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    冷休克诱导斑鳜三倍体的条件优化研究

    Optimization of cold shock conditions for triploid induction in Siniperca scherzeri

      摘要
    • 摘要:
      目的 本实验旨在探索冷休克法诱导斑鳜（Siniperca scherzeri）三倍体的最佳诱导条件。
      方法 实验分别设置3个诱导时机（TP）（授精后2.5、4.0、5.5 min）、3个诱导温度（T）（2、4、6 ℃）和3个诱导持续时间（D）（10、12、14 min），使用正交实验表L9(34)获得9个实验组，分别记为P1~P9组，以22 ℃下正常发育的斑鳜为对照组（P0）。
      结果 1）冷休克对斑鳜受精卵孵化率的影响极显著，P0组的孵化率极显著高于其他组（P<0.01）。2）P4组（TP 4.0 min、T 2 ℃、D 12 min）诱导率最高，为（83.33±5.77）%；其次为P7组（TP 5.5 min、T 2 ℃、D 14 min）和P5组（TP 4.0 min、T 4 ℃、D 14 min），分别为（73.33±5.77）%和（70.00±0.00）%，P7组与P4组差异不显著（P>0.05），除外P5和P7组均显著高于其他实验组（P<0.05）。P9组（TP 5.5 min、T 6 ℃、D 12 min）斑鳜三倍体诱导率最低，仅有（3.33±5.77）%。3）TP、T和D三者之间交互作用不显著（P>0.05），但两两之间存在着显著的交互作用（P<0.05），三者之间不存在主次关系。4）单因素方差分析结果显示，TP为授精后4.0 min时斑鳜三倍体平均诱导率显著高于TP为授精后2.5和5.5 min时（P<0.05）；T为2、4 ℃时斑鳜三倍体平均诱导率显著高于T为6 ℃时（P<0.05）；D为12、14 min时斑鳜三倍体平均诱导率显著高于D为10 min时（P<0.05）。5）6月龄斑鳜三倍体的全长和体质量与二倍体均无显著性差异（P>0.05）。
      结论 采用冷休克法可成功诱导出斑鳜三倍体，以诱导率为评价指标，可得冷休克法诱导斑鳜三倍体的适宜条件：诱导时机为授精后4.0 min、诱导温度为2或4 ℃、诱导持续时间为12或14 min， 以授精后4.0 min、2 ℃、14 min为最佳。

       

      Abstract:
      Objective The purpose of this experiment is to explore the optimal induction conditions for triploidy in Siniperca scherzeri using cold shock method.
      Methods The experiment set three time points of induction (TP) (2.5, 4.0, 5.5 min after insemination), three temperatures of induction (T) (2, 4, 6 ℃), and three durations of induction (D) (10, 12, 14 min) , respectively. Nine experimental groups were obtained using the orthogonal experimental table L9(34), and denoted as P1−P9, respectively, with P0 (S. scherzeri normally developed at 22 ℃) serving as the control (untreated) group.
      Results 1) Cold shock had a significant effect on the hatching rate of fertilized eggs in S. scherzeri, with the hatching rate of group P0 significantly higher than those of the other groups (P<0.01). 2) Group P4 (TP 4.0 min, T 2 ℃, D 12 min) exhibited the highest triploid induction rate in S. scherzeri at (83.33±5.77)%, followed by group P5 (TP 4.0 min, T 4 ℃, D 14 min) at (73.33±5.77)% and group P7 (TP 5.5 min, T 2 ℃, D 14 min) at (70.00±0.00)%. The induction rate of group P7 was not significantly different from that of group P4 (P>0.05), but the rates of P5 and P7 groups were significantly higher than those of the other experimental groups besides group P4 (P<0.05). While group P9 (TP 5.5 min, T 6 ℃, D 12 min) had the lowest induction rate at only (3.33±5.77)%. 3) The interaction among TP, T and D was not significant (P>0.05), but significant interactions existed between each pair of the three factors (P<0.05), and no primary-secondary order existed among the three factors. 4) One-way ANOVA showed that the mean triploid induction rate in S. scherzeri at 4.0 min after insemination (TP) was significantly higher than that at 2.5 and 5.5 min after insemination (P<0.05). The mean triploid induction rate in S. scherzeri at 2, 4 ℃ (T) was significantly higher than that at 6 ℃ (P<0.05). The mean triploid induction rate in S. scherzeri at 12, 14 min (D) was significantly higher than that at 10 min (P<0.05). 5) The total length and body mass of 6-month-old triploid S. scherzeri show no significant differences compared with those of diploids (P>0.05).
      Conclusion Triploidy can be successfully induced in S. scherzeri by the cold shock method. Using the induction rate as the evaluation index, the suitable conditions for inducing triploidy in S. scherzeri by the cold shock method are as follows: TP at 4.0 min after insemination, T at 2 or 4 ℃, and D of 12 or 14 min. The optimal conditions are TP at 4.0 min after insemination, T at 2 ℃, and D of 14 min.

       

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