Vibrio cholerae biofilm growth program and architecture revealed by single-cell live imaging
Vibrio cholerae biofilm growth program and architecture revealed by single-cell live imaging
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DOI：
10.1073/pnas.1611494113
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发表时间：
2016-09-06
影响因子：
11.1
通讯作者：
Bassler, Bonnie L.
中科院分区：
文献类型：
--
作者：
Yan, Jing;Sharo, Andrew G.;Bassler, Bonnie L.
生物膜是表面相关的细菌群落，在自然界和感染过程中至关重要。尽管进行了大量的工作来鉴定生物膜成分并发现它们是如何被调节的，但在单个细胞水平上对生物膜结构知之甚少。在这里，我们使用最先进的显微镜技术来实现霍乱弧菌生物膜的单细胞分辨率成像，因为它从一个单一的初始细胞发展到10,000个细胞的成熟生物膜，并发现支撑结构进化的力量。诱变、基质标记和模拟表明，表面粘附介导的压缩导致霍乱弧菌生物膜从二维分支形态转变为致密有序的三维簇。我们发现杆状细菌的定向增殖在形成霍乱弧菌生物膜的生物膜结构中起主导作用，这种生长模式由一个基因rbmA控制。竞争分析表明，密集生长方式具有为生物膜提供优越力学性能的优势。我们的单细胞技术可以广泛地将基因与生物膜精细结构联系起来，并为评估细胞对外部刺激的异质性提供了一条途径。
Biofilms are surface-associated bacterial communities that are crucial in nature and during infection. Despite extensive work to identify biofilm components and to discover how they are regulated, little is known about biofilm structure at the level of individual cells. Here, we use state-of-the-art microscopy techniques to enable live single-cell resolution imaging of a Vibrio cholerae biofilm as it develops from one single founder cell to a mature biofilm of 10,000 cells, and to discover the forces underpinning the architectural evolution. Mutagenesis, matrix labeling, and simulations demonstrate that surface adhesion-mediated compression causes V. cholerae biofilms to transition from a 2D branchedmorphology to a dense, ordered 3D cluster. We discover that directional proliferation of rod-shaped bacteria plays a dominant role in shaping the biofilm architecture in V. cholerae biofilms, and this growth pattern is controlled by a single gene, rbmA. Competition analyses reveal that the dense growth mode has the advantage of providing the biofilm with superior mechanical properties. Our single-cell technology can broadly link genes to biofilm fine structure and provides a route to assessing cell-to-cell heterogeneity in response to external stimuli.