本文選題：河涌沉積物 + 硝酸鹽還原硫氧化細(xì)菌(NR-SOB)��； 參考：《江西農(nóng)業(yè)大學(xué)》2015年碩士論文

【摘要】：最近的研究發(fā)現(xiàn),沉積物中分布著具有硝酸鹽還原硫氧化功能的微生物(NR-SOB)。這些微生物可以通過硝酸鹽異化還原成銨、硝酸鹽還原成亞硝酸鹽和反硝化等過程實現(xiàn)硫氧化。其中,尤其是具有硫氧化反硝化功能的NR-SOB可以在將硝酸鹽轉(zhuǎn)化為氮氣的同時,使具有毒害性的硫化物氧化為單質(zhì)硫或者硫酸鹽,實現(xiàn)氮硫共脫除。甚至,所氧化生成的硫酸鹽能一步作為硫酸鹽還原菌的電子受體使沉積物中的有機(jī)物氧化為二氧化碳,從而實現(xiàn)碳氮硫的共脫除,在沉積物修復(fù)中發(fā)揮重要作用。珠江三角洲黑臭河涌沉積物中存在著大量含氮、含硫的污染物,但目前有關(guān)這類沉積物中NR-SOB的種類及其功能活性特點了解極少。本研究利用前期從珠江三角洲典型黑臭河涌沉積物中獲得了具有硝酸鹽還原硫氧化功能的菌群F1,分析了該菌群的硝酸鹽還原硫氧化功能活性特征,并探索了影響該菌群硝酸鹽還原硫氧化功能活性的關(guān)鍵因素。進(jìn)一步對該菌群進(jìn)行傳代富集,得到了三個不同的NR-SOB菌群(分別命名為F2、F3和F4),分析比較四個菌群的硝酸鹽還原硫氧化功能及菌群結(jié)構(gòu)組成,闡明菌群中微生物與菌群功能活性的關(guān)系。并進(jìn)一步分離鑒定了這四個菌群中的關(guān)鍵功能菌株。獲得如下主要研究結(jié)果:⑴研究從黑臭河涌沉積物環(huán)境獲得的菌群F1主要經(jīng)過反硝化作用將硝酸鹽還原為氮氣,而硫代硫酸鹽主要被氧化為硫酸鹽,產(chǎn)生的亞硝酸鹽和單質(zhì)硫較少,是能實現(xiàn)氮硫的共去除NR-SOB菌群。pH、Fe~(2+)和Mg~(2+)都對菌群F1的硝酸鹽還原硫氧化功能活性有顯著的影響。pH為7.5時,菌群F1硝酸鹽還原硫氧化功能活性最高,產(chǎn)生更少的亞硝酸鹽,同時菌群F1能有較好的pH耐受性,能在pH為5.5時進(jìn)行硫氧化反硝化作用。Mg~(2+)的存在提高了菌群F1亞硝酸鹽還原活性;而Fe~(2+)的存在則抑制了菌群F1亞硝酸鹽還原活性。在無氨氮存在時仍然進(jìn)行硫氧化反硝化作用,與其他沉積物中進(jìn)行DNRA作用的微生物功能差異很大。⑵高通量測序結(jié)果表明,Thiobacillus是菌群F1的優(yōu)勢菌屬,占38.63±2.25%的豐度,Alicyclobacillus豐度為約8%。以O(shè)TU34034代表的γ-變形菌門豐度約為10.8%。因此,黑臭河涌沉積物中大部分的NR-SOB為未培養(yǎng)的微生物。硝酸鹽還原硫氧化功能指標(biāo)與菌群結(jié)構(gòu)組成相關(guān)性分析表明,以O(shè)TU34034為代表的Gammaproteobacteria中的微生物、OTU44535和OTU14246為代表的Betaproteobacteria中的微生物及Alicyclobacillus與菌群F1硝酸鹽還原硫氧化活性強(qiáng),產(chǎn)生亞硝酸鹽氮及單質(zhì)硫少有極顯著的相關(guān)性。而Thiobacillus其豐度的升高會導(dǎo)致菌群F1硝酸鹽還原硫氧化活性減弱。而科水平的Rhizobiaceae豐度的升高也會導(dǎo)致菌群F1硝酸鹽還原硫氧化功能活性減弱。⑶分離鑒定了129株具有硫氧化反硝化功能的NR-SOB功能菌株。通過16S rRNA基因序列分析結(jié)果表明這些菌株分屬于5種類型的微生物,分別是固氮菌、卡斯特蘭尼氏菌、脫硫弧菌、根瘤菌和硫桿菌。菌群F1中僅分離到了根瘤菌,其他NR-SOB分別在富集菌群中分離得到。因此,人工富集培養(yǎng)過程提高了黑臭河涌沉積物中NR-SOB的可培養(yǎng)性。硫桿菌、固氮菌和根瘤菌屬的硝酸鹽還原硫氧化產(chǎn)物中亞硝酸鹽均較低,其中以硫桿菌的活性最強(qiáng)。而卡斯特蘭尼氏菌和脫硫弧菌則產(chǎn)生較多的亞硝酸鹽。固氮菌和根瘤菌的硝酸鹽還原硫氧化功能在國內(nèi)外均鮮見報道,極有可能是一些新型的微生物。相關(guān)研究結(jié)果將為進(jìn)一步發(fā)揮NR-SOB的功能活性,加速黑臭河涌沉積物修復(fù)提供科學(xué)理論指導(dǎo)。
[Abstract]:Recent studies have found that microorganisms (NR-SOB) with nitrate reduction oxidation function are distributed in the sediments. These microorganisms can be oxidized by nitrate dissimilation reduction, nitrate reduction to nitrite and denitrification. In particular, NR-SOB with sulfur oxidation denitrification function can be used in the removal of nitrate. When it is converted into nitrogen, the toxic sulphides are oxidized into elemental sulfur or sulphate to achieve nitrogen and sulfur removal. Even the oxidized sulfate can be used as the electron acceptor of the sulfate reducing bacteria to oxidize the organic matter in the sediments to carbon dioxide, and the CO removal of carbon, nitrogen and sulfur is achieved, and in the restoration of sediment There are a large number of nitrogen and sulfur containing pollutants in the sediments of the black and smelly river in the Pearl River Delta, but there is little understanding of the species and functional activity of NR-SOB in this kind of sediments. The bacterial group F1, analyzed the characteristics of the functional activity of the nitrate reduction oxidation of the bacteria group, and explored the key factors affecting the functional activity of the nitrate reduction oxidation of the bacteria group. Further, three different NR-SOB bacteria groups (named F2, F3 and F4) were obtained, and the nitrate of the bacteria group was analyzed and compared. The relationship between the oxidation function of the original sulfur and the structure of the bacteria group, clarifying the relationship between the microbial community and the functional activity of the bacteria