Abstract Pseudomonas plecoglossicida is an important pathogen of maricultured fish in China and caused great loss. In this study, the mRNA level of pvdE gene of wild-type strain of P. plecoglossicida (NZBD9) was significantly decreased by 5 shRNAs and the mutant with the best silencing efficiency (89.4%) were chosen for subsequent research. Injection of pvdE-RNAi strain at a dose of 105 cfu/fish did not cause death in orange-spotted grouper and there were only a few less obvious white spots on the surface of infected spleens. Although the injection of pvdE-RNAi strain of P. plecoglossicida induced antibody production of orange-spotted grouper, the premonition of mutant infected grouper did not produce good immune protection against subsequent wild strain infection. The results of RNA-seq revealed the significant differences between the transcriptome of orange-spotted grouper infected by pvdE-RNAi strain of P. plecoglossicida and the counterpart fish infected by stain NZBD9 as well as those of orange-spotted grouper injected by PBS. Th1 and Th2 cell differentiation, Intestinal immune network for IgA production, Th17 cell differentiation, Natural killer cell mediated cytotoxicity, Chemokine signaling pathway, Hematopoietic cell lineage and T cell receptor signaling pathway were the seven immune pathways most affected by silence of pvdE gene.

中文翻译：

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RNA-seq 和 RNAi 的整合为 pvdE 基因对 Pseudomonas plecoglossicida 的致病作用和对橙斑石斑鱼 (Epinephelus coioides) 免疫反应的影响提供了新的见解

摘要 Pseudomonas plecoglossicida 是我国海水养殖鱼类的重要病原体，造成了巨大损失。在本研究中，野生型 P. plecoglossicida (NZBD9) 的 pvdE 基因的 mRNA 水平显着降低了 5 个 shRNA，并选择了具有最佳沉默效率 (89.4%) 的突变体进行后续研究。以 105 cfu/鱼的剂量注射 pvdE-RNAi 菌株不会导致橙斑石斑鱼死亡，并且在受感染的脾脏表面只有少数不太明显的白点。虽然注射P. plecoglossicida 的pvdE-RNAi 菌株诱导了橙斑石斑鱼的抗体产生，但突变感染石斑鱼的预感并没有对随后的野生菌株感染产生良好的免疫保护。RNA-seq的结果揭示了被P. plecoglossicida pvdE-RNAi菌株感染的橙斑石斑鱼的转录组与被NZBD9染色的对应鱼以及PBS注射的橙斑石斑鱼的转录组之间的显着差异。Th1和Th2细胞分化、IgA产生的肠道免疫网络、Th17细胞分化、自然杀伤细胞介导的细胞毒性、趋化因子信号通路、造血细胞谱系和T细胞受体信号通路是受pvdE基因沉默影响最大的七种免疫通路。