文章摘要
体外培养条件下三角帆蚌胚胎早期发育观察及胚胎发育生物学零度和有效积温分析
Continuous observation on early embryonic development of Hyriopsis cumingii under in vitro culture condition and analysis of biological zero point and effective accumulated temperature
投稿时间:2020-12-08  修订日期:2021-03-07
DOI:
中文关键词: 三角帆蚌,胚胎发育,体外培养,生物学零度,有效积温
英文关键词: Hyriopsis cumingii  embryonic development  in vitro culture  biological zero point  developmental accumulated temperature
基金项目:国家重点研发计划(2018YFD0901406);现代农业产业技术体系建设专项(CARS-49);国家自然科学基金项目(31872565);江苏省苏北科技专项(SZ-SQ2019045)
作者单位邮编
胡宏辉 上海海洋大学农业农村部淡水水产种质资源重点实验室 201306
白志毅 上海海洋大学农业农村部淡水水产种质资源重点实验室水产动物遗传育种中心上海市协同创新中心 201306
郭思鹏 上海海洋大学农业农村部淡水水产种质资源重点实验室 
冯上乐 上海海洋大学农业农村部淡水水产种质资源重点实验室 
孙田洋 上海海洋大学农业农村部淡水水产种质资源重点实验室 
陈卫民 浙江武义伟民水产养殖有限公司 
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中文摘要:
      为了探寻三角帆蚌胚胎体外培养方法,本研究以解剖获得的雌雄配子为材料,使用与三角帆蚌体液等渗的平衡盐溶液(BSS)进行人工体外受精和体外培养,观察各阶段胚胎的形态特征,并分析计算不同胚胎发育阶段的生物学零度和有效积温。结果表明,在温度为25±1 ℃的BSS盐溶液培养条件下,受精卵发育至第二极体排放、二细胞期、四细胞期、八细胞期、十六细胞期和桑椹期的时间分别为1.8 h、2.8 h、5.6 h、10.5 h、13.9 h和17.3 h,胚胎最长发育至桑椹期后停止发育。另外,池塘平均水温分别为26.5 ℃、28.1 ℃和29.2 ℃时,三角帆蚌胚胎发育至成熟钩介幼虫分别需要10 d、9d和8 d,计算得出三角帆蚌胚胎发育的生物学零度为14.81 ℃,根据生物学零度计算胚胎发育至卵裂期、囊胚期、原肠期、膜内钩介幼虫期和成熟钩介幼虫的有效积温分别为12.95 ℃×d、25.99 ℃×d、42.27 ℃×d、69.21 ℃×d和118.14 ℃×d。研究结果可为突破三角帆蚌全人工繁育技术奠定基础,对淡水贝类现代生物育种技术研发和种质资源保护具有重要意义。
英文摘要:
      In this study, in order to do research on the progress of Hyriopsis cumingii embryos and explore the in vitro culture its early embryos, male and female gametes obtained by anatomy were used as materials for in vitro fertilization and in vitro culture with isoosmotic balance salt solution (BSS) suitable for freshwater mussels. And the morphological characteristics of each stage of embryo were observed with the required time for development recorded. Meanwhile, the embryos in its nurturing pouch of outer gill were continuously observed and calculate the embryonic developmental biological zero point and effective accumulated temperature of embryonic development at the same time. Results show that when the water temperature is 25 ± 1 ℃, the anatomy of the sperms and eggs were mixed after diluting by BSS saline and observed fertilized eggs with polar body emissions at 1.8 h, 2-cell stage at 2.8 h, 4-cell stage at 5.6 h, 8-cell stage at 10.5 h, 16-cell stage at 13.9 h, and morula stage at 17.3 h under optical microscope. And then embryos did not change, and some of embryos were deformed. Meanwhile, when the average water temperature in the pond was 26.54 ℃, 28.08 ℃ and 29.51 ℃, it took respectively 10, 9 and 8 d for embryos to develop into mature glochidium. And the biological zero point for embryonic development of H. cumingii was 14.81℃, and the effective accumulated temperature were 12.95 ℃×d, 25.99 ℃×d, 42.27 ℃×d, 69.21 ℃×d and 118.14 ℃×d for embryos developing to cleavage stage, blastocyst stage, gastrulation stage, glochidium stage, mature mlochidium stage, respectively. The results of this study can provide reference for the researches on artificial breeding and development of modern biological genetic breeding technology of H. cumingii.
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