文章摘要
董忠典,黎学友,黄承勤,张海瑞,黄顺楷,张宁,郭昱嵩,王中铎.湖栖鳍虾虎鱼性腺转录组比较分析[J].水产学报,2021,45(3):365~380
湖栖鳍虾虎鱼性腺转录组比较分析
Comparative transcriptome analysis of the gonad of the lacustrine goby (Gobiopterus lacustris)
投稿时间:2020-03-30  修订日期:2020-06-01
DOI:10.11964/jfc.20200312214
中文关键词: 湖栖鳍虾虎鱼  RNA-Seq  配子发生  繁殖
英文关键词: Gobiopterus lacustris  RNA-Seq  gametogenesis  reproduction
基金项目:国家自然科学基金(41806195,31201996);广东省科技厅项目(2017A030303075);广东海洋大学创新强校项目(230419069,230419055);广东海洋大学博士启动项目
作者单位E-mail
董忠典 广东海洋大学水产学院南海水产经济动物增养殖重点实验室广东 湛江 524088  
黎学友 广东海洋大学水产学院南海水产经济动物增养殖重点实验室广东 湛江 524088  
黄承勤 广东海洋大学水产学院南海水产经济动物增养殖重点实验室广东 湛江 524088  
张海瑞 广东海洋大学水产学院南海水产经济动物增养殖重点实验室广东 湛江 524088  
黄顺楷 广东海洋大学水产学院南海水产经济动物增养殖重点实验室广东 湛江 524088  
张宁 广东海洋大学水产学院南海水产经济动物增养殖重点实验室广东 湛江 524088  
郭昱嵩 广东海洋大学水产学院南海水产经济动物增养殖重点实验室广东 湛江 524088  
王中铎 广东海洋大学水产学院南海水产经济动物增养殖重点实验室广东 湛江 524088 aduofa@hotmail.com 
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中文摘要:
      为获得湖栖鳍虾虎鱼性腺基因表达谱,筛选性别决定、配子发生和繁殖相关基因,实验对其卵巢和精巢分别进行了RNA-Seq分析。测序原始数据经de novo拼接组装共获得62 573个单基因(unigene),N50和平均长度分别为3 082 bp和1 869 bp。在NR、NT、SwissProt、PFAM、KOG、GO及KEGG数据库中对上述获得的unigene进行注释,分别有41 480、32 848、37 500、35 394、18 318、35 394和27 009个unigene获得注释。基因表达差异分析显示,在湖栖鳍虾虎鱼卵巢和精巢中存在10 954个差异基因(different expression gene,DEG),其中2 383个DEG表达水平在精巢中上调,8 571个DEG在卵巢中上调。对14个DEG进行实时荧光定量PCR (qPCR) 验证,结果显示,qPCR结果与RNA-Seq分析一致。在差异基因中,筛选到了部分参与配子发生与成熟的基因,包括foxl2、dmrt1、cyp19a1ainhainhbsycp2、zglp1、tdrpzpsesra。差异基因富集结果中包含“泛素介导蛋白水解”、“卵母细胞减数分裂”、“孕酮介导卵母细胞成熟”、“p53信号通路”、“类固醇激素合成通路”和“PI3/AKT信号通路”等参与性腺发育及繁殖的KEGG通路。最后,从湖栖鳍虾虎鱼性腺转录组中共获得38 550个简单重复序列标记(simple sequence repeats,SSR)和192 450个单核苷酸多态性(single nucleotide polymorphism,SNP)标记。研究结果有助于对湖栖鳍虾虎鱼性别相关基因的功能研究和性别相关分子标记的开发。
英文摘要:
      Lacustrine goby (Gobiopterus lacustris) belongs to a genus of gobies that are small in size and endemic to freshwater, brackish waters or coastal environments around the Indian and Pacific oceans. G. lacustris has almost transparent skin and clearly visible internal organs, making it an ideal fish model for potential research in development, reproduction and other fields. Here, we constructed gonadal transcriptomes of G. lacustris using Illumina sequencing for the first time and identified genes that may be involved in gonadal development, gametogenesis and reproduction. Row reads were assembled into 62 573 unigenes with N50 value of 3 082 bp and a mean length of 1 869 bp. 41 480, 32 848, 37 500, 35 394, 18 318, 35 394 and 27 009 unigenes were successfully annotated in NR, NT, SwissProt, PFAM, KOG, GO and KEGG databases, respectively. Gene expression patterns in the ovary and testis were compared, and 10 954 differentially expressed genes (DEGs) were identified. Among these genes, 8 571 were up-regulated in the ovary, and 2 383 were up-regulated in the testis. qPCR analysis of 14 selected genes showed patterns consistent with the transcriptome results. Numerous DEGs involved in gonadal development and gametogenesis were identified, including foxl2, dmrt1, cyp19a1a, inha, inhb, sycp2, zglp1, tdrp, zps and esra. Using GO and KEGG enrichment analyses, pathways involved regulation of gonadal development and gametogenesis, such as "ubiquitin mediated proteolysis", "oocyte meiosis", "progesterone-mediated oocyte maturation", "p53 signaling pathway" and "PI3K-Akt signaling pathway", were also identified. In addition, 38 550 simple sequence repeats were identified from 20 517 SSR containing sequences, and 192 450 single nucleotide polymorphisms were detected. This study denotes the first gonadal transcriptomic analysis of G. lacustris and provides a valuable dataset for future functional analysis of sex-associated and molecular marker-assisted selections in G. lacustris.
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