文章摘要
胡  娟 1 ,林芳妹 1 ,吕晴霁 1 ,王  庆 1 ,杨慧荣 1 ,石和荣 2,赵会宏.Dmrt 基因在斜带石斑鱼胚胎发育 不同阶段的表达分析[J].广东农业科学,2019,46(3):119-126
Dmrt 基因在斜带石斑鱼胚胎发育 不同阶段的表达分析
Analysis on the Expression of Dmrt Gene in Different Stages of Embryonic Development of Epinephelus coioides
  
DOI:10.16768/j.issn.1004-874X.2019.03.019
中文关键词: 斜带石斑鱼  Ecdmrt1  Ecdmrt2  Ecdmrt3  胚胎发育
英文关键词: Epinephelus coioides  Ecdmrt1  Ecdmrt2  Ecdmrt3  embryonic development
基金项目:广东省自然科学基金(2017A030313099);广东省科技计划项目(2015A020208002);广东省海洋和 渔业发展专项资金(技术推广)项目(2017A0002);广东省鱼港建设和渔业产业发展资金(技术推广)项目(A201600A02)
作者单位
胡  娟 1 ,林芳妹 1 ,吕晴霁 1 ,王  庆 1 ,杨慧荣 1 ,石和荣 2,赵会宏 1. 华南农业大学海洋学院广东 广州 5106402. 广东省海洋渔业试验中心广东 惠州 516081 
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中文摘要:
      【目的】对斜带石斑鱼(Epinephelus coioides)胚胎发育过程及 Dmrts 基因在此过程的表达情况进 行研究,为以后深入研究 Dmrts 基因在石斑鱼中的功能提供基础。【方法】用自然产卵方式取得受精卵在室内 恒温(24±0.5℃)、静水中孵化,在盐度 31.0 的海水中培育,整个胚胎发育过程历时约 29 h 50 min,之后进入 胚后发育阶段。利用光学显微镜观察斜带石斑鱼胚胎发育过程,分别取分裂期、囊胚期、原肠期、神经胚期、 器官形成期、初孵仔鱼期的胚胎保存于 RNAlater 中且每组 5 个平行。【结果】通过荧光定量分析 Dmrt 在不同 胚胎发育时期的相对表达量,结果表明,斜带石斑鱼 Dmrt 基因在胚胎发育过程中,都从卵裂期开始表达。其 中 Ecdmrt1(E. coioides Dmrt1)基因在囊胚期表达水平达到峰值,卵裂期和初孵仔鱼期表达量较低,Ecdmrt1 基 因在囊胚期表达是分裂期的 5.4 倍。Ecdmrt2(E. coioides Dmrt2) 基因在胚胎初期相对表达量比较平稳,而在神 经胚期表达量显著上升,是分裂期的 9.2 倍;Ecdmrt2 基因在初孵仔鱼中表达水平最高,是分裂期的 17.6 倍。 Ecdmrt2b 基因在胚胎发育过程中表达量逐渐增加,在器官形成期表达量达到峰值(P < 0.05),在初孵仔鱼中 表达水平又下降。Ecdmrt3(E. coioides Dmrt3)基因在胚胎发育初期表达量相对较低,在器官形成期表达量显著 上升,是分裂期 7.1 倍,在初孵仔鱼中表达量达到峰值。【结论】Ecdmrt 基因在胚胎发育过程中均从卵裂期开 始表达,且可能参与胚胎的组织器官发育及仔鱼发育生长。
英文摘要:
      【Objective】The embryonic development process of the Epinephelus coioides and the expression of Dmrts gene in this process were studied to provide a basis for further study on the function of Dmrts gene in E. coioides. 【Method】The fertilized eggs obtained by natural spawning method were incubated in a constant temperature (24±0.5℃ ) and in still water, and cultured in seawater with a salinity of 31.0. The whole embryonic development process was completed in about 29 h 50 min and entered the post-embryonic development stage. The embryonic development process of the E. coioides was observed by light microscopy. The embryos in the cleavage stage, blastocyst stage, gastrula stage, neurula stage, organogenesis stage and newly hatched larvae stage were stored in RNAlater with 5 parallels in each group. 【Result】The relative expression of Dmrt in different embryonic development stages were analyzed by fluorescence quantitative analysis. The results showed that the Dmrt gene of the E. coioides began to express from the cleavage stage during embryonic development. The expression level of Ecdmrt1 (E. coioides Dmrt1) reached the peak at the blastocyst stage, and the expression level of Ecdmrt1 gene was relatively low at the cleavage stage and larvae stage. The expression level of Ecdmrt1 gene at the blastocyst stage was 5.4 times that of cleavage stage. The relative expression of Ecdmrt2 (E. coioides Dmrt2) gene was relatively stable at the early embryonic stage but increased significantly at the neurula stage (P<0.05), which was 9.2 times that of cleavage stage. The Ecdmrt2 gene had the highest expression level in the newly hatched larvae, which was 17.6 times that of the cleavage stage. The expression level of Ecdmrt2b gene increased gradually during embryonic development and reached the peak at the organogenesis stage (P<0.05) while the expression level decreased at the newly hatched larvae stage. The expression level of Ecdmrt3 (E. coioides Dmrt3) was relatively low in the early stage of embryonic development, and significantly increased at the organogenesis stage (P<0.05), which was 7.1 times that of the cleavage stage, and reached the peak at the newly hatched larvae stage. 【Conclusion】Ecdmrt gene expressed from the cleavage stage during embryonic development, and may be involved in the development of embryonic organs and larvae.
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