Platform for high-throughput sequencing and culturing of microbial biodiversity

在此提案中，我们申请资金用于购买高通量测序和培养微生物多样性的平台。所要求的设备包括制备用于DNA和RNA测序的分子文库所需的工具，通过测序来量化微生物的生物多样性，并在实验室培养中成功培养微生物种群。该设备包括用于高速处理生物样品以提取和纯化DNA和RNA的自动化仪器，制备用于高通量测序的生物样品文库，以及在不同环境条件下在实验室中培养微生物种群。该设备将使从生物样本快速转移到实验室培养的序列和菌株，然后以目前可用设备无法实现的速度获得生物学见解成为可能。所要求的设备对于支持我们研究陆地地下和植物相关栖息地微生物生物多样性的研究项目至关重要。拉扎尔的研究重点是地球的地下，它从地表延伸到地下基岩层，是地球上包含我们大部分饮用水的部分。地球的这一地区也是在地球化学恶劣条件下发挥作用的新型酶的来源，这些酶在生物修复等生物技术领域具有极其重要的意义。Kembel的研究重点是了解生活在植物上的微生物种群和群落的生物多样性，特别是在叶子（层层）上，以及量化植物相关微生物对植物健康和生态系统功能的重要性。了解植物与微生物的关系在林业和农业中具有重要的应用。我们越来越多地使用基于测序和基于培养的综合方法来量化微生物多样性，从而有可能超越量化多样性模式，直接测试微生物多样性与宿主和生态系统功能之间的因果关系。该设备将使众多用户能够快速量化从不同环境中收集的数千个样品中的微生物生物多样性，并在实验室中培养这些栖息地的微生物，从而使实验能够了解它们的生态、进化以及彼此之间、与宿主以及与环境的相互作用。该设备将大大加快我们量化微生物多样性的步伐。这将为我们如何管理和保护微生物生物多样性以帮助人类和生态系统适应不断变化的气候提供关键见解。它还将使培训学生和研究人员掌握分子生物学的尖端方法成为可能，这是加拿大和全球生物多样性和健康研究以及工业非常需要的一套技能。

In this proposal we are requesting funding to purchase a platform for high-throughput sequencing and culturing of microbial biodiversity. The requested equipment consists of the tools required for the preparation of molecular libraries for DNA and RNA sequencing to quantify microbial biodiversity by sequencing, and to successfully grow microbial populations in culture in the laboratory. This equipment includes automated instruments for high--speed processing of biological samples to extract and purify DNA and RNA, prepare libraries of biological samples for high--throughput sequencing, and to grow microbial populations in the laboratory under differing environmental conditions. The equipment will make it possible to rapidly move from biological samples to sequences and strains in culture in the lab, and then to biological insights at a rate that is impossible using currently available equipment. The requested equipment is essential to support our research programs studying microbial biodiversity in terrestrial subsurface and plant-associated habitats. Lazar's research focuses on the terrestrial subsurface, which extends from the surface to underground bedrock layers and is the part of the Earth that contains most of our drinking water. This region of the Earth is also a source of novel enzymes functioning in geochemically harsh conditions, which are of extreme importance in biotechnology fields such as bioremediation. Kembel's research focuses on understanding the biodiversity of microbial populations and communities living on plants, in particular on leaves (the phyllosphere), and on quantifying the importance of plant-associated microbes for plant health and ecosystem function. Understanding plant-microbe associations has important applications in forestry and agriculture. Increasingly, we use combined sequencing-based and culturing-based approaches to quantify microbial diversity that make it possible to move beyond quantifying patterns of diversity, towards direct tests of the causality of links between microbial diversity and host and ecosystem function. The equipment will make it possible for numerous users to rapidly quantify microbial biodiversity in thousands of samples collected from different environments, and to grow microbes from these habitats in the lab to allow experiments to understand their ecology, evolution, and interactions with each other, with their hosts, and with the environment. The equipment will greatly accelerate the pace at which we can quantify microbial biodiversity. This will lead to key insights into how we can manage and protect microbial biodiversity to help humans and ecosystems adapt to a changing climate. It will also make it possible to train students and researchers in cutting--edge methods in molecular biology, a skill set that is in high demand in biodiversity and health research and industry within Canada and globally.