厌氧氨氧化和反硝化厌氧甲烷氧化细菌在土壤中的分布、 活性及其应用
文献类型：学位论文
| 作者 | 夏超 |
| 学位类别 | 硕士 |
| 答辩日期 | 2015-05 |
| 授予单位 | 中国科学院研究生院 |
| 授予地点 | 北京 |
| 导师 | 祝贵兵 |
| 关键词 | 厌氧氨氧化细菌，反硝化厌氧甲烷氧化细菌，土壤，分布，活性，anammox bacteria, n-damo bacteria, soil, distribution, activity |
| 其他题名 | Distribution, Activity and Application of Anaerobic Ammonia Oxidizing Bacteria and Nitrite-Dependent Anaerobic Methane Oxidizing Bacteria in Soil |
| 学位专业 | 环境工程 |
| 中文摘要 | 厌氧氨氧化（anaerobic ammonium oxidation，anammox）和反硝化厌氧甲烷氧化（nitrite-dependent anaerobic methane oxidation，n-damo）的发现打破了人们长久以来对生物氮、碳循环的传统认识。厌氧氨氧化细菌（anammox bacteria）和反硝化厌氧甲烷氧化细菌（ n-damo bacteria）在水生态系统中均有分布，并且在全球氮循环和缓解温室效应中发挥重要作用。 海洋沉积物、海水、湿地沉积物和稻田土壤中普遍存在厌氧氨氧化细菌，但是从陆地的草原土壤、森林土壤和旱地土壤中无法检测到厌氧氨氧化细菌。土壤的含水率，特别是土壤中是否存在着上覆水、重力水和地下水等自由水，可能是决定厌氧氨氧化细菌在土壤中分布规律的重要生态因子。反硝化厌氧甲烷氧化细菌在稻田、河流、湖泊、泥炭沼泽等湿地生态系统中有分布，但是其在陆地土壤中的分布规律仍不甚清楚，决定其分布的关键生态因子也是未知的。 故本研究以不同类型土壤（河口潮间带、湖泊岸边带、稻田土壤、旱地土壤和极端环境沉积物）作为研究对象，应用分子生物学、环境理化因子分析、稳定同位素示踪和人工模拟培养等方法，对厌氧氨氧化细菌和反硝化厌氧甲烷氧化细菌进行研究，以期探明这两类微生物在土壤中的分布规律、活性和重要生态因子。 河口潮间带表层土壤在潮汐的作用下，普遍分布着厌氧氨氧化细菌（1.86×103 ~8.60×106 copies g-1）。湖泊岸边带表层土壤在未被水淹的状态下无法检测到厌氧氨氧化细菌，但是在被湖水长期淹没后能从中检测到厌氧氨氧化细菌（1.97×104~4.34×106 copies g-1）。湖泊岸边带垂向土壤受地下水的作用，其表层无厌氧氨氧化细菌分布，其分布的热区位于地下水位附近及以下深层土壤（1.75×107~6.10×107 copies g-1）。稻田垂向土壤受到灌溉的影响，其厌氧氨氧化细菌分布热区位于地下水位水位线以上的区域（ 6.90×105~7.66×106 copiesg-1）。结果说明厌氧氨氧化细菌在土壤中的存在需要特定的生态位，土壤是否与地下水或者地表水等自由水接触对厌氧氨氧化细菌的分布有决定性影响。 另外，厌氧氨氧化细菌存在于一些典型极端水环境中并且发生厌氧氨氧化反应，其在温度、pH和底物浓度方面有着广阔的生态位，这些因子均不是决定厌氧氨氧化细菌存在与否的关键生态因子。未检测到厌氧氨氧化细菌的旱地、林地表层土壤经过加水长期人工模拟培养后，厌氧氨氧化细菌成功得到激活、富集。结果证明决定厌氧氨氧化细菌在土壤分布的关键限制性生态因子是土壤自由水。并不是所有河口潮间带表层土壤中均存在M.oxyfera-like 细菌（0~3.26×106 copies g-1）。湖泊岸边带表层土壤长期处于落干状态时无法从中检测到M.oxyfera-like细菌，但被湖水长期淹没后可以检测到M.oxyfera-like细菌（1.28×104~2.28×105 copies g-1）。湖泊岸边带不同深度的垂向土壤都存在M.oxyfera-like细菌，其分布热区位于地下水位线附近及以下的深层土壤（1.05×105~1.49×106 copies g-1）。稻田土壤 M. oxyfera-like细菌丰度在表面两层较低，其分布的热区位于地下水位以上的区域（ 1.15×103~5.56×106 copies g-1）。 结果说明 M. oxyfera-like在土壤中的存在需要特定的生态位，土壤自由水状态是影响其分布的重要但非唯一限制性生态因子。 研究结果修正和完善了微生物氮循环过程理论体系，为厌氧氨氧化反应和反硝化厌氧甲烷氧化在土壤中的研究起到向导作用。 |
| 英文摘要 | The discoveries of anaerobic ammonium oxidation (anammox) and nitrite-dependent anaerobic methane oxidation (n-damo) changed the traditional knowledge about biological nitrogen and carbon cycle. Both anaerobic ammonium oxidation bacteria (anammox bacteria) and nitrite-dependent anaerobic methane oxidation bacteria (n-damo bacteria) occur in water ecosystem, and play important roles in the global nitrogen cycle and the remission of greenhouse effect. Anammox bacteria have widespread occurrence in ocean sediments, seawater, wetland sediments and paddy field. But they can’t be found in steppe soil, forest soil and dry land in terrestrial ecosystem. Soil moisture, whether the soil has free water such as overlying water, gravity water and groundwater, to be specific, may be the important ecological factor that could determine the occurrence of anammox bacteria in soil. N-damo bacteria have been found in wetland ecosystem, such as paddy fields,rivers, lakes and peat bogs. But the distribution and the key ecological factors, which determine the occurrence, of n-damo bacteria in soil are still unclear. So this paper took typical soil (estuarine intertidal zone, lake riparian zone, paddy soil, dry