Jiang Hanjun, Lei Qi, Ma Wenhao, Wang Junru, Gong Jing, Guo Xusheng, Cao Xiaojuan
School of Fisheries, Xinyang Agriculture and Forestry University, Xinyang 464000, China.
Fishery Biological Engineering Technology Research Center of Henan Province, Xinyang 464000, China.
Life (Basel). 2025 Aug 2;15(8):1223. doi: 10.3390/life15081223.
(1) Background: Polyploid fish are highly important in increasing fish production, improving fish quality, and breeding new varieties. The loach (), as a naturally polyploid fish, serves as an ideal biological model for investigating the mechanisms of chromosome doubling; (2) Methods: In this study, tetraploidization in diploid loach was induced by heat shock treatment, and, for the first time, the role of the key cell cycle gene (cyclin-dependent kinase 1) in chromosome doubling was investigated; (3) Results: The experimental results show that when eggs are fertilized for 20 min and then subjected to a 4 min heat shock treatment at 39-40 °C, this represents the optimal induction condition, resulting in a tetraploid rate of 44%. Meanwhile, the results of the knockout model (2n ) constructed using CRISPR/Cas9 showed that the absence of significantly increased the chromosome doubling efficiency of the loach. The qPCR analysis revealed that knockout of significantly upregulated cyclin genes (, and ), while inhibiting expression of the separase gene ( < 0.05); (4) Conclusions: During chromosome doubling in diploid loaches induced by heat shock, knocking out the gene can increase the tetraploid induction rate. This effect may occur through downregulation of the gene. This study offers novel insights into optimizing the induced breeding technology of polyploid fish and deciphering its molecular mechanism, while highlighting the potential application of integrating gene editing with physical induction.
(1) 背景：多倍体鱼类在提高鱼类产量、改善鱼肉品质和培育新品种方面具有重要意义。泥鳅作为一种天然多倍体鱼类，是研究染色体加倍机制的理想生物学模型；(2) 方法：本研究通过热休克处理诱导二倍体泥鳅四倍体化，并首次研究关键细胞周期基因（细胞周期蛋白依赖性激酶1）在染色体加倍中的作用；(3) 结果：实验结果表明，卵子受精20分钟后，在39 - 40°C下进行4分钟热休克处理，这是最佳诱导条件，四倍体率达44%。同时，利用CRISPR/Cas9构建的敲除模型（2n）结果显示，的缺失显著提高了泥鳅的染色体加倍效率。qPCR分析表明，的敲除显著上调了细胞周期蛋白基因（、和），同时抑制了分离酶基因的表达（<0.05）；(4) 结论：在热休克诱导二倍体泥鳅染色体加倍过程中，敲除基因可提高四倍体诱导率。这种效应可能通过下调基因发生。本研究为优化多倍体鱼类诱导育种技术、解读其分子机制提供了新见解，同时突出了基因编辑与物理诱导相结合的潜在应用。
