Griffiths Robert I, Whiteley Andrew S, O'Donnell Anthony G, Bailey Mark J
Molecular Microbial Ecology Laboratory, CEH-Oxford, Oxford OX1 3SR, United Kingdom.
Appl Environ Microbiol. 2003 Dec;69(12):6961-8. doi: 10.1128/AEM.69.12.6961-6968.2003.
The effects of water stress upon the diversity and culturable activity of bacterial communities in the rhizosphere of an established upland grassland soil have been investigated. Intact monoliths were subjected to different watering regimens over a 2-month period to study community adaptation to moisture limitation and subsequent response to stress alleviation following rewetting. Genetic diversity was analyzed with 16S-based denaturing gradient gel electrophoresis (DGGE) of total soil-extracted DNA (rRNA genes) and RNA (rRNA transcripts) in an attempt to discriminate between total and active communities. Physiological response was monitored by plate counts, total counts, and BIOLOG-GN2 substrate utilization analyses. Controlled soil drying decreased the total number of CFU on all the media tested and also decreased the substrate utilization response. Following rewetting of dried soil, culture-based analyses indicated physiological recovery of the microbial population by the end of the experiment. In contrast, DGGE analyses of community 16S rRNA genes, rRNA transcripts and cultured communities did not reveal any changes relating to the moisture regimens, despite the observed physiological effects. We conclude that the imposed moisture regimen modulated the physiological status of the bacterial community and that bacterial communities in this soil are resistant to water stress. Further, we highlight the need for a reexamination of rRNA transcript-based molecular profiling techniques as a means of describing the active component of soil bacterial communities.
研究了水分胁迫对已开垦的旱地草原土壤根际细菌群落多样性和可培养活性的影响。在两个月的时间里,对完整的土柱进行不同的浇水方案处理,以研究群落对水分限制的适应性以及再湿润后对应激缓解的后续反应。通过对从土壤中提取的总DNA(rRNA基因)和RNA(rRNA转录本)进行基于16S的变性梯度凝胶电泳(DGGE)分析遗传多样性,试图区分总群落和活性群落。通过平板计数、总计数和BIOLOG-GN2底物利用分析监测生理反应。控制土壤干燥降低了所有测试培养基上的CFU总数,也降低了底物利用反应。干燥土壤再湿润后,基于培养的分析表明在实验结束时微生物种群实现了生理恢复。相比之下,尽管观察到了生理效应,但对群落16S rRNA基因、rRNA转录本和培养群落的DGGE分析并未揭示与水分方案相关的任何变化。我们得出结论,施加的水分方案调节了细菌群落的生理状态,并且该土壤中的细菌群落对水分胁迫具有抗性。此外,我们强调需要重新审视基于rRNA转录本的分子谱分析技术,将其作为描述土壤细菌群落活性成分的一种手段。