本文選題：厭氧氨氧化 + 水旱輪作�。� 參考：《浙江大學(xué)》2017年碩士論文

【摘要】：目前,厭氧氨氧化(Anammox)過(guò)程作為公認(rèn)的脫氮途徑,廣泛存在缺氧的自然環(huán)境中,但它在稻田土壤氮素循環(huán)的作用和地位仍缺乏深入了解。本項(xiàng)研究在借鑒國(guó)內(nèi)外在陸地生態(tài)系統(tǒng)中Anammox活性等方面的研究成果的基礎(chǔ)上,以水旱輪作稻田土壤Anammox過(guò)程為主線,聯(lián)合室內(nèi)培養(yǎng)和長(zhǎng)期定位肥料試驗(yàn)基礎(chǔ),運(yùn)用15N同位素示蹤技術(shù)、熒光定量PCR、DNA-SIP、克隆文庫(kù)和高通量測(cè)序等先進(jìn)技術(shù),探究水旱輪作體系土壤Anammox對(duì)氮?dú)猱a(chǎn)生的貢獻(xiàn)、Anammox微生物豐度和群落結(jié)構(gòu)及其影響因素等。主要研究結(jié)果如下:1)結(jié)果表明:無(wú)論是小麥季還是水稻季均檢測(cè)到了 Anammox活性,而且種植季節(jié)之間存在顯著差異,其中水稻季Anammox速率為0.9nmolN2·g-1-h-1,小麥季為0.46 nmol N2·g-1·h-1,不同施肥處理之間差異不顯著。從全年來(lái)看,水旱輪作過(guò)程中土壤中通過(guò)Anammox反應(yīng)流失的氮含量占總氮?dú)馍闪康?.15%-9.62%。進(jìn)一步對(duì)Anammox細(xì)菌豐度進(jìn)行分析,發(fā)現(xiàn)水稻和小麥季均測(cè)檢到了高豐度的Anammox細(xì)菌,長(zhǎng)期不同施肥處理?xiàng)l件下Anammox細(xì)菌的豐度表現(xiàn)為:豬糞堆肥配施化學(xué)肥料(PMCF)秸稈還田配施化學(xué)肥料(SRCF)和化學(xué)肥料(CF)不施肥(CK)。Anammox肼合成酶β亞基(hzs-β)基因序列分析表明,在水稻種植季節(jié),Anammox 細(xì)菌與 Ca.Brocadia,Ca.Scalindua 和 Caa.Jettenia 屬同源性較高。相比之下,所有的Anammox肼氧化酶(hzo)基因僅與Ca.Brocadia屬表現(xiàn)出較高的同源性,表明hzs基因比hzo基因有更高的敏感性。PMCF處理中的序列與Ca.Jettenia和Ca.Brocadia有較高的同源性,并顯示出最高的多樣性。2)通過(guò)實(shí)驗(yàn)室培養(yǎng)實(shí)驗(yàn)研究了不同有機(jī)碳源(無(wú)外源碳施用(CK)、施用尿素(UR)、施用秸稈(SR)、施用豬糞(PM)、施用淀粉(ST)和施用葡萄糖(GL))對(duì)Anammox過(guò)程和活性的影響。結(jié)果顯示,添加不同有機(jī)碳源進(jìn)行培養(yǎng)之后,Anammox過(guò)程的氮?dú)猱a(chǎn)生速率和對(duì)氮?dú)獾呢暙I(xiàn)率在1.04-3.19 nmol N2·g-1·h-1和4.22%-22.25%。雖然所有處理中均能檢測(cè)到Anammox活性,但各處理產(chǎn)生氮?dú)饬坎煌?CK處理Anammox速率最高,其他添加有機(jī)碳源之后的Anammox速率較低,差異顯著,說(shuō)明有機(jī)碳源的添加對(duì)Anammox活性具有顯著的抑制作用,同時(shí)有機(jī)碳對(duì)反硝化活性具有顯著的促進(jìn)作用。碳源添加對(duì)Anammox速率的影響如下:GLST,URSR。定量PCR結(jié)果表明,不同有機(jī)碳源均對(duì)Anammox微生物豐度具有一定的抑制作用,但PM處理對(duì)Anammox豐度的影響最小。選取活性最高的PM和CK處理的13C-Anammox肼合成酶β亞基(hzs-β)功能基因序列進(jìn)行DNA-SIP分析,發(fā)現(xiàn)活性厭氧氨氧化細(xì)菌與Ca.Brocadia屬具有較高的同源性。3)利用長(zhǎng)期定位試驗(yàn)土壤,通過(guò)室內(nèi)培養(yǎng)試驗(yàn),研究了不同水分狀態(tài)及溶解氧濃度(70%FC、干濕交替(淹水7天,落干7天)、淹水Ⅰ:溶解氧濃度為5.8 mg·L-1、淹水Ⅱ:溶解氧濃度為2.6 mg·L-1)對(duì)土壤Anammox過(guò)程的影響。結(jié)果表明,Anammox速率范圍在0.56-1.47nmolN2·g-1·h-1之間。70%田間持水量(70%FC)處理中的Anammox速率最低(0.61 nmol N2.g 1·h-1),淹水Ⅱ處理最高(].14 nmol N2·g-1·h-1);說(shuō)明溶解氧含量較高時(shí)會(huì)降低Anammox速率。水分處理會(huì)明顯增加Anammox細(xì)菌數(shù)量,不論是干濕交替還是淹水處理,Anammox細(xì)菌豐度較高,不同施肥處理中,CF和SRCF處理中Anammox細(xì)菌hzs-β基因拷貝數(shù)較高。Anammox細(xì)菌群落結(jié)構(gòu)在不同水分處理后差異并不明顯,絕大部分Anammox細(xì)菌與Ca.Brocadia屬具有較高的同源性,Ca.Jettenia屬數(shù)量較少。
[Abstract]:At present, the anaerobic ammonia oxidation (Anammox) process for nitrogen removal routes recognized, widespread anoxic environment, but its role and status in the paddy soil nitrogen cycle is still poorly understood. In this study, on the basis of research results of Anammox activity at home and abroad in the terrestrial ecosystem area, drought and flood the rotation of paddy soil Anammox process as the main line, combined with indoor culture and long-term base fertilizer experiment using 15N isotope tracer technique, fluorescence quantitative PCR, DNA-SIP, clone libraries and high-throughput sequencing and other advanced technology, explore the rotation system of soil Anammox to produce nitrogen contribution, Anammox microbial abundance and community structure and its influencing factors. The main results are as follows: 1) the results show that both the wheat or rice season were detected in the Anammox activity, but there are significant differences between the planting season, The rice season Anammox rate was 0.9nmolN2 g-1-h-1, 0.46 nmol N2 wheat season g-1 H-1, there was no significant difference between different treatments. From the view of the rotation process, soil total nitrogen content by 3.15%-9.62%. nitrogen Anammox reaction loss further