Bomar Lindsey, Graf Joerg
Department of Molecular and Cell Biology, University of Connecticut, Storrs, Connecticut 06269, USA.
Biol Bull. 2012 Aug;223(1):155-66. doi: 10.1086/BBLv223n1p155.
Host-associated microbial communities are widespread in nature and vital to the health and fitness of the host. Deciphering the physiology of the microbiome in vivo is critical to understanding the molecular basis of the symbiosis. Recently, the development and application of high-throughput sequencing techniques, particularly RNA-seq, for studying microbial communities has enabled researchers to address not only which microbes are present in a given community but also how the community functions. For microbes that can also be cultivated in the laboratory, RNA-seq provides the opportunity to identify genes that are differentially expressed during symbiosis by comparing in vitro to in vivo transcriptomes. In the current study, we used RNA-seq to identify genes expressed by the digestive-tract microbiome of the medicinal leech, Hirudo verbana, and by one of the two dominant symbionts, Aeromonas veronii, in a rich medium. We used a comparative approach to identify genes differentially expressed during symbiosis and gain insight into the symbiont's physiology in vivo. Notable findings include evidence for the symbionts experiencing environmental stress, performing arginine catabolism, and expressing noncoding RNAs that are implicated in stationary phase survival, a state in which A. veronii persists for months within the host.
与宿主相关的微生物群落广泛存在于自然界中,对宿主的健康和适应性至关重要。解析体内微生物组的生理学对于理解共生的分子基础至关重要。最近,用于研究微生物群落的高通量测序技术,特别是RNA测序的发展和应用,使研究人员不仅能够确定特定群落中存在哪些微生物,还能了解群落的功能。对于那些也能在实验室中培养的微生物,RNA测序提供了通过比较体外和体内转录组来鉴定共生过程中差异表达基因的机会。在当前的研究中,我们使用RNA测序来鉴定药用蛭(Hirudo verbana)消化道微生物组以及两种主要共生菌之一的维氏气单胞菌(Aeromonas veronii)在丰富培养基中表达的基因。我们采用比较方法来鉴定共生过程中差异表达的基因,并深入了解共生菌在体内的生理学。值得注意的发现包括共生菌经历环境压力、进行精氨酸分解代谢以及表达与稳定期存活相关的非编码RNA的证据,在稳定期维氏气单胞菌可在宿主体内持续存在数月。