Elucidation of Microbial Response to Temperature Variation and the Diurnal Cycle in the Yellowstone Octopus Hot Spring: A Proteomic Study

关于自然界中的微生物如何适应和改变它们对环境变化的反应，人们知之甚少。与实验室培养的分离细菌不同，“野外”的微生物生活在复杂的群落中。这些群落由各种各样的物种组成，其中大多数仍然是科学界所未知的。一个引人注目的例子是黄石国家公园章鱼泉中以喜热生物叠层垫的形式存在的群落。一天中光线的变化会影响草席中微生物之间的相互作用，比如交叉进食。除了光线的变化，温泉的温度也在变化。推动这项工作的假设是，特定蛋白质的存在将反映微生物席群落居民在光和温度变化期间的活动、分布和变化。质谱法将被用于鉴定这些蛋白质样品从垫子在一个新的方法被称为环境蛋白质组学。将使用标记方法来量化在不同采样条件下产生的蛋白质的数量。基因组和转录组数据将与蛋白质数据进行比较，所有数据都是在相同条件下收集的。这种方法是一种确定环境对微生物群落影响的新方法。目前有很多关于不同喜热微生物菌株的基因组信息，但对这些微生物产生的蛋白质的表征才刚刚开始。在一个特征良好的微生物席中，微生物产生的蛋白质的知识将为微生物群落提供宝贵的资源，并有助于解释微生物化石记录。在这个项目过程中收集的蛋白质组学数据将被存入黄石国家公园的数据库。这将使该提案的发现与公园中正在进行的基因组工作相协调。少数民族本科生将通过暑期本科生研究计划参与这项研究。这些暑期学生将在一个令人兴奋的环境中进行严格的实验室工作和科学实地考察。这个吸引人的环境也将用于教育公众，通过外展演讲计划和黄石国家公园的展览。因此，该项目将提高公众的科学素养，并提供有关微生物群落适应的新方法和信息。

Little is known about how microorganisms in nature adapt and change in reaction to changes in their surroundings. Unlike the isolated bacteria grown in laboratories, microbes in 'the wild' live in complex communities. These communities are comprised of diverse species, most of which are still unknown to science. One fascinating example is the community that exists as laminate mats of heat-loving organisms in Octopus Spring in Yellowstone National Park. Changes in light throughout the day affect interactions such as cross-feeding between microbes in the mats. In addition to changes in light, the hot spring changes in temperature. The hypothesis driving this work is that the presence of particular proteins will reflect the activities, distribution, and changes in the inhabitants of the microbial mat community during changes in light and temperature. Mass Spectrometry will be used to identify these proteins in samples from the mat in a new approach known as environmental proteomics. A labeling method will be used to quantify the amounts of the proteins produced under different sampling conditions. Genomic and transcriptomic data will be compared to protein data, all collected under the same conditions.This methodology is a new approach to identifying the influences of the environment on a microbial community. There is currently much genomic information about the different strains of heat-loving microbes, but characterization of the proteins produced by these organisms is just beginning. Knowledge of the proteins produced by the microorganisms in a well-characterized microbial mat will provide a valuable resource to the microbiological community and will aid in interpreting the microbial fossil record. Proteomic data collected during the course of this project will be deposited in Yellowstone National Park databases. This will coordinate findings from this proposal with the ongoing genomic work in the park. Minority undergraduate students will be involved in this research through a Summer Undergraduate Research Program. These summer students will be exposed to both rigorous labwork and scientific fieldwork in an exciting setting. This attractive setting will also be used to educate the general public, both through an outreach speaking program and displays in Yellowstone National Park. Thus this project will both improve the scientific literacy of the public and provide new methods and information about adaptation in microbial communities.