温度渐变和骤变下凡纳滨对虾ddit4l基因的表达分析  

Expression Analysis of the ddit4l Gene in Litopenaeus vannamei Under Sudden and Gradual Change of Temperature

作　　者：程琳 李旭鹏 曹宝祥[2,3] 盖超伟 栾生[2,3] 孔杰 孟宪红 CHENG Lin;LI Xupeng;CAO Baoxiang;GAI Chaowei;LUAN Sheng;KONG Jie;MENG Xianhong(National Demonstration Center for Experimental Fisheries Science Education,Shanghai Ocean University Shanghai 201306,China;Laboratory for Marine Fisheries Science and Food Production Processes,Pilot National Laboratory for Marine Science and Technology(Qingdao),Qingdao,Shandong 266071,China;Key Laboratory for Sustainable Utilization of Marine Fisheries Resources,Ministry of Agriculture and Rural Affairs,Yellow Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences,Qingdao,Shandong 266071,China)

机构地区：[1]上海海洋大学、水产科学国家级实验教学示范中心,上海201306 [2]青岛海洋科学与技术试点国家实验室海洋渔业科学与食物产出过程功能实验室,山东青岛266071 [3]中国水产科学研究院黄海水产研究所、农业农村部海洋渔业可持续发展重点实验室,山东青岛266071

出　　处：《渔业科学进展》2023年第2期87-97,共11页Progress in Fishery Sciences

基　　金：国家自然科学基金联合基金项目(U1706203);山东省农业重大应用技术创新项目(SD2019YY001);山东省农业良种工程项目(2019LZGC014);中国水产科学研究院创新团队项目(2020TD26,2021CG01)共同资助。

摘　　要：为探索凡纳滨对虾(Litopenaeusvannamei)DNA损伤诱导因子-4(DNAdamageinducible transcript 4-like, ddit4l)的结构及功能,以转录组测序获得的unigene序列为基础,采用直接PCR扩增方法,获得ddit4l完整的ORF序列(Lv-ddit4l)。该基因的ORF长495 bp,编码164个氨基酸,分子量为18.51 kDa,理论等电点为5.35,包含1个RTP801C超基因家族功能结构域。多序列比对结果和系统进化树显示,Lv-ddit4l与北太平洋雪蟹(Chionoecetes opilio)的同源性最高,为59.76%,且与北太平洋雪蟹先聚为一支。组织表达结果显示,Lv-ddit4l在眼柄、鳃、肝胰腺、神经、肌肉5种组织中均表达,且表达量基本相同。在温度渐变和温度骤变条件下,Lv-ddit4l在不同组织中表达变化模式有较大差别。与26℃水温(对照组)相比,水温渐变至32℃时,眼柄中Lv-ddit4l表达量显著上调(P<0.05)。之后水温缓慢升至最高温38℃再以同样的速度回温。回温至32℃时,鳃、肝胰腺中Lv-ddit4l表达量显著上调(P<0.05)。在环境水温从26℃骤变至36℃后10min时,鳃、肝胰腺、神经中Lv-ddit4l表达量显著上调(P<0.05);24 h时,肌肉中Lv-ddit4l表达量显著上调(P<0.05)。当环境温度分别从38℃、36℃恢复回26℃后,凡纳滨对虾体内原先上调表达的Lv-ddit4l表达量呈下调恢复趋势。研究表明,Lv-ddit4l可能与凡纳滨对虾响应高温胁迫过程相关。Temperature is an important environmental factor that affects the growth, development, and metabolism of shrimp. High temperatures tend to cause a decrease in the immunity and production performance of shrimp, resulting in a significant increase in shrimp disease and mortality. Based on transcriptome sequencing data, we screened for the involvement of DNA damage-inducible transcript 4like(ddit4l) in the temperature stress process of Litopenaeus vannamei. Ddit4l(also known as Redd2 and Rtp801L) is involved in regulating cell survival, proliferation, and apoptosis by affecting DNA damage repair factors, hypoxia-inducible factors, and other signaling pathways when stimulated by DNA damage,hypoxia, ischemia, and oxidative stress, among others. Ddit4l may be an important transducer of pathological stress in autophagy through mammalian target of rapamycin(mTOR) signaling. Autophagy is a process in which cells use lysosomes to degrade their own damaged organelles and macromolecules, and the process plays an important role in maintaining the balance of protein metabolism and the stability of the intracellular environment, promoting cell growth and development. An important regulator of autophagy is the mTOR, which responds to various stimuli, such as growth factors, cellular energy status,oxygen concentrations, and stress, to regulate cell metabolism and growth. Ddit4l and its homolog ddit4are upstream mTOR inhibitors in several tissues and cell models. We set up sudden temperature change and gradual temperature change experiments to induce a stress response in L. vannamei. In the gradual change experiment, the water temperature was increased from 26℃ to 38℃ at a rate of 4℃ per day, and cooled back to 26℃ at the same rate. Two individuals were randomly selected from each of the three parallel experiments in the experimental group at each time point. The eyestalk, gill, hepatopancreas,nerve, and muscle were used for subsequent RNA extraction and gene expression quantification. RNA quality and concentration were al

关 键 词：凡纳滨对虾 高温 Lv-ddit4l 基因表达 

分 类 号：S917.4[农业科学—水产科学]