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中国东北大豆和稻田土壤中厌氧氨氧化菌和氨氧化古菌的逆转录基因表达。

Reverse-transcriptional gene expression of anammox and ammonia-oxidizing archaea and bacteria in soybean and rice paddy soils of Northeast China.

出版信息

Appl Microbiol Biotechnol. 2014 Mar;98(6):2675-86. doi: 10.1007/s00253-013-5242-x.

Abstract

The relative gene expression of hydrazine oxidoreductase encoding gene (hzo) for anaerobic ammonium oxidizing bacteria (anammox) and ammonia monooxygenase encoding gene (amoA) for both ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB) in Sanjiang Plain soybean and rice paddy soils of Northeast China was investigated by using real-time reverse-transcriptional quantitative PCR. Metabolically active populations of anammox, AOA, and AOB in rice paddy soils were evident by the presence and successful quantification of hzo mRNA and amoA mRNA genes. The expression ratio of amoA gene for both AOA and AOB varied between soybean soils and different rice paddy soils while the expression of hzo gene for anammox was detectable only in rice paddy soils by showing a diverse relative expression ratio in each soil sample. Gene expression of both archaeal and bacterial amoA genes in rice paddy soils differed among the three sampling depths, but that of hzo was not. Both archaeal and bacterial amoA genes showed an increase trend of expression level with continuation of rice paddy cultivation, but the low expression ratio of hzo gene indicated a relatively small contribution of anammox in overall removal of inorganic nitrogen through N2 even under anoxic and high nitrogen input in agriculture. Bacterial amoA gene from two soybean fields and three rice paddy fields were also analyzed for community composition by denaturing gradient gel electrophoresis fingerprint. Community shift was observed between soybean and paddy fields and within each of them. The consistent occurrence of three bands 5, 6, and 7 in all samples showed their high adaptability for both arid cultivation and continuous rice paddy cultivation. Our data suggest that AOA and AOB are playing a more important role in nitrogen transformation in agricultural soils in oxic or anoxic environment and anammox bacteria may also contribute but in a less extent to N transformation in these agricultural soils under anoxic condition.

摘要

采用实时反转录定量 PCR 法,研究了中国东北三江平原大豆和水稻田土壤中厌氧氨氧化菌(anammox)肼氧化还原酶编码基因(hzo)和氨氧化古菌(AOA)和氨氧化菌(AOB)氨单加氧酶编码基因(amoA)的相对基因表达。通过 hzo mRNA 和 amoA mRNA 基因的存在和成功定量,证明了水稻田土壤中 anammox、AOA 和 AOB 的代谢活性种群的存在。在不同的水稻田土壤中,AOA 和 AOB 的 amoA 基因表达比例存在差异,而仅在水稻田土壤中检测到 anammox 的 hzo 基因表达,且在每个土壤样本中表现出不同的相对表达比例。水稻田土壤中古菌和细菌 amoA 基因的表达在三种采样深度之间存在差异,但 hzo 基因的表达则不然。古菌和细菌 amoA 基因的表达水平均随水稻田的持续种植而呈上升趋势,但 hzo 基因的低表达比例表明,即使在农业中缺氧和高氮输入的情况下,anammox 通过 N2 对无机氮的去除作用也相对较小。还对来自两个大豆田和三个水稻田的细菌 amoA 基因进行了变性梯度凝胶电泳指纹图谱分析,以研究其群落组成。在大豆田和水稻田之间以及它们各自内部都观察到群落的转移。所有样本中均一致出现 5、6 和 7 三个条带,表明它们对旱地和连续水稻种植具有高度的适应性。我们的数据表明,在有氧或缺氧环境下,AOA 和 AOB 在农业土壤中的氮转化中发挥着更为重要的作用,而 anammox 细菌也可能会有所贡献,但在缺氧条件下,其对这些农业土壤中的氮转化的贡献较小。

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