Transcriptomic and metabolomic insights into the role of the &lt;i&gt;flgK&lt;/i&gt; gene in the pathogenicity of &lt;i&gt;Pseudomonas plecoglossicida&lt;/i&gt; to orange-spotted grouper (&lt;i&gt;Epinephelus coioides&lt;/i&gt;)

Yuan, Bin; Zhao, Lingmin; Zhuang, Zhixia; Wang, Xiaoru; Fu, Qi; Huang, Huabin; Huang, Lixing; Qin, Yingxue; Yan, Qingpi

doi:10.24272/j.issn.2095-8137.2022.216

Cited by 27 publications

(17 citation statements)

References 66 publications

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“…The determination of adhesion ability of wild-type and znuA, znuB, znuC -RNAi strains of A. salmonicida SRW-OG1 were carried out according to the methods of Huang et al ( 37 , 44 ) with slight modifications. Briefly, 30 μL of sterile healthy E. coioides epidermal mucus was spread evenly on the glass slides, incubated overnight at 25°C in the dark, subsequently fixed with 4% methanol at 25°C for 30 min.…”

Section: Methodsmentioning

confidence: 99%

Virulence regulation of Zn2+ uptake system znuABC on mesophilic Aeromonas salmonicida SRW-OG1

et al. 2023

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Psychrophilic Aeromonas salmonicida could not grow above 25°C and therefore thought unable to infect mammals and humans. In our previous study, a mesophilic A. salmonicida SRW-OG1 was isolated from Epinephelus coioides with furunculosis. Through the analysis of preliminary RNA-seq, it was found that the Zn2+ uptake related genes znuA, znuB and znuC might be involved in the virulence regulation of A. salmonicida SRW-OG1. Therefore, the purpose of this study was to explore the effect of znuABC silencing on the virulence regulation of A. salmonicida SRW-OG1. The results showed that the growth of the znuA-RNAi, znuB-RNAi, and znuC-RNAi strains was severely restricted under the Fe2+ starvation, but surprisingly there was no significant difference under the Zn2+ restriction. In the absence of Zn2+ and Fe2+, the expression level of znuABC was significantly increased. The motility, biofilm formation, adhesion and hemolysis of the znuA-RNAi, znuB-RNAi, and znuC-RNAi strains were significantly reduced. We also detected the expression of znuABC under different growth periods, temperatures, pH, as well as Cu2+ and Pb2+ stresses. The results showed that znuABC was significantly up-regulated in the logarithmic phase and the decline phase of A. salmonicida. Interestingly, the trend of expression levels of the znuABC at 18, 28, and 37°C was reversed to another Zn2+ uptake related gene zupT. Taken together, these indicated that the znuABC was necessary for A. salmonicida SRW-OG1 pathogenicity and environmental adaptability, and was cross regulated by iron starvation, but it was not irreplaceable for A. salmonicida SRW-OG1 Zn2+ uptake in the host.

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Section: Methodsmentioning

confidence: 99%

Virulence regulation of Zn2+ uptake system znuABC on mesophilic Aeromonas salmonicida SRW-OG1

et al. 2023

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“…Sterile skin mucus of E. coioides was spread on a cover slide and fixed with 4% methanol [52]. Bacterial suspension (OD 600nm = 0.3) was added onto the mucus and incubated at 28°C for 2 h. After washing gently with PBS thrice, the cover slide was air‐dried, fixed with 4% methanol, and stained with 0.1% (w/v) crystal violet.…”

Section: Methodsmentioning

confidence: 99%

Effect of the zinc transporter ZupT on the virulence mechanisms of mesophilic Aeromonas salmonicida SRW‐OG1

