Adams J L, McLean R J
Department of Biology, Southwest Texas State University, San Marcos, Texas 78666-4616, USA.
Appl Environ Microbiol. 1999 Sep;65(9):4285-7. doi: 10.1128/AEM.65.9.4285-4287.1999.
Slow growth has been hypothesized to be an essential aspect of bacterial physiology within biofilms. In order to test this hypothesis, we employed two strains of Escherichia coli, ZK126 (DeltalacZ rpoS(+)) and its isogenic DeltarpoS derivative, ZK1000. These strains were grown at two rates (0.033 and 0.0083 h(-1)) in a glucose-limited chemostat which was coupled either to a modified Robbins device containing plugs of silicone rubber urinary catheter material or to a glass flow cell. The presence or absence of rpoS did not significantly affect planktonic growth of E. coli. In contrast, biofilm cell density in the rpoS mutant strain (ZK1000), as measured by determining the number of CFU per square centimeter, was reduced by 50% (P < 0.05). Deletion of rpoS caused differences in biofilm cell arrangement, as seen by scanning confocal laser microscopy. In reporter gene experiments, similar levels of rpoS expression were seen in chemostat-grown planktonic and biofilm populations at a growth rate of 0.033 h(-1). Overall, these studies suggest that rpoS is important for biofilm physiology.
生长缓慢被认为是生物膜内细菌生理学的一个重要方面。为了验证这一假设,我们使用了两株大肠杆菌,ZK126(DeltalacZ rpoS(+))及其同基因的DeltarpoS衍生物ZK1000。这些菌株在葡萄糖限制的恒化器中以两种速率(0.033和0.0083 h(-1))生长,该恒化器与一个装有硅橡胶导尿管材料塞子的改良Robbins装置或一个玻璃流动池相连。rpoS的存在与否对大肠杆菌的浮游生长没有显著影响。相比之下,通过测定每平方厘米的CFU数量来衡量,rpoS突变株(ZK1000)中的生物膜细胞密度降低了50%(P < 0.05)。通过扫描共聚焦激光显微镜观察,rpoS的缺失导致生物膜细胞排列的差异。在报告基因实验中,在生长速率为0.033 h(-1)的恒化器培养的浮游和生物膜群体中观察到相似水平的rpoS表达。总体而言,这些研究表明rpoS对生物膜生理学很重要。