Monitoring Spatial Segregation in Surface Colonizing Microbial Populations.
Monitoring Spatial Segregation in Surface Colonizing Microbial Populations.
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DOI：
10.3791/54752
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发表时间：
2016-10-29
期刊：
影响因子：
--
通讯作者：
Kovács ÁT
中科院分区：
文献类型：
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作者：
Hölscher T;Dragoš A;Gallegos-Monterrosa R;Martin M;Mhatre E;Richter A;Kovács ÁT
微生物提供了一个有趣的系统来研究群体中个体之间的社会互动。微生物的短世代和相对简单的遗传修饰程序促进了社会微生物学领域的发展。为了评估某些微生物物种的适应性，可以使用适合于适当荧光滤光器的显微镜对一个种群内的选定菌株或其遗传修饰衍生物进行荧光标记和跟踪。在琼脂培养基上扩增不同微生物物种的菌落可用于监测产生独特荧光蛋白的细胞的空间分布。在这里，我们提出了一个详细的协议，使用绿色和红色荧光蛋白产生细菌菌株，以遵循表面殖民化过程中的空间排列，包括鞭毛驱动的社区运动（群集），胞外多糖和疏水蛋白依赖的生长介导的传播（滑动），和复杂的菌落生物膜形成。革兰氏阳性菌枯草芽孢杆菌的非驯化分离株可用于仔细检查某些表面铺展特性及其对琼脂固化培养基上二维分布的影响。通过改变用于引发集落生物膜的细胞的数量，分类水平可以在连续规模上变化。延时荧光显微镜可用于见证不同表型和基因型之间的相互作用，在一定的分类水平，并确定任何一个相对成功。
Microbes provide an intriguing system to study social interaction among individuals within a population. The short generation times and relatively simple genetic modification procedures of microbes facilitate the development of the sociomicrobiology field. To assess the fitness of certain microbial species, selected strains or their genetically modified derivatives within one population, can be fluorescently labelled and tracked using microscopy adapted with appropriate fluorescence filters. Expanding colonies of diverse microbial species on agar media can be used to monitor the spatial distribution of cells producing distinctive fluorescent proteins. Here, we present a detailed protocol for the use of green- and red-fluorescent protein producing bacterial strains to follow spatial arrangement during surface colonization, including flagellum-driven community movement (swarming), exopolysaccharide- and hydrophobin-dependent growth mediated spreading (sliding), and complex colony biofilm formation. Non-domesticated isolates of the Gram-positive bacterium, Bacillus subtilis can be utilized to scrutinize certain surface spreading traits and their effect on two-dimensional distribution on the agar-solidified medium. By altering the number of cells used to initiate colony biofilms, the assortment levels can be varied on a continuous scale. Time-lapse fluorescent microscopy can be used to witness the interaction between different phenotypes and genotypes at a certain assortment level and to determine the relative success of either.