低氧胁迫下中华绒螯蟹血淋巴细胞的转录组学  

Transcriptome analysis of Eriocheir sinensis hemocytes in response to hypoxia

作　　者：候利波 陆银月 任秋霖 孔祥会 朱雷 顾伟 孟庆国 HOU Libo;LU Yinyue;REN Qiulin;KONG Xianghui;ZHU Lei;GU Wei;MENG Qingguo(College of Fisheries,Henan Normal University,Xinxiang 453007,China;Jiangsu Key Laboratory for Aquatic Crustacean Diseases,College of Marine Science and Engineering,Nanjing Normal University,Nanjing 210046,China)

机构地区：[1]河南师范大学水产学院,河南新乡453007 [2]南京师范大学海洋科学与工程学院,江苏省水生甲壳动物病害重点实验室,江苏南京210046

出　　处：《水产学报》2023年第4期25-39,共15页Journal of Fisheries of China

基　　金：国家重点研发计划(2018YFD0900602);国家自然科学基金(31870168);江苏现代农业产业技术体系专项[JATS(2020)303];河南省自然科学基金(212300410175)。

摘　　要：为了探究低氧胁迫对中华绒螯蟹血淋巴细胞影响的分子机制,实验通过使用Nova Seq 6000测序技术,检测了低氧组(1.5±0.5)mg/L和对照组(6.0±0.5)mg/L分别处理24 h后的中华绒螯蟹血淋巴细胞的转录组数据变化。首先,对测序得到的原始数据进行拼接、注释以及筛选,分析获得的128614个转录本和128614条基因,平均长度为2634 bp(N50),Q30>92%。其次,以1.2倍为阈值,共筛选出1687个差异表达基因,其中上调基因899个,下调基因788个。GO和KEGG分析发现,低氧胁迫对中华绒螯蟹血细胞的三羧酸循环、糖酵解途径、ECM-受体相互作用、间隙连接、细胞凋亡、酚氧化酶系统以及其他免疫相关基因等产生了显著影响。最后,随机挑选转录组数据中的10个基因进行实时荧光定量PCR(qRT-PCR)验证,结果显示与转录组数据分析相一致,其中包括6个上调的基因:整合素1、铜/锌超氧化物歧化酶、磷酸肌醇3激酶、磷酸甘油酸突变酶2、PDGF/VEGF相关因子1和relish;4个下调的基因:无脊椎连接蛋白7、热休克蛋白90、线粒体ATP合成酶α和天冬氨酸转氨酶。本研究初步阐明了中华绒螯蟹血淋巴细胞短时间内应对低氧胁迫的分子机制,同时该结果也可为今后研究其他甲壳动物在应对低氧胁迫时的生理机制和分子机制提供参考。To understand the effect of hypoxia on Chinese mitten crab Eriocheir sinensis,the transcriptomic profiles of hemocytes in the hypoxia(1.5±0.5)mg/L and the normoxia groups[(6.0±0.5)mg/L of DO]were obtained using Nova Seq 6000.Firstly,by splicing,annotating and screening the original data,in total,128614 transcripts were obtained with an average length of 2634 bp.Meanwhile,using a 1.2-fold change(P<0.05)in expression as a physiologically significant benchmark.A total of 1687 differentially expressed genes were identified,including 899 up-regulated and 788 down-regulated genes.GO and KEGG analysis showed that many processes or pathways are involved in the crab against hypoxia stress,such as tricarboxylic acid cycle(TCA cycle),glycolysis,ECM-receptor interaction,gap junction,cell apoptosis,prophenoloxidase system(proPO),etc.Finally,ten selected genes from these processes or pathways were verified by qRT-PCR analysis.The results showed 6 up-regulated genes(integrin-β1,extracellular Cu/Zn-superoxide dismutase,phosphoinositide-3 kinase,phosphoglycerate mutase 2,PDGF/VEGF-related factor 1,and relish)and 4 down-regulated genes(innexin-7,90 ku heat shock,mitochondrial ATP synthase subunitα,and aspartate aminotransferase)corresponded with the transcriptome analysis.This study preliminarily elucidated the molecular mechanism of hemolymph response to short-time hypoxic stress,and provided some valuable information for the further analysis of the mechanism under hypoxia stress in crabs.

关 键 词：中华绒螯蟹 血淋巴细胞 低氧胁迫 转录组分析 

分 类 号：Q786[生物学—分子生物学] S917.4[农业科学—水产科学]