本文關(guān)鍵詞： 施肥 氨氧化細(xì)菌 氨氧化古菌 亞硝酸鹽氧化細(xì)菌 群落結(jié)構(gòu)　出處：《沈陽(yáng)農(nóng)業(yè)大學(xué)》2017年碩士論文　論文類(lèi)型：學(xué)位論文

【摘要】：在土壤生態(tài)系統(tǒng)中,硝化作用是土壤氮素轉(zhuǎn)化的關(guān)鍵環(huán)節(jié),在土壤質(zhì)量可持續(xù)發(fā)展、植物生長(zhǎng)、氮肥利用率、環(huán)境污染以及食品安全等方面有重要的影響。硝化作用是由硝化微生物驅(qū)動(dòng)逐步將氨鹽轉(zhuǎn)化為硝酸鹽的過(guò)程。參與此過(guò)程的微生物統(tǒng)稱(chēng)為硝化菌群,主要包括氨氧化細(xì)菌、氨氧化古菌和亞硝酸鹽氧化細(xì)菌等。施肥主要通過(guò)改變土壤中微生物多樣性和群落結(jié)構(gòu)來(lái)對(duì)硝化作用產(chǎn)生影響。本研究以沈陽(yáng)農(nóng)業(yè)大學(xué)長(zhǎng)期定位施肥試驗(yàn)菜地土壤為試驗(yàn)材料,基于Biolog技術(shù)和高通量測(cè)序技術(shù)研究有機(jī)肥配施氮肥對(duì)設(shè)施菜地土壤肥力和土壤硝化菌群的影響,旨在明確硝化菌群與土壤肥力之間的互作關(guān)系,以期為研究硝化菌群尤其是土壤生態(tài)系統(tǒng)NOB補(bǔ)充施肥和設(shè)施栽培方面的數(shù)據(jù)。同時(shí)為完善設(shè)施菜地肥力評(píng)價(jià)與設(shè)施土壤健康管理提供微生物參數(shù),為農(nóng)田N20的減排以及氮肥利用率的提高提供科學(xué)基礎(chǔ)。本研究主要結(jié)果如下:1.有機(jī)肥與氮肥配施可以提高土壤肥力和土壤綜合酶活性。特別是有機(jī)肥配施氮肥可以顯著提高土壤有機(jī)質(zhì)、全氮、速效磷、速效鉀含量;土壤銨態(tài)氮、硝態(tài)氮、堿解氮含量和電導(dǎo)率在不施有機(jī)肥或有機(jī)肥投入量一定時(shí)均隨施氮量的增加呈現(xiàn)遞增趨勢(shì),而pH隨氮肥投入量的增加呈遞減趨勢(shì)。有機(jī)肥施入可以提升土壤脲酶、轉(zhuǎn)化酶、纖維素分解酶、熒光素二乙酸酯酶活性。中性磷酸酶活性對(duì)施肥最為敏感,各處理之間差異較大;長(zhǎng)期氮肥單施對(duì)過(guò)氧化氫酶有一定的抑制作用,僅高倍量氮肥(N2、MN2)對(duì)多酚氧化酶活性影響顯著。2.氮肥單施抑制了土壤微生物功能多樣性,而配施一定量的有機(jī)肥有利于提高微生物功能多樣性,但是配施過(guò)量氮肥仍然會(huì)導(dǎo)致微生物功能多樣性的下降。長(zhǎng)期偏氮施肥促使氨基酸類(lèi)碳源成為最主要碳源利用種類(lèi),并且施有機(jī)肥處理的碳源利用率高于氮肥單施或不施肥的處理。綜合比較,以長(zhǎng)期有機(jī)肥配施一倍量氮肥(MN1)表現(xiàn)最佳,N2表現(xiàn)最差。3.基于高通量測(cè)序技術(shù),對(duì)設(shè)施菜地細(xì)菌與古菌概況進(jìn)行初步分析。共鑒別出來(lái)包括unclassfied群體在內(nèi)的門(mén)水平下37類(lèi),種水平下1507種細(xì)菌和門(mén)水平下7類(lèi),屬水平下26類(lèi),種水平下40種古菌。細(xì)菌中變形菌門(mén)(Proteobacteria)是設(shè)施菜地土壤中優(yōu)勢(shì)群體;奇古菌門(mén)(Thaumarchaeota)是古菌中的優(yōu)勢(shì)菌群。施肥對(duì)各處理細(xì)菌古菌群落結(jié)構(gòu)影響較大,在門(mén)水平上,N2與其它處理細(xì)菌群落結(jié)構(gòu)差異較大,MN1與其它處理古菌群落結(jié)構(gòu)差異較大。4.本研究在設(shè)施菜地中共檢測(cè)到4個(gè)AOB類(lèi)群,共15種AOB,分別隸屬于亞硝化球菌屬(Nitrosococcus),亞硝化單胞菌屬(Nitrosomonas),亞硝化螺菌屬(Nitrosospira)和Nitrosomonadaceaeuncultured,Nitrosomonadaceae uncultured是菜地中 AOB 的優(yōu)勢(shì)菌群;檢測(cè)出 3 屬共 5 種 AOA,分別是 Nitrosoarchaeum、Nitrososphaera和Nitrosotalea,Nitrososphaera為菜地中AOA中的優(yōu)勢(shì)菌群;共檢測(cè)到4屬共14種NOB,分別隸屬于硝化螺菌屬(Nitrospira)、硝化球菌屬(Nitrococcus)、硝化刺菌屬(Nitrospina)和Nitrolancea,Nitrospira為設(shè)施菜地NOB中的優(yōu)勢(shì)菌群。整體上,AOA在各處理中相對(duì)豐度均在80%以上,表明AOA可能在本施肥條件下氨氧化作用中發(fā)揮主導(dǎo)作用。5.在本研究中,土壤養(yǎng)分均隨著有機(jī)肥的投入而表現(xiàn)出升高的趨勢(shì),但是硝化菌群豐度卻整體表現(xiàn)出下降的趨勢(shì),因此土壤養(yǎng)分大多與硝化菌群不同程度地呈顯著負(fù)相關(guān)。三類(lèi)硝化菌群僅NOB與多種土壤理化性質(zhì)、多種酶活性和土壤功能多樣性相關(guān)性顯著,表明NOB尤其是硝化螺菌屬NOB對(duì)土壤環(huán)境變化較為敏感,受土壤環(huán)境影響較大,而同時(shí)在土壤生化反應(yīng)和微生物代謝活性潛在作用更大。綜合起來(lái),氮肥單施有利于古菌和硝化菌群數(shù)量(豐度)的增長(zhǎng),而有機(jī)肥投入有利于豐富二者的物種多樣性。整體上,二倍量氮肥單施(N2)對(duì)硝化菌群群落結(jié)構(gòu)影響最大。
[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.

【學(xué)位授予單位】：沈陽(yáng)農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：S626;S154.3

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