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Isolation of isoprene degrading bacteria from soils, development of isoA gene probes and identification of the active isoprene-degrading soil community using DNA-stable isotope probing

机译:从土壤中分离异戊二烯降解细菌,isoA基因探针的开发以及使用DNA稳定同位素探测技术鉴定降解异戊二烯的活性土壤群落

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摘要

© 2016 The Authors. Environmental Microbiology published by Society for Applied Microbiology and John Wiley & Sons Ltd. Emissions of biogenic volatile organic compounds (bVOCs), are an important element in the global carbon cycle, accounting for a significant proportion of fixed carbon. They contribute directly and indirectly to global warming and climate change and have a major effect on atmospheric chemistry. Plants emit isoprene to the atmosphere in similar quantities to emissions of methane from all sources and each accounts for approximately one third of total VOCs. Although methanotrophs, capable of growth on methane, have been intensively studied, we know little of isoprene biodegradation. Here, we report the isolation of two isoprene-degrading strains from the terrestrial environment and describe the design and testing of polymerase chain reaction (PCR) primers targeting isoA, the gene encoding the active-site component of the conserved isoprene monooxygenase, which are capable of retrieving isoA sequences from isoprene-enriched environmental samples. Stable isotope probing experiments, using biosynthesized (13) C-labelled isoprene, identified the active isoprene-degrading bacteria in soil. This study identifies novel isoprene-degrading strains using both culture-dependent and, for the first time, culture-independent methods and provides the tools and foundations for continued investigation of the biogeography and molecular ecology of isoprene-degrading bacteria.
机译:©2016作者。应用微生物学协会和John Wiley&Sons Ltd.出版的环境微生物学。生物挥发性有机化合物(bVOC)的排放是全球碳循环中的重要元素,占固定碳的很大比例。它们直接或间接地导致全球变暖和气候变化,并对大气化学产生重大影响。植物向大气中排放的异戊二烯的量与所有来源的甲烷排放量相似,每种植物约占总VOC的三分之一。尽管对能够在甲烷上生长的甲烷营养生物进行了深入研究,但我们对异戊二烯的生物降解知之甚少。在这里,我们报告了从陆地环境中分离出的两种异戊二烯降解菌株,并描述了针对isoA的聚合酶链反应(PCR)引物的设计和测试,isoA是编码保守的异戊二烯单加氧酶活性位点成分的基因,能够从富含异戊二烯的环境样品中检索isoA序列的方法。使用生物合成的(13)C标记异戊二烯进行的稳定同位素探测实验,确定了土壤中降解异戊二烯的活性细菌。这项研究确定了新的异戊二烯降解菌株,同时使用了依赖于培养物的方法,并且首次使用了不依赖培养物的方法,并为继续研究降解异戊二烯的细菌的生物地理学和分子生态学提供了工具和基础。

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