达氏鲟HSP70-4基因序列特征及其饥饿与病原菌感染下的差异表达模式分析  

Sequence Characterization and Differential Expression Profiles After Starvation and Bacterial Infection of HSP70-4 inAcipenser dabryanus

作　　者：龚全[1] 李青芝 孙甲华 林珏[1] 肖宇 苏旭涛 刘光迅[1] 杜军[1] GONG Quan;LI Qing-Zhi;SUN Jia-Hua;LIN Jue;XIAO Yu;SU Xu-Tao;LIU Guang-Xun;DU Jun(Fisheries Institute,SichuanAcademy ofAgricultural Sciences,Chengdu 611713,China)

机构地区：[1]四川省农业科学院水产研究所,成都611731

出　　处：《农业生物技术学报》2021年第8期1584-1593,共10页Journal of Agricultural Biotechnology

基　　金：四川省农业科学院前言学科研究基金项目(2019QYXK021);四川省科技计划资助项目(2021YJ0278);四川省科技计划资助项目(2021YFYZ0015);四川省淡水鱼创新团队。

摘　　要：热休克蛋白(heat shock protein,HSPs)是一类在不同生命形式间高度保守的蛋白质,参与蛋白质的折叠、生物基质的稳定、大分子装配、多肽的降解及转录调控等。为研究达氏鲟(Acipenser dabryanus)热休克蛋白70-4(HSP70-4)基因组织差异表达谱及其在饥饿和细菌侵染状态下的表达变化,本研究通过达氏鲟全长转录组测序数据库,获得HSP70-4基因序列,其cDNA全长为4029 bp,ORF为2550 bp(GenBank No.MZ285757),编码849个氨基酸。其中5'UTR为234 bp,3'UTR为1245 bp。系统发育树显示,达氏鲟HSP70-4与小体鲟(A.ruthenu)HSP70-4聚在一支,置信度为100%。组织分布结果表明,达氏鲟肝脏、性腺、肌肉、脑、眼睛、皮肤、头肾、心脏、肠、脾脏和鳃11个组织中均有HSP70-4表达,其中,眼睛中的表达量最高,其次为脑和肌肉,肝脏、头肾、皮肤和肠道中的表达量较低。在饥饿胁迫下,达氏鲟HSP70-4 mRNA出现了上调表达,饥饿14 d后,肠道及肌肉中HSP70-4基因均出现显著上调表达;感染迟缓爱德华氏菌(Edwardsiella tarda)(1×10^(7) CFU/mL)12 h后,皮肤、脾脏和头肾中的HSP70-4与对照组相比显著上调表达(P<0.05);24 h后,鳃组织中的表达量开始显著上调(P<0.05)。研究结果表明达氏鲟HSP70-4参与了机体对抗饥饿及致病菌侵染的生物学过程,为进一步研究达氏鲟HSP70-4基因的诱导差异表达机制以及抗逆机理提供理论依据。Heat shock proteins(HSPs)are highly conserved among different species,which are involved in protein folding,stabilization of biological matrix,assembly of macromolecules,degradation of polypeptides and transcriptional regulation.To study the expression profiles of HSP70-4 in different tissues or under the stress of fasting and bacteria invasion,The sequence of HSP70-4 gene were identified based on full-length transcriptome sequencing of Acipenser dabryanus.The full-length cDNA sequence of HSP70-4 was 4029 bp,which included 234 bp of the 5'-UTR,1245 bp of the 3'-UTR and 2550 bp of the ORF(GenBank No.MZ285757)that encoded 849 amino acids.Phylogenetic analysis showed that the A.dabryanus HSP70-4 gene clustered mostly close to the A.ruthenus HSP70-4,supported with a significant bootstrap value(100).HSP70-4 was universally expressed in all 11 of the tested tissues including liver,gonad,muscle,brain,eye,skin,head kidney,heart,intestine,spleen,and gill.The expression of HSP70-4 was the highest in the eye and was also detected at relatively high levels in the brain and muscle,and relatively weakly expressed in the liver,head kidney,skin,and intestine.Under starvation stress,the expression of HSP70-4 was upregulated,and significantly upregulated at 14 d post-fasting in intestine and muscle.HSP70-4 in the skin,spleen and head kidney was significantly upregulated(P<0.05)at 12 h post-challenge towards Edwardsiella tarda(1×10^(7) CFU/mL)infection and the upregulation of HSP70-4 in the gill started at 24 h post-challenge.The results showed that short-term bacteria infection resulted in significantly upregulated expression of HSP70-4 gene in the gill,skin,spleen and kidney of A.dabryanus.This study demonstrated that HSP70-4 was involved in the biological process of fighting hunger and infection by pathogenic bacteria in A.dabryanus.This study demonstrated that HSP70-4 was involved in the biological process of fighting hunger and infection by pathogenic bacteria and provided a theoretical basis for the further study on the mech

关 键 词：达氏鲟 HSP70-4 饥饿胁迫 细菌侵染 

分 类 号：S917.4[农业科学—水产科学]