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

微生物群落结构和功能的分子遗传学分析可以为评估生态系统的压力、影响和恢复提供直接而有力的工具。这些信息可以改善生物修复系统的设计和运行，监测自然衰减（MNA）的评价和资源配置，并将在决策和监管监督中增加科学依据。知识价值。我们开发了一套用于检测和枚举芳香加氧酶基因的引物和实时PCR协议，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，即芳香烃有氧生物降解的发生。我们的引物已经在纯培养物、土壤微观环境和两个现场样品的控制良好的实验中进行了测试。跟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。目前还没有可靠的原位方法来评估用于生物修复曝气的组件的有效性，例如真空泵、空气喷射器或氧气释放剂。直接测量编码芳香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。选择芳香加氧酶基因作为指标，是因为它们在汽油污染场所主要关注的污染物BTEX的生物降解中起关键作用。现代生物技术的发展可以大大优化昂贵的曝气技术和氧释放剂的利用。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学特征的可行性，以在实地规模的大学-工业合作中推进最先进的LUST场地修复。在三年的时间里，我们的跨学科大学-工业团队建议进行一个研究项目，将获得关于这个最近开发的监测石油污染地点的工具的现场性能数据。我们预计不同“年龄”、地球化学和地质的地点将具有特征和可检测的分子特征。

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.