本文選題：氮循環(huán) + 反硝化�。� 參考：《大連海洋大學》2017年碩士論文

【摘要】：隨著工業(yè)現(xiàn)代化程度的提高以及沿海地區(qū)人口的逐漸增多,大量的生活污水以及工業(yè)、農(nóng)業(yè)廢水被排入海洋,由此導致了陸源無機氮的過量輸入,已成為近海富營養(yǎng)化的主要原因之一,由此引起的環(huán)境危害亦呈現(xiàn)出逐年增加的趨勢。反硝化過程可以有效地去除河口中的硝酸鹽與亞硝酸鹽,對水體中多余氮素的去除有著十分積極的意義。為深入了解遼河口的沉積物中反硝化過程與環(huán)境因子之間的關系,本研究于2014年5月采集該地區(qū)沉積物樣品,測定其理化性質(TN、TOC、Sal、NO2-、NO3-、NH4+、ORP和DO);以亞硝酸鹽還原酶基因(nirS、nirK)以及一氧化二氮還原酶基因(nosZ)作為分子標記,采用實時定量PCR和illumina Miseq高通量測序技術測定了采集沉積物中亞硝酸鹽還原酶基因和一氧化二氮還原酶基因的豐度和nirK、nirS和nosZ 3種基因型反硝化細菌菌群多樣性;分別分析了這三種功能基因的豐度與TN、TOC、Sal、NO2-、NO3-、NH4+、ORP和DO的相關性,同時分析了3種基因型反硝化細菌與上述理化因子的相關性。結果表明:從岸向海方向,鹽度以及溶解氧逐漸升高,無機氮營養(yǎng)鹽(硝酸鹽、亞硝酸鹽和銨鹽)的濃度以及總有機碳的含量沿河口向海方向整體呈現(xiàn)下降的趨勢;遼河口沉積物中nirS型基因的豐度為5.61×105copies/g~3.67×107copies/g,nirK的基因豐度為1.47×105copies/g~5.61×106copies/g,nosZ的基因豐度為1.44×103 copies/g~1.37×106copies/g。不同功能基因豐度受到不同環(huán)境理化因子的影響,其中nirK基因豐度與鹽度及其他環(huán)境因素無明顯的相關性;nirS基因的豐度與鹽度成顯著負相關(P=0.008,r=0.924),與NO2-呈顯著正相關(P=0.007,r=0.933),與TOC也呈顯著正相關的關系(P=0.007,r=0.93),與其他理化因子無顯著相關性。nosZ基因的豐度與鹽度呈顯著負相關(P=0.044,r=-0.823),而與NO2-呈顯著正相關(p=0.013,r=0.905),與TOC也呈顯著正相關關系(P=0.031,r=0.852)與其他理化因子無顯著相關性;nirK基因型反硝化菌的屬水平樣品聚類樹與柱狀圖組合分析得到其優(yōu)勢屬為中華根瘤菌屬(Sinorhizobium),所占比例達到了23.55%,與根瘤菌屬含量以及NO3-濃度顯著負相關(P=0.019,r=-0.885,P=0.09,r=-0.921),推測中華根瘤菌屬多樣性的影響因子可能為NO3-。對nirS基因型反硝化菌屬水平樣品聚類樹與柱狀圖組合分析得到其優(yōu)勢屬為假單胞菌屬(Pseudomonas),所占比例達到了19.64%,其與慢生根瘤菌屬呈現(xiàn)顯著負相關性(P=0.000,r=-0.985)。nosZ基因反硝化菌屬水平樣品聚類樹與柱狀圖組合分析圖得到其優(yōu)勢屬為固氮螺菌屬(Azospirillum)所占比例達25.69%,為主要優(yōu)勢菌;nirK、nir S以及nosZ基因型反硝化細菌的主要環(huán)境因子分別為NH4+和NO3-、Sal和NH4+、Sal和NO2-。nirK的OUT數(shù)量同環(huán)境因子并無顯著相關性,而nirS的OUT數(shù)量與ORP、DO和NH4+呈顯著相關性(P0.05),說明nirS在環(huán)境梯度下的變化比nirK更明顯。而nosZ的多樣性則與pH、ORP以及DO呈顯著相關性。結果表明:1.遼河口由陸向海方向,鹽度、無機氮營養(yǎng)鹽和有機碳的含量整體呈現(xiàn)下降趨勢。2.不同功能基因豐度受到不同環(huán)境理化因子的影響。3.遼河口的反硝化細菌由陸向海方向,含有nirK、nir S以及nosZ基因的反硝化細菌的多樣性逐漸降低。4.NH4+、NO3-、NO2-以及鹽度與含有這三種功能基因的細菌的關系最為密切。
[Abstract]:With the improvement of industrial modernization and the increasing population of coastal areas, a large number of domestic sewage and industrial wastewater have been discharged into the ocean, resulting in excessive input of inorganic nitrogen from land source, which has become one of the main causes of eutrophication in the coastal areas, and the resulting environmental hazards are also increasing year by year. Nitrification process can effectively remove nitrate and nitrite in estuaries. It is of great significance to remove the excess nitrogen in the water body. In order to understand the relationship between the denitrification process and environmental factors in the sediments of the Liaohe estuary, this study collected the sediment samples in this area in May 2014 and measured its physical and chemical properties (TN, TOC,) Sal, NO2-, NO3-, NH4+, ORP and DO), using the nitrite reductase gene (nirS, nirK) and the two nitrogen reductase gene (nosZ) as molecular markers, the abundance and the abundance of the nitrite reductase gene and the two N reductase gene in the sediments were measured by real-time quantitative PCR and Illumina Miseq high throughput sequencing technology. The diversity of 3 genotypes of denitrifying bacteria in nosZ, and the correlation between the abundances of the three functional genes and the TN, TOC, Sal, NO2-, NO3-, NH4+, ORP and DO were analyzed respectively. The correlation between the 3 genotypic denitrifying