Knief Claudia, Frances Lisa, Cantet Franck, Vorholt Julia A
Institute of Microbiology, ETH Zurich, Wolfgang-Pauli-Strasse 10, 8093 Zurich, Switzerland.
Appl Environ Microbiol. 2008 Apr;74(7):2218-28. doi: 10.1128/AEM.02532-07. Epub 2008 Feb 8.
Bacteria of the genus Methylobacterium are widespread in the environment, but their ecological role in ecosystems, such as the plant phyllosphere, is not very well understood. To gain better insight into the distribution of different Methylobacterium species in diverse ecosystems, a rapid and specific cultivation-independent method for detection of these organisms and analysis of their community structure is needed. Therefore, 16S rRNA gene-targeted primers specific for this genus were designed and evaluated. These primers were used in PCR in combination with a reverse primer that binds to the tRNA(Ala) gene, which is located upstream of the 23S rRNA gene in the 16S-23S intergenic spacer (IGS). PCR products that were of different lengths were obtained due to the length heterogeneity of the IGS of different Methylobacterium species. This length variation allowed generation of fingerprints of Methylobacterium communities in environmental samples by automated ribosomal intergenic spacer analysis. The Methylobacterium communities on leaves of different plant species in a natural field were compared using this method. The new method allows rapid comparisons of Methylobacterium communities and is thus a useful tool to study Methylobacterium communities in different ecosystems.
甲基杆菌属细菌在环境中广泛分布，但其在生态系统（如植物叶际）中的生态作用尚未得到很好的理解。为了更好地了解不同甲基杆菌属物种在不同生态系统中的分布情况，需要一种快速且特异的非培养方法来检测这些微生物并分析其群落结构。因此，设计并评估了针对该属的16S rRNA基因特异性引物。这些引物与结合到tRNA(Ala)基因的反向引物一起用于PCR，tRNA(Ala)基因位于16S - 23S基因间隔区（IGS）中23S rRNA基因的上游。由于不同甲基杆菌属物种IGS的长度异质性，获得了不同长度的PCR产物。这种长度变异使得通过自动核糖体基因间隔区分析能够生成环境样品中甲基杆菌群落的指纹图谱。使用该方法比较了自然田间不同植物物种叶片上的甲基杆菌群落。这种新方法能够快速比较甲基杆菌群落，因此是研究不同生态系统中甲基杆菌群落的有用工具。
