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摘要：农药在农业的生产与发展中起着重要的作用，但同时造成了严重的环境污染，而甲拌磷是其中毒性最大的农药之一。在治理甲拌磷产生的污染方面，生物降解成为一种重要的途径。为探讨受甲拌磷污染底泥中的优势菌群，我们采用宏基因组学方法，基于Illumina公司高通量测序技术，并以SSU rRNA基因序列作为分类指标，在农药厂排污口的底泥里获得19291个SSU rRNA基因片段。研究结果表明，Bacteria占66.44%，Eukaryota和Archaea分别占32.98%、0.58%。其中，最丰富的门是Proteobacteria(27.09%)，其次是Actinobacteria(14.78%)、Firmicutes(4.48%)。在纲等级上，16.82%，12.74%和5.66%分别属于γ-proteobacteria，no rank Actinobacteria和α-proteobacteria。此外，也存在许多低丰富度的其他纲。我们还发现，基于16S rRNA基因序列的分类结果表明41.07%，36.28%和7.34%分别属于Proteobacteria，其次是Actinobacteria和Firmicutes。很明显，这和基于SSU rRNA基因序列分析分类的结果一致。总之，这项研究表明SSU rRNA基因序列分析也可作为物种分类方法之一。本研究表明了Proteobacteria这一优势菌群，具有降解受甲拌磷污染底泥的潜力。38545
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毕业论文关键词：土壤宏基因组;SSU rRNA;16S rRNA;微生物群落;生物降解;甲拌磷污染
Study of bacterial persity in bottom sludge contaminated with phorate based on SSU rRNA gene sequence
Abstract:Pesticide plays an important role in the production and development of agriculture, but also caused serious environmental pollution. Phorate is the most toxic among these pesticides. The treatment of phorate pollution, in terms of biodegrading it, is probably today’s most important biotechnological process. To explore the superior strains in phorate-contaminated bottom sludge, we adopted metagenomic analysis based on Illumina high-throughput sequencing and used SSU rRNA as an indicator of taxonomy. Then 19291 SSU rRNA gene reads were derived from the land around a pesticide plant. The results showed that approximately 66.44% are Bacteria, and the Eukaryota and Archaea are 32.98%,0.58%, respectively. Among these, the most abundant phyla were Proteobacteria (27.09%), followed by Actinobacteria(14.78%), Firmicutes(4.48%). At the class level, about 16.82%, 12.74%, 5.66% were affiliated to class Gammaproteobacteria, no rank Actinobacteria, Alphaproteobacteria, respectively. Additionally, many other classes are also present in low abundances. Besides, we also found that the taxonomy based on 16S rRNA gene sequence showed us that about 41.07%, 36.28% and 7.34% were affiliated to Proteobacteria, followed by Actinobacteria, Firmicutes, respectively. These results were consistent with the taxonomy by SSU rRNA gene sequence analysis. In conclusion, this study demonstrated SSU rRNA gene sequence analysis can be used as one of taxonomy methods. In the present study, it showed the superior strains, proteobacteria may have the potential for use in bioremediation of phorate-contaminated bottom sludge.
Keywords: Soil metagenome; SSU rRNA; 16S rRNA; Microbial community; Biodagradation; Phorate contamination
目录
引言 1
1 材料与方法 2
1.1 底泥样品采集 2
1.2 DNA提取与纯化 2
1.3 Illumina公司(Solexa)测序 3
1.4 生物信息学分析 3
1.4.1 SSU rRNA基因序列分析 3
1.4.2 16S rRNA基因序列分析 3
2 结果与分析 4
2.1 基于SSU rRNA基因序列的底泥微生物群落的分类学分析 4
2.2 基于16S rRNA基因序列的底泥微生物群落的分类学分析 4
3 讨论 8
3.1 基于SSU rRNA和16S rRNA基因序列的分类学比较 8
3.2微生物降解作用 8
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