bacteria and the above physicochemical factors was analyzed. The results showed that the salinity and the dissolved oxygen increased gradually from the shore to the sea, and the inorganic nitrogen was gradually increased. The concentration of nutrients (nitrate, nitrite and ammonium salt) and the total organic carbon content declined along the direction of the estuary to the sea. The abundance of nirS gene in the Liaohe deposit was 5.61 x 105copies/g~3.67 x 107copies/g, the gene abundance of nirK was 1.47 x 105copies/g~5.61 * 106copies/g, and the gene abundance of nosZ was 1.44 * 103. The abundance of different functional genes in copies/g~1.37 x 106copies/g. was affected by different environmental physical and chemical factors, and there was no significant correlation between the abundance of nirK gene and salinity and other environmental factors, and the abundance of nirS gene was negatively correlated with salinity (P=0.008, r=0.924), and was positively correlated with NO2- (P=0.007, r=0.933), and was also positively correlated with TOC. P=0.007 (r=0.93) and no significant correlation with other physical and chemical factors, the abundance of.NosZ genes was negatively correlated with salinity (P=0.044, r=-0.823), but positively correlated with NO2- (p=0.013, r=0.905), and there was a significant positive correlation with TOC (P=0.031, r=0.852) without significant correlation with other physical and chemical factors; the genera of the nirK genotypic denitrifying bacteria. The combination of the horizontal sample cluster tree and the histogram analysis obtained that the dominant genus was Sinorhizobium, the proportion of which was 23.55%, which was negatively correlated with the content of rhizobia and NO3- (P=0.019, r=-0.885, P=0.09, r=-0.921). The influence factor of the diversity of Rhizobium in China may be NO3-. against nirS genotypes. The dominant genus is Pseudomonas (Pseudomonas), which has a dominant genus of 19.64%. It has a significant negative correlation with the genus chronic rhizobia (P=0.000, r=-0.985).NosZ gene denitrifying bacteria in the level of the group tree and histogram analysis map to obtain the advantage of nitrogen fixation. The proportion of Azospirillum is 25.69%, which is the dominant dominant bacteria. The main environmental factors of nirK, NIR S and nosZ genotype denitrifying bacteria are NH4+ and NO3- respectively. Sal and NH4+, Sal and NO2-.nirK OUT quantities have no significant correlation with environmental factors. The variation of the environmental gradient is more obvious than that of nirK, while the diversity of nosZ has a significant correlation with pH, ORP and DO. The results show that 1. Liaohe estuary is from land to sea, salinity, inorganic nitrogen, and organic carbon content as a whole is decreasing,.2. of different functional genes abundances are affected by different environmental physical and chemical factors in the.3. Liaokou estuary. The diversity of denitrifying bacteria containing nirK, NIR S and nosZ genes gradually decreased.4.NH4+, NO3-, NO2-, and salinity were most closely related to the bacteria containing these three functional genes.

【學位授予單位】：大連海洋大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：X52;Q933

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