紫外线照射使精子遗传物质失活,用细胞松弛素B(CB)处理受精卵抑制第二极体释放,诱导刺参(Apostichopusjaponicus)雌核发育二倍体。用强度为2561μW/(cm2.s)的紫外线(254 nm)照射不同时间的精子与正常卵子受精。结果发现随照射时间的增加,卵裂率、早期胚胎存活率和小耳幼虫发生率逐渐降低,照射30 s时小耳幼虫发生率降为0。受精后35 min,显微镜下观察到有20%~30%受精卵排出第1极体时,用浓度0.5μg/mL的CB持续处理受精卵20 min,诱导出刺参第二极体抑制型雌核发育二倍体,雌核发育二倍体发育速度低于正常二倍体。微卫星分析表明,雌核发育二倍体诱导成功率为93.3%。
Gynogenetic diploids in sea cucumber (Apostichopus japonicus) were induced by means of ultraviolet (UV) radiation and cytochalasin B (CB) treatment for the inhibition of the second polar body release. The sperms irradiated by UV at an intensity of 2561μW/(cm2·s) for various durations were fertilized with the normal eggs. With increasing irradiation time, cleavage rate, survival rate at early embryonic phase and developmental rate of early auricularia decreased, and none of early auricularia occurred at 30 s. Thirty-five minutes later after fertilization, it could be observed under the optical microscope that 20~30~ of eggs released the first polar body. Then the fertilized eggs were exposed to 0. 5/lg/mL CB for 20 min. Compared with those in the control group, the developmental speed in the gynogenetic diploid group was slower. Microsatellite analysis demonstrated that 93.3% of gynogenetic diploids were success- fully induced.