Advanced Understanding of Prokaryotic Biofilm Formation through Use of a Cost-Effective and Versatile Multipanel Adhesion (mPAD) Mount
Advanced Understanding of Prokaryotic Biofilm Formation through Use of a Cost-Effective and Versatile Multipanel Adhesion (mPAD) Mount
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通过使用经济高效且多功能的多面板粘附 (mPAD) 安装座深入了解原核生物膜的形成
DOI：
10.1128/aem.02283-21
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发表时间：
2022
影响因子：
4.4
通讯作者：
Pohlschroder, Mechthild
中科院分区：
文献类型：
--
作者：
Schulze, Stefan;Schiller, Heather;Solomonic, Jordan;Telhan, Orkan;Costa, Kyle;Pohlschroder, Mechthild
大多数微生物存在于生物膜中，生物膜包括被细胞外基质包围的细胞聚集体，细胞外基质提供保护免受外部应力。基于它们形成的条件，生物膜结构以显著的方式变化。例如，当微生物在静态条件下孵育时形成的生物膜不同于当微生物遇到流动液体的剪切力时形成的生物膜。此外，生物膜随时间动态地发展。在这里，我们描述了一种具有成本效益的盖玻片保持器，用三维（3D）打印机打印，有利于在广泛的站立和摇动培养条件下的表面粘附测定。这种多板粘附（mPAD）安装进一步允许在多个时间点对培养物进行采样，确保样品之间的一致性和可比性，并能够分析生物膜形成的动力学。作为原理证明，使用mPAD底座进行振荡、好氧培养，我们证实了先前的流动室实验，该实验表明铜绿假单胞菌野生型菌株和吩嗪缺失突变株（Δphz）菌株形成具有相似结构但突变株密度降低的生物膜。将该分析扩展到缺氧条件下，我们揭示了微菌落形成和生物膜形成只能在振荡条件下观察到，并且与野生型培养物相比，Δ phz突变体中的微菌落形成和生物膜形成减少，表明如果氧气作为电子受体不可用，则吩嗪对于生物膜的形成至关重要。此外，虽然模型古菌Haloferax volcanii在静态条件下不需要古菌的表面附着，我们证明了一个H。缺乏古细菌的volcanii突变体在摇动条件下生物膜形成的早期阶段受到损害。重要性由于mPAD安装的多功能性，我们预期它将有助于分析广泛范围的细菌和古细菌中的生物膜形成。因此，它有助于回答有关生物膜形成的调控和结构成分的关键生物学问题，并在广泛的环境，生物技术和医学背景下理解这一过程。
Most microorganisms exist in biofilms, which comprise aggregates of cells surrounded by an extracellular matrix that provides protection from external stresses. Based on the conditions under which they form, biofilm structures vary in significant ways. For instance, biofilms that develop when microbes are incubated under static conditions differ from those formed when microbes encounter the shear forces of a flowing liquid. Moreover, biofilms develop dynamically over time. Here, we describe a cost-effective coverslip holder, printed with a three-dimensional (3D) printer, that facilitates surface adhesion assays under a broad range of standing and shaking culture conditions. Thismultipaneladhesion (mPAD) mount further allows cultures to be sampled at multiple time points, ensuring consistency and comparability between samples and enabling analyses of the dynamics of biofilm formation. As a proof of principle, using the mPAD mount for shaking, oxic cultures, we confirm previous flow chamber experiments showing that the Pseudomonas aeruginosa wild-type strain and a phenazine deletion mutant (Δphz) strain form biofilms with similar structure but reduced density in the mutant strain. Extending this analysis to anoxic conditions, we reveal that microcolony formation and biofilm formation can only be observed under shaking conditions and are decreased in the Δphzmutant compared to wild-type cultures, indicating that phenazines are crucial for the formation of biofilms if oxygen as an electron acceptor is unavailable. Furthermore, while the model archaeon Haloferax volcanii does not require archaella for surface attachment under static conditions, we demonstrate that an H. volcanii mutant that lacks archaella is impaired in early stages of biofilm formation under shaking conditions.IMPORTANCEDue to the versatility of the mPAD mount, we anticipate that it will aid the analysis of biofilm formation in a broad range of bacteria and archaea. Thereby, it contributes to answering critical biological questions about the regulatory and structural components of biofilm formation and understanding this process in a wide array of environmental, biotechnological, and medical contexts.
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影响因子：
4.8
作者：
Schiller H;Schulze S;Mutan Z;de Vaulx C;Runcie C;Schwartz J;Rados T;Bisson Filho AW;Pohlschroder M
通讯作者：
Pohlschroder M
Haloferax volcanii 鞭毛是运动所必需的，但不参与 PibD 依赖性表面粘附
影响因子：
3.2
作者：
M. Tripepi;Saheed Imam;M. Pohlschröder
通讯作者：
M. Pohlschröder
影响因子：
3.6
作者：
O'Toole, GA;Kolter, R
通讯作者：
Kolter, R
影响因子：
2.8
作者：
de Silva, Roshali T.;Abdul-Halim, Mohd F.;Duggin, Iain G.
通讯作者：
Duggin, Iain G.
影响因子：
4.4
作者：
Holten，Matthew P.;Fonseca，Dallas R.;Costa，Kyle C.
通讯作者：
Costa，Kyle C.