Metagenetic analysis of microbial community behavior

描述（由申请人提供）：该申请涉及社会行为研究模式生物的广泛挑战领域：识别和发展模式生物，允许对社会行为的遗传，生化，生理和环境成分进行综合分析。rfa - od - 09 - 00401 -通用- 102。最近的科学进展表明，微生物群落与广泛的人类疾病有关。这种新出现的理解提示了涉及操纵微生物群落的治疗策略。然而，目前对微生物群落行为的理解是原始的，不支持战略干预的设计。对微生物群落进行有针对性的管理需要对个体和整个群落的社会行为的遗传、生化、生理和环境因素有一个综合的了解。这样的理解，反过来，需要一个简单的模型系统，可以遗传地操纵。该系统将为微生物社会行为的遗传、生化、生理和环境成分的综合分析提供基础，包括微生物群落入侵和群落如何排斥入侵者等现象。拟议的工作将开发一个系统来解决这种昆虫肠道群落的知识差距，Manduca sexta。在这个项目中，我们引入了“元遗传学”的概念，这是研究群体表型的遗传基础。我们将确定参与粪肠球菌入侵群落的基因，并确定当粪肠球菌是社区居民时，是否有相同的基因参与排斥入侵者。我们开发了一种光学成像技术来筛选E. faecalis菌株OGR1F的发光突变体，该菌株是人类和昆虫组织的强大菌落。这项工作的长期目标是了解微生物群落稳健性的本质。我们将开始用元遗传学来剖析这一特征，即群体行为的遗传分析。我们将使用的模型系统是烟草角虫（Manduca sexta）肠道中的微生物群落。Manduca是一个完善的模型系统，许多与人类相关的生理发现（如心跳逆转）已经完成。该系统提供了一个简单的模型，在这个模型中可以阐明支配个人和社区行为的原则。因此，我们的具体目标是：鉴定在肠道群落入侵中有缺陷的粪肠球菌突变体。2. 在Aim 1中鉴定的入侵突变体的遗传特征。3. 描述Manduca肠道微生物群落中入侵突变体的行为。创新：该项目的创新源于概念框架，这是一个旧概念的新表述-基因分析在社区中的应用。该模型系统也是创新的，可以通过活体动物的发光成像直接量化微生物种群。通过分析得出的原则最终将在更复杂的群落中进行测试，从而根据经验数据为群落中的微生物行为制定一个一般的理论框架。研究者：研究者是Howard Hughes医学研究所教授，研究微生物生态学遗传学已有30年。她在功能宏基因组学（并创造了这个词）、宿主-微生物相互作用、微生物相互作用和多微生物疾病方面发表了重要的研究成果。除了她的科学专业知识外，Handelsman还是一位熟练的导师，她在指导方面发表了文章并提供了国家领导。环境:。Handelsman实验室位于威斯康星大学细菌学系，有足够的设备、空间和智力来有效地完成这个项目。该部门最近搬到了一个设备齐全的现代化建筑，里面有40个微生物研究实验室，为威斯康星大学麦迪逊分校100多名微生物学教师提供了一个中心。影响:。这项工作有可能深刻地改变微生物群落生态学领域，因为它为在群落水平上分析基因型-表型关系提供了一个新的框架。该模型系统可适用于其他应用，并可为导出面对扰动时群落行为的预测模型提供基础。这种模式对于操纵社区以造福人类健康这一最终目标至关重要。这项工作为在群落水平上分析基因型-表型关系提供了一个新的框架，有可能深刻地改变微生物群落生态学领域。该模型系统可适用于其他应用，并可为导出面对扰动时群落行为的预测模型提供基础。这种模式对于操纵社区以造福人类健康这一最终目标至关重要。

DESCRIPTION (provided by applicant): The application addresses broad challenge area Model organisms for social behavior studies: Identification and development of model organisms that allow for integrative analyses of the genetic, biochemical, physiological, and environmental components of social behavior. RFA-OD-09-00401- GM-102. Recent scientific advances have implicated microbial communities in a broad range of human diseases. This emerging understanding suggests treatment strategies that involve manipulating microbial communities. However, current understanding of the behavior of microbial communities is primitive and does not support the design of strategic interventions. Targeted management of microbial communities will require an integrated understanding of the genetic, biochemical, physiological, and environmental components of social behavior of individuals and of the community as a whole. Such an understanding, in turn, requires a model system that is simple and can be manipulated genetically. This system will provide the foundation for integrative analyses of the genetic, biochemical, physiological, and environmental components of social behavior of microorganisms, including phenomena such as invasion of microbial communities and how communities exclude invaders. The proposed work will develop a system for addressing this knowledge gap in the gut community of the insect, Manduca sexta. In this project, we introduce the concept of "metagenetics," which is the study of the genetic basis for community phenotypes. We will identify genes involved in invasion of the community by Enterococcus faecalis, and determine whether the same genes are involved in exclusion of invaders when E. faecalis is a community resident. We have developed an optical imaging technique to screen luminescent mutants of E. faecalis strain OGR1F, which is a powerful colonist of both human and insect tissue. The long-term goal of this work is to understand the nature of robustness of microbial communities. We will begin to dissect this characteristic using metagenetics, the genetic analysis of community behavior. The model system we will use is the microbial community in the gut of Manduca sexta, the tobacco hornworm. Manduca is a well-established model system in which many physiological discoveries relevant to humans have been made (such as heartbeat reversal). The system provides a simple model in which the principles governing individual and community behavior can be elucidated. As such, our specific aims are to: 1. Identify mutants of Enterococcus faecalis that are defective in gut community invasion. 2. Genetically characterize invasion mutants identified in Aim 1. 3. Characterize behavior of invasion mutants in the Manduca gut microbial community. Innovation: The project's innovation derives from the conceptual framework, which is a new formulation of an old concept - application of genetic analysis to communities. The model system is also innovative, enabling direct quantification of microbial populations with luminescence imaging in live animals. The principles developed through this analysis will be tested ultimately in more complex communities to formulate a general theoretical framework for microbial behavior in communities based on empirical data. Investigator: The investigator is a Howard Hughes Medical Institute Professor who has studied the genetics of microbial ecology for 30 years. She has published significant work in functional metagenomics (and coined the word), host-microbe interactions, microbial interactions, and polymicrobial disease. In addition to her scientific expertise, Handelsman is a skilled mentor who has published and provided national leadership on mentoring. Environment:. Situated in the Department of Bacteriology at the University of Wisconsin, the Handelsman lab has access to sufficient equipment, space, and intellectualism to complete this project effectively. The department recently moved to a well-equipped, modern building that contains 40 microbiology research labs, providing a hub for the more than 100 microbiology faculty at the University of Wisconsin-Madison. Impact:. The work has the potential to profoundly alter the field of microbial community ecology because it provides a new framework for analysis of genotype-phenotype relationships at the community level. The model system may be adapted to other applications and it may provide the basis for deriving predictive models about community behavior in the face of perturbation. Such models are essential to the ultimate goal of manipulating communities for the benefit of human health. PHS 416-1/416-9 (Rev. 9/08) Page Continuation Format Page The work has the potential to profoundly alter the field of microbial community ecology because it provides a new framework for analysis of genotype-phenotype relationships at the community level. The model system may be adapted to other applications and it may provide the basis for deriving predictive models about community behavior in the face of perturbation. Such models are essential to the ultimate goal of manipulating communities for the benefit of human health.