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基于皮肤转录组的半滑舌鳎无眼侧黑化相关基因的筛选鉴定

赵娜 何晓旭 贾磊 朱春华 张博

赵娜,何晓旭,贾磊,等. 基于皮肤转录组的半滑舌鳎无眼侧黑化相关基因的筛选鉴定[J]. 海洋学报,2022,44(2):57–66 doi: 10.12284/hyxb2022044
引用本文: 赵娜,何晓旭,贾磊,等. 基于皮肤转录组的半滑舌鳎无眼侧黑化相关基因的筛选鉴定[J]. 海洋学报,2022,44(2):57–66 doi: 10.12284/hyxb2022044
Zhao Na,He Xiaoxu,Jia Lei, et al. Identification of genes related to blind side hypermelanosis of Cynoglossus semilaevis based on skin transcriptome sequencing[J]. Haiyang Xuebao,2022, 44(2):57–66 doi: 10.12284/hyxb2022044
Citation: Zhao Na,He Xiaoxu,Jia Lei, et al. Identification of genes related to blind side hypermelanosis of Cynoglossus semilaevis based on skin transcriptome sequencing[J]. Haiyang Xuebao,2022, 44(2):57–66 doi: 10.12284/hyxb2022044

基于皮肤转录组的半滑舌鳎无眼侧黑化相关基因的筛选鉴定

doi: 10.12284/hyxb2022044
基金项目: 天津市自然科学基金(17JCQNJC15000);国家海水鱼产业技术体系项目(CARS-47-Z01);天津市水产产业技术体系项目(ITTFRS2017011)。
详细信息
    作者简介:

    赵娜(1984—),女,河北省保定市人,副研究员,博士,研究方向为水生动物免疫及代谢调节。E-mail:penguinzn@163.com

    通讯作者:

    张博,副研究员,博士,研究方向为遗传育种与生物技术。E-mail:zb611273@163.com

    朱春华,教授,博士,研究方向为水产养殖学。E-mail:hu860025@163.com

  • 中图分类号: S965.3

Identification of genes related to blind side hypermelanosis of Cynoglossus semilaevis based on skin transcriptome sequencing

  • 摘要: 半滑舌鳎(Cynoglossus semilaevis)作为比目鱼(Pleuronectiformes)中的代表物种,是我国沿海地区重要的海水经济鱼类。半滑舌鳎体色异常的问题长期以来一直困扰着从业者。导致体色变化的原因多种多样,遗传因素是最根本原因。针对半滑舌鳎无眼侧黑化的研究多在环境、营养、生理、常见色素基因克隆等方面展开,新的功能基因挖掘工作欠缺。本研究选取半滑舌鳎不同体色皮肤进行转录组测序,通过差异基因GO和KEGG富集比对分析,在可能的6条黑色素相关的KEGG通路中筛查差异基因并对差异表达最为显著的10个基因进行了实时荧光定量聚合酶链式反应的验证。本研究找出了半滑舌鳎无眼侧黑化表达显著变化的5个功能基因,分别为txndc、alox15b、ptgs2、ptgisPTGIS、atp1a2ap<0.05)。其中txndc、alox15b、ptgs2、ptgis基因在黑化组中表达水平高于对照组;功能方面,有3个基因均与花生四烯酸(AA)有一定的相关性,这为多不饱和脂肪酸参与半滑舌鳎体色异常的分子调控机制提供了一定的理论支持。
  • 图  1  半滑舌鳎无眼侧黑化鱼体色特征

    Fig.  1  The body color feature of the Cynoglossus semilaevis with no eye side blackened fish

    图  2  测序样品相关性分析热图

    Fig.  2  Correlation analysis heat map of sequencing samples

    图  3  半滑舌鳎正常皮肤与黑化皮肤样品间差异表达基因火山图和差异表达基因丰度热图

    a. 红色点代表差异基因,黑色点代表被过滤掉的数据,Count为测得计量数;b. 纵向代表基因类型

    Fig.  3  Volcano map of the number of differentially expressed genes and heat map of abundance of differentially expressed gene between normal and hypermelanotic skin samples of Cynoglossus semilaevis

    a. The red dots represent differential expressed genes, the black dots represent the filtered date, Counts is the measured quantity; b. the vertical one represents genes

    图  4  半滑舌鳎正常与黑化皮肤转录组测序差异基因富集注释的GO通路和KEGG通路

    Fig.  4  GO pathways and KEGG pathways analysis of differential expressed genes enrichment annotations of transcriptome sequencing of normal and hypermelanotic skin of Cynoglossus semilaevis

    图  5  6个候选体色相关mRNA在半滑舌鳎6个测序样品中的qPCR表达检测结果箱线图

    蓝色CSC代表半滑舌鳎皮肤正常对照组定量结果,白色CSH代表半滑舌鳎皮肤黑化实验组相对表达结果

    Fig.  5  Box plot of qPCR expression results of 6 candidate body color-related mRNAs in 6 sequencing samples of Cynoglossus semilaevis

    The CSC represents the relative expression in skin of normal C. semilaevis in the control group, and the CSH represents the relative expression in the skin of hypermelanotic C. semilaevis in the experimental group

    表  1  6个半滑舌鳎皮肤转录组测序样品过滤后数据产出及质控分析

    Tab.  1  Clean reads output and quality control analysis of six Cynoglossus semilaevis skin transcriptome sequencing samples

    样本
    名称
    总有效读数碱基
    数目
    Q20质量
    分值
    Q30质量
    分值
    GC含量
    百分比/%
    N碱基
    含量
    csc165 031 0729 72597.0392.2047.680
    csc273 612 17010 98196.8391.9348.190
    csc366 089 0049 88696.9591.9648.810
    csh179 487 97011 89797.2592.5547.610
    csh270 698 40610 57997.3192.7248.380
    csh361 545 5229 20897.0292.0448.220
    下载: 导出CSV

    表  2  6个半滑舌鳎皮肤转录组测序样品基因组比对统计表

    Tab.  2  Genomic comparison statistics of six Cynoglossus semilaevis skin transcriptome sequencing samples

    样品名称总数目唯一比对到参考基因组上的
    测得条目数目及占比
    比对到参考基因组多处的
    测得条目数目及占比
    不能比对到参考基因组的
    测得条目数目及占比
    csh1 39 743 985 32 335 924(81.36%) 1 897 171(4.77%) 5 510 890(13.87%)
    csc1 32 515 536 24 786 485(76.23%) 1 145 579(3.52%) 6 583 472(20.25%)
    csh2 35 349 203 28 571 030(80.83%) 1 563 099(4.42%) 5 215 074(14.75%)
    csc2 36 806 085 28 143 927(76.47%) 1 482 619(4.03%) 7 179 539(19.51%)
    csc3 33 044 502 25 549 970(77.32%) 1 369 439(4.14%) 6 125 093(18.54%)
    csh3 30 772 761 24 553 311(79.79%) 1 291 583(4.20%) 4 927 867(16.01%)
    下载: 导出CSV
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出版历程
  • 收稿日期:  2021-08-19
  • 修回日期:  2021-11-02
  • 网络出版日期:  2021-12-06
  • 刊出日期:  2022-02-01

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