高温胁迫下黑龙江茴鱼幼鱼肝脏组织结构变化及转录组表达特征  

Histological structure and transcriptome characteristics in liver of juvenile Amur grayling(Thymallus arcticus grubei)under high temperature stress

作　　者：丰超杰 刘霞飞 张颖[2] 吕伟华[2] 韩世成[2] 张永泉[2] 许式见 马波[2] FENG Chaojie;LIU Xiafei;ZHANG Ying;Lü Weihua;HAN Shicheng;ZHANG Yongquan;XU Shijian;MA Bo(College of Fisheries and Life Science,Shanghai Ocean University,Shanghai 201306,China;Heilongjiang River Fisheries Research Institute,Chinese Academy of Fishery Sciences,Harbin 150076,China;College of Fisheries and Life Science,Dalian Ocean University,Dalian 116023,China;Hangzhou Qiandaohu Xunlong Sci-tech Company Limited,Quzhou 324000,China)

机构地区：[1]上海海洋大学水产与生命学院,上海201306 [2]中国水产科学研究院黑龙江水产研究所,黑龙江哈尔滨150076 [3]大连海洋大学水产与生命学院,辽宁大连116023 [4]杭州千岛湖鲟龙科技股份有限公司,浙江衢州324000

出　　处：《大连海洋大学学报》2023年第4期603-614,共12页Journal of Dalian Ocean University

基　　金：农业农村部农业财专项“东北地区重点水域渔业资源与环境调查”;中国水产科学研究院基本科研业务费资助(2020TD07)。

摘　　要：为探究急性高温胁迫下黑龙江茴鱼(Thymallus arcticus grubei)的响应调控机制,采用组织病理学和高通量测序技术(RNA-Seq)分析了高温组(17℃)和对照组(11℃)随胁迫时间(1、6、12、24、48 h)的延长,黑龙江茴鱼幼鱼(体质量为1.73 g±0.23 g)肝脏组织结构变化及转录组表达特征。结果表明:17℃高温下,随胁迫时间的延长,黑龙江茴鱼幼鱼肝脏组织结构出现明显的病理损伤,于胁迫12 h后肝脏组织出现不同程度的核萎缩变形及空泡化现象,且在胁迫48 h时肝脏细胞损伤最为严重,甚至出现细胞溶解现象;转录组测序共获得214.44 Gbp有效数据,平均GC含量为49.45%,Q30碱基分布为93.48%~96.06%;筛选到差异基因(DEGs)数共9144个,且DEGs数量随胁迫时间的延长呈先升高后降低的趋势,在胁迫24 h时差异基因数量最多,有7148个,其中3653个上调,3495个下调;进一步将差异基因进行GO功能注释显示,被注释的差异基因主要与代谢、催化和结合等功能有关;KEGG富集分析显示,差异基因在糖酵解/糖异生、乙醛酸与二羧酸代谢、果糖与甘露糖代谢、内质网中的蛋白加工、核糖体及PPAR信号等通路上显著富集;随机选择6个DEGs进行实时荧光定量PCR验证,基因表达变化趋势与转录组数据一致。研究表明,在急性高温胁迫下,黑龙江茴鱼幼鱼肝脏组织在12 h后出现不同程度的应激损伤,通过转录组表达分析发现,差异基因主要富集在免疫应激和能量代谢等相关通路上。In order to explore the response mechanism of Amur grayling(Thymallus arcticus grubei)under acute high temperature stress,the changes in liver histopathological structure and transcriptome expression characteristics were investigated in juvenile Amur grayling with body weight of(1.73±0.23)g exposed to 17℃(high temperature group,at warming rate of 1℃/h)and 11℃(control group)for 1,6,12,24 and 48 h by histopathological method and high-throughput RNA-Seq sequencing technology.The results showed that the liver tissue structure of Amur grayling showed obvious pathological damage with the extension of stress time at 17℃in high temperature group.The liver tissue had varying degrees of nuclear atrophy,deformation and vacuolization in 12 hours of stress,and the hepatocytes were most severely damaged,even cell dissolution at 48 hours of stress.A total of 214.44 Gbp of valid data were obtained by transcriptome sequencing,with an average GC content of 49.45%and a Q30 base distribution of 93.48%-96.06%.A total of 9144 differentially expression genes(DEGs)were screened,and the number of DEGs was found to be increased first and then decreased with the extension of temperature stress.The maximal differential expression genes(7148)were observed at 24 h of stress,3653 of which were up-regulated and 3495 of which were down regulated.The GO function annotation of DEGs showed that the annotated DEGs were primarily involved in metabolism,catalysis and binding functions;KEGG enrichment analysis showed that DEGs were significantly enriched in glycolysis/gluconeogenesis,glyoxylic acid and dicarboxylic acid metabolism,fructose and mannose metabolism,protein processing in endoplasmic reticulum,ribosome and PPAR signaling pathways.Six DEGs were randomly selected for real-time fluorescent quantitative PCR validation,and the trend of gene expression was consistent with the transcriptome data.It can be seen that under the acute temperature stress,the liver tissue of Amur grayling appeared different degrees of stress damage 12 hours lat

关 键 词：黑龙江茴鱼 高温胁迫 肝脏 转录组 组织病理 

分 类 号：S964[农业科学—水产养殖]