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双链 RNA 测序揭示了转录调控因子基因在鮰爱德华氏菌与青石斑鱼互作中的作用。

Dual RNA-Seq reveals the role of a transcriptional regulator gene in pathogen-host interactions between Pseudomonas plecoglossicida and Epinephelus coioides.

机构信息

College of Environment and Public Health, Xiamen Huaxia University, Xiamen, Fujian, 361024, China; Fisheries College, Key Laboratory of Healthy Mariculture for the East China Sea, Ministry of Agriculture, Jimei University, Xiamen, Fujian, 361021, China.

College of Environment and Public Health, Xiamen Huaxia University, Xiamen, Fujian, 361024, China.

出版信息

Fish Shellfish Immunol. 2019 Apr;87:778-787. doi: 10.1016/j.fsi.2019.02.025. Epub 2019 Feb 16.

DOI:10.1016/j.fsi.2019.02.025
PMID:30776540
Abstract

Pseudomonas plecoglossicida is a highly pathogenic bacterium for maricultured fish and causes serious losses. A transcriptional regulator gene RK21_RS10315 was found up-regulated during the whole infection process, which was confirmed by qRT-PCR. Five shRNA were designed to silence RK21_RS10315 gene, and the gene expression was reduced up to 96.1%. Compared with the counterpart infected with wild type strain, the infection of RK21_RS10315-RNAi strain resulted in the death time delay, and 90% reduction in mortality of Epinephelus coioides, as well as the alleviation in the symptoms of E. coioides spleen. Moreover, compared with the fish infected with wild type strain, the infection of RK21_RS10315-RNAi strain of P. plecoglossicida resulted in a significant change both in transcriptome of spleen of infected E. coioides and P. plecoglossicida. The KEGG analysis showed that genes of 16 immune pathways in E. coioides were affected by the silence of RK21_RS10315 of P. plecoglossicida. Among them, intestinal immune network for IgA production pathway and leukocyte transendothelial migration pathway were more prominent than other pathways. 19 euk-DEMs in these immune pathways had varying degrees of correlation with 19 pro-DEMs, and the expression of ipxA, grpE, yhbJ, truD and suhB from 19 pro-DEMs were predicted more related to RK21_RS10315 in P. plecoglossicida.

摘要

鮰爱德华氏菌是一种对海水养殖鱼类具有高度致病性的细菌,会造成严重的损失。我们发现一个转录调控基因 RK21_RS10315 在整个感染过程中上调,qRT-PCR 进一步证实了这一点。我们设计了 5 条 shRNA 来沉默 RK21_RS10315 基因,基因表达降低了 96.1%。与野生型菌株感染的对照相比,RK21_RS10315-RNAi 菌株的感染导致死亡时间延迟,死亡率降低了 90%,同时石斑鱼脾脏的症状也得到了缓解。此外,与野生型菌株感染的鱼相比,RK21_RS10315-RNAi 菌株的感染导致感染石斑鱼的脾脏转录组发生了显著变化。KEGG 分析显示,16 条免疫途径中的基因受到了鮰爱德华氏菌 RK21_RS10315 的沉默的影响。其中,肠道免疫网络 IgA 产生途径和白细胞跨内皮迁移途径比其他途径更为显著。在这些免疫途径中,19 个真核 DEGs 与 19 个原核 DEGs 有不同程度的相关性,并且这些原核 DEGs 中的 ipxA、grpE、yhbJ、truD 和 suhB 的表达与鮰爱德华氏菌 RK21_RS10315 的关系更为密切。

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