冷、热休克法诱导黄颡鱼三倍体的比较研究  

Comparative studies in the induction of triploidy for Pelteobagrus fulvidraco by cold and heat shocks

作　　者：宋立民[1,2] 王卫民[1] 周小云[1] 阎里清 王玉芬[2] 

机构地区：[1]华中农业大学水产学院,农业动物遗传育种与繁殖教育部重点实验室,湖北武汉430070 [2]中国水产科学研究院北戴河中心实验站,河北秦皇岛066100 [3]湖北省漳河水库水产公司,湖北荆门448156

出　　处：《水产学报》2010年第5期768-776,共9页Journal of Fisheries of China

基　　金：国家科技支撑计划项目(2006BAD03B07)

摘　　要：分别采用冷、热休克抑制第二极体释放的方法诱导黄颡鱼三倍体。结果表明,在卵受精后2min,5℃处理20min,胚胎时期的三倍体率达70%左右,孵化率50%左右,幼鱼时期三倍体(含嵌合体)的检出率为25%,此条件为冷休克处理的优化参数;在卵受精后2min,40℃处理2min,胚胎时期的三倍体诱导率达58%,孵化率为39%,幼鱼时期三倍体(含嵌合体)的检出率为40%,此条件为热休克处理的优化参数。正交分析得出,冷休克条件下起始休克时间是原肠期三倍化率和孵化率的重要影响因子,温度对畸形个体的产生有重要影响;热休克条件下,参考三倍体率、畸形率、孵化期相对存活率三者而言,休克温度均是重要因素。比较观察到冷休克处理组的胚胎受损情况严重,后期的成活率较热休克处理组要低,总体诱导效果逊于热休克处理组。The yellow catfish Pelteobagrus fulvidraco has become a valuable aquaculture species in China and the possible use of sterile triploids is an interesting option for its culture. The optimal conditions for the induction of triploidy for P. fulvidraco by means of cold and heat shocks were investigated and compared in the present study. Three treatment variables were considered： the time after fertilization when the shock was applied,the temperature and the duration of the shock. Ploidy was determined by chromosome counting and flow cytometric analysis. The diploid,triploid,haploid and aneuploid （or heteroploid） were all identified in the P. fulvidraco eggs （or juvenile individuals） after cold and heat shocks. The survival rate in stages of gastrulae and post hatch,the rates of triploidy induction and deformity under different treatment methods were evaluated. The results indicated that optimal treatment conditions for cold shocks were 5 ℃ for 20 min at 2 min after fertilization,which resulted in about 70% triploidy embryos,50% relative survival at hatching stage,and 25% triploidy individuals in juvenile period. The optimal treatment conditions for heat shocks were 40 ℃ for 2 min at 2 min after fertilization,which results in 58% triploidy embryos,39% relative survival at hatching stage,and 40% triploidy individuals in juvenile period. The orthogonal design analysis showed that the time after fertilization when the shock was applied was the most important factor to the rates of hatching and triploid induction at embryonic stage in cold-shock groups,while in heat-shock groups temperature was the most important factor to the inducing rates of triploids,deformities and relative survival at hatching stage. The results demonstrated that cold shock was less effective in inducing triploidy in P. fulvidraco for its serious injury to embryos and lower survival rate after hatching compared with heat shock. In conclusion,this paper presents the optimal conditions for triploidy induction in P. fulvidraco using cold

关 键 词：黄颡鱼 冷休克 热休克 人工诱导 三倍体 

分 类 号：Q343.2[生物学—遗传学] S917[农业科学—水产科学]