有机肥配施氮肥条件下设施菜地硝化菌群多样性研究

发布时间：2018-02-12 16:57

本文关键词： 施肥氨氧化细菌氨氧化古菌亚硝酸盐氧化细菌群落结构　出处：《沈阳农业大学》2017年硕士论文　论文类型：学位论文

【摘要】：在土壤生态系统中,硝化作用是土壤氮素转化的关键环节,在土壤质量可持续发展、植物生长、氮肥利用率、环境污染以及食品安全等方面有重要的影响。硝化作用是由硝化微生物驱动逐步将氨盐转化为硝酸盐的过程。参与此过程的微生物统称为硝化菌群,主要包括氨氧化细菌、氨氧化古菌和亚硝酸盐氧化细菌等。施肥主要通过改变土壤中微生物多样性和群落结构来对硝化作用产生影响。本研究以沈阳农业大学长期定位施肥试验菜地土壤为试验材料,基于Biolog技术和高通量测序技术研究有机肥配施氮肥对设施菜地土壤肥力和土壤硝化菌群的影响,旨在明确硝化菌群与土壤肥力之间的互作关系,以期为研究硝化菌群尤其是土壤生态系统NOB补充施肥和设施栽培方面的数据。同时为完善设施菜地肥力评价与设施土壤健康管理提供微生物参数,为农田N20的减排以及氮肥利用率的提高提供科学基础。本研究主要结果如下:1.有机肥与氮肥配施可以提高土壤肥力和土壤综合酶活性。特别是有机肥配施氮肥可以显著提高土壤有机质、全氮、速效磷、速效钾含量;土壤铵态氮、硝态氮、碱解氮含量和电导率在不施有机肥或有机肥投入量一定时均随施氮量的增加呈现递增趋势,而pH随氮肥投入量的增加呈递减趋势。有机肥施入可以提升土壤脲酶、转化酶、纤维素分解酶、荧光素二乙酸酯酶活性。中性磷酸酶活性对施肥最为敏感,各处理之间差异较大;长期氮肥单施对过氧化氢酶有一定的抑制作用,仅高倍量氮肥(N2、MN2)对多酚氧化酶活性影响显著。2.氮肥单施抑制了土壤微生物功能多样性,而配施一定量的有机肥有利于提高微生物功能多样性,但是配施过量氮肥仍然会导致微生物功能多样性的下降。长期偏氮施肥促使氨基酸类碳源成为最主要碳源利用种类,并且施有机肥处理的碳源利用率高于氮肥单施或不施肥的处理。综合比较,以长期有机肥配施一倍量氮肥(MN1)表现最佳,N2表现最差。3.基于高通量测序技术,对设施菜地细菌与古菌概况进行初步分析。共鉴别出来包括unclassfied群体在内的门水平下37类,种水平下1507种细菌和门水平下7类,属水平下26类,种水平下40种古菌。细菌中变形菌门(Proteobacteria)是设施菜地土壤中优势群体;奇古菌门(Thaumarchaeota)是古菌中的优势菌群。施肥对各处理细菌古菌群落结构影响较大,在门水平上,N2与其它处理细菌群落结构差异较大,MN1与其它处理古菌群落结构差异较大。4.本研究在设施菜地中共检测到4个AOB类群,共15种AOB,分别隶属于亚硝化球菌属(Nitrosococcus),亚硝化单胞菌属(Nitrosomonas),亚硝化螺菌属(Nitrosospira)和Nitrosomonadaceaeuncultured,Nitrosomonadaceae uncultured是菜地中 AOB 的优势菌群;检测出 3 属共 5 种 AOA,分别是 Nitrosoarchaeum、Nitrososphaera和Nitrosotalea,Nitrososphaera为菜地中AOA中的优势菌群;共检测到4属共14种NOB,分别隶属于硝化螺菌属(Nitrospira)、硝化球菌属(Nitrococcus)、硝化刺菌属(Nitrospina)和Nitrolancea,Nitrospira为设施菜地NOB中的优势菌群。整体上,AOA在各处理中相对丰度均在80%以上,表明AOA可能在本施肥条件下氨氧化作用中发挥主导作用。5.在本研究中,土壤养分均随着有机肥的投入而表现出升高的趋势,但是硝化菌群丰度却整体表现出下降的趋势,因此土壤养分大多与硝化菌群不同程度地呈显著负相关。三类硝化菌群仅NOB与多种土壤理化性质、多种酶活性和土壤功能多样性相关性显著,表明NOB尤其是硝化螺菌属NOB对土壤环境变化较为敏感,受土壤环境影响较大,而同时在土壤生化反应和微生物代谢活性潜在作用更大。综合起来,氮肥单施有利于古菌和硝化菌群数量(丰度)的增长,而有机肥投入有利于丰富二者的物种多样性。整体上,二倍量氮肥单施(N2)对硝化菌群群落结构影响最大。
[Abstract]:In soil ecosystem, nitrification is a key link of soil nitrogen transformation in soil quality, plant growth, sustainable development, nitrogen utilization rate has an important influence on environmental pollution and food safety. Nitrification is driven by microbial nitrification will be gradually transformed into ammonium nitrate. The microbial processes involved in the process referred to as nitrifying bacteria, including ammonia oxidizing bacteria and ammonia oxidizing bacteria and nitrite oxidizing bacteria. Fertilization mainly by changing the soil microbial diversity and community structure of nitrification effect. This research takes Shenyang Agricultural Uinversity long-term fertilization soil as the experimental materials, Biolog technology research and high-throughput sequencing technology fertilizer nitrogen fertilizer on greenhouse soil fertility and soil nitrifying bacteria based on impact, aims to clear the nitrifying bacteria and soil fertility between The interaction, in order to study the nitrifying bacteria especially soil ecosystem NOB fertilization and cultivation of the data. At the same time provide facilities for microbial parameters of vegetable and soil fertility evaluation facilities for health management, soil N20 emissions and increase the utilization ratio of nitrogen fertilizer to provide a scientific basis. The main results of this study are as follows: 1. organic fertilizer and nitrogen fertilizer could improve soil fertility and soil enzyme activity. Especially the organic fertilizer nitrogen fertilizer can increase soil organic matter, total nitrogen, available phosphorus, available potassium content; soil ammonium nitrogen, nitrate nitrogen, nitrogen content and conductivity in no organic fertilizer or organic fertilizer a certain amount of time with the increase of nitrogen increased, while pH with N inputs increase. Organic fertilizer can improve soil urease, invertase, cellulose