Integration of RNA-seq and RNAi provides a novel insight into the immune responses of Epinephelus coioides to the impB gene of Pseudomonas plecoglossicida
Integration of RNA-seq and RNAi provides a novel insight into the immune responses of Epinephelus coioides to the impB gene of Pseudomonas plecoglossicida
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DOI：
10.1016/j.fsi.2020.06.023
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发表时间：
2020-10-01
影响因子：
4.7
通讯作者：
Yan, Qingpi
中科院分区：
文献类型：
--
作者：
Liu, Zixu;Zhao, Lingmin;Yan, Qingpi
杀舌假单胞菌是引起斜带石斑鱼内脏白色斑点病的一种革兰氏阴性细菌，给水产养殖业造成严重的经济损失。先前研究的RNA-seq结果表明，在感染期间，杀舌类疫霉中impB基因的表达显著上调。设计并合成了4条shRNA，对杀多舌类青霉impB基因进行沉默，最大沉默效率为95.2%。腹腔注射杀舌片吸虫impB-RNAi菌株不会引起大肠杆菌感染。coioides死亡，感染鱼的脾脏没有表现出明显的临床症状。虽然注射突变株可提高E. coioides血清不能有效保护E. coioides对野生株感染。与E.用野生型菌株感染的coioides，E.感染impB-RNAi菌株的coioides差异很大。KEGG富集分析表明，大肠杆菌趋化因子信号通路的关键基因。通过沉默P. plecoglossicida中的impB，coioides被下调。通过注射用杀舌片吸虫的impB-RNAi菌株感染没有产生针对E. coioides。本研究为了解大肠杆菌的免疫反应提供了新的视角。coioides与P. plecoglossicida的impB基因的同源性。
Pseudomonas plecoglossicida is a Gram-negative bacterium that causes visceral white spot disease in Epinephelus coioides and leads to severe aquatic economic losses. The RNA-seq results of a previous study showed that the expression of the impB gene in P. plecoglossicida was significantly upregulated during infection. Four shRNAs were designed and synthesized to silence the impB gene in P. plecoglossicida, and the maximum silencing efficiency was 95.2%. Intraperitoneal injection of the impB-RNAi strain of P. plecoglossicida did not cause E. coioides death, and the spleens of infected fish did not show significant clinical symptoms. Although the injection of the mutant strain increased the antibody titer in E. coioides serum, it could not effectively protect E. coioides against wild strain infection. Compared with E. coioides infected with the wild type strain, the RNA-seq results for E. coioides infected with the impB-RNAi strain differed greatly. The KEGG enrichment analysis showed that key genes of the chemokine signalling pathway of E. coioides were downregulated by the silencing of impB in P. plecoglossicida. Infection with the impB-RNAi strain of P. plecoglossicida through injection did not produce good immune protection against E. coioides. The present study provides a novel insight into the immune responses of E. coioides to the impB gene of P. plecoglossicida.