【摘要】：湖泊濕地是甲烷氣體的重大排放源。產(chǎn)甲烷菌在地球生物化學(xué)碳素循環(huán)過(guò)程中起著關(guān)鍵作用,它是產(chǎn)生溫室氣體的主要因素之一。探究湖泊沉積物中產(chǎn)甲烷菌可為甲烷氣體的排放研究提供生物學(xué)支撐。本文以杭州西湖為研究對(duì)象,采用限制性片段長(zhǎng)度多態(tài)性分析及分子克隆等技術(shù),研究了西湖春夏兩季不同區(qū)域沉積物中產(chǎn)甲烷菌群落結(jié)構(gòu)和多樣性,并分析水生植物、沉積物營(yíng)養(yǎng)水平等對(duì)菌落的影響。主要結(jié)果如下:1、通過(guò)產(chǎn)甲烷菌16SrDNA克隆文庫(kù)的酶切圖譜分析,冬季西湖共得到38種酶切類型。測(cè)序結(jié)果表明,西湖共存在著4種菌目:甲烷微菌目、甲烷八疊球菌目、甲烷桿菌目和甲烷球菌目,前兩種菌目均隸屬于甲烷微菌綱,該菌綱在西湖各子湖中作為優(yōu)勢(shì)種存在,部分比例高達(dá)68%。隸屬于各甲烷菌目的甲烷菌科檢測(cè)出7種,分別為M.sarcinaceae、M.caldococacceae、M.regulaceae、M.microbiaceae、M.bacteriaceae、M.thermaceae和M.saetaceae。2、夏季西湖共得到21種酶切類型,檢測(cè)出3種菌目:甲烷微菌目、甲烷八疊球菌目和甲烷桿菌目,其中,同屬于甲烷微菌綱的甲烷微菌目、甲烷八疊球菌目的比例差異性較大;前者作為優(yōu)勢(shì)菌存在,含量明顯高于后者。隸屬于各甲烷菌目的甲烷菌科有5種:M.microbiaceae、M.bacteriaceae、M.saetaceae、M.regulaceae和M.saricinaceae,甲烷微菌科仍作為優(yōu)勢(shì)菌科存在于各子湖中,后三者所占比例較少,皆低于15%。3、冬季西湖湖西區(qū)和外湖區(qū)沉積物中,產(chǎn)甲烷菌16SrDNA克隆文庫(kù)的酶切類型分別是23、24種。反映湖西區(qū)沉積物產(chǎn)甲烷菌群落結(jié)構(gòu)多樣性的Shannon指數(shù)介于1.978-2.323,而外湖區(qū)該指數(shù)最大值僅為2.002,大部分均低于1.978,Simpson多樣性指數(shù)規(guī)律與其類似,說(shuō)明湖西區(qū)各子湖產(chǎn)甲烷菌結(jié)構(gòu)的多樣性高于外湖區(qū)各子湖。另外,冬季湖西區(qū)各子湖的甲烷菌科種類高于外湖區(qū)各子湖,M.sarcinaceae和M.caldococaceae僅在湖西區(qū)發(fā)現(xiàn)。4、冬季湖西區(qū)各子湖水生植物恢復(fù)區(qū)的產(chǎn)甲烷菌菌落均勻度高于匱乏區(qū),但產(chǎn)甲烷菌種類規(guī)律不明顯,茅家埠的恢復(fù)區(qū)和匱乏區(qū)的產(chǎn)甲烷菌綱數(shù)量等同;烏龜潭匱乏區(qū)的菌綱種類高于恢復(fù)區(qū)1種;而浴湖灣的結(jié)論與之相反。可能因?yàn)楦髯雍謴?fù)區(qū)的水生植物覆蓋率和生長(zhǎng)周期不同,外加沉積物本底營(yíng)養(yǎng)物濃度的綜合作用所致。5、在冬夏季節(jié),甲烷微菌科均作為優(yōu)勢(shì)菌科存在,比例高達(dá)50%。但冬季西湖產(chǎn)甲烷菌結(jié)構(gòu)和種類多樣性高于夏季,M.thermaceae和M.caldococcaceae僅在冬季檢測(cè)到�？赡苡捎诖蟛糠炙参镌谙募咎幱谏L(zhǎng)旺盛期,根系泌氧作用對(duì)于沉積物中局部氧化還原狀態(tài)的貢獻(xiàn)較大,一定程度上抑制厭氧產(chǎn)甲烷菌的生長(zhǎng)。
[Abstract]:Lake wetland is an important source of methane gas emission. Methanogenic bacteria play a key role in the process of biogeochemical carbon cycle, and it is one of the main factors to produce greenhouse gases. To explore the methanogenic bacteria in lake sediments can provide biological support for the study of methane gas emissions. In this paper, the community structure and diversity of methanogenic bacteria in sediments of different regions of West Lake in spring and summer of Hangzhou were studied by restriction fragment length polymorphism analysis and molecular cloning, and aquatic plants were analyzed. Effects of sediment nutrition level on colony. The main results are as follows: 1. A total of 38 restriction endonuclease types were obtained in West Lake in winter by restriction endonuclease analysis of methanogenic bacteria 16SrDNA clone library. The results of sequencing showed that there were four species of bacteria in the West Lake: methane microflora, methane octacoccus, methanomycetes and methanomycetes. The first two bacteria belonged to methane microflora, which existed as dominant species in each lake of the West Lake. Some of them are as high as 68%. Seven species of methanomycetes belonging to various methanogens were