decomposing enzyme, two fluorescein Acetic acid esterase activity. Neutral phosphatase activity was most sensitive to fertilization, differences between each treatment; long-term single application of nitrogen fertilizer has certain inhibitory effect on catalase, only the high amount of N times (N2, MN2) had significant effects on polyphenol oxidase activity of.2. inhibited the single application of nitrogen fertilizer on soil microbial diversity, and organic fertilizer when a certain amount of help to improve microbial functional diversity, but excessive nitrogen fertilizer will still lead to a decline in microbial functional diversity. The long-term nitrogen fertilization to partial amino acid carbon becomes the main carbon source utilization rate is higher than that of single species, nitrogen fertilizer or fertilizer treatment using organic fertilizer and carbon source comprehensive comparison, the long-term organic fertilizer as the nitrogen content (MN1) were the best, the worst performance of N2.3. high-throughput sequencing technology based on protected vegetable bacteria and archaea of preliminary analysis identified. Out of the gate level including unclassfied people under 37, a level 1507 level 7 kinds of bacteria and the door, belonging to level 26, level 40 Archaea bacteria. Bacteria in door deformation (Proteobacteria) is the predominant population in greenhouse soil; odd bacteria (Thaumarchaeota) is the door of Archaea the dominant bacteria. Fertilization has great effect on the bacterial archaeal community structure in each treatment, at the gate level, the bacterial community structure between N2 and other large, MN1 and other archaeal community structure differences in the vegetable field.4. this study detected 4 AOB groups, a total of 15 AOB, respectively. Belonging to Nitrosococcus (Nitrosococcus), Nitrosomonas and nitrosospira (Nitrosomonas) and Nitrosomonadaceaeuncultured (Nitrosospira), Nitrosomonadaceae uncultured AOB is a vegetable dominant bacteria; detection of 3 genus and 5 species of AOA, divided Don't be Nitrosoarchaeum, Nitrososphaera and Nitrosotalea, Nitrososphaera for vegetable in AOA dominant bacteria; 4 genus and 14 species of NOB were detected respectively belong to nitrification Spirillum (Nitrospira), nitrococcus (Nitrococcus), nitrospina (Nitrospina) and Nitrolancea, Nitrospira for NOB in the greenhouse dominant bacteria. On the whole, AOA in each treatment in relative abundance were above 80%, suggesting that AOA is involved in the fertilization under the conditions of ammonia oxidation play a leading role in.5. in this study, the soil nutrient with organic fertilizer and showed increasing trend, but the overall abundance of nitrifying bacteria was demonstrated the downward trend, so the soil nutrient and most of nitrifying bacteria in different degrees were significantly negatively correlated. Three kinds of nitrifying bacteria is only NOB with a variety of physicochemical properties of soil, a significant variety of enzyme activity and soil functional diversity correlation showed that N OB especially NOB is sensitive to the changes of soil nitrification Spirillum, impact on soil environment, while soil biochemical and microbial metabolic activity greater potential role. Together, the single application of nitrogen fertilizer is beneficial to Archaea and nitrifying bacteria number (abundance) growth, and the input of organic manure in favor of the rich the two species diversity. On the whole, two times the amount of nitrogen fertilizer (N2) the greatest impact on the nitrifying bacteria community structure.

【学位授予单位】：沈阳农业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S626;S154.3

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