Department of Microbiology, University of Washington, Seattle, WA 98195, USA.
ISME J. 2010 Mar;4(3):388-98. doi: 10.1038/ismej.2009.117. Epub 2009 Nov 19.
Methylotrophs, organisms able to gain energy and carbon from compounds containing no carbon-carbon bonds, such as methane, methanol and methylated amines, are widespread in nature. However, knowledge of their nutrient preference and their metabolism is mostly based on experiments with cultures grown in defined laboratory conditions. Here, we use transcriptomics to explore the activity of one methylotroph, Methyotenera mobilis in its natural environment, lake sediment from which it has been previously isolated. Cells encapsulated in incubation cassettes were exposed to sediment conditions, with or without supplementation with a carbon/energy source (methylamine), and gene-expression patterns were compared for those cells to patterns for cells incubated in a defined medium supplemented with methylamine. A few specific trends in gene expression were observed at in situ conditions that may be of environmental significance, as follows. Expression of genes for the linear formaldehyde oxidation pathway linked to tetrahydromethanopterin increased, suggesting an important role for this pathway in situ, in contrast to laboratory condition culture, in which the cyclic ribulose monophosphate pathway seemed to be the major route for formaldehyde oxidation. Along with the ribulose monophosphate cycle that is also a major pathway for assimilating C(1) units, the methylcitric acid cycle seemd to be important in situ, suggesting that multicarbon compounds may be the natural carbon and/or energy substrates for M. mobilis, challenging the notion of an obligately methylotrophic lifestyle for this bacterium. We also detected a major switch in expression of genes responsible for the mode of motility between different conditions: from flagellum-enabled motility in defined medium to in situ expression of pili known to be involved in twitching motility and adherence. Overall, this study offers a novel approach for gaining insights into the lifestyle of individual microbes in their native environments.
能够从不含碳碳键的化合物(如甲烷、甲醇和甲基化胺)中获取能量和碳的甲基营养菌在自然界中广泛存在。然而,关于它们的营养偏好和代谢的知识主要是基于在定义明确的实验室条件下培养的实验。在这里,我们使用转录组学来探索一种甲基营养菌 Methyotenera mobilis 在其自然环境中的活性,该菌先前从其分离的湖泊沉积物中分离出来。将细胞封装在孵育盒中,暴露于沉积物条件下,无论是否添加碳/能源源(甲胺),并将这些细胞的基因表达模式与在补充有甲胺的定义培养基中孵育的细胞的基因表达模式进行比较。在原位条件下观察到一些特定的基因表达趋势,这些趋势可能具有环境意义,如下所示。与实验室条件培养相比,与四氢甲酰喋呤相连的线性甲醛氧化途径的基因表达增加,表明该途径在原位具有重要作用,而在实验室条件培养中,环状核酮糖 1-磷酸途径似乎是甲醛氧化的主要途径。与也是同化 C1 单位的主要途径的核酮糖 1-磷酸循环一起,甲基柠檬酸循环在原位似乎很重要,这表明多碳化合物可能是 M. mobilis 的天然碳和/或能源底物,这对该细菌的严格甲基营养生活方式的概念提出了挑战。我们还检测到与不同条件下的运动模式相关的基因表达的主要开关:从定义培养基中的鞭毛运动到已知参与蠕动运动和粘附的纤毛的原位表达。总的来说,这项研究为深入了解个体微生物在其原生环境中的生活方式提供了一种新方法。