施肥至农田土壤的抗生素抗性基因的污染特征和传播扩散机制研究

项目名称： 施肥至农田土壤的抗生素抗性基因的污染特征和传播扩散机制研究

项目编号： No.31270542

项目类型： 面上项目

立项/批准年度： 2013

项目学科： 生物科学

项目作者： 毛大庆

作者单位： 天津大学

项目金额： 85万元

中文摘要： 抗生素在医疗和养殖业的滥用及在制药业的无标准排放，严重污染了环境，并造成了ARGs泛滥。针对普遍采用畜禽粪便和污泥施肥农田土壤这一现状，以及这一现状下抗生素抗性基因在土壤中增殖和扩散所造成的潜在风险，本研究拟采用分子生物学研究手段RTQ-PCR和最新的Metagenomics高通量测序等技术，野外采样和土壤微宇宙室内模拟相结合，研究土壤中ARGs的消长规律，以及增殖和扩散的分子机制。通过对携带ARGs的质粒特异序列分析，包括细胞接合装置结构蛋白编码基因、复制相关基因和连锁基因等，结合抗性基因存在形态分析，以及土壤细菌种群多态性比较分析，揭示抗生素抗性基因经施肥至土壤并增殖和传播扩散的分子机制。项目研究成果将有助于了解我国土壤环境抗生素和抗性基因污染现状，为标准制定提供基础参考数据；并基于土壤介质中抗性基因增殖和扩散机制的揭示，为实际控制和治理土壤环境抗性基因提供参考。

中文关键词： 抗生素；抗性基因；重金属；纳米材料；质粒

英文摘要： The abuse of antibiotics in medical and breeding industry, and the emissions without criterion of pharmaceutical industry, have greatly promoted antibiotics and ARGs pollution in environment. Meanwhile, It is the widespread use of fertilization with poultry/animal feces and sludge, that cause a potential risk of rapid proliferation and spread of ARGs in soil. Therefore, the methods of RTQ-PCR and metagenomics high-throughput sequencing are used in this study, and field-investigation combining soil-microcosm simulation experiment are objective to grasp pollution situation and dynamic changes of ARGs in soil, also to explore the molecular mechanism of ARGs proliferation and spread in soil bacteria. The specific sequences of plasmids are sequenced and analyzed with bioinformatics methods, which includes those genes encoding a key structure protein assembling the cell conjugation, a start replication protein and flank linked genes. The extracellular ARGs and intracellular ARGs are then analyzed on the basis of DNA extraction, and population polymorphism comparison of soil bacteria are also conducted. Finally, the mechanism of ARGs proliferation and spread can be deduced on the basis of these results. This study will provide us the first-hand data of antibiotics and ARGs pollution in soil, and provide us the basis fo

英文关键词： antibiotics；antibiotic resistance gene；heavy metals；Nanoparticles；plasmid