池蝶蚌急性高温和菌刺激胁迫应答HSP70 B2-like的表达特征  

Analysis of expression characteristics of HSP70 B2-like in response to acute thermal stress and pathogenic bacteria in the freshwater pearl mussel,Hyriopsis schlegelii

作　　者：陈凯欣 赵溪 邱齐骏 吴娣[1,2] 彭扣 CHEN Kaixin;ZHAO Xi;QIU Qijun;WU Di;PENG Kou(School of Life Sciences,Nanchang University,Nanchang 330031,China;Key Laboratory of Aquatic animal resources and utilization of Jiangxi,Nanchang University,Nanchang 330031,China;Fuzhou Agricultural Science Institute of Jiangxi,Fuzhou 344000,China)

机构地区：[1]南昌大学生命科学学院,南昌330031 [2]江西省水产动物资源与利用重点实验室,南昌330031 [3]江西省抚州市农业科学研究所,抚州344000

出　　处：《生物学杂志》2023年第6期74-80,共7页Journal of Biology

基　　金：国家自然科学基金项目(31860737);江西省现代农业产业技术体系-特种水产产业技术体系项目(XARS-10)。

摘　　要：为探究热激蛋白70(Heat shock protein 70,HSP70)对池蝶蚌(Hyriopsis schlegelii)在应对不利环境胁迫时的作用,研究结合转录组测序和RACE-PCR获得池蝶蚌HSP70家族一个成员(暂命名:HSP70 B2-like)cDNA全长序列(2209 bp),同源分析显示,该基因编码的蛋白含有HSP70家族的签名序列,C末端存在细胞质特异性调控信号EEMD。与其他物种的HSP70具有很高的相似性。进化树分析表明,获得序列与褶纹冠蚌HSP70亲缘关系最近,并和其他软体动物HSP70 B2亚家族聚为同一进化支。实时荧光定量PCR结果表明,池蝶蚌HSP70 B2-like在检测的多个组织中均有表达,且闭壳肌中表达量最高。经热刺激(37℃)后,池蝶蚌闭壳肌、肝胰腺、外套膜和肾中的HSP70 B2-like mRNA表达快速上调,6 h达到峰值(P<0.01),随后逐渐下降。池蝶蚌鳃和血细胞中的HSP70 B2-like mRNA表达量则是先降后升再回落。但血细胞、外套膜和肾中的HSP70 B2-like在48 h再次出现表达上调。注射嗜水气单胞菌后,HSP70 B2-like在肝胰腺、血细胞和闭壳肌的表达量均显著上升,6 h开始上调(P<0.05),持续到24 h,到48 h回落。而鳃中的HSP70 B2-like基因的表达则是先降后升再回落。研究结果表明,池蝶蚌HSP70 B2-like是一个应激蛋白基因,可能在机体耐受高温胁迫和抗病原菌浸染时发挥重要作用。To explore the role of heat shock protein 70(HSP70)of Hyriopsis schlegelii in response to adverse environmental stress,transcriptome sequencing and rapid amplification of cDNA ends(RACE)were used to amplify full length cDNA of one member in HSP70 family genes from H.schlegelii(designated as HSP70 B2-like).H.schlegelii HSP70 B2-like has 2209 nucleotides.The homologous analysis indicated that the deduced amino acid sequence contained the signature sequences of HSP70 family and the cytoplasm characteristic motif(EEMD)in the C-terminal domain.The deduced amino acid sequence shared high identities with previously reported HSP70s.Phylogenetic tree based on some known HSP70 amino acids showed that HSP70 B2-like in H.schlegelii was clustered closely with that of C.plicata and grouped with HSP70 B2-like subfamily in other molluscs.The results of real-time fluorescence quantitative PCR showed that the HSP70 B2-like of H.schlegelii was expressed in various tissues,of which the highest expression was in adductor muscles.When H.schlegelii adults exposed to acute high temperature of 37℃,the expression of HSP70 B2-like in adductor muscle,hepatopancreas,mantle and kidney was up-regulated rapidly,reached maximum peak at 6 h(P<0.01),and then decreased gradually.But,the expression level of HSP70 B2-like mRNA in gill and haemocytes decreased first,then increased and then decreased.And its expression in haemocytes,mantle and kidney was up-regulated again at 48 h.After the injection of Aeromonas hydrophila,the expression levels of HSP70 B2-like in hepatopancreas,haemocytes and adductor muscle were significantly increased,which began to be up-regulated at 6 h(P<0.05),continued to 24 h,and fell back at 48 h.However,the expression of HSP70 B2-like gene in the gill decreased first,then increased and last decreased.All these results indicated that HSP70 B2-like in H.schlegelii was a stress-expressed protein gene which might be involved in the response to acute heat stress and resistance to pathogen infection.

关 键 词：池蝶蚌 热激蛋白70 克隆 环境胁迫 组织表达 

分 类 号：Q786[生物学—分子生物学] Q959.215