Establishment of an inducing medium for type III effector secretion in Xanthomonas campestris pv. campestris

Jiang, Guo-Feng; Tang, Ji-Liang; Yang, Mei; Liu, San; Liu, Jiao; Liang, Xiaoxia; Bai, Xian-Fang; Tang, Dong-Jie; Lu, Guang-Tao; He, Yong-Qiang; Yu, Diqiu

doi:10.1590/s1517-83822013000300045

Cited by 15 publications

(14 citation statements)

References 25 publications

(44 reference statements)

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“…XAM-M is a variant of XAM-1 medium (Facincani et al, 2014) and includes MgSO 4 and casamino acids, similar to the original XVM2 (Wengelnik et al, 1996). Casamino acids have been shown previously to be necessary for the induction of some hrp genes, such as hrpF (Jiang et al, 2013;Schulte and Bonas, 1992).…”

Section: Xac Pathogenicity: Plant-mimicking Medium and Periplasmic Prmentioning

confidence: 99%

Comparative proteomic analysis ofXanthomonas citrissp.citriperiplasmic proteins reveals changes in cellular envelope metabolism duringin vitropathogenicity induction

Artier

Zandonadi

Carvalho

et al. 2017

Molecular Plant Pathology

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Citrus canker is a plant disease caused by Gram-negative bacteria from the genus Xanthomonas. The most virulent species is Xanthomonas citri ssp. citri (XAC), which attacks a wide range of citrus hosts. Differential proteomic analysis of the periplasm-enriched fraction was performed for XAC cells grown in pathogenicity-inducing (XAM-M) and pathogenicity-non-inducing (nutrient broth) media using two-dimensional electrophoresis combined with liquid chromatography-tandem mass spectrometry. Amongst the 40 proteins identified, transglycosylase was detected in a highly abundant spot in XAC cells grown under inducing condition. Additional up-regulated proteins related to cellular envelope metabolism included glucose-1-phosphate thymidylyltransferase, dTDP-4-dehydrorhamnose-3,5-epimerase and peptidyl-prolyl cis-trans-isomerase. Phosphoglucomutase and superoxide dismutase proteins, known to be involved in pathogenicity in other Xanthomonas species or organisms, were also detected. Western blot and quantitative real-time polymerase chain reaction analyses for transglycosylase and superoxide dismutase confirmed that these proteins were up-regulated under inducing condition, consistent with the proteomic results. Multiple spots for the 60-kDa chaperonin and glyceraldehyde-3-phosphate dehydrogenase were identified, suggesting the presence of post-translational modifications. We propose that substantial alterations in cellular envelope metabolism occur during the XAC infectious process, which are related to several aspects, from defence against reactive oxygen species to exopolysaccharide synthesis. Our results provide new candidates for virulence-related proteins, whose abundance correlates with the induction of pathogenicity and virulence genes, such as hrpD6, hrpG, hrpB7, hpa1 and hrpX. The results present new potential targets against XAC to be investigated in further functional studies.

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Section: Xac Pathogenicity: Plant-mimicking Medium and Periplasmic Prmentioning

confidence: 99%

Comparative proteomic analysis ofXanthomonas citrissp.citriperiplasmic proteins reveals changes in cellular envelope metabolism duringin vitropathogenicity induction

Artier

Zandonadi

Carvalho

et al. 2017

Molecular Plant Pathology

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“…Previous study suggested that the hrp genes of Xanthomonas express in defined minimal media but fails to express when grown in complex rich media (Schulte and Bonas, ). Hrp encoding genes of Xcc expresses most in a defined minimal medium XCM2 [20 mM succinic acid, 0.15 g/L casamino acids, 60.34 mM K 2 HPO 4 , 7.57 mM (NH 4 ) 2 SO 4 , 0.01 mM MgSO4, 33.07 mM KH 2 PO 4 , pH 6.6] compared to the other chemically defined media (Jiang et al ., ). We performed expression analysis of hrcU operon and hrpG using promoter probe strains Xcc 8004 P hrcU :: gusA and Xcc 8004 P hrpG :: gusA , respectively, in XCM2 media and XCM2 + 50 µM FeSO 4 .…”

Section: Resultsmentioning

confidence: 97%

Low-iron conditions induces the hypersensitive reaction and pathogenicityhrpgenes expression inXanthomonasand is involved in modulation of hypersensitive response and virulence

