The Effect of tonB Gene on the Virulence of Pseudomonas plecoglossicida and the Immune Response of Epinephelus coioides.
The Effect of tonB Gene on the Virulence of Pseudomonas plecoglossicida and the Immune Response of Epinephelus coioides.
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DOI：
10.3389/fmicb.2021.720967
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发表时间：
2021
影响因子：
5.2
通讯作者：
Yan Q
中科院分区：
文献类型：
--
作者：
Hu L;Zhao L;Zhuang Z;Wang X;Fu Q;Huang H;Lin L;Huang L;Qin Y;Zhang J;Yan Q
杀褶舌假单胞菌是养殖鱼类“内脏白色斑点病”的病原，造成了严重的经济损失。tonB基因在革兰氏阴性菌从外膜摄取营养物质的过程中起着至关重要的作用。本实验室前期的研究结果表明，杀舌类青霉tonB基因在感染后的斜带石斑鱼脾脏中表达显著上调。探讨tonB基因对杀多舌青霉毒力和大肠杆菌免疫反应的影响。利用RNAi技术敲除多舌拟青霉tonB基因，并比较了野生型菌株与tonB-RNAi菌株的差异。结果表明：4株杀多舌拟青霉tonB基因突变体的表达量均显著降低，最高敲除效率为94.0%;感染tonB-RNAi菌株的coioides比感染P. plecoglossicida野生菌株的对应物高20%。同时，E.与野生型相比，tonB-RNAi株感染的大肠杆菌在脾脏中携带的病原体数量减少，脾脏表面的白色斑点减少，运动性、趋化性、粘附性和生物膜形成能力均明显减弱。tonB-RNAi株感染的大肠杆菌与野生型感染的大肠杆菌不同，抗原加工和呈递途径以及补体和凝血级联途径是最丰富的免疫途径。结果表明，tonB基因是杀多舌青霉的毒力基因，参与了运动、趋化、粘附和生物膜形成的调控，影响了大肠杆菌的免疫应答。coioides到P. plecoglossicida感染。
Pseudomonas plecoglossicida is the causative agent of “visceral white spot disease” in cultured fish and has resulted in serious economic losses. tonB gene plays a crucial role in the uptake of nutrients from the outer membranes in Gram-negative bacteria. The previous results of our lab showed that the expression of tonB gene of P. plecoglossicida was significantly upregulated in the spleens of infected Epinephelus coioides. To explore the effect of tonB gene on the virulence of P. plecoglossicida and the immune response of E. coioides, tonB gene of P. plecoglossicida was knocked down by RNAi; and the differences between the wild-type strain and the tonB-RNAi strain of P. plecoglossicida were investigated. The results showed that all of the four mutants of P. plecoglossicida exhibited significant decreases in mRNA of tonB gene, and the best knockdown efficiency was 94.0%; the survival rate of E. coioides infected with the tonB-RNAi strain was 20% higher than of the counterpart infected with the wild strain of P. plecoglossicida. Meanwhile, the E. coioides infected with the tonB-RNAi strain of P. plecoglossicida carried less pathogens in the spleen and less white spots on the surface of the spleen; compared with the wild-type strain, the motility, chemotaxis, adhesion, and biofilm formation of the tonB-RNAi strain were significantly attenuated; the transcriptome data of E. coioides infected with the tonB-RNAi strain were different from the counterpart infected with the wild strain of P. plecoglossicida; the antigen processing and presentation pathway and the complement and coagulation cascade pathway were the most enriched immune pathways. The results indicated that tonB was a virulence gene of P. plecoglossicida; tonB gene was involved in the regulation of motility, chemotaxis, adhesion, and biofilm formation; tonB gene affected the immune response of E. coioides to P. plecoglossicida infection.
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