| 摘要: |
| 采用紫外线照射对大黄鱼(Pseudosciaena crocea Richardson)精子遗传物质进行完全灭活,灭活后精子与卵子受精获得雌核发育单倍体受精卵,再通过冷休克对单倍体受精卵进行倍性恢复,成功诱导出大黄鱼雌核发育二倍体。实验结果表明:(1)在紫外照射过程中照射剂量与大黄鱼精子和卵子受精后其成活率和孵化率呈现出明显的Hertwig效应,紫外照射时间超过2 min(剂量>2 400 ergs/mm2),可使大黄鱼精子遗传物质完全灭活。大黄鱼雌核发育诱导中精子遗传灭活的最适紫外照射时间为2.5 min(剂量为3 000 ergs/mm2);(2)冷休克法可成功抑制大黄鱼受精卵第二极体排放,诱导效果受处理时刻、处理时间和处理温度三因素的影响。综合雌核发育二倍体的诱导率、处理后受精卵早期胚胎存活率和仔鱼孵化率等因素,较适合的染色体加倍条件为23℃培育水温下授精后2 min,在3-4℃海水中处理10 min。 |
| 关键词: 大黄鱼(Pseudosciaena crocea) 雌核发育 人工诱导 紫外照射 冷休克 |
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| 基金项目:中国科学院合作种子资金-宁波市科技计划项目(2002D21004);宁波市科技攻关项目(01N40111) |
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| Preliminary study on artificial induction of diploid gynogenesis in the large yellow croaker Pseudosciaena crocea |
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| Abstract: |
| Gynogenetic diploids of Pseudosciaena crocea were induced by activating egg development with UV-irradiated sperm combined with cold shock to prevent extrusion of the second polar body. The typical Hertwig’s effect was observed when the sperm was exposed to UV light and the effective UV intensity for sperm irradiation covered a wide range from 2 400 to 8 400 erg/mm2(i.e. exposing time from 2 to 7 min), and the optimal UV dosage for gynogenetic haploid induction was 3 000 erg/mm2(2.5 min). Cold shock could inhibit the extrusion of second polar body successfully and the induction rates of gynogenetic diploids in the experimental groups were mainly affected by the starting time, intensity and duration of shocks. Treatment optimal parameters are follows: treatment temperature was 3-4 ℃ for 10 mins and timing was 3 min after fertilization. |
| Key words: Pseudosciaena crocea diploid gynogenesis artificial induction UV-irradiation cold shock |
