Interrogating laboratory-adapted strains of Bacillus subtilis to elucidate the selective pressures of laboratory conditions on multicellular bacterial behaviors

研究实验室适应的枯草芽孢杆菌菌株，以阐明实验室条件对多细胞细菌行为的选择压力

• 批准号: 10577916
• 负责人: Anna Lenora McLoon
• 金额: $ 30.92万
• 依托单位: SIENA COLLEGE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-01-01 至 2025-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10577916
• 关键词: Accounting; Anabolism; Atomic Force Microscopy; Bacillus subtilis; Bacteria; Bacterial Physiology; Behavior; Binding; Cell Aggregation; Cells; Cellular Structures; Cellularity; Chemotaxis; Coculture Techniques; Costs and Benefits; Data; Environment; Environmental Health; Evolution; Face; Filament; Flagella; Flagellin; Generations; Genes; Genetic; Goals; Growth; Homeostasis; Human; Human body; Infection; Invaded; Ions; Iron; Laboratories; Laboratory culture; Life; Mediating; Metabolic Pathway; Metabolism; Metals; Microbial Biofilms; Microscopy; Modeling; Modification; Molecular; Molecular Cloning; Monitor; Multicellular Process; Mutation; Natural Products; Nutrient availability; Organism; Outcome; Pathogenesis; Pathway interactions; Pattern; Phenotype; Pigments; Play; Point Mutation; Population; Probability; Process; Production; Property; Proteins; Research; Resources; Role; Scientist; Siderophores; Students; Swimming; System; Techniques; Testing; Time; Trace Elements; Work; antimicrobial; bacillibactin; cell motility; college; cost; design; fitness; genome sequencing; improved; innovation; insight; interest; metal chelator; microbial; mutant; novel; opportunistic pathogen; pressure; prevent; reuptake; secondary metabolite; small molecule; social; trait; undergraduate student; uptake; whole genome

Abstract Bacteria carry out a variety of multicellular processes that influence their pathogenesis and environmental roles in the natural environment. However, it has become apparent that when these organisms are studied in the laboratory, they undergo significant genetic modification over time. The long-term goal of this project is to understand how the laboratory environment itself selects against and/or changes the fitness effects of multicellular bacterial behaviors including biofilm formation, motility, and the secretion of pigmented secondary metabolites. These features, however, are critical due to their influence on bacterial pathogenesis and their positive or negative environmental effects. The PI will characterize laboratory adapted strains of B. subtilis isolated from populations that grew in the common laboratory medium LB for approximately 300 generations. These strains have distinctive changes in motility, biofilm formation, and pigment production. The goals of this project are to use these laboratory-adapted strains to: 1) Identify the molecular mechanism(s) that causes an unusual “social swimming” behavior in one laboratory adapted strain, as surprisingly, this strain actively forms large, multicellular aggregates in broth culture. 2) Quantify the costs and benefits of motility in laboratory culture, as preliminary data suggest many laboratory-adapted strains have reduced or altered motility. 3) Identify the pigments produced by a laboratory-adapted strain and the wildtype B. subtilis strain NCIB3610 under distinct laboratory conditions, and quantify the effects of the production of these pigments on fitness in the laboratory. 4) Identify the environmental and metabolic pathways responsible for the triggering of production of pigmented natural products by Bacillus subtilis, focusing on pulcherrimin; and determine the relationship between pigment production and biofilm formation. This research is innovative because social swimming is a novel phenotype that could inform understanding of the evolution of multicellularity. Also, preliminary data identify discrepancies between our observations and the current model explaining the role of the pigment pulcherrimin in Bacillus subtilis, suggesting that additional research on the role of this pigment in B. subtilis is needed. This is important due to the role of pulcherrimin in biofilm formation and its antimicrobial properties. Additionally, this work will provide insights to scientists studying multicellular processes like motility in the laboratory, as this work will identify probable effects of the laboratory environment itself. Furthermore, this proposed project will enhance the research environment at Siena College significantly by providing support to involve more undergraduate students in research, increasing research capacity, and due to Siena’s student population, will aid in the larger goal of increasing diversity in STEM.

抽象的 细菌进行多种影响其发病机理和环境作用的多细胞过程 在自然环境中。但是，很明显，当这些生物在 实验室，随着时间的流逝，他们进行了重大的遗传修饰。该项目的长期目标是 了解实验室环境本身如何选择和/或改变适应性的影响 多细胞细菌行为，包括生物膜形成，运动性和分泌的次要的次要行为 代谢物。但是，这些特征由于它们对细菌发病机理的影响及其 积极或负面的环境影响。 PI将表征实验室改编的枯草芽孢杆菌菌株 从在普通实验室中LB中生长的人群分离约300代。 这些菌株的运动性，生物膜形成和色素产生有明显的变化。目标的目标 项目将使用这些实验室适应的菌株进行：1）确定导致的分子机制 在一个实验室适应的压力中，异常的“社交游泳”行为，令人惊讶的是，这种压力会积极形成 肉汤培养中的大型多细胞聚集体。 2）量化运动文化运动的成本和收益， 由于初步数据表明，许多实验室适应的菌株降低或改变了运动性。 3）确定 由实验室适应的菌株和野生型枯草芽孢杆菌菌株NCIB3610产生的颜料 实验室条件，并量化这些色素生产对实验室健身的影响。 4） 确定负责触发生产的环境和代谢途径 枯草芽孢杆菌的天然产物，专注于粉末蛋白；并确定色素之间的关系 生产和生物膜形成。这项研究具有创新性，因为社交游泳是一种新颖的表型 可以告知人们对多细胞的演变的理解。另外，初步数据确定差异 在我们的观察结果与当前模型之间，解释了色素脉搏素在芽孢杆菌中的作用 枯草脂，表明需要对这种色素在枯草芽孢杆菌中的作用进行进一步的研究。这很重要 由于粉细胞霉素在生物膜形成及其抗菌特性中的作用。此外，这项工作还将 为研究多细胞过程（如实验室中的运动能力）提供见解，因为这项工作将 确定实验室环境本身的有问题影响。此外，这个提议的项目将增强 锡耶纳学院的研究环境可以通过提供更多的支持来参与更多的本科生 在研究，提高研究能力以及由于锡耶纳的学生人数，将有助于更大的目标 茎的多样性增加。