虾肝肠胞虫4个孢壁蛋白基因的鉴定、序列特征及表达分析  

Identification,sequence characteristics and expression analyses of four spore wall protein genes of Enterocytozoon hepatopenaei(EHP)in Litopenaeus vannamei

作　　者：李枝敏 王元[1] 房文红[1] 周俊芳[1] 李新苍[1] LI Zhimin;WANG Yuan;FANG Wenhong;ZHOU Junfang;LI Xincang(East China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Shanghai 200090, China;National Demonstration Center for Experimental Fisheries Science Education, National Pathogen Collection Center for Aquatic Animals, Shanghai Ocean University, Shanghai 201306, China)

机构地区：[1]农业农村部东海渔业资源开发利用重点实验室,中国水产科学研究院东海水产研究所,上海200090 [2]上海海洋大学,水产科学国家级实验教学示范中心,国家水生动物病原库,上海201306

出　　处：《海洋渔业》2021年第1期81-92,共12页Marine Fisheries

基　　金：中央级公益性科研院所基本科研业务费专项资金项目(2019T01);国家自然科学基金面上项目(31772886);东海水产研究所基本科研业务费(2019M03)。

摘　　要：对上海某养殖场凡纳滨对虾(Litopenaeus vannamei)开展了虾肝肠胞虫(Enterocytozoon hepatopenaei,EHP)检测,发现部分对虾存在EHP感染。随后对EHP感染阳性对虾肝胰腺组织进行转录组测序和数据分析,从中筛选到4个孢壁蛋白基因,分别命名为EhEnP1、EhSWP7、EhSWP12和EhSWP26。获取并比较4个基因cDNA序列和基因组序列,发现这4个基因均不含内含子;进一步比较氨基酸序列,发现这4个基因编码的氨基酸序列相似性很低,但它们分别与泰国当地EHP相应的4个假设孢壁蛋白氨基酸序列一致,提示上海本地EHP与泰国当地EHP很可能属于同一个种。qRT-PCR实验结果显示,这4个孢壁蛋白基因均在肝胰腺中表达量最高,并且EhSWP12表达量显著高于其他3个基因(P<0.05)。研究结果表明,EHP主要感染肝胰腺,EhSWP12是EHP孢壁蛋白中的主要结构蛋白组分。鉴定了EHP的4个孢壁蛋白基因,并明确其序列特征及表达情况,研究结果可为EHP生物学特性和侵染机制研究提供基础数据。Enterocytozoon hepatopenaei(EHP)is a newly emerged Microsporidian parasite that causes retarded growth of shrimp,which severely affects the normal development of shrimp industry.By now the mechanism that how this pathogen infects the shrimp has remained unclear,which severely impairs the effectiveness of the prevention and control of EHP.Some spore wall proteins(SWPs)of Microsporidia,as the major structure components,appear to be involved in the invasions in certain hosts.Therefore,identification and characterization of EHP SWPs are of great significance to the study of pathogenic biology and infection mechanism of EHP mediated by SWPs,which would facilitate the control of this pathogen.However,the reports on the EHP SWPs are few,and the potential functions of these molecules are largely unknown.In order to understand the biological characteristics and infection mechanism of local EHP in Shanghai,farmed shrimp(Litopenaeus vannamei)in the suburban farm of Shanghai confirmed to be infected by EHP were sampled to the following assays.The data from transcriptome sequencing with the hepatopancreas of EHP-infected shrimp revealed that four putative SWP-like protein genes were found and they were designated as EhEnP1,EhSWP7,EhSWP12,and EhSWP26.Sequence similarity analyses revealed that the amino acid sequences of these four molecules were identical with the ones reported in Thailand,which were obtained from the data of the genome sequence of EHP.Then the corresponding DNA sequences of these four SWP genes were cloned and sequenced.The results showed that the cDNA sequences of four SWP genes contained non-coding sequences which were the same as their respective DNA sequences.These results revealed that these four SWP genes had no introns.We also found that only EhSWP7 of these four proteins had a putative signal peptide and a functional domain MICSWaP was found in EhSWP26.Quantitative real-time PCR(qRT-PCR)was performed to analyze the expression levels.All four SWP genes expressed the highest in hepatopancreas,moderately

关 键 词：凡纳滨对虾 EHP 孢壁蛋白基因 序列特征 表达分析 

分 类 号：S917[农业科学—水产科学]