analysis of Anammox found that the abundance of bacteria. Rice and wheat season were the detection to the high abundance of Anammox bacteria, long-term abundance performance under different fertilizer treatments of Anammox bacteria: pig manure combined with chemical fertilizer (PMCF) straw fertilizer chemical fertilizer (SRCF) and chemical fertilizer (CF) fertilizer (CK) hydrazine synthase beta subunit (.Anammox hzs- beta) gene sequence analysis showed that the planting season in rice, Anammox bacteria and Ca.Brocadia and Caa.Jettenia belong to Ca.Scalindua, the homology is high. In contrast, Anammox (HZO) of all the hydrazine oxidase gene and Ca.Broc The genus adia showed high homology, suggesting that the HZS gene has higher sensitivity to.PMCF treatment in Ca.Jettenia and Ca.Brocadia and has higher sequence homology than HZO gene, and showed the highest diversity of.2) the experimental study on the cultivation of different organic carbon sources through the laboratory (without exogenous application of carbon (CK), application urea (UR), straw (SR), pig manure (PM), the application of starch (ST) and application of glucose (GL)) influence on the Anammox process and activity. The results showed that after training with different organic carbon sources, nitrogen Anammox process for nitrogen production rate and the contribution rate of 1.04-3.19 in nmol N2 - g-1 - H-1 and 4.22%-22.25%. although all treatments were detected in Anammox activity, but the generation of nitrogen content is different, CK Anammox rate is the highest, the other after the addition of organic carbon source Anammox rate is low, indicating significant difference, organic carbon source Add has a significant inhibitory effect on the activity of Anammox, while organic carbon has significant effect on the denitrification activity. Adding carbon source effect on the rate of Anammox is as follows: GLST, URSR. quantitative PCR results showed that different organic carbon source has certain inhibitory effect on Anammox microbial abundance, but the effect of PM treatment on Anammox the abundance of selected minimum. The highest activity PM and CK 13C-Anammox hydrazine synthase beta subunit (hzs-) gene sequence by DNA-SIP analysis, found that the activity of anammox bacteria and the genus Ca.Brocadia has high homology of.3) using soil long-term experiment, through laboratory incubation experiment of different moisture state and dissolved oxygen concentration (70%FC, alternate (7 days of flooding, dry 7 days), flooding I: dissolved oxygen concentration of 5.8 mg L-1, flooding II: dissolved oxygen concentration of 2.6 mg L-1) on soil Anammox process Effect. The results show that the Anammox rate in the range between 0.56-1.47nmolN2 - g-1 - H-1.70% of field capacity (70%FC) in the treatment of Anammox was the lowest (0.61 nmol N2.g 1 - h-1), flooding treatment was the highest (].14 nmol II N2 - g-1 - h-1); it showed that the higher the content of dissolved oxygen will reduce the rate of Anammox water. Anammox treatment significantly increased the number of bacteria, whether or waterlogged dry wet alternation Anammox bacterial abundance was higher in different treatments, CF and SRCF in Anammox gene of bacterial hzs- beta copy bacterial community structure in a higher number of.Anammox in different water treatments after the difference is not obvious, the majority of Anammox bacteria and the genus Ca.Brocadia have high homology Ca.Jettenia, is a smaller number.

【學(xué)位授予單位】：浙江大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S153

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本文編號(hào)：1742294