| 摘要: |
| 采用 ISSR-PCR 技术对曼氏无针乌贼基因组 DNA 进行扩增, 筛选分析了 ISSR-PCR 扩增时的主要影响因子, 包括 Mg2+浓度、 引物浓度、 dNTPs 浓度、 Taq 酶浓度以及退火温度, 并用 8 条 ISSR引物对舟山养殖群体进行了遗传多样性分析, 最终确立的适用于曼氏无针乌贼 ISSR 扩增的 25?l 反应体系为: Mg2+ 1.5mmol/L, Taq DNA 聚合酶 1.0U, 引物 0.15?mol/L, dNTPs 2.0mmol/L, 退火温度为52.2℃。结果表明, 扩增得到 70 个清晰的扩增位点, 其中多态性位点 60 个, 多态位点率为 85.71%, Nei's 基因多样性为 0.3105±0.1760, Shannon's 多样性指数为 0.4610±0.2397。 本研究结果表明, 舟山曼氏无针乌贼养殖群体遗传多样性较丰富, 因此舟山曼氏无针乌贼养殖群体可能有比较高的适应生存的能力, 蕴藏着比较大的进化潜能及比较丰富的育种潜力。 |
| 关键词: 曼氏无针乌贼, ISSR, 体系优化, 遗传多样性 |
| DOI：10.11693/hyhz201104011011 |
| 分类号: |
| 基金项目:国家高技术研究发展计划(863计划)项目: 名贵头足类苗种规模繁育关键技术, 2010AA10A404号; 国家海洋公益性行业科研专项, 201005013 号; 海洋渔业科学与技术浙江省重中之重学科开放课题, 20100105 号; 浙江海洋学院人才引进启动项目, 21135011509 号; 国家科技部科技支撑计划: 东海区优势种类扩繁及高效健康养殖技术集成与示范, 2011BAD13B08 号 |
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| OPTIMIZATION OF THE ISSR SYSTEM FOR SEPIELLA MAINDRONI AND ANALYSIS OF GENETIC DIVERSITY OF THE CULTURED POPULATION |
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XU Mei-Ying1,2, YE Ying-Ying1, GUO Bao-Ying1, QI Peng-Zhi2, WU Chang-Wen1
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1.Marine Science College of Zhejiang Ocean University, Key Laboratory of Mariculture Equipments and Engineering Technology of Zhejiang Province;2.Zhejiang Dahaiyang Science and Technology Co., Ltd.
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| Abstract: |
| In this study, ISSR-PCR amplification technology was adopt on Sepiella maindroni genome DNA, and the effect of Mg2+ concentration, Taq DNA polymerase, dNTPs, primer and the annealing temperature on the ISSR-PCR amplification was analyzed. 8 ISSR primers were used to assess the genetic diversity of Zhoushan cultured S. maindroni, and established the suitable ISSR-PCR system with a total volume of 25?l was as followed: Mg2+ 1.5mmol/L, Taq DNA polymerase 1.0U, primer 0.15?mol/L, dNTPs 2.0mmol/L, and the annealing temperature is 52.2℃. Of the 70 ISSR loci tested, 60 (85.71%) were polymorphic, Nei's genetic diversity is 0.3105±0.1760, Shannon's information index is 0.4610±0.2397. The results of this study showed that there was a higher genetic diversity level in the Zhoushan cultured population of S. maindroni. Therefore, Zhoushan cultured S. maindroni maybe have higher adapt ability to survive, containing larger evolution potential and relatively rich breeding potential. |
| Key words: Sepiella maindroni, ISSR, System optimization, Genetic Diversity |
