【摘要】 2004年5~6月,用紫外线照射法使精子遗传物质失活,结合用6-二甲基氨基嘌呤(6-dime-thylaminopurine,6-DMAP)处理受精卵抑制第2极体释放,诱导栉孔扇贝雌核发育二倍体。采用中心波长254nm、光照强度800μw/cm2·s的紫外线照射栉孔扇贝精子50s,然后立即与正常卵子授精,在20℃水温下,受精后35~40min光镜下观察到20%~30%卵子排出第1极体时,分别以40、50、60、70、80mg/L的6-DMAP持续处理受精卵10、15、20min,染色体计数法检测各处理组二倍体率,从而筛选诱导栉孔扇贝雌核发育二倍体两因素的最佳组合。结果表明,6-DMAP的浓度和持续处理时间对二倍体诱导率和D形幼虫率影响较显著,6-DMAP浓度以60、70mg/L较为适宜,最佳处理时间为20min,综合考虑,60mg/L6-DMAP处理20min组得到了最佳效果,二倍体率为57.6%、D形幼虫率22.25%。随后对筛选出的最佳条件组担轮幼虫进行了大量的染色体统计和流式细胞仪分析,结果基本一致,二倍体率分别为53.7%和57.3%。更多还原

【Abstract】 Gynogenetic diploids in scallop (Chlamys farreri) were induced by means of ultraviolet (UV) light irradiation and 6-DMAP treatment from May to June in 2004. After being treated by the UV rays irradiation (254 nm) at the intensity of 800 μw/cm~2·s for 50 s, the genetically inactivated sperms were used for fertilizing normal eggs to produce haploid gynogenesis. 35~40 min later after fertilization at 20 ℃ water temperature, 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 different concentrations of 6-DMAP (40、50、60、70、80 mg/L) for 10, 15, and 20 min. respectively. The diploid percentage and survival rate of D-larvae were determined by means of chromosome distributions and optical microscope for the sake of finding the optical combination of concentration and exposure time. The results showed that the optimal 6-DMAP concentration and exposure time were 60~70 mg/L and 20 min. In general, the optimal condition could result in 57.6% diploid rate and 22.25% survival rate of D-larvae. And then, the diploid rates of trochophore in the groups of the optimal conditions were examined by means of both counting chromosome methods and flow cytometer, the results were approximately similar, with the diploid rates being 57.3% and 53.7%, respectively.更多还原