摘要:通过对运城盐湖水样提取总DNA,之后进行PCR,再对16SrRNA进行454高通量测序,测出其序列,从而获得运城盐湖细菌群落的组成和多样性特征。经过测序得出,细菌的有效序列有27597条,优化序列为25938条,59个OTU。与该地的古菌群落相比,无论是在物种的数量和多样性方面,都没有太大的差别。从细菌组成分析得知,在门的水平上,主要有5个门,其中蓝藻门(Cyanobacteria)最占优势,大约为41.86%,其次是拟杆菌门(Bacteroidetes),大约为37.86%,厚壁菌门(Firmicutes)10.31%,变形菌门(Proteobacteria)7.61%。在属的水平上,有29个属,仍然是Chloroplast最占优势,其次为Salinibacter28.07%,乳球菌属(Lactococcus)9.53%,假单胞菌属(Pseudomonas)1.68%,未分类和不可培养的占了15.95%。在种的水平上,共有23个种,最占优势的为Dunaliella_tertiolecta,Salinibacter_sp._CB7约有11.59%,乳球菌属(Lactococcus_piscium)8.73%,uncultured_Salinibacter_sp 8.47%,不可培养微生物和非分类的大约占22.97%。运城盐湖细菌的研究很有意义,可以为微生物方面的研究提供很好的资源。32733 毕业论文关键词:盐湖、16S rRNA、454高通量测序、细菌、多样性。
Bacterial community composition of Yuncheng salt lake in the Shanxi,China
Abstract: Through extracting total DNA of Yuncheng Salt Lake water sample for PCR, then 454 high-throughput sequencing of 16S rRNA, measuring their sequences, Yuncheng Salt Lake bacterial community composition and persity of the characteristics can be obtained. From sequencing results, we can see that there are 27597 pieces of effective sequences of bacteria, 25938 pieces of optimized sequences, 59 OTU. Compared with the archaeal community, both in the number and persity of species, there are not too many differences. From analysis of bacterial composition , at the level of the gates, there are five doors. In which Cyanobacteria most dominant, about 41.86%, followed by Bacteroidetes, about 37.86%, Firmicutes10.31%, Proteobacteria7.61%. In the genus level, there are 29 genera, Chloroplast is still the most dominant, followed by Salinibacter28.07%, Lactococcus9.53%, Pseudomonas 1.68% and the unclassified and uncultivated account for 15.95%. At the species level, there are 23 species, the most dominant one is Dunaliella tertiolecta, Salinibacter_sp._CB7 about 11.59%, Lactococcus piscium8.73%, uncultured salinibacter sp 8.47%, the unclassified and uncultivated account about 22.97%. It is meaningful to study the Yuncheng Salt Lake bacteria, it will provide rich resources for our further research.
Keywords: Salt Lake、16S rRNA、454 high-throughput sequence、bacteria、persity.
目录
前言 1
1.材料 2
1.1 采样 2
1.2采样地 2
1.3 仪器 2
2.方法 2
2.1 总体方法流程 2
2.2 具体过程 3
2.2.1 基因组DNA提取 3
2.2.2 基因组DNA的电泳 —— 1%琼脂糖凝胶电泳 3
2.2.3 PCR扩增和胶回收 3
2.2.4 荧光定量 3
2.2.5 高通量测序 4
2.2.6 数据分析 4
3.结果与分析 4
3.1 454高通量测序数据 4
3.2 细菌的丰度 5
3.3.细菌的多样性分析 6
3.4 细菌群落组成分析 7
3.4.1 细菌在门的水平上的群落组成 7
3.4.2 细菌在纲的水平上的群落组成 9
3.4.3 细菌在属的水平上的群落组成 9
4.结论 11
致谢 12
参考文献 12
前言