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制和环境作用的多细胞过程 然而，当在自然环境中研究这些生物体时，情况就变得很明显了。 随着时间的推移，它们会在实验室中进行重大的基因改造。该项目的长期目标是 了解实验室环境本身如何选择和/或改变健康效应 多细胞细菌行为，包括生物膜形成、运动和色素次生分泌 然而，这些特征由于它们对细菌发病机制及其代谢的影响而至关重要。 PI 将表征实验室适应的枯草芽孢杆菌菌株。 从在普通实验室培养基 LB 中生长大约 300 代的群体中分离出来。 这些菌株在运动性、生物膜形成和色素产生方面具有独特的变化。 该项目将使用这些实验室适应菌株来： 1) 确定导致 一种实验室适应性菌株中不寻常的“社交游泳”行为，令人惊讶的是，这种菌株积极形成 肉汤培养中的大型多细胞聚集体 2) 量化实验室培养中运动性的成本和收益。 初步数据表明许多实验室适应菌株的运动性已降低或改变 3) 识别。 实验室适应菌株和野生型枯草芽孢杆菌菌株 NCIB3610 在不同条件下产生的色素 实验室条件，并量化这些颜料的生产对实验室健康的影响 4)。 确定负责触发色素产生的环境和代谢途径 天然产物枯草芽孢杆菌，重点关注普切明并确定色素之间的关系； 这项研究具有创新性，因为社交游泳是一种新的表型。 此外，初步数据还发现了差异。 我们的观察结果与当前模型之间的差异解释了色素 pulcherrimin 在芽孢杆菌中的作用 枯草芽孢杆菌，表明需要对这种色素在枯草芽孢杆菌中的作用进行更多研究，这很重要。 由于普切明在生物膜形成中的作用及其抗菌特性，这项工作还将。 为研究实验室运动等多细胞过程的科学家提供见解，因为这项工作将 此外，该拟议项目将增强实验室环境本身的可能影响。 锡耶纳学院的研究环境通过提供支持让更多本科生参与而得到显着改善 在研究方面，提高研究能力，并且由于锡耶纳的学生人数，将有助于实现更大的目标 增加 STEM 的多样性。