人工诱导泥鳅雌核发育的细胞学研究和诱导参数优化  

CYTOLOGICAL STUDY ON ARTIFICIALLY INDUCED GYNOGENESIS AND OPTIMIZATION OF INDUCED PARAMETERS IN LOACH,MISGURNUS ANGUILLICAUDATUS

作　　者：钟汶蓉 陶彬彬 徐闻 谭娟[4] 罗红瑞 宋焱龙[2] 陈戟[2] 胡炜[2,3] ZHONG Wen-Rong;TAO Bin-Bin;XU Wen;TAN Juan;LUO Hong-Rui;SONG Yan-Long;CHEN Ji;HU Wei(Dalian Ocean University,Dalian 116023,China;State Key Laboratory of Freshwater Ecology and Biotechnology,Hubei Hongshan Laboratory,Institute of Hydrobiology,Chinese Academy of Sciences,Wuhan 430072,China;University of Chinese Academy of Sciences,Beijing 100049,China;Huaihua University,Huaihua 418000,China)

机构地区：[1]大连海洋大学,大连116023 [2]中国科学院水生生物研究所,淡水生态与生物技术国家重点实验室,湖北洪山实验室,武汉430072 [3]中国科学院大学,北京100049 [4]怀化学院,怀化418000

出　　处：《水生生物学报》2024年第2期334-341,共8页Acta Hydrobiologica Sinica

基　　金：国家现代农业产业技术体系项目(CARS-46);湖北省重点研发计划(2021BBA088)资助。

摘　　要：为培育泥鳅(Misgurnus anguillicaudatus)全雌品种,研究通过细胞荧光染色后进行细胞学观察确定人工诱导泥鳅雌核发育的热休克起始时间,并进一步利用核型和流式细胞分析等方法对人工诱导雌核发育泥鳅进行鉴定。结果显示,二倍体泥鳅受到灭活的鲤(Cyprinus carpio)精子刺激后,第二极体与卵核分开的时间在人工授精后3—5min;卵子在人工授精后3.5min后再热休克2min的诱导孵化率达到10.26%。通过分析野生型泥鳅胚胎、人工诱导雌核发育泥鳅胚胎、泥鳅×鲤杂交胚胎和单倍体泥鳅胚胎的发育,发现部分雌核发育的胚胎可以正常发育,而杂交胚胎和单倍体胚胎会出现明显的发育障碍。核型和流式细胞检测分析表明利用本研究获得的热休克参数进行处理可得到二倍体雌核发育子代,二倍率达64.71%。研究从细胞学层面获得泥鳅雌核发育的诱导参数,可为利用人工诱导雌核发育开展全雌泥鳅育种提供指导。The female loach(Misgurnus anguillicaudatus)generally have a larger size compared to males.The application of artificially induced gynogenesis in the cultivation of strains presents great potential.In this study,the starting time of heat shock of artificially induced gynogenesis was determined by cytological observation after cell fluorescence staining.The artificially induced gynogeny embryos were further identified by using karyotype and flow cytometric analysis.The results showed that artificial gynogenesis of loach was successfully induced by inactivated common carp sperm.Cytological observation of fertilization showed that the second polar body was observed at 3—5min after insemination.The optimal starting time of heat shock treatment at 41℃was found to be 3.5min after insemination,followed by a 2min heat shock,resulting in an optimal hatchability rate of 10.26%.By analyzing the development of wild-type loach embryos,artificially induced gynogenetic loach embryos,loach×carp hybrid embryos and haploid loach embryos,it was found that some gynogenetic embryos developed normally,while the hybrid and haploid embryos showed obvious developmental disorders.Karyotype and flow cytometric analysis of the gynogenetic loach embryos further demonstrated that treatment with the heat shock parameters obtained in this study resulted in 64.71%diploid gynogenetic offspring.In this study,the optimized induction parameters of loach meiogynogenesis were obtained through the cytological analysis,which could provide a technical basis for the artificially induced meiogynogenesis in the cultivation of all-female loach strains.

关 键 词：人工诱导 减数分裂雌核发育 热休克 受精细胞学 泥鳅 

分 类 号：S961.6[农业科学—水产养殖]