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为探索诱导鱼类雄核发育单倍体的新方法及形成机制,以大鳞副泥鳅为研究对象,无须卵子失活,仅用冷休克诱导雄核发育单倍体,并对其后代的早期胚胎发育、染色体数目、分裂胚细胞学观察等进行了系统分析。早期胚胎发育结果显示,0℃与3℃处理组的孵化率分别为0.83%和57.42%,二者之间孵化率差异显著(P<0.05),0℃和3℃处理组均表现出单倍体综合征。染色体倍性鉴定结果显示,0℃处理组染色体分布为18~27,单倍体率为81.40%,3℃处理组染色体分布为20~30,单倍体率为68.97%。分裂胚细胞学观察结果显示,3℃处理组受精后60 min,卵核和第二极体位于胚表面,有一同释放迹象。结果表明,对大鳞副泥鳅受精卵进行3℃冷休克处理60 min,可以成功诱导雄核发育单倍体,研究推测冷休克诱导大鳞副泥鳅雄核发育的细胞学机制是雌核和第二极体一同释放。
In order to explore a new method and mechanism of inducing haploid development in fish and its male nucleus, Paramruhot sp. Was studied without any inactivation of egg, only the cold shock was used to induce haploid development in male nucleus, Embryonic development, the number of chromosomes, the observation of dividing embryo cytology and so on were systematically analyzed. The results of early embryo development showed that hatching rates at 0 ℃ and 3 ℃ were 0.83% and 57.42%, respectively (P <0.05), and the hatching rates at 0 ℃ and 3 ℃ were all single Body syndrome. Chromosome ploidy identification results showed that the chromosomal distribution of 0 ℃ treatment group was 18-27, the haploid rate was 81.40%, the chromosomal distribution was 20-30 at 3 ℃, the haploid rate was 68.97%. The results of the blastocyst assay showed that the egg nuclei and the second polar body were located on the embryo surface at 60 min after fertilization at 3 ℃, showing signs of release. The results showed that cryopreservation of Paramisgurnus dabryanus eggs by cold shock treatment at 3 ℃ for 60 min could successfully induce the development of the male haplogroups. The cytological mechanism of cold shock-induced male nucleus development in Paramisgurnus dabryanus is gynoecious Diode released together.