cyclic AMP Receptor-Like Protein (Clp) in Xanthomonas oryzae pv. oryza

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