Dissecting the virulence‐related functionality and cellular transcription mechanism of a conserved hypothetical protein in <i>Xanthomonas oryzae</i> pv. <i>oryzae</i>

Wang, Bo; Wu, Guichun; Zhang, Yuqiang; Qian, Guoliang; Liu, Fengquan

doi:10.1111/mpp.12664

Cited by 30 publications

(30 citation statements)

References 54 publications

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“…Biofilm formation assays were performed as described previously (Wang et al ., ). Bacterial cells were cultured in NB medium to OD 600 ~ 1.0 and 4 mL of cell suspensions were transferred to sterilized tubes and incubated at 28 °C for 7 days without shaking.…”

Section: Methodsmentioning

confidence: 97%

Contribution of the cold shock protein CspA to virulence in Xanthomonas oryzae pv. oryzae

et al. 2018

Molecular Plant Pathology

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Summary Xanthomonas oryzae pv. oryzae ( Xoo ) causes a damaging bacterial leaf blight disease in rice. Cold shock proteins (Csps) are highly conserved nucleic acid‐binding proteins present in various bacterial genera, but relatively little is known about their functions in Xanthomonas . Herein, we identified four Csps (CspA–CspD) in the Xoo PXO99 A strain. Deletion of cspA decreased cold adaptation and a few known pathogenic factors, including bacterial pathogenicity, biofilm formation and polysaccharide production. Furthermore, we performed transcriptomic and chromosome immunoprecipitation (ChIP) experiments to identify direct targets of CspA and to determine its DNA‐binding sequence. Integrative data analysis revealed that CspA directly regulates two genes, PXO_RS11830 and PXO_RS01060 , by binding to a conserved CCAAT sequence in the promoter region. We generated single‐deletion mutants of each gene and the results indicate that both are responsible for Xanthomonas pathogenicity. In addition, quantitative real‐time polymerase chain reaction and western blotting showed that CspA suppressed the expression of its direct targets. In summary, our study clarifies the characteristics of Csps in Xanthomonas and greatly advances our understanding of the mechanisms underlying the contribution of CspA to bacterial virulence.

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Section: Methodsmentioning

confidence: 97%

Contribution of the cold shock protein CspA to virulence in Xanthomonas oryzae pv. oryzae

et al. 2018

Molecular Plant Pathology

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“…Bacterial one-hybrid assays were performed as previously described (41,42). In brief, the bacterial one-hybrid reporter system contains three components: the plasmids pBXcmT and pTRG, which are used to clone the target DNA and to express the target protein, respectively, and the E. coli XL1-Blue MRFˊ kan strain, which is the host strain for the propagation of pBXcmT and pTRG recombinants (43).…”

Section: Bacterial One-hybrid Assaysmentioning

confidence: 99%

Two Functional Fatty Acyl Coenzyme A Ligases Affect Free Fatty Acid Metabolism To Block Biosynthesis of an Antifungal Antibiotic in Lysobacter enzymogenes

