RII Track-4: Physiology of Sulfate-Reducing Bacteria under Energy-Limiting conditions

RII Track-4：能量限制条件下硫酸盐还原菌的生理学

• 批准号: 2033286
• 负责人: Aude Amelie Picard
• 金额: $ 17.6万
• 依托单位: University of Nevada Las Vegas
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-02-01 至 2024-01-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2033286&HistoricalAwards=false
• 关键词: RII; Track; Physiology; Sulfate; Reducing

Bacteria have the ability to survive long periods of time under energy-limiting conditions, common in natural environments. To understand the fundamental aspects of energy limitation, bacteria can be studied in cultures for months to years without re-supplying the medium with energy sources. This project aims to study long-term cultures of sulfate-reducing bacteria (SRB), which drive the sulfur and carbon biogeochemical cycles in anoxic environments. The proposed research will investigate the integrity of the cell and outer membranes of SRB and quantify metabolic processes to gain physiological insights into the long-term survival of SRB. This project will advance fundamental knowledge about long-term survival of microorganisms in anoxic environments. The project is also of importance for the economically vital processes that are influenced by SRB, such as the corrosion of iron infrastructure, wastewater treatment, and bioremediation of metal contaminated environments. This fellowship will improve 1) STEM education through the delivery of lectures on high-resolution imaging to graduate courses, 2) the research capacity of the PI, and 3) the research infrastructure at the University of Nevada, Las Vegas. The goals of this project are to characterize the extended life cycle of sulfate-reducing bacteria and to decipher the role of sulfide mineral crusts in preserving the viability of SRB in their extended life cycle. Our preliminary experiments have indicated that mineral-encrusted bacteria can be preserved alive for several years and regrow in fresh medium when non-encrusted cells of similar age do not regrow. The proposed research will characterize the metabolic and physiological changes in mineral-free and encrusted sulfate-reducing bacteria in the long-term stationary phase. It will also identify the early steps of mineral crust formation at the surface of cells and the steps of mineral crust removal when cells regrow in fresh medium. Transmission electron microscopy (TEM) will be used to image the integrity of the cells and their membranes and cell wall at high resolution, while electron diffraction will be used to gain information about the mineral phases forming around cells. Super resolution fluorescence microscopy (iPALM) will be used to localize and quantify metabolic processes such as new protein synthesis, proton gradient formation and adenosine triphosphate synthesis during the extended life cycle of mineral-free and encrusted SRB. This project will lead to a better understanding of long-term survival of sulfate-reducing bacteria and of the role of sulfide minerals in preserving long-term viability of SRB, which contribute to the formation of those minerals.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

细菌有能力在自然环境中常见的能量限制条件下长时间存活。为了了解能量限制的基本方面，可以在培养物中研究细菌数月至数年，而无需向培养基中重新提供能量来源。本项目旨在研究硫酸盐还原菌（SRB）的长期培养，SRB在缺氧环境中驱动硫和碳的生物地球化学循环。拟议的研究将调查SRB的细胞和外膜的完整性，并量化代谢过程，以获得对SRB长期生存的生理学见解。该项目将推进有关微生物在缺氧环境中长期生存的基础知识。该项目对于受SRB影响的经济上至关重要的过程也很重要，例如铁基础设施的腐蚀，废水处理和金属污染环境的生物修复。该奖学金将通过向研究生课程提供高分辨率成像讲座来改善1）STEM教育，2）PI的研究能力，以及3）拉斯维加斯的内华达州大学的研究基础设施。该项目的目标是确定硫酸盐还原菌的延长生命周期的特征，并破译硫化物矿物结壳在维持硫酸盐还原菌在其延长生命周期中的生存能力方面的作用。我们的初步实验表明，矿物结壳的细菌可以存活数年，并在新鲜培养基中再生，而类似年龄的非结壳细胞不会再生。拟议的研究将表征无矿物质和结壳硫酸盐还原菌在长期稳定期的代谢和生理变化。它还将确定在细胞表面形成矿物质结壳的早期步骤，以及当细胞在新鲜培养基中再生时去除矿物质结壳的步骤。透射电子显微镜（TEM）将用于以高分辨率对细胞及其膜和细胞壁的完整性进行成像，而电子衍射将用于获得有关细胞周围形成的矿物相的信息。超分辨率荧光显微镜（iPALM）将被用来定位和量化代谢过程，如新的蛋白质合成，质子梯度形成和腺苷三磷酸合成在延长的生命周期的无矿物质和结壳SRB。该项目将使人们更好地了解硫酸盐还原菌的长期生存以及硫化物矿物在维护硫酸盐还原菌长期生存能力方面的作用，这有助于这些矿物的形成。该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。