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采用紫外线照射的方式使精子遗传物质失活,通过细胞松弛素B(CB)的处理来抑制受精卵第二极体的释放,人工诱导紫贻贝(Mytilus gallopovincialis)雌核发育二倍体。将精子在强度为2561μW/(cm~2·s)的紫外线(254 nm)下进行时间梯度的照射,之后与正常的卵子受精,实验发现随照射时间的增加,卵裂率、早期胚胎存活率和D形幼虫发生率总体呈下降趋势,在照射55 s时D形幼虫发生率降为0。受精后25 min,用浓度0.5μg/mL的细胞松弛素B(CB)持续处理受精卵20 min,诱导出紫贻贝第二极体抑制型雌核发育二倍体。通过微卫星分析鉴定所得的子代个体均为雌核发育二倍体。
UV irradiation was used to inactivate the genetic material of sperm. The release of the second polar body of the fertilized egg was inhibited by treatment with cytochalasin B (CB), and the diploid of gynogenetic Mytilus gallopovneumis was induced. The sperm was irradiated with time gradient under ultraviolet (254 nm) intensity of 2561μW / (cm ~ 2 · s) and then fertilized with normal eggs. It was found that with the increase of irradiation time, the cleavage rate, early embryo survival rate And D larvae generally showed a decreasing trend. The incidence of D larvae dropped to 0 at 55 s. At 25 min after fertilization, fertilized eggs were treated with cytochalasin B (CB) at a concentration of 0.5 μg / mL for 20 min to induce the second polar inhibitory-type gynogenetic diploid. The progeny individuals identified by microsatellite analysis were all gynogenetic diploids.