Molecular Genetic Characterization of Contaminated Environments as a Tool for the Assessment of Remediation Technology

0302645微生物群落结构和功能的NIES分子遗传分析可以为评估受压力、受影响和恢复的生态系统提供直接和强大的工具。这些信息可以改进生物修复系统的设计和运行、监测自然衰减(MNA)的评估和资源分配，并将增加合理科学在决策和监管监督中的使用。我们已经开发了一套检测和计数芳香族加氧酶基因的引物和实时PCR方法，并将它们与PCR-DGGE群落分析并行使用，以评估将该生物技术用作生物修复评估和监测工具的可行性。加氧酶基因的检测可以直接了解当地生态系统的同化功能，即芳香烃的好氧生物降解发生。我们的引物已经在纯培养、土壤微观世界和来自两个野外地点的样本上进行了良好控制的实验。追踪参与优先污染物生物降解的分解代谢基因可用于优化修复系统和评估MNA。目前还没有可靠的现场方法来评估用于生物修复曝气的组件的有效性，如真空泵、空气喷雾器或氧气释放改进剂。直接测量编码芳香族加氧酶的基因将为监测工程曝气系统和MNA提供有效的工具。芳香族加氧酶基因之所以被选为指标，是因为它们在BTEX的生物降解过程中发挥着关键作用，BTEX是汽油污染场地主要关注的污染物。通过开发现代生物技术工具，可以大大优化昂贵的曝气技术和氧气释放改进剂的利用。我们建议评估利用微生物群落结构和分解代谢功能的分子遗传特征来推进现场规模的大学-工业合作中性爱场所修复的最新技术的可行性。在三年的时间里，我们的跨学科大学-行业团队提议进行一个研究项目，该项目将获得关于这个最近开发的用于监测石油污染场地的工具的现场性能数据。我们预计，不同‘年代’、不同地球化学和不同地质学的地点将具有特征和可检测到的分子特征。

0302645 Nies Molecular genetic analysis of microbial community structure and function can provide a direct and powerful tool to assess stressed, impacted, and recovering ecosystems. This information can improve bioremediation system design and operation, evaluation of monitored natural attenuation (MNA) and resource allocation, and it will increase the use of sound science in decision making and regulatory oversight.Intellectual Merit. We have developed a set of primers and a real-time PCR protocol to detect and enumerate aromatic oxygenase genes, and have used them in parallel with PCR-DGGE community analysis, to assess the feasibility of using this biotechnology as a bioremediation assessment and monitoring tool. The detection of oxygenase genes yields direct insight into the assimilative function of the local ecosystem, i.e. the occurrence of aerobic biodegradation of aromatic hydrocarbons. Our primers have been tested in well controlled experiments with pure cultures, in soil microcosms, and on samples from two field sites. Tracking catabolic genes involved in the biodegradation of priority pollutants can be used to optimize remediation systems and evaluate MNA. Reliable in-situ methods currently do not exist to evaluate the effectiveness of the components used for bioremediation aeration, such as vacuum pumps, air spargers, or oxygen releasing amendments. The direct measurement of genes encoding aromatic oxygenases would provide an effective tool for monitoring engineered aeration systems and for MNA. Aromatic oxygenase genes were chosen as indicators because they play a key role in the biodegradation of BTEX, the pollutants of principal concern at gasoline-contaminated sites. The utilization of expensive aeration technologies and oxygen releasing amendments could be significantly optimized by the development of modern biotechnology tools. We propose to evaluate the feasibility of using molecular genetic characterization of the microbial community structure, and catabolic function, to advance the state-of-the-art of LUST site remediation in a field-scale university-industry collaboration. Over a three year period, our interdisciplinary university-industry team proposes to conduct a research project that will obtain field performance data about this recently developed tool for monitoring petroleum contaminated sites. We expect that sites of different 'ages,' geochemistry and geology will have characteristic and detectable molecular signatures.