污水生物强化除磷体系中菌群网络关系及其环境响应机理的微生物组学研究

Enhanced biological phosphorus removal (EBPR) systems have been widely applied to remove and recycle phosphorus from wastewater due to its high automation level and low operation cost. Polyphosphate accumulating organisms (PAOs) are dominant functional bacteria in the system. Their microbial and functional structures affect the EBPR system operation directly, thus are catching more and more academic attention. However, the major PAOs named Accumulibacter remain recalcitrant from isolation and pure cultivation, and their microbial response to environmental changes and microbial relationships remain unclear, preventing the EBPR system from precise control and stable operation. In this study, high throughput sequencing based metagenomic and metatranscriptomic approaches, together with molecular biological and bioinformatic analyses, will be applied to set up a quantification assay for identifying Accumulibacter clades; to screen environmental conditions for Accumulibacter isolation through genome extraction and comparative genomics; to investigate the variation of microbial structures and gene expression in response to environmental selection pressures, thus to reveal and validate the microbial metabolic networks and relationships. The results of this study will provide significant information and technical support for EBPR operation and optimization.

生物强化除磷体系（EBPR）具有自动化程度高、运行费用低等技术优势，广泛应用于污水中磷的去除和回收。聚磷菌是该工艺中主要的功能细菌，其菌群结构和功能优化直接影响到体系的除磷效果，是当前的研究热点。然而，主要聚磷菌Accumulibacter尚未实现纯培养，且体系中微生物应对环境变化的瞬间响应机制和菌间作用关系仍不明确，导致EBPR工艺无法实现精准调控和出水稳定达标。本项目拟采用基于高通量测序技术的宏基因组学和宏转录组学方法，结合分子生物学和生物信息学分析，建立聚磷菌快速检测方法，为研究其菌群结构提供技术手段；解析EBPR微生物的基因组全谱图，利用比较基因组分析，筛选出富集纯化聚磷菌的条件；探究微生物在不同环境压力下的菌群更替规律和基因表达特征，从而揭示体系中微生物对环境变化的响应机制和主要代谢网络关系。本项目研究将为EBPR工艺优化与稳定运行提供重要的理论依据和技术支撑。

生物强化除磷体系（EBPR）具有自动化程度高、运行费用低等技术优势，广泛应用于污水中磷的去除和回收。聚磷菌是该工艺主要的功能细菌，其菌群结构和功能优化直接影响到体系的除磷效果，是当前的研究热点。然而，主要聚磷菌Accumulibacter尚未实现纯培养，且体系中微生物应对环境变化的瞬间响应机制和菌间作用关系仍不明确，导致EBPR工艺无法实现精准调控和出水稳定达标。本项目采用基于高通量测序技术的微生物组学方法，结合分子生物学和生物信息学分析，（1）建立了针对Accumulibacter多聚磷酸盐激酶ppk1基因的实时荧光定量PCR快速检测方法，显著提高了聚磷菌检测技术的分辨率；（2）构建了Accumulibacter在厌氧和好氧阶段的碳素、电子和能量流通图，提出了一套富集未可培养微生物的诊断和优化方法；（3）解析了以亚硝酸盐为电子受体的EBPR系统微生物基因组全谱图，利用比较基因组分析，揭示温室气体N2O的生成机理和菌群代谢网络关系；（4）探究了微生物在不同环境压力下的菌群更替规律，揭示体系中微生物应对环境变化的响应机制。本项目研究结果将为EBPR工艺优化与稳定运行提供重要的理论依据和技术支撑。