group. And further separating and identifying the key functional strains of the four bacteria groups. The main results are as follows: (1) the study of the bacterial group F1 obtained from the sediment environment of the black odor river is mainly reduced to the nitrate by denitrification. Nitrogen, while thiosulfate is mainly oxidized to sulphate, producing nitrite and sulfur less, is a common removal of NR-SOB bacteria.PH, Fe~ (2+) and Mg~ (2+) have significant effect on the functional activity of nitrate reduction and oxidation of the bacterial group F1, when.PH is 7.5, the functional activity of the microbial group F1 nitrate reduction sulfur oxidation is the highest. Less nitrite, at the same time, the bacteria group F1 can have better pH tolerance, and the presence of sulfur oxidation denitrification.Mg~ (2+) can increase the reduction of nitrite reduction activity of bacteria group F1 at the time of pH 5.5, while Fe~ (2+) inhibits the F1 nitrite reduction activity of the bacteria group. The microbial function of the DNRA action in the sediments is very different. (2) high throughput sequencing results show that Thiobacillus is the dominant genus of bacteria group F1, accounting for 38.63 + 2.25% abundance. The abundance of Alicyclobacillus is about 8%., the abundance of gamma deformable bacteria is about 10.8%., and most of the NR-SOB in the black and smelly river sediments is not cultured. The correlation analysis between the functional indexes of nitrate reduction sulfur oxidation and the structural composition of bacteria group showed that microbes in Gammaproteobacteria, OTU44535 and OTU14246 represented by OTU34034, and Alicyclobacillus and Alicyclobacillus and microbial group F1 nitrate reduced sulfur oxidation activity and produced nitrite. There was a very significant correlation between nitrogen and sulfur, but the increase in the abundance of Thiobacillus could lead to the reduction of the oxidation activity of the microbial group F1 nitrate reduction, and the increase of the Rhizobiaceae abundance of the family level also resulted in the decrease of the functional activity of the reduction sulfur oxidation of the bacteria group F1 nitrate. (3) 129 strains of NR-SOB with the oxidation denitrification function of the NR-SOB were identified. Functional strains. The results of 16S rRNA gene sequence analysis showed that these strains were divided into 5 types of microorganisms, namely nitrogen fixing bacteria, Custer ranibe, Vibrio desulphuri, Rhizobium and Thiobacillus. The bacteria group F1 isolated only the rhizobia, and the other NR-SOB were separated in the enriched bacteria group respectively. Therefore, the artificial enrichment culture process was improved. The culturability of NR-SOB in the sediments of the black and smelly rivers. The nitrites in the nitrate reduction oxidation products of Thiobacillus, nitrogen fixing bacteria and Rhizobium are low, and the activity of Thiobacillus is the strongest, while the Custer and the Vibrio desulphuri produce more nitrite. The nitrate reduction and oxidation function of nitrogen fixing bacteria and Rhizobium bacteria There are few reports at home and abroad. It is likely to be some new microbes. The results of the study will provide scientific theoretical guidance for further exerting the functional activity of NR-SOB and accelerating the restoration of black and smelly river sediments.
【學(xué)位授予單位】：江西農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：X172;X52

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