低渗诱导太平洋牡蛎三倍体以及与其他诱导方法的比较  

Triploid induction in Pacific oyster (Crassostrea gigas) by hypotonic treatment and comparison with other induction methods

作　　者：孔静[1] 王昭萍[1] 于瑞海[1] 刘剑[1] 张跃环[1] 

机构地区：[1]中国海洋大学海水养殖教育部重点实验室,山东青岛266003

出　　处：《中国水产科学》2011年第3期581-587,共7页Journal of Fishery Sciences of China

基　　金：国家自然科学基金项目(30771622);国家高技术研究发展规划项目(2006AA10A401)

摘　　要：通过低渗方法抑制受精卵第二极体（PB2）的释放，诱导太平洋牡蛎（Crassostreα gigas）三倍体。在水温为23～25℃的条件下，分别采用不同低渗海水（盐度分别为4、6、8、10、12、14、16），在不同处理时机，即第1个第一极体（PBll出现、30％的PBl出现、40％的PBl出现、50％的PBl出现以及第1个第二极体（PB2）出现时，对太平洋牡蛎受精卵进行10min、15min、20min、25min持续处理。处理后，于正常盐度海水中孵化，收集D形幼虫，通过流式细胞仪进行倍性测定，确定最佳诱导条件。结果表明，当太平洋牡蛎受精卵PBl出现40％时，在盐度为8的低渗海水中，持续处理15min，得到最高三倍体诱导率为（89．16±1．39）％。与对照组相比，低渗诱导获得的三倍体幼虫表现出明显的生长优势。将低渗方法同高温（32℃）、低温（2℃）和6-DMAP（450μmol／L）处理等多倍体诱导方法进行比较，结果显示，低渗组的卵裂率和孵化率显著高于高温、低温和6-DMAP诱导的太平洋牡蛎三倍体（P〈0．05）。6-DMAP诱导的三倍体率虽略高于低渗诱导，但差异不显著（P〉0．05）。通过综合评价指数（Ie）比较，认为低渗诱导多倍体的方法比高温、低温和6-DMAP诱导具有明显的优势。Triploid Pacific oysters （Crassostrea gigas） were induced by inhibiting polar body II （PB2） releasing with hypotonic treatment. Different hypotonic treatments, including salinity （4, 6, 8, 10, 12, 14 and 16）, initial treatment time （at the time when the first PB1, 30 percents of PB1, 40 percents of PB1, 50 percents of PBl and the first PB2 were observed） and duration time （10, 15, 20 and 25 min） were tested at 23-25℃ water temperature. Flow cytometry analysis showed that the highest triploidy rate was （89.16±1.39）% when the fertilized eggs were treated for 15 min with hypotonic solution with salinity of 8 at the time when 40 percents of PB1 appeared. The results also showed that shell height of triploids were larger than that of diploids. And triploid larvae appeared much superiority in growth. Then four different triploid induction methods were compared including hypotonic treatment, cold shock （2℃）, heat shock （32℃） and 6-DMAP （450 μtmol/L） treatment. The triploids induced by hypotonic treatment were superior over those by cold shock, heat shock and 6-DMAP, and the difference in hatching success and D-larval size was significant （P〈0.05） The induction ratio of 6-DMAP treatment was higher than that by hypotonic treatment, but the difference was not significant （P〉0.05）. General evaluation index （Ie） of hypotonic treatment is superior over those of other three triploid induction treatments. Because of the low price, the high induction ratio of triploids and security, it is feasible and worthwhile to induce triploid by hypotonic treatment.

关 键 词：太平洋牡蛎 三倍体 低渗诱导 

分 类 号：S917[农业科学—水产科学]