东海陆架沉积物古菌多样性研究【毕业论文】.docVIP

本科毕业论文 （20 届） 东海陆架沉积物古菌多样性研究 专业：生物科学 目录 摘要 i Abstract ii 引言 1 1.材料与方法 2 1.1实验材料 2 1.1.1实验样品 2 1.1.2 主要生化试剂（盒） 2 1.1.3 主要试剂及培养基配制 3 1.1.4 实验仪器 4 1.2实验方法 5 1.2.1样品DNA的提取 5 1.2.2 16S rDNA基因的PCR扩增 6 1.2.3 PCR产物的纯化 6 1.2.4 重组载体的构建 7 1.2.5 重组载体的转化 7 1.2.6 重组子的克隆 8 1.2.7 基因文库克隆的鉴定 8 1.2.8 系统发育分析 8 1.2.9 温度梯度凝胶电泳(TGGE)分析 8 2.结果与分析 10 2.1 样品DNA的提取 10 2.2 16S rDNA基因扩增与纯化 10 2.3 克隆文库的构建与鉴定 11 2.4 系统发育分析 12 2.4.1古菌多样性分析 12 2.4.2 古菌各类群分布分析 14 2.4.3 古菌系统发育分析 16 2.5 温度梯度凝胶电泳（TGGE）结果 16 3.讨论 17 3.1 实验方法的改进 17 3.1.1 16S rDNA基因扩增体系的优化 17 3.1.2 核酸序列分析方法研究古菌多样性的优缺点 18 3.2 古菌群落特征分析 18 3.2.1 古菌群落垂直分布特征分析 18 3.2.2古菌多样性分析 19 参考文献 20 附译文：南海西沙海槽表层沉积物微生物多样性 22 附译文原文 32 致谢 44 摘要 [摘要]海底沉积物中蕴含大量的微生物类群，它们是海洋生态系统的重要组成部分。通过对海洋沉积物古菌多样性的研究，有助于人们更深入地了解海洋。泉古生菌(Crenarchaeota)和广古生菌(Euryarchaeota) 。以Marine Crenarchaeotic Group I(MG I，%）和Miscellaneous Crenarchaeotic Group (MCG，%)为主要类群，其余还有少量Marine Benthic Group C (MBGC，4.3%)、Marine Benthic Group A (MBGA，1.1%)和Anaerobic Methanotrophic Euryarchaeota（ANME，1.1%）。 [关键词东海陆架；沉积物；16S rDNA；古菌多样 Abstract [Abstract]Marine sediments were estimated to contain a huge amount of microbe, which was of great importance to the marine ecology. It would help us explore the ocean by the analysis of archaeal diversity in sediments. We used molecular technology based on 16S rDNA to study the archaeal community of marine sediments from the East China Sea. The result showed that all the archaea were uncultivable and could be divided into two groups, Crenarchaeota and Euryarchaeota, respectively. Two archaea groups, Marine Crenarchaeotic GroupⅠ(MG I，5.1%)and Miscellaneous Crenarchaeotic Group (MCG，3.2%), were the dominate groups. The remaining were Marine Benthic Group C (MBGC，4.3%)、Marine Benthic Group A (MBGA，1.1%) and Anaerobic Methanotrophic Euryarchaeota（ANME，1.1%）. Irevealed that the sediments of the East China Sea contain a large amount of archaea. Meanwhile it would lay a foundation for the deep research. [Key words] the East China Sea; Sediments; 16S rDNA; archaeal dive