detected, which were M. sarcinaceae, M. caldococacceae, M. microbiaceae, M. thermaceae and M. saetaceae.2. A total of 21 enzyme digestion types were obtained in the West Lake in summer. Three kinds of bacteria were detected: methane microbacteria, methanosporium and methanomycetes, among which, the proportion of methane microbacteria belonging to methane microflora and methanosporium was quite different. The former exists as the dominant bacteria, and the content of the former is significantly higher than that of the latter. There are five species of methanomycetes belonging to each order: M. microbiaceae, M. bacteriaceae, M. saetaceae, M. regulaceae and M. saricinaceae. methane microbacteria still exist as dominant bacteria in each lake, and the proportion of the latter three is less than 15%. In winter, the restriction endonuclease types of methanogenic bacteria 16SrDNA clone libraries in the sediments of West Lake and Outer Lake were 23 and 24 species, respectively. The Shannon index, which reflects the diversity of methanogenic bacteria community in the western part of the lake, is between 1.978 and 2.323, while the maximum value of the index is only 2.002, most of which are lower than 1.978. the law of Shannon diversity index is similar to that in the outer lake area. The results showed that the diversity of methanogenic bacteria structure in the western lake was higher than that in the outer lake. In addition, the methanogens of each lake in winter were higher than those in the lake, and M.sarcinaceae and M.caldococaceae were only found in the west of the lake. 4. The uniformity of methanogenic bacteria in the recovery area of aquatic plants in the western part of the lake was higher than that in the scarce area. However, the species of methanogenic bacteria was not obvious, and the number of methanogenic bacteria in the convalescent and scarce areas of Maojiabu was the same. The species of bacteria in Wuqiutan fatigue area were higher than those in restoration area, but the conclusion in Bath Lake Bay was opposite. It may be due to the different coverage and growth cycle of aquatic plants in each sub-lake restoration area, and the comprehensive effect of sediment background nutrient concentration. 5. In winter and summer, methane microflora exists as the dominant family, the proportion is as high as 50%. However, the structure and species diversity of methanogenic bacteria in West Lake in winter were higher than those in summer, and M.thermaceae and M.caldococcaceae were only detected in winter. It may be that most aquatic plants are in the peak growth period in summer, and the root oxygen secretion contributes greatly to the local redox state in sediments, and to a certain extent, it can inhibit the growth of anaerobically methanogenic bacteria.
【學(xué)位授予單位】：武漢理工大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：X172

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