The Pseudomonas fluorescens SBW25 wrinkly spreader biofilm requires attachment factor, cellulose fibre and LIPS interactions to maintain strength and integrity
The Pseudomonas fluorescens SBW25 wrinkly spreader biofilm requires attachment factor, cellulose fibre and LIPS interactions to maintain strength and integrity
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DOI：
10.1099/mic.0.27984-0
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发表时间：
2005-09-01
期刊：
影响因子：
2.8
通讯作者：
Rainey, PB
中科院分区：
文献类型：
--
作者：
Spiers, AJ;Rainey, PB
荧光假单胞菌SBW25的皱褶扩散（WS）分离物在气液界面形成实质性的生物膜。该生物膜由细胞外部分乙酰化的纤维素纤维基质组成，以前用mini-Tn5诱变WS已经确定了调控和纤维素生物合成操纵子。一个未被鉴定的WS突变体WS-5仍然表达纤维素，但产生非常弱的生物膜。在这项工作中，WS-5中的mini-Tn5插入位点已被确定为直接位于to/-pal操纵子的上游。与其他革兰氏阴性菌的toll - pal突变体一样，WS-5表现出“漏膜”表型，包括偶然利用蔗糖的能力，对亲水性染料碘化丙啶的吸收增加，以及脂多糖（LPS）表达的丧失。WS-5细胞的相对疏水性发生改变，在生物膜中的募集和维持能力较WS差。WS-5生物膜在发展期间对化学干扰也不太敏感。而WS-5与WS-5的生长速率、纤维素表达量和附着性无显著差异。最后，WS-5生物膜可以与表达LPS的生物膜和附着缺陷突变体WS-4部分互补，从而形成强度显著提高的混合生物膜。这些发现表明，WS气液生物膜强度的一个主要组成部分来自LPS与生物膜的纤维素基质之间的相互作用，并且在WS生物膜中，纤维素纤维、附着因子和LPS是生物膜发育、强度和完整性所必需的。
The wrinkly spreader (WS) isolate of Pseudomonas fluorescens SBW25 forms a substantial biofilm at the air-liquid interface. The biofilm is composed of an extracellular partially acetylated cellulose-fibre matrix, and previous mutagenesis of WS with mini-Tn5 had identified both the regulatory and cellulose-biosynthetic operons. One uncharacterized WS mutant, WS-5, still expressed cellulose but produced very weak biofilms. In this work, the mini-Tn5 insertion site in WS-5 has been identified as being immediately upstream of the to/-pal operon. Like Tol-Pal mutants of other Gram-negative bacteria, WS-5 showed a 'leaky-membrane' phenotype, including the serendipitous ability to utilize sucrose, increased uptake of the hydrophilic dye propidium iodide, and the loss of lipopolysaccharide (LPS) expression. WS-5 cells were altered in relative hydrophobicity, and showed poorer recruitment and maintenance in the biofilm than WS. The WS-5 biofilm was also less sensitive to chemical interference during development. However, growth rate, cellulose expression and attachment were not significantly different between WS and WS-5. Finally, WS-5 biofilms could be partially complemented with WS-4, a biofilm- and attachment-deficient mutant that expressed LPS, resulting in a mixed biofilm with significantly increased strength. These findings show that a major component of the WS air-liquid biofilm strength results from the interactions between LPS and the cellulose matrix of the biofilm - and that in the WS biofilm, cellulose fibres, attachment factor and LPS are required for biofilm development, strength and integrity.