land soil and sediments from extreme environments) as research objects to explore the distribution, activity and important environmental factor of anammox bacteria and n-damo bacteria in soil, using molecular biology techniques, physicochemical factors analysis, isotope tracing techniques and artificial cultivation. Anammox bacteria widely occurred in estuarine intertidal zone (1.86×103~8.60×106 copies g-1), with influences of tide. Anammox bacteria didn’t occur in surface soil in lake riparian when the soil wasn’t flooded. But when the soil was flooded by lake water for long time, anammox bacteria could be detected (1.97×104~4.34×106 copies g-1). Anammox bacteria were not detected in surface soil in lake riparian zone, and the hotspots were located nearby and below groundwater level (1.75×107~ 6.10×107 copies g-1) because of the effect of ground water. The hotspots of anammox bacterial abundance were located above the water level in paddy field (2.01×104~7.66×106 copies g-1) with the influences of irrigation. Anammox bacteria not ubiquitously occur in soil ecosystems reflects their specific ecological requirements, that is, the free water such as overlying water and groundwater in soil is the decisive factor that stimulate the occurrence of anammox bacteria. Moreover, anammox bacteria exist in some typical extreme environments (6.94×104~8.05×106 copies g-1), such as high and low temperature, high and pH and eutrophic condition. This result indicates that environmental temperature, pH and nutrition conditions are not the important ecological factor that determine the occurrence of anammox bacteria in soil. And in the dry land soil and the woodland surface soil where anammox bacteria weren’t detected before, anammox bacteria were successfully activated and enriched after adding water for long time, indicating that the key restriction factor is the condition of free water in soil. M. oxyfera-like bacteria could be found in every samples from estuarine intertidal zone (0~3.26×106 copies g-1). M. oxyfera-like bacteria didn’t occur in surface soil in lake riparian when the soil wasn’t flooded. But when the soil was flooded by lake water for long time, M. oxyfera-like bacteria could be detected (1.28×104~2.28×105 copies g-1). M. oxyfera-like bacteria occurred in all depths of soil in lake riparian zone and the hotspots were located nearby and below groundwater (1.05×105~1.49×106 copies g-1). The abundances of M. oxyfera-like bacteria of paddy soil were low in the surface layers and the hotspots were located above the groundwater (1.15×103~5.56×106 copies g-1). The results indicate that M. oxyfera-like bacteria need specific ecological requirements to occur in soil., and the free water in soil is an important but not unique factor. Hopefully, our research results will revise and improve the theoretical system of the nitrogen cycle, and play the guide role for the research on anammox and n-damo in soil. |
| 源URL | [http://ir.rcees.ac.cn/handle/311016/34446]  |
| 专题 | 生态环境研究中心_环境水质学国家重点实验室 |
| 推荐引用方式 GB/T 7714 | 夏超. 厌氧氨氧化和反硝化厌氧甲烷氧化细菌在土壤中的分布、 活性及其应用[D]. 北京. 中国科学院研究生院. 2015. |
入库方式： OAI收割
来源：生态环境研究中心
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