流速胁迫对美国红鱼的转录特性研究  

Hepatic Transcriptome Characterization of Flow Velocity Stress in Sciaenops ocellatus

作　　者：娄宇栋 冯建 何娇娇 晁帅 王萍 LOU Yu-dong;FENG Jian;HE Jiao-jiao(School of Ocean Science and Technology of Zhejiang Ocean University, Zhejiang Key Laboratory of Marine Aquaculture Facilities and Engineering Technology, Zhoushan 316022, China)

机构地区：[1]浙江海洋大学海洋科学与技术学院浙江省海洋养殖装备与工程技术重点实验室,浙江舟山316022 [2]宁波大学海洋学院,浙江宁波315211

出　　处：《浙江海洋大学学报（自然科学版）》2019年第1期13-22,29,共11页Journal of Zhejiang Ocean University：Natural Science

基　　金：国家自然科学基金(31602205);浙江省自然科学重点基金项目(Z16E090006);国家海洋公益性行业科研专项号(201505025);舟山市海洋专项(2015C41001)

摘　　要：水流对鱼类生长发育有着重要的影响,为了在分子水平上研究美国红鱼在流速胁迫下的机理,在给定最大续航游泳速度0.9 m·s^(-1)的条件下,利用转录组测序的方法研究了流速胁迫对美国红鱼肝脏转录组的影响。实验结果如下:测序总共获9.32 Gb Clean Data,各样品Clean Data均达到4.54 Gb,Q30碱基百分比在92.12%及以上。De novo组装后共获得70148条Unigene,其中长度在1 kb以上的Unigene有12 465条。基于Unigene库的基因结构分析,其中SSR分析共获得10214个SSR标记;SNP分析试验组T01和对照组T02的纯合型数目分别为38 020和29 627,杂合型数目分别为57 835和66 088个。经过筛选后得到显著性差异表达的基因有1 173个,其中上调的基因数目为204个,下调的基因数目为969个。通过GO富集分析,有424个富集到生物过程(biological process),有90个富集到胞组分(cellular component),有310个富集到分子功能(molecular function)。差异表达基因的富集分析结果显示,能够注释的差异表达基因数目有1096个,注释到KEGG通路的有423个:富集到新陈代谢通路(metabolic pathways)、环境信息处理(environmental information processing)和细胞过程(cellular processes)的差异表达基因相对较多,分别为136、69和67个;有4条为显著性富集通路,分别为甘氨酸、丝氨酸和苏氨酸代谢、色氨酸代谢、视黄醇代谢和类固醇类的生物合成;应对流速胁迫的相关基因,如Hedgehog信号通路(Hedgehog signaling pathway)中Hh、PKA、CK1、Wnt等基因表现为上调、RNA降解(RNA degradation)中的Dcp1、EDC3基因表现为下调等。为了验证RNA-seq分析的表达谱,对11个基因进行了相关mRNA水平的qPCR测定,经过Spearman的rho测试发现qPCR的数据显著相关(R^2=0.979 7),对比qPCR和RNA测序的数据,上调和下调基因均非常吻合。Fish live in the water all their life. They cannot live without water. Their growth, development, reproduction and migration can’t be separated from water. Water flow speed directly affects the water environment;it can determine the growth environment of fish At present, most of the studies of water flow on fish are focus on swimming mechanisms and swimming rates. However, the study of water flow affecting fish on molecular scale is still rare. Study of the interaction on flow velocity and energy at the molecular level is of special value in Sciaenops ocellatus. The study improving the research of the flow stress on the liver transcriptome of S. ocellatus. In this study, under given maximum endurance swimming speed of 0.9 m·s^-1, we used transcriptome sequencing method to study the effect of water flow speed stress on S. ocellatus hepatic transcriptome. The corresponding experimental results were listed as follows: a total of 9.32 Gb Clean Data were sequenced, all Clean Data has reached 4.54 Gb, Q30 base percentage 92.12% and more. A total of 70 148 genes were obtained after De novo assembly, there were 12 465 unigene in length above 1 kb. Based on the unigene Library of gene structure analysis, in which SSR analysis, 10 214 SSR markers were obtained;SNP analysis we get that the number of homozygous types of T01 in the experimental group and the control group were 38 020 and 29 627, and the number of T02 in the experimental group was higher than that in the control group, The number of heterozygous types was 57 835 and 66 088. After screening, there were 1 773 differentially expressed genes. Among them, the number of up-regulated genes was 204, and the number of down regulated genes was 969. Through GO enrichment analysis, 424 were enriched into biological processes, with 90 enriched into cellular component, and 310 enriched to molecular function. The enrichment analysis of differentially expressed genes showed that there are 1 096 annotated differentially expressed genes, 423 of which are annotated to the KEG

关 键 词：流速 胁迫 美国红鱼 转录 Q-PCR 

分 类 号：S917.4[农业科学—水产科学] Q344[生物学—遗传学]