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摘要：【目的】本文针对废弃多年生产甲拌磷的农药厂对其周围土壤环境的影响问题，通过对农药厂出水口所排放的废水流经的2个底泥样地取样（一处位于出水口底泥NYC，另一处位于出水流经的底泥NYCDN，距离出水口2km），将细菌的群落结构及其多样性进行对比，探究甲拌磷对土壤环境细菌多样性赋存特征的影响，研究结果有助于阐明环境污染与生态效应之间的关系。【方法】采用环境微生物的免培养方法，通过提取底泥样品宏基因组，PCR后利用454焦磷酸测序技术，探究废弃多年的农药厂排放的甲拌磷废水对土底泥中细菌多样性的影响。【结果】根据97%的序列相似性水平划分OTU，NYC有1023个OTU，NYCDN有1031个OTU。其中在NYC中，细菌菌群主要由80个属构成，归属于已知的15 个门，且以变形菌门(Proteobacteria)为主(51.89%)，其次为浮霉菌门(Planctomycetes, 10.59%)；在NYCDN中，菌群主要由156个属细菌构成，归属于已知的21个门，且以厚壁菌门(Firmicutes)为主(44%)，其次为变形菌门(Proteobacteria, 35.58%)。在属分类水平上，在NYC中的细菌菌群主要由Urania-1B-19海洋沉积物(Urania-1B-19_marine_sediment_group, 4.38%)、水单胞菌属(Aquimonas, 3.27%)、腈基降解菌属(Nitriliruptor, 2.98%)和浮霉菌属(Planctomycetaceae_uncultured, 1.47%)等组成。在NYCDN中的细菌菌群主要由瘤胃球菌属(Ruminococcaceae, 10.12%)、(Sulfurimonas, 9.38%)、(Anoxynatronum, 4.71%)和(Alkaliflexus, 4.42%)等组成。【结论】NYCDN的细菌多样性要比NYC的细菌多样性要高。甲拌磷废水从NYC流至NYCDN，随着距离出水口距离的增加，土壤细菌多样性升高，这可能与NYC在甲拌磷出水口处，其甲拌磷污染物的浓度要比NYCDN处的浓度要高有关。这一结果表明，农药厂排的废水，严重影响了土壤细菌的多样性，也说明了土壤细菌具有处理废水的能力。37390
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毕业论文关键词：甲拌磷；细菌；454焦磷酸测序；生物多样性。
The bacterial persity in phorate contaminated sediment based on 16S rRNA gene sequences
YU Hong Supervisor: WEN Hongyu
(School of Life Sciences, Jiangsu Normal University, Xuzhou, Jiangsu, 221116, China)
Abstract:【Object】The aim of the study was to evaluate the bacterial persity in the contaminated soil by the sewage released by a pesticide factory( mainly produced phorate). Two soils near the pesticide factory devoted in the past to sewage discharging activities were selected: one(NYC) situated near the sewage outlet and the other one(NYCDN) placed in approximately 2 km away from the sewage outlet. We will compare the bacterial persity in two sites(NYC, NYCDN). It helps to understand the interaction between the environmental pollution and ecological change.【Methods】The persity of bacterial communities were assessed by the culture-independent methods (the 454 pyrosequencing technology). 【Results】The Chao1, ACE richness estimator, the Shannon and Simpson persity indices were used to estimate the bacterial species richness at a 3% dissimilarity level in each sample. There're a total of 1023 and 1031 OTU in the NYC and NYCDN, respectively.A total of 80 genera in NYC were detected by pyrosequencing, while there were almost there were almost twice as many genera in NYCDN.Furthermore, the most predominant bacterial phyla in NYC was Proteobacteria (51.89%)and Firmicutes(44%), respectively. Besides, the most predominant bacterial phyla in NYCDN was Firmicutes(44%) and Proteobacteria(35.58%), respectively. In addition, the most prevalent genera observed were Urania-1B-19 marine sediment group(Phycisphaerae class, Planctomycetes phylum) in NYC and Ruminococcaceae (Bacilli class, Firmicutes phylum)in NYCDN among all the 199 different bacterial genera, respectively.【Conclusion】It was possible to conclude that bacterial species richness in NYC was significantly lower than that of bacteria in NYCDN. With the flowing of the phorate sewage from NYC to NYDCN, soil bacterial persity was increased in NYDCN. This may be related to the higher phorate concentrations in NYC.The results showed that sewage from the pesticide factory had been seriously affected the persity of soil bacteria. It also showed the positive influence of the soil bacteria , which means there existed certain bacteria that have the ability to deal with the phorate pollutants.
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