Pandey

Patnana

Padhi

et al. 2018

Environmental Microbiology Reports

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Expression of hrp (hypersensitive reaction and pathogenicity) genes inside the host is crucial for virulence of phytopathogenic bacteria. The hrp genes encode components of type3 secretion system (T3SS), HR elicitors and several regulators, which are involved in the co-ordinated expression of hrp genes in the host environment and in hrp inducing chemically defined medium. However, little is known about specific host or environmental factors which may play a role in the induction of hrp gene expression. In this study, we show that iron-limiting condition elicits induced expression of hrp genes, including type3 secretion system (T3SS) and effectors (T3E). Expression analysis using qRT-PCR and promoter probe strains suggest significant induction in the expression of Hrp and T3S-associated genes of Xanthomonas campestris pv. campestris (Xcc) under low-iron condition, and is suppressed by exogenous supplementation of iron. Furthermore, we show that with exogenous iron supplementation, wild type Xcc exhibited reduced disease symptoms in host-plant, and exhibited significant reduction in HR and callose deposition in the non-host plants. Xanthomonas oryzae and oryzicola pathovars also exhibited the iron affect, albeit to a lesser extend compared with the Xcc. Overall, our results suggest that low-iron condition inside the host may play a crucial role in pathogenicity.

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“…Xcc strains were grown at 28 °C in nutrient rich medium NYG [24] or minimal media MMX (23.8 mM glucose, 3.87 mM sodium citrate, 15.1 mM (NH 4 ) 2 SO 4 , 0.81 mM MgSO 4 , 23 mM K 2 HPO 4 , 44 mM KH 2 PO 4 , pH 7.0) [24] and XCM1 (20 mM succinic acid, 0.15 g/l casamino acids, 7.57 mM (NH 4 ) 2 SO 4 , 1 mM MgSO 4 , 60.34 mM K 2 HPO 4 , 33.07 mM KH 2 PO 4 , pH 6.6) [25]. Antibiotics were used at the following final concentrations as required: ampicillin (Amp), 100 μg/ml; gentamycin (Gm), 10 μg/ml; kanamycin (Kan), 25 μg/ml; rifampicin (Rif), 50 μg/ml; and tetracycline (Tc), 15 μg/ml for Escherichia coli and 5 μg/ml for Xcc .…”

Section: Methodsmentioning

confidence: 99%

RpfC regulates the expression of the key regulator hrpX of the hrp/T3SS system in Xanthomonas campestris pv. campestris

et al. 2018

Self Cite

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BackgroundThe Gram-negative phytopathogenic bacterium Xanthomonas campestris pv. campestris recruits the hrp/T3SS system to inject pathogenicity effector proteins into host cells and uses the rpf/DSF cell-cell signaling system to regulate the expression of virulence factors such as extracellular enzymes and polysaccharide. Whether these two systems have any connection is unknown.MethodsPositive regulator candidates affecting hrpX expression were identified by sacB strategy. The transcriptional expression was determined by qRT-PCR and GUS activity analysis. Transcriptome analysis was performed by RNA deep-sequencing. The hypersensitive response (HR) was determined in the nonhost plant pepper ECW-10R and electrolyte leakage assay.ResultsMutation of the gene encoding the sensor RpfC of the rpf/DSF system significantly reduced the expression of hrpX, the key regulator of the hrp/T3SS system, all of the genes in the hrp cluster and most reported type III effector genes. Mutation of rpfG did not affect the expression of hrpX. The rpfC mutant showed a delayed and weakened HR induction.ConclusionsRpfC positively regulates the expression of hrpX independent of RpfG, showing a complex regulatory network linking the rpf/DSF and hrp/T3SS systems.Electronic supplementary materialThe online version of this article (10.1186/s12866-018-1233-5) contains supplementary material, which is available to authorized users.

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Establishment of an inducing medium for type III effector secretion in Xanthomonas campestris pv. campestris

Cited by 15 publications

References 25 publications

Comparative proteomic analysis ofXanthomonas citrissp.citriperiplasmic proteins reveals changes in cellular envelope metabolism duringin vitropathogenicity induction

Comparative proteomic analysis ofXanthomonas citrissp.citriperiplasmic proteins reveals changes in cellular envelope metabolism duringin vitropathogenicity induction

Low-iron conditions induces the hypersensitive reaction and pathogenicityhrpgenes expression inXanthomonasand is involved in modulation of hypersensitive response and virulence

RpfC regulates the expression of the key regulator hrpX of the hrp/T3SS system in Xanthomonas campestris pv. campestris

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