• 中文核心期刊要目总览
  • 中国科技核心期刊
  • 中国科学引文数据库(CSCD)
  • 中国科技论文与引文数据库(CSTPCD)
  • 中国学术期刊文摘数据库(CSAD)
  • 中国学术期刊(网络版)(CNKI)
  • 中文科技期刊数据库
  • 万方数据知识服务平台
  • 中国超星期刊域出版平台
  • 国家科技学术期刊开放平台
  • 荷兰文摘与引文数据库(SCOPUS)
  • 日本科学技术振兴机构数据库(JST)
王路英, 刘子旭, 赵玲敏, 黄力行, 覃映雪, 苏永全, 郑炜强, 王凡, 鄢庆枇. 2020: 基于Dual RNA-seq研究dksA基因在变形假单胞菌与大黄鱼的病原体-宿主相互作用中的功能. 动物学研究, 41(4): 410-422. DOI: 10.24272/j.issn.2095-8137.2020.048
引用本文: 王路英, 刘子旭, 赵玲敏, 黄力行, 覃映雪, 苏永全, 郑炜强, 王凡, 鄢庆枇. 2020: 基于Dual RNA-seq研究dksA基因在变形假单胞菌与大黄鱼的病原体-宿主相互作用中的功能. 动物学研究, 41(4): 410-422. DOI: 10.24272/j.issn.2095-8137.2020.048
Lu-Ying Wang, Zi-Xu Liu, Ling-Min Zhao, Li-Xing Huang, Ying-Xue Qin, Yong-Quan Su, Wei-Qiang Zheng, Fan Wang, Qing-Pi Yan. 2020: Dual RNA-seq provides novel insight into the roles of dksA from Pseudomonas plecoglossicida in pathogen-host interactions with large yellow croakers (Larimichthys crocea). Zoological Research, 41(4): 410-422. DOI: 10.24272/j.issn.2095-8137.2020.048
Citation: Lu-Ying Wang, Zi-Xu Liu, Ling-Min Zhao, Li-Xing Huang, Ying-Xue Qin, Yong-Quan Su, Wei-Qiang Zheng, Fan Wang, Qing-Pi Yan. 2020: Dual RNA-seq provides novel insight into the roles of dksA from Pseudomonas plecoglossicida in pathogen-host interactions with large yellow croakers (Larimichthys crocea). Zoological Research, 41(4): 410-422. DOI: 10.24272/j.issn.2095-8137.2020.048

基于Dual RNA-seq研究dksA基因在变形假单胞菌与大黄鱼的病原体-宿主相互作用中的功能

Dual RNA-seq provides novel insight into the roles of dksA from Pseudomonas plecoglossicida in pathogen-host interactions with large yellow croakers (Larimichthys crocea)

  • 摘要: 变形假单胞菌(Pseudomonas plecoglossicida)是革兰氏阴性菌,呈杆状,有鞭毛,该菌在大黄鱼(Larimichthys crocea)养殖过程中引起内脏白点病并有非常高的死亡率,可造成严重的经济损失。经互作转录组数据分析和实时定量PCR验证,dksA基因在变形假单胞菌感染斜带石斑鱼(Epinephelus coioides)48小时后的表达量上调显著(log2FC=3.12 ,P<0.001)。利用RNAi技术显著降低了变形假单胞菌中基因dksA的表达量,最佳沉默效率达到96.23%。分别用变形假单胞菌野生株与效果最好的沉默菌株注射感染健康大黄鱼,观察大黄鱼的存活情况,并于注射后48小时取脾脏进行dual RNA-seq。与野生型毒株相比,感染沉默菌株的大黄鱼内脏白点病症状减轻,死亡时间延迟了2 天且死亡率降低20%。经dual RNA-seq 得知人工感染后病原和宿主的转录组发生了显著变化。dksA基因的沉默导致病原菌的细胞组分,鞭毛和核糖体组装相关基因大量下调,而fliC基因的显著上调可能是变形假单胞菌毒力维持的一种途径。GO和KEGG结果表明沉默菌株感染的大黄鱼的多个与免疫相关通路中炎性因子基因大量下调,在这些免疫途径中,受影响的基因与多个免疫应答过程相关。研究结果表明,dksA是变形假单胞菌的毒力基因,相比于野生株感染,沉默菌株感染的大黄鱼免疫反应较弱。

     

    Abstract: Pseudomonas plecoglossicida is a rod-shaped, gram-negative bacterium with flagella. It causes visceral white spot disease and high mortality in Larimichthys crocea during culture, resulting in serious economic loss. Analysis of transcriptome and quantitative real-time polymerase chain reaction (PCR) data showed that dksA gene expression was significantly up-regulated after 48 h of infection with Epinephelus coioides (log2FC=3.12, P<0.001). RNAi of five shRNAs significantly reduced the expression of dksA in P. plecoglossicida, and the optimal silencing efficiency was 96.23%. Compared with wild-type strains, the symptoms of visceral white spot disease in L. crocea infected with RNAi strains were reduced, with time of death delayed by 48 h and mortality reduced by 25%. The dksA silencing led to a substantial down-regulation in cellular component-, flagellum-, and ribosome assembly-related genes in P. plecoglossicida, and the significant up-regulation of fliC may be a way in which virulence is maintained in P. plecoglossicida. The GO and KEGG results showed that RNAi strain infection in L. crocea led to the down-regulation of inflammatory factor genes in immune-related pathways, which were associated with multiple immune response processes. Results also showed that dksA was a virulence gene in P. plecoglossicida. Compared with the wild-type strains, RNAi strain infection induced a weaker immune response in L. crocea.

     

/

返回文章
返回