Metabolic Interactions Supporting Effective TCE Bioremediation under Various Biog

描述（由申请人提供）：基于dehalococoides的生物修复系统的优化，以处理三氯乙烯（TCE）污染的超级基金场地，依赖于对复杂微生物相互作用的知识密集型理解，这些相互作用塑造了暴露于各种地球化学条件下的TCE脱氯微生物群落的结构和功能健壮性。先前的研究表明，不同的地球化学参数，如pH值的降低或替代终端电子受体的存在可导致不完全脱氯。我们建议采用分子、生化和分析相结合的方法来评估连续流化学恒温器中可持续和确定的多菌株TCE-脱氯菌群之间的相互作用，以研究群落相互作用和生物地球化学条件对脱氯活性的影响。具体而言，我们建议分析社区水平的转录组学、细胞间代谢组学和先进的随机矩阵理论网络分析，以了解地球化学胁迫对tce脱氯菌群的影响。该项目旨在提供网络基因调控和代谢物交换的基本理解，这些基因调控和代谢物交换影响厌氧dehalococides -含微生物群落并控制其TCE脱氯能力。在提议的工作中，将研究各种与环境相关的地球化学应力，包括pH值，盐度和引入系统的潜在竞争电子受体（例如硫酸盐离子）。这些地球化学压力的影响将通过相互作用和相互依赖的脱氯群体之间交换的代谢物、细胞过程和遗传调控进行监测和跟踪。由此产生的知识将提高设计和优化前瞻性工程解决方案的能力，从而减少与成功的地下水生物修复相关的时间和成本。在这个拟议的研究中要调查的具体目标是：构建可持续的tce脱氯微生物群落，包括在强大的tce脱氯群落中代表基本功能的多种确定的微生物物种，并在趋化剂中建立它们的持续增长。2. 识别生理变化、遗传调控变化和细胞间代谢变化，这些变化发生在tce脱氯联合体中，以响应生物地球化学条件的变化，如pH值、盐度和替代电子受体的存在。超转录组学和代谢组学分析将用于获得对地球化学压力的复杂反应的系统级理解。3. 建立生物地球化学条件、微生物遗传调控和代谢相互作用之间的相关性，以阐明响应各种生物地球化学条件的联盟成员之间的网络相互作用。这些结果将用于研究在变化的地球化学条件下稳定脱氯群落功能的刺激或增强策略。

DESCRIPTION (provided by applicant): The optimization of Dehalococcoides-based bioremediation systems to treat trichloroethene (TCE)-contaminated Superfund sites relies upon knowledge-intensive understanding of complex microbial interactions that shape the structural and functional robustness of TCE-dechlorinating microbial communities exposed to a variety of geochemical conditions. Previous studies have shown that varied geochemical parameters, such as decreased pH or the presence of alternative terminal electron acceptors can result in incomplete dechlorination. We propose to use a combination of molecular, biochemical and analytical tools to evaluate interactions within sustainable and defined multi-strain microbial TCE- dechlorinating consortia in continuous-flow chemostats to study community interactions and the effects of biogeochemical conditions on dechlorination activity. Specifically, we propose to analyze community-level transcriptomics, intercellular metabolomics, and advanced Random Matrix Theory network analysis to understand the impact of geochemical stresses to the TCE-dechlorinating consortia. This project seeks to deliver a fundamental understanding of networked gene regulations and metabolite exchanges that impact anaerobic Dehalococcoides- containing microbial communities and control their TCE dechlorination capabilities. In the proposed work, a variety of environmentally-relevant geochemical stresses will be investigated including pH, salinity and the introduction of potential competitive electron acceptors to the system (e.g., sulfate ions). Effects of these geochemical stresses will be monitored and tracked through the exchanged metabolites, cellular processes and genetic regulations occurred amongst the interactive and interdependent populations in dechlorinating consortia. The generated knowledge will lead to an improved ability to design and optimize proactive engineering solutions to decrease the time and cost associated with successful groundwater bioremediation. The specific objectives to be investigated in this proposed study are, 1. Construct sustainable TCE-dechlorinating microbial consortia with multiple defined microbial species that represent the essential functions within robust TCE-dechlorinating communities, and establish their sustained growth in chemostats. 2. Identify the physiological changes, genetic regulatory changes, and the intercellular metabolic changes that occur in the TCE-dechlorinating consortia in response to changes in biogeochemical conditions, such as pH, salinity, and the presence of alternative electron acceptors. Metatranscriptomics and metabolomics analyses will be used to obtain a systems-level understanding of the complex responses of the consortia to geochemical stresses. 3. Establish correlations among biogeochemical conditions, microbial genetic regulations and metabolic interactions in order to elucidate the networked interactions among the members of the consortia that respond to various biogeochemical conditions. The results will be used to investigate stimulation or augmentation strategies that can stabilize the functions of dechlorination communities under changing geochemical conditions.