School of Oceanography, University of Washington, Seattle, WA, USA.
ISME J. 2013 Oct;7(10):2023-33. doi: 10.1038/ismej.2013.75. Epub 2013 May 9.
Archaeal ammonia oxidizers (AOAs) are increasingly recognized as prominent members of natural microbial assemblages. Evidence that links the presence of AOA with in situ ammonia oxidation activity is limited, and the abiotic factors that regulate the distribution of AOA natural assemblages are not well defined. We used quantitative PCR to enumerate amoA (encodes α-subunit of ammonia monooxygenase) abundances; AOA amoA gene copies greatly outnumbered ammonia-oxidizing bacteria and amoA transcripts were derived primarily from AOA throughout the water column of Hood Canal, Puget Sound, WA, USA. We generated a Michaelis-Menten kinetics curve for ammonia oxidation by the natural community and found that the measured Km of 98±14 nmol l(-1) was close to that for cultivated AOA representative Nitrosopumilus maritimus SCM1. Temperature did not have a significant effect on ammonia oxidation rates for incubation temperatures ranging from 8 to 20 °C, which is within the temperature range for depths of measurable ammonia oxidation at the site. This study provides substantial evidence, through both amoA gene copies and transcript abundances and the kinetics response, that AOA are the dominant active ammonia oxidizers in this marine environment. We propose that future ammonia oxidation experiments use a Km for the natural community to better constrain ammonia oxidation rates determined with the commonly used (15)NH4(+) dilution technique.
古菌氨氧化菌 (AOA) 日益被认为是自然微生物组合中的重要成员。将 AOA 的存在与原位氨氧化活性联系起来的证据有限,并且调节 AOA 自然组合分布的非生物因素尚未明确定义。我们使用定量 PCR 来计数 amoA(编码氨单加氧酶的α亚基)丰度;在整个水柱中,AOA amoA 基因拷贝数大大超过氨氧化菌,并且 amoA 转录本主要来自 AOA。我们为自然群落的氨氧化生成了米氏动力学曲线,发现测量的 Km 值为 98±14 nmol l(-1),接近培养的 AOA 代表 Nitrosopumilus maritimus SCM1 的 Km 值。温度对 8 至 20°C 范围内的孵育温度的氨氧化速率没有显著影响,这在该地点可测量氨氧化深度的温度范围内。这项研究通过 amoA 基因拷贝和转录本丰度以及动力学反应提供了充分的证据,表明 AOA 是该海洋环境中主要的活性氨氧化菌。我们建议未来的氨氧化实验使用自然群落的 Km 值来更好地约束使用常用的 (15)NH4(+) 稀释技术确定的氨氧化速率。