长蛸不同群体ISSR分析【毕业论文】.doc

本科毕业论文 （20 届） 长蛸不同群体ISSR分析 专业：生物科学 目录 摘要 I Abstract II 引言 1 1 材料与方法 3 1.1 实验材料 3 1.2 实验仪器 3 1.3 实验方法 3 1.3.1 长蛸基因组DNA的提取 3 1.3.2 检测长蛸基因组DNA 4 1.3.3 长蛸ISSR-PCR 4 1.3.4 长蛸数据统计分析 4 2 实验结果与分析 6 2.1 不同居群的遗传多样性 6 2.2 遗传变异与分化 7 3 讨论 9 3.1 长蛸的遗传多样性 9 小结 10 参考文献 11 致谢 29 摘要 [摘要] 本文采用ISSR-PCR技术对长蛸基因组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等，建立了ISSR优化体系。最终确立的25 μl优化体系为：Mg2+，1.5 m mol/L，Taq DNA聚合酶1.0 U引物0.15μmol/L，dNTPs 2.0 m mol/L，退火温度为52.2℃。并用条ISSR引物对舟山群体进行遗传多样性分析，共得到118个清晰的扩增位点，其中多态性位点55个，多态位点率为46.59%46.61%，Neis基因多样性为0.1633Shannons多样性指数为0.241744.07%。Neis基因多样性为0.1510，Shannons多样性指数为0.2253，表明群体遗传多样性。ISSR；体系优化；遗传多样性。 Abstract [Abstract] This study adopted ISSR-PCR amplification technology on Octopus variabilis genome DNA, and analyzed the main influencing factors, including the concentration of Mg2+, Taq DNA polymerase, dNTPs, the primers, as well as the annealing temperature, and established the suitable ISSR-PCR system for Octopus variabilis. The reaction system with a total volume of 25μl was as the following: Mg2+ 1.5 mmol/L, Taq DNA polymerase 1.0 U, primer 0.15 mmol/L, dNTPs 2.0 mmol/L, the annealing temperature is 52.2℃. And the ISSR technique was used to assess the genetic diversity of Zhoushan population and Wenzhou population, 7 ISSR primers was used to PCR amplification. Of the 118 ISSR loci tested, 55 (46.59%) were polymorphic, the results of Zhoushan population showed that the proportion of polymorphic loci was 46.61%, Neis genetic diversity is 0.1633±0.2044, Shannons information index is 0.2417±0.2916. The results of Wenzhou population showed that the proportion of polymorphic loci was 44.07%. Neis genetic diversity is 0.1510± 0.1971, Shannons information index is 0.2253±0.2830. The results showed that the genetic diversity of Zhoushan population is higher than that of Wenzhou population. [Key words] Octopus variabilis; ISSR; System Optimization; Genetic. 引言 长蛸(Octopus variabilis)，隶属软体动物门，头足纲，八腕目，蛸科(Octopodidae)，俗称长八带鱼短蛸俗称八带鱼、鲅须1