二次正交旋转组合设计优化人工诱导翘嘴鲌雌核发育 [中文引用][英文引用]

采用紫外线照射鲤鱼(Cyprinus carpio)精子使其遗传物质失活,并用冷休克方法抑制翘嘴鲌(Culter alburnus)卵第二极体的排出,诱导出翘嘴鲌雌核发育二倍体。运用3因素5水平二次正交旋转组合设计方法,分别建立冷休克起始时间(x₁)、处理时间(x₂)和处理温度(x₃) 3个因素与翘嘴鲌雌核发育率(y₁)和孵化率(y₂)两个指标之间的二次回归模型,探究翘嘴鲌减数分裂雌核发育的最优诱导条件组合。并采用响应面分析法(RSM),讨论各因素水平的单因素效应、互作效应对响应值的影响。结果表明,受精后6 min进行冷休克,处理时间为18 min,休克温度为7℃时,雌核发育率达最大(18.60%);受精后6 min进行冷休克,处理时间为23 min,休克温度为5℃时,孵化率达最大(33.00%)。各因素对响应值的影响均是呈开口向下的抛物面关系。根据因素贡献率得到,起始时间对翘嘴鲌雌核发育影响极显著,冷休克处理温度次之,而冷休克处理时间的影响最小。

Culter alburnus gynogenetic diploids were created by activating eggs with ultraviolet-irradiated common carp sperm and preventing extrusion of the second polar body using cold shock to induce chromosome duplication in the eggs. A three-factor five-level quadratic orthogonal rotation combined design was used to determine the optimal combination of three experimental factors, including time after fertilization (x₁), treatment duration (x₂), and shock temperature (x₃) to induce artificial gynogenesis in C. alburnus, using gynogenesis rate (y₁) and hatching rate (y₂) as indicators in the quadratic regression model. Furthermore, the response surface method was used to determine the effect of single factors and their interactive effect on the response value. We found that the optimal combination of factors to induce gynogenesis (18.00%) was cold shock for 18 min at 7℃, applied 6 min after fertilization, whereas that for maximal egg hatch (33.00%) was cold shock for 23 min at 5℃, applied 6 min after fertilization. The relationship between the various factors on the response values was parabolic with a downward opening. According to the factor contribution rates, cold shock start time had a highly significant effect on cold shock, followed by shock temperature and treatment duration. This is the first study to use a quadratic orthogonal rotation combined design to determine the optimal combination of factors to induce gynogenesis in C. alburnus. These results provide a basis to explore the sex-determining mechanism in C. alburnus.