Analysis of Nucleic Acids from Difficult Media: Method Optimization for Bentonite Clays and Rock Cores Associated with Deep Geological Repositories for Nuclear Waste

核能是加拿大无碳能源生产的重要组成部分。虽然核能不会排放温室气体，但它确实会产生“用过的核燃料”，这是一种高放射性的危险废物，需要谨慎管理，以保护人类和环境。国际共识已确定，深层地质储存库是管理用过核燃料的最安全和最有效的手段。核废料管理组织(NWMO)正在为加拿大所有用过的核燃料选址和设计一个深层地质库。拟议的NSERC CRD研究将开发分子和生物信息学工具，用于表征深层地质储存库中的微生物及其编码功能，以更好地预测和沟通它们的代谢活动可能如何长期影响地球化学环境。拟议的研究将涉及以元基因组和元转录方法以及相关的生物信息学分析为基准，具体目标是(1)将16S rRNA基因功能预测与元基因组和元转录数据进行比较，以确定在编码微生物过程中以负担得起的价格和可靠的方式分析大量样本的最佳做法，以及(2)开发用于元基因组分析和基因组到细胞可视化的软件应用程序，最终以更有效和自动化的方式交流亚表面微生物基因组的功能潜力。这项研究中提出的见解将为NWMO提供所需的工具，通过开发适当的基因组方法，为加拿大DGR设计和地点特征提供信息。此外，开发的评估低生物量样品中微生物群落的方法将进一步扩大我们对地球上地下生命的极限、能力和生存的理解。该提案还将培训HQP与正在实施和改进科学知识有关的专门知识，以支持和改善加拿大的能源和环境部门。

Nuclear energy is an important component of carbon-free energy production in Canada. Although nuclear energy does not emit greenhouse gases, it does produce "used nuclear fuel", a hazardous waste that is highly radioactive and requires careful management to protect people and the environment. International consensus has determined that a deep geological repository is the safest and most effective means of managing used nuclear fuel. The Nuclear Waste Management Organization (NWMO) is siting and designing a deep geological repository for all of Canada's used nuclear fuel. The proposed NSERC CRD research will develop molecular and bioinformatic tools for characterization of microorganisms, and their encoded functions, in a deep geological repository to better predict and communicate how their metabolic activity may impact the geochemical environment in the long-term. The proposed research will involve benchmarking metagenomic and metatranscriptomic methods, and associated bioinformatic analyses, with the specific objectives of (1) comparing 16S rRNA gene functional predictions to metagenomic and metatranscriptomic data to identify best practices for analyzing large numbers of samples affordably and reliably for encoded microbial processes, and (2) developing software applications for metagenome analysis and genome to cell visualizations, ultimately for communicating the functional potential of subsurface microbial genomes more effectively and in an automated way. The insights developed in this research will provide NWMO with tools required to inform Canadian DGR design and site characterization by developing appropriate genomic methodologies. In addition, the methods developed to assess microbial communities in low biomass samples will further expand our understanding of the limits, capability, and survival of subsurface life on Earth. This proposal will also train HQP in expertise relevant to ongoing implementation and improvement of scientific knowledge to support and improve Canada's energy and environmental sectors.