Hou

et al. 2020

Appl Environ Microbiol

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In Lysobacter enzymogenes OH11, RpfB1 and RpfB2 were predicted to encode acyl coenzyme A (CoA) ligases. RpfB1 is located in the Rpf gene cluster. Interestingly, we found an RpfB1 homolog (RpfB2) outside this canonical gene cluster, and nothing is known about its functionality or mechanism. Here, we report that rpfB1 and rpfB2 can functionally replace EcFadD in the Escherichia coli fadD mutant JW1794. RpfB activates long-chain fatty acids (n-C16:0 and n-C18:0) for the corresponding fatty acyl-CoA ligase (FCL) activity in vitro, and Glu-361 plays critical roles in the catalytic mechanism of RpfB1 and RpfB2. Deletion of rpfB1 and rpfB2 resulted in significantly increased heat-stable antifungal factor (HSAF) production, and overexpression of rpfB1 or rpfB2 completely suppressed HSAF production. Deletion of rpfB1 and rpfB2 resulted in increased L. enzymogenes diffusible signaling factor 3 (LeDSF3) synthesis in L. enzymogenes. Overall, our results showed that changes in intracellular free fatty acid levels significantly altered HSAF production. Our report shows that intracellular free fatty acids are required for HSAF production and that RpfB affects HSAF production via FCL activity. The global transcriptional regulator Clp directly regulated the expression of rpfB1 and rpfB2. In conclusion, these findings reveal new roles of RpfB in antibiotic biosynthesis in L. enzymogenes. IMPORTANCE Understanding the biosynthetic and regulatory mechanisms of heat-stable antifungal factor (HSAF) could improve the yield in Lysobacter enzymogenes. Here, we report that RpfB1 and RpfB2 encode acyl coenzyme A (CoA) ligases. Our research shows that RpfB1 and RpfB2 affect free fatty acid metabolism via fatty acyl-CoA ligase (FCL) activity to reduce the substrate for HSAF synthesis and, thereby, block HSAF production in L. enzymogenes. Furthermore, these findings reveal new roles for the fatty acyl-CoA ligases RpfB1 and RpfB2 in antibiotic biosynthesis in L. enzymogenes. Importantly, the novelty of this work is the finding that RpfB2 lies outside the Rpf gene cluster and plays a key role in HSAF production, which has not been reported in other diffusible signaling factor (DSF)/Rpf-producing bacteria.

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“…The Xanthomonas oryzae pv. oryzae (Xoo) strain PXO99 A was kindly provided by Prof. G. L. Qian (Nanjing Agricultural University) [37]. Xoo was cultivated at 28℃ on nutrient agar (NA) medium in plates or in nutrient broth (NB) medium in flasks.…”

Section: Bacterial Strains and Culture Conditionsmentioning

confidence: 99%

Growth inhibition and metabolomic analysis of Xanthomonas oryzae pv. oryzae treated with resveratrol

Jia

Luo

Guan

et al. 2020

Preprint

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Background Xanthomonas oryzae pv. oryzae ( Xoo ) can cause destructive bacterial blight in rice. As an antibacterial, resveratrol may inhibit Xoo growth. This study focused on the potential structural-activity relationship of resveratrol and its derivatives against Xoo growth, and 1 H-NMR-based metabolomic analysis was applied to investigate the global metabolite changes in Xoo after resveratrol treatment. Results Resveratrol showed the strongest inhibitory effects on Xoo growth compared with its derivatives, which lacked double bonds (compounds 4 – 6 ) or hydroxyls were substituted with methoxyls (compounds 7 – 9 ). The IC 50 of resveratrol against Xoo growth was 11.67 ± 0.58 μg/mL. Results indicated that the double bond of resveratrol contributed to its inhibitory effects on Xoo growth, and hydroxyls were vital for this inhibition. Interestingly, resveratrol also significantly inhibited Xoo flagellum growth. Based on 1 H-NMR global metabolic analysis, a total of 30 Xoo metabolites were identified, the changes in the metabolic profile indicated that resveratrol could cause oxidative stress as well as disturb energy, purine, amino acid, and NAD + metabolism in Xoo , resulting in the observed inhibitory effects on growth. Conclusions This study showed that the double bond of resveratrol contributed to its inhibitory effects on Xoo growth, and hydroxyls were also the important active groups. Resveratrol could cause oxidative stress of Xoo cells, and disturb the metabolism of energy, purine, amino acid and NAD +, thus inhibit Xoo growth.

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Dissecting the virulence‐related functionality and cellular transcription mechanism of a conserved hypothetical protein in Xanthomonas oryzae pv. oryzae

Cited by 30 publications

References 54 publications

Contribution of the cold shock protein CspA to virulence in Xanthomonas oryzae pv. oryzae

Contribution of the cold shock protein CspA to virulence in Xanthomonas oryzae pv. oryzae

Two Functional Fatty Acyl Coenzyme A Ligases Affect Free Fatty Acid Metabolism To Block Biosynthesis of an Antifungal Antibiotic in Lysobacter enzymogenes

Growth inhibition and metabolomic analysis of Xanthomonas oryzae pv. oryzae treated with resveratrol

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