Indirect Enrichment of Desirable, but Less Fit Phenotypes, from a Synthetic Microbial Community Using Microdroplet Confinement.
Indirect Enrichment of Desirable, but Less Fit Phenotypes, from a Synthetic Microbial Community Using Microdroplet Confinement.
复制标题
使用微滴限制从合成微生物群落中间接富集所需但不太适合的表型。
DOI：
10.1021/acssynbio.3c00008
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发表时间：
2023
影响因子：
4.7
通讯作者：
Shamoo,Yousif
中科院分区：
文献类型：
--
作者：
GanigaPrabhakar,Ramya;Fan,Gaoyang;Alnahhas,RazanN;Hirning,AndrewJ;Bennett,MatthewR;Shamoo,Yousif
微生物群落内的空间结构可以为社会互动提供几乎无限的机会，是进化的重要驱动力。由于代谢物通常是分子信号，代谢物在微生物群落内的扩散可以影响群落的组成和动力学，其方式可能具有挑战性。我们使用微滴内的合成微生物群落的封装来研究空间结构和代谢物扩散对种群动态的影响，并研究由该社区的一个成员作弊的影响。合成群落由三种菌株组成：制造可扩散的群体感应分子（N-（3-氧代十二烷酰基）-l-高丝氨酸内酯，C12-氧代-HSL）或阿勒的“生产者”;被阿勒杀死的“接收者”;以及从接收者的灭绝中受益但没有与阿勒合成相关的成本的非生产者或“作弊者”。我们表明，尽管阿勒微滴之间的快速扩散，微滴施加的空间结构允许更有效的，但短暂的富集更罕见的和生长缓慢的生产者亚群。最终，非生产者人口将生产者推向灭绝。通过包括荧光激活的微滴分选，并提供持续的竞争，由接收器菌株，我们证明了一种策略，间接富集一种罕见的和未标记的生产者。在快速扩散的条件下，从更大的群体中筛选和富集代谢物生产者的能力为合成生态学和生物技术的应用开发提供了重要的框架。
Spatial structure within microbial communities can provide nearly limitless opportunities for social interactions and are an important driver for evolution. As metabolites are often molecular signals, metabolite diffusion within microbial communities can affect the composition and dynamics of the community in a manner that can be challenging to deconstruct. We used encapsulation of a synthetic microbial community within microdroplets to investigate the effects of spatial structure and metabolite diffusion on population dynamics and to examine the effects of cheating by one member of the community. The synthetic community was composed of three strains: a “Producer” that makes the diffusible quorum sensing molecule (N-(3-oxododecanoyl)-l-homoserine lactone, C12-oxo-HSL) or AHL; a “Receiver” that is killed by AHL; and a Non-Producer or “cheater” that benefits from the extinction of the Receivers, but without the costs associated with the AHL synthesis. We demonstrate that despite rapid diffusion of AHL between microdroplets, the spatial structure imposed by the microdroplets allows a more efficient but transient enrichment of more rare and slower-growing Producer subpopulations. Eventually, the Non-Producer population drove the Producers to extinction. By including fluorescence-activated microdroplet sorting and providing sustained competition by the Receiver strain, we demonstrate a strategy for indirect enrichment of a rare and unlabeled Producer. The ability to screen and enrich metabolite Producers from a much larger population under conditions of rapid diffusion provides an important framework for the development of applications in synthetic ecology and biotechnology.