Integration of RNA‐seq and RNAi reveals the contribution of znuA gene to the pathogenicity of Pseudomonas plecoglossicida and to the immune response of Epinephelus coioides

He, Le; Wang, Luying; Zhao, Lingmin; Zhuang, Zhixia; Wang, Xiaoru; Huang, Huabin; Fu, Qi; Huang, Lixing; Qin, Yingxue; Wang, Pan; Yan, Qingpi

doi:10.1111/jfd.13502

Cited by 20 publications

(8 citation statements)

References 70 publications

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“…It was found that ZnuC is an important protein for Zn uptake in Pseudomonas aeruginosa ( Huang et al, 2021 ). Similarly, the authors found that silencing the znuA gene in this strain could achieve an 89.2% reduction of deaths and that RNAi-treated strains also induced antibody production in grouper ( He et al, 2021 ). We have summarized some genes that we expect to enhance host defense through RNAi technology ( Table 1 ).…”

Section: Rnai-mediated Inhibition Of Pseudomonasmentioning

confidence: 77%

Potential dsRNAs can be delivered to aquatic for defense pathogens

Nie

Chen

Tang

et al. 2022

Front. Bioeng. Biotechnol.

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The use of antibiotics to facilitate resistance to pathogens in aquatic animals is a traditional method of pathogen control that is harmful to the environment and human health. RNAi is an emerging technology in which homologous small RNA molecules target specific genes for degradation, and it has already shown success in laboratory experiments. However, further research is needed before it can be applied in aquafarms. Many laboratories inject the dsRNA into aquatic animals for RNAi, which is obviously impractical and very time consuming in aquafarms. Therefore, to enable the use of RNAi on a large scale, the methods used to prepare dsRNA need to be continuously in order to be fast and efficient. At the same time, it is necessary to consider the issue of biological safety. This review summarizes the key harmful genes associated with aquatic pathogens (viruses, bacteria, and parasites) and provides potential targets for the preparation of dsRNA; it also lists some current examples where RNAi technology is used to control aquatic species, as well as how to deliver dsRNA to the target hydrobiont.

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Section: Rnai-mediated Inhibition Of Pseudomonasmentioning

confidence: 77%

Potential dsRNAs can be delivered to aquatic for defense pathogens

Nie

Chen

Tang

et al. 2022

Front. Bioeng. Biotechnol.

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“…Bacterial growth curves were drawn according to He et al. (2021) with some modifications. The bacteria were cultured overnight to exponential phase.…”

Section: Methodsmentioning

confidence: 99%

Comparative transcriptome analysis explored the molecular mechanisms of a luxR‐type regulator regulating intracellular survival of Aeromonas hydrophila

Chen,

Mao,

Yan

et al. 2024

Journal of Fish Diseases

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Aeromonas hydrophila is not a traditional intracellular bacterium. However, previous studies revealed that pathogenic A. hydrophila B11 could temporarily survive for at least 24 h in fish phagocytes, and the regulation of intracellular survival in bacteria was associated with regulators of the LuxR‐type. The mechanisms of luxR08110 on the A. hydrophila's survival in macrophages were investigated using comprehensive transcriptome analysis and biological phenotype analysis in this study. The results showed that after luxR08110 was silenced, the intracellular survival ability of bacteria was significantly diminished. Comparative transcriptome analysis revealed that luxR08110 was a critical regulator of A. hydrophila, which regulated the expression of over 1200 genes, involving in bacterial flagellar assembly and chemotaxis, ribosome, sulphur metabolism, glycerolipid metabolism, and other mechanisms. Further studies confirmed that after the inhibition of expression of luxR08110, the motility, chemotaxis and adhesion of A. hydrophila significantly decreased. Moreover, compared with the wild‐type strain, the survival rates of silencing strain were all considerably reduced under both H2O2 and low pH stress conditions. According to both transcriptome analysis and phenotypic tests, the luxR08110 of A. hydrophila could act as global regulator in bacteria intracellular survival. This regulator regulated intracellular survival of A. hydrophila mainly through two ways. One way is to regulate bacterial flagellar synthesis and further affects the motility, chemotaxis and adhesion of bacteria. The other way is to regulate sulphur and glycerolipid metabolisms, thus affecting bacterial energy production and the ability to resist environmental stress.

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“…Knocked-down strains of A. salmonicida SRW-OG1 znuA, znuB and znuC were constructed using RNAi, according to the method described by Bittel et al ( 34 )and Girard et al ( 35 , 36 ) with slight modifications. The sequence of A. salmonicida SRW-OG1 znuA, znuB and znuC genes were introduced into Invitrogen Block-iT TM RNAi Designer ( https://rnaidesigner.thermofisher.com/rnaiexpress/design.do ) to design shRNA online ( Supplementary Table 1 ).…”

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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Integration of RNA‐seq and RNAi reveals the contribution of znuA gene to the pathogenicity of Pseudomonas plecoglossicida and to the immune response of Epinephelus coioides

Cited by 20 publications

References 70 publications

Potential dsRNAs can be delivered to aquatic for defense pathogens

Potential dsRNAs can be delivered to aquatic for defense pathogens

Comparative transcriptome analysis explored the molecular mechanisms of a luxR‐type regulator regulating intracellular survival of Aeromonas hydrophila

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

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