Wang

Huang

2023

Animal Research and One Health

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As a typical psychrophilic bacterial pathogen, Aeromonas salmonicida causes furunculosis in wild and farmed freshwater and marine fish, leading to substantial economic losses in the global aquaculture industry. Previous studies have shown that A. salmonicida is unable to grow above 25°C, hence limiting its infection to cold‐water fish. However, we isolated A. salmonicida SRW‐OG1, a mesophilic pathogenic strain from the warm‐water fish Epinephelus coioides. Through RNA‐seq analysis, we observe significant upregulation of the zupT gene at 28°C. ZupT is a member of zinc‐regulated transporters and iron‐regulated transporter‐like proteins (ZIP family) and is closely associated with transcriptional regulation of virulence in certain pathogens. Consequently, our study aimed to examine the role of zupT during A. salmonicida SRW‐OG1 infection at high temperatures. Our findings demonstrate that the zupT‐RNAi strain exhibits severe growth restriction under limited Zn2+ and Fe2+ conditions. Notably, this strain shows significantly reduced mortality rates and colonization abilities. Moreover, its motility, biofilm formation, adhesion, and hemolytic activities are significantly diminished. Confocal laser scanning microscopy reveals earlier and accelerated biofilm dissociation in the zupT‐RNAi strain. Analysis of extracellular products at 36 h indicates a considerable reduction in relative extracellular protein content in the zupT‐RNAi strain. Taken together, our results highlight the vital role of the zupT gene in zinc transport and the fitness of A. salmonicida SRW‐OG1 within the host.

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“…RNAi strains were constructed according to the methods described before (Tokunaga et al, 2015; Yang et al, 2023; Yuan et al, 2022). Briefly, three short hairpin RNA sequences targeting different mRNA coding regions of rstA were designed, respectively, and synthesized by Tsingke Biotechnology Co., Ltd. (Table 2).…”

Section: Methodsmentioning

confidence: 99%

“…RNAi strains were constructed according to the methods described before (Tokunaga et al, 2015;Yang et al, 2023;Yuan et al, 2022).…”

Section: Construction Of Strain Rsta-rnaimentioning

confidence: 99%

The role and mechanisms of rstA in the intracellular survival of fish pathogenic Aeromonas hydrophila

Zhang

Chen

Yan

et al. 2023

Journal of Fish Diseases

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In this study, RNAi technology was used to silence the gene rstA in Aeromonas hydrophila. The strain rstA‐RNAi displayed significant decrease in intracellular survival compared with that of the wild‐type strain B11. Transcriptome analysis explored that the expression of some important anti‐stress protein genes was significantly upregulated in rstA‐RNAi compared with the wild‐type strain, while the expression of the genes related to iron acquisition and type VI secretion system was significantly downregulated. Further study found that under low pH and H2O2 stress, the anti‐stress protein genes were expressed at a low level in rstA‐RNAi, the growth ability of rstA‐RNAi was also significantly lower than that of wild‐type strain. The results also displayed that with the fluctuation of iron concentration, the expression of some genes related to iron acquisition remained at a low level in rstA‐RNAi, and the growth ability of rstA‐RNAi was lower than that of the wild‐type strain under the same culture conditions, indicating rstA can regulate iron acquisition and further affect the bacteria growth. The adhesion ability of rstA‐RNAi to fish macrophages was reduced, suggesting rstA may be also affect the formation of type VI secretion system of A. hydrophila.

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Transcriptomic and metabolomic insights into the role of the <i>flgK</i> gene in the pathogenicity of <i>Pseudomonas plecoglossicida</i> to orange-spotted grouper (<i>Epinephelus coioides</i>)

Cited by 27 publications

References 66 publications

Virulence regulation of Zn2+ uptake system znuABC on mesophilic Aeromonas salmonicida SRW-OG1

Virulence regulation of Zn2+ uptake system znuABC on mesophilic Aeromonas salmonicida SRW-OG1

Effect of the zinc transporter ZupT on the virulence mechanisms of mesophilic Aeromonas salmonicida SRW‐OG1

The role and mechanisms of rstA in the intracellular survival of fish pathogenic Aeromonas hydrophila

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