MO Collaborative Research: A Genomics and Cultivation-Based Study on Novel Candidate Divisions OD1, SR1, OP11, and TM7 in a Sulfur-Rich Spring (Zodletone Spring, OK)

在过去的二十年中，用于识别微生物的技术的进步表明，有许多新的、以前未知的细菌在多种环境中似乎非常丰富。这些群体中的许多可能具有重要的生态作用，但尚未在实验室中进行研究。 该项目的目标是研究这四个重要细菌群（称为候选菌群 OD1、SR1、OP11 和 TM7）的生理特征、代谢能力和潜在生态作用。 这四个类群已被证明在佐德尔顿泉（俄克拉荷马州西南部的低温（20摄氏度）硫化物和富含硫磺的泉水）中相对丰富，并且它们被认为是促进该泉水中发生的生物过程的群落的重要成员。这些过程包括碳氢化合物的降解、矿物质的形成和光合作用。 Zodletone泉水将被用作微生物观测站来研究这些群体。将使用一套基因组技术，其中包括对单个微生物细胞基因组的 DNA 进行测序，这些微生物细胞实际上是使用流式细胞仪从泉水中捕获的。结果将允许鉴定可用于上述过程的基因和代谢途径的组装。最终，这些数据将用于了解化学反应以及每个群体成员所发挥的生态作用。随着基因组研究，这些细菌群将在实验室中培养和研究。这些细菌来自很少存在氧气的环境，因此氧气可能对这些细胞有毒。 复杂的分离策略将用于分离和培养通常不会在实验室中生长的细菌，并且这些细菌将适用于对氧敏感的微生物。这些培养研究将使我们能够验证细菌是否能够催化为它们提出的反应，并使我们能够研究单个反应以了解每个过程是如何发生的。 该提案中获得的结果将被纳入俄克拉荷马州立大学和俄克拉荷马大学提供的本科生和研究生课程中。该提案还将为科学领域代表性不足的群体的成员（尤其是美洲原住民）提供一些研究生和本科生研究机会。最后，该项目将通过俄克拉荷马大学的 K20 中心和俄克拉荷马州立大学的青少年科学与人文研讨会 (JSHS) 项目，帮助继续并扩大对俄克拉荷马州高中的推广工作。

During the last two decades, advances in techniques used to identify microorganisms have demonstrated that there are many groups of novel, previously unknown bacteria that appear to be very abundant in multiple environments. It is likely that many of these groups have important ecological roles and yet they have not been studied in the laboratory. The goal of this project is to investigate the physiological characteristics, metabolic abilities, and potential ecological roles of four of these important bacterial groups, termed candidate divisions, OD1, SR1, OP11, and TM7. These four groups have been shown to be relatively abundant in Zodletone spring, a low temperature (20 degrees C) sulfide and sulfur-rich spring in southwestern Oklahoma and they are thought to be important members of the community in terms of facilitating the biological processes that are occurring in this spring. These processes include degradation of hydrocarbons, formation of minerals and photosynthesis. Zodletone spring will be used as a microbial observatory to study these groups. A set of genomic techniques will be used, which involve sequencing DNA from the genome of one individual microbial cell, literally fished from the spring using a flow cytometer. The results will allow identification of the genes and assembly of the metabolic pathways that may be used for the above processes. Ultimately the data will be used to understand the chemical reactions and the ecological roles performed by members of each these groups. Along with genomic studies, these groups of bacteria will be cultivated and studied in the laboratory. These bacteria come from an environment in which oxygen is rarely present and as a result, oxygen is likely toxic to these cells. Sophisticated isolation strategies will be used for isolating and growing bacteria that would not normally grow in the laboratory and these will be adapted to work with microorganisms that are oxygen sensitive. These cultivation studies will allow us to verify that the bacteria are able to catalyze the reactions that have been proposed for them and will allow us to study individual reactions to understand how each process occurs. The results obtained in this proposal will be integrated into undergraduate and graduate courses offered at Oklahoma State University and the University of Oklahoma. The proposal will also provide several graduate and undergraduate research opportunities to members (especially Native Americans)of underrepresented groups in science. Finally, the project will help continuing and expanding outreach efforts to Oklahoma high schools through the K20 Center at the University of Oklahoma and the Junior Science and Humanities Symposia (JSHS) Program at Oklahoma State University.