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Diversity of methanotrophs in Zoige wetland soils under both anaerobic and aerobic conditions
Authors:Juanli Yun  Anzhou M  Yaoming Li  Guoqiang Zhuang  Yanfen Wang and Hongxun Zhang
Institution:1. Graduate University of Chinese Academy of Sciences,Beijing 100049,China
2. Research Center for Eco-Environment Sciences,Chinese Academy of Sciences,Beijing 100085,China
Abstract:Zoige wetland is one of the most important methane emission centers in China. The oxidation of methane in the wetland a ects global warming, soil ecology and atmospheric chemistry. Despite their global significance, microorganisms that consume methane in Zoige wetland remain poorly characterized. In this study, we investigated methanotrophs diversity in soil samples from both anaerobic site and aerobic site in Zoige wetland using pmoA gene as a molecular marker. The cloning library was constructed according to the pmoA sequences detected. Four clusters of methanotrophs were detected. The phylogenetic tree showed that all four clusters detected were a liated to type I methanotrophs. Two novel clusters (cluster 1, cluster 2) were found to relate to none of the recognized genera of methanotrophs. These clusters have no cultured representatives and reveal an ecological adaptation of particular uncultured methanotrophs in Zoige wetland. Two clusters were belonging to Methylobacter and Methylococcus separately. Denaturing gradient gel electrophoresis gel bands pattern retrieved from these two samples revealed that the community compositions of anaerobic soil and aerobic soil were di erent from each other while anaerobic soil showed a higher metanotrophs diversity. Real-time PCR assays of the two samples demonstrated that aerobic soil sample in Zoige wetland was 1.5 times as much copy numbers as anaerobic soil. These data illustrated that methanotrophs are a group of microorganisms influence the methane consumption in Zoige wetland.
Keywords:cloning library  denaturing gradient gel electrophoresis  methanotrophs  pmoA gene  real-time PCR  Zoige wetland
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