热休克法诱导虹鳟三倍体技术研究

Technical Research on Induced Triploid of Rainbow Trout by Heat Shock

本文以虹鳟(Oncorhynchus mykiss)为实验对象,采用热休克法诱导虹鳟三倍体,利用流式细胞仪(CytoFLEX S)以及比较红细胞长径对虹鳟倍性进行鉴定;在热休克法诱导虹鳟三倍体的两个正交重复实验中,主要研究了处理的起始时间、温度、持续时间三个因素对于虹鳟三倍体诱导效果的影响,结果显示,在实验设定的起始时间、温度、以及持续时间内,温度大小对发眼率、诱导率以及综合评分影响程度都是最大(R值最大),在发眼率结果中,25℃时有最高的发眼率;在诱导率结果中,25℃、27℃明显高于29℃组的诱导率;在综合评分结果中25℃、27℃明显高于29℃组的综合评分。在起始时间为10~20 min内,起始时间对诱导率没有显著影响,但起始时间为20 min时诱导效果最优。在起始时间与处理强度双因素实验结果中,综合评分高于70.00%的共两组,分别为第一组、第二组,热休克的起始时间都是25 min,作用参数分别为25℃持续20 min、27℃持续15 min,是较为理想的诱导三倍体处理强度参数。

In this study,the rainbow trout(Oncorhynchus mykiss)was used as the experimental object.The heat shock method was used to induce the triploid of rainbow trout.The ploidy of rainbow trout was identified by using flow cytometry(CytoFLEX S)and comparison of the long axis of erythrocyte.In the two orthogonal replicate experiments induced by heat shock induced rainbow trout triploid,the effects of three factors,including start time,temperature and duration,on the induction of rainbow trout triploid were studied.The results showed that the temperature had the greatest impact on the eye rate,induction rate and overall score(maximum R value)at the start time,temperature,and duration of the experiment.In the eye rate results,the highest eye rate was obtained at 25?C;in the induction rate results,induction rate of the 25?C,27?C group was significantly higher than that of the 29?C group;in the comprehensive scoring results;the comprehensive scores of the group at 25?C and 27?C were significantly higher than those at 29?C.In the initial time of 10~20 min,the initial time had no significant effect on the induction rate,but the induction effect was optimal when the starting time was 20 min.In the two-factor experiment results of starting time and treatment intensity,there are two groups whose comprehensive score is higher than 70.00%:the first group and the second group,respectively.The starting time of heat shock is 25 min,and the action parameter is 25?C for 20 min and 27?C for 15 min respectively,which is an ideal parameter for inducing triploid treatment intensity.