基于高通量测序技术的宏基因组学与宏转录组学在环境样品抗生素抗性基因全谱表征和宿主溯源中的应用研究

项目名称： 基于高通量测序技术的宏基因组学与宏转录组学在环境样品抗生素抗性基因全谱表征和宿主溯源中的应用研究

项目编号： No.51508302

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 其他

项目作者： 李炳

作者单位： 清华大学

项目金额： 20万元

中文摘要： 抗生素抗性基因（ARGs）在环境中的分布特征和扩散机制已成为国内外研究热点。本项目选取抗生素生产废水处理系统中活性污泥作为模式环境样品，率先使用基于高通量测序技术的宏基因组学和宏转录组学手段，深入开展抗生素制药厂废水处理系统中ARGs全谱图表征和宿主溯源的相关研究，包括：1）ARGs数据库的构建；2）复杂环境样品中ARGs全谱图定量检测；3）ARGs携带细菌的基因型鉴定；4）高浓度抗生素胁迫条件下ARGs的表达特性；5）ARGs所在遗传元件的定位和 6）不同加氯消毒条件下ARGs在胞内/胞外DNA，死细菌/活细菌胞内DNA中的分布规律表征。通过建立复杂样品中ARGs定量检测标准方法，为ARGs在大尺度不同环境中的分布特征和扩散机制研究提供技术支持，同时展示了宏基因组学/宏转录组学手段在环境样品ARGs宿主溯源研究中的可行性，并为制订ARGs在优控污染行业废水/废物中的排放标准提供科学依据。

中文关键词： 抗生素抗性基因；高通量测序；宏基因组学；宏转录组学；加氯消毒

英文摘要： The occurrence, spread and transformation of antibiotic resistance genes (ARGs) have recently drawn great attention all over the world. In this study, activated sludge in the antibiotic production wastewater treatment plant will be selected as the model of environmental sample to conduct the ARGs full-spectrum characterization and ARGs microbial source tracking study using the high-throughput sequencing based metagenomics approach as well as metatranscriptomics approach. This study mainly focuses on (1) comprehensive integration and establishment of antibiotic resistance genes database, (2) full-spectrum characterization and quantification of ARGs in complex environmental samples,(3) identification of antibiotic resistant bacteria, (4) full-spectrum characterization of ARGs expression under high antibiotic stress, (5) the location of ARGs on genetic elements, and (6) the occurrence profiles of ARGs on extracellular/intracellular DNA, dead/live cell intracellular DNA under different chlorination conditions. The successful establishment of a set of standard ARGs detection and quantification approach will provide technique support for the following study of ARGs occurrence pattern and their fate within large-scale environments. Meanwhile, the success of this study will also demonstrate the feasibility of utilizing both high-throughput sequencing based metagenomics approach and metatranscriptomics approach to investigate the microbial source tracking of ARGs in complex environmental samples. This study could provide technical reference for the similar studies in the future. Additionally, the conclusions and results of this project could also be used as scientific evidences to formulate the discharge standard of ARGs in wastewater or solid waste from the ARGs pollution industry which needs the highest priority control.

英文关键词： Antibiotic resistance genes;High-throughput sequencing;Metagenomics;Metatranscriptomics;Chlorination