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
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
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DOI：
10.1111/1462-2920.13345
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发表时间：
2016-08-01
影响因子：
5.1
通讯作者：
Murrell, J. Colin
中科院分区：
文献类型：
--
作者：
El Khawand, Myriam;Crombie, Andrew T.;Murrell, J. Colin
生物源性挥发性有机化合物（bVOCs）的排放是全球碳循环的重要组成部分，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。植物向大气排放异戊二烯的数量与所有来源的甲烷排放量相似，各占挥发性有机化合物总量的约三分之一。虽然能够在甲烷上生长的甲烷氧化菌已经被深入研究，但我们对异戊二烯的生物降解知之甚少。本文报道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（编码保守异戊二烯单加氧酶活性位点成分的基因）的聚合酶链反应（PCR）引物的设计和测试，该引物能够从富含异戊二烯的环境样品中检索isoA序列。利用生物合成的c -13标记异戊二烯进行稳定同位素探测实验，鉴定了土壤中具有活性的异戊二烯降解菌。本研究首次利用培养依赖性和非培养依赖性方法鉴定出新的异戊二烯降解菌株，为进一步研究异戊二烯降解细菌的生物地理学和分子生态学提供了工具和基础。
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 C-13-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.