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太平洋暖池区深海沉积物中微生物分子生态学研究【作者】曾润颖 【导师】徐洵

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太平洋暖池区深海沉积物微生物分子生态学研究水利论文Ed {S1~+MM'[v bk
Molecular Ecology Study of Microorganisms in the Pacific Ocean "Warm Pool" Deep Sea Sediments

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作者】曾润颖 【导师徐洵水利论文ue;j&sUW;SN
【作者基本信息】厦门大学,动物学,2003年,博士

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SW7\l+iQ+Q(S0【中文摘要】 西太平洋“暖池”在全球气候变迁过程中起着举足轻重的作用,是目前气候变迁机制研究的热点,同时也是地球科学研究的焦点之一。本论文首次对“暖池”区海底沉积物中的微生物进行了分子生态学研究,分析了微生物多样性,对由微生物参与的物质能量循环特征进行了探讨,为研究深海生物圈在地球圈层相互作用中的地位提供了参考资料。此外,本文还对采集自太平洋的深海沉积物和南极、北极区域的样品进行了低温微生物的分离培养及低温酶的筛选与性质分析。 通过构建沉积物中微生物16S rDNA克隆文库,采用PCR-RFLP分析、DNA-DNA杂交、16S rDNA序列测定以及系统发育分析的方法,研究了两太平洋“暖池”区和东太平洋“结核”区两个不同海区深海沉积物中的微生物多样性和群落结构特征及其与环境的相互关系,得到了一些与“暖池”区环境特点紧密相关的新发现,新认识。不同海区深海沉积物中的细菌都以紫细菌(Proteobacteria)为主,属于γ-和α-紫细菌亚群的细菌种类和数量均最为丰富;而属于β-亚群的细菌很少。不同海区沉积物中紫细菌类群的差别在于δ-亚群和ε-亚群,这两个亚群的细菌大部分都和硫代谢相关。在“结核”区沉积物中...更多没有检测到属于这两个亚群的细菌,而在“暖池”区沉积物中属于这两个亚群的细菌数量较多(最高可达29%),并且δ-亚群的细菌呈现随沉积物深度增加而减少的趋势。除了紫细菌之外,CFB类群在“暖池”区沉积物中也是一类较重要的细菌,主要集中在表层沉积物;而在“结核”区没有检测到属于CFB类群的细菌,表明“暖池”区表层存在较多的有机物。古菌的多样性分析结果表明该海区沉积物中的古菌绝大多数属于嗜泉古菌界(Crenarchaeota),其中包含许多未知的古菌遗传信息。 在“暖池”区沉积物中发现了丰富的与硫及甲烷代谢相关的细菌,对这些细菌的种类组成及其在各层次沉积物中的分布、数量进行了分析,发现该海区沉积物中存在一条完整的硫和甲烷代谢循环途径,它们的代谢在物质能量代谢中占据重要的地位。而16S rDNA克隆文库分析和DNA-DNA杂交的结果表明“结核”区沉积物中这两类细菌数目很少,说明“暖池”区沉积物中的微生物群落结构特征是对环境因素的一种响应,同时也可能是影响该海区深海及海洋环境的一个重要因素。 对深海沉积物和极地环境样品进行了低温微生物的分离和培养,得到了300多株嗜(耐)冷细菌,并对其进行了生长特性的分析。结果表明所有的细菌都能够在低温下(4℃~15℃)正常生长,而在中温范围内(25℃~30℃)的生长状况较差,仅有少数细菌能够在37℃缓慢生长。与深海沉积物相比,嗜冷菌在南极样品中所占的比例较大。采用16S rDNA序列比较的方法对部分低温菌株进行了分子分类与鉴定,结果表明所鉴定的菌株中假交替单胞菌属(Pseudoalteromonas sp.)和假单胞菌属(Pseudomonas sp.)的细菌占较大比例。摘要 通过所分离出的嗜(耐)冷细菌的165 rDNA序列的比较分析,对嗜冷海洋细菌的特异165 rDNA序列进行了探索,并设计了一对引物。初步的分析表明,所设计的PCR引物能够较好地区分海洋性细菌和陆源性细菌,并且可以用于嗜冷海洋细菌的区分。 采用选择性培养基对深海和极地细菌进行了产低温酶菌株的筛选,最终得到了16株具有低温蛋白酶或低温几丁质酶活性的菌株,对其中5株活力最高的低温酶产生菌进行了菌种的分子鉴定,并分析了温度对这五种低温酶的活力、产酶和热稳定性的影响,为进一步的研究与开发奠定基础。  还原

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【英文摘要】 The west Pacific Ocean "Warm Pool" plays an important role in global climate change, which makes it become one of the most important focuses of geoscience research. In this study, we investigate the microbial diversity of deep sea sediments and analyze the characteristic of substance and energy conversion driven by microorganism in "Warm Pool" area for the first time.Samples including deep sea sediments of the Pacific Ocean and other environmental samples from polar region were collected and analyzed. The profile of microbial community structures in different deep sea sediments was evaluated and the interaction between microorganisms and environment was analyzed by culture-independent molecular phylogenetic methods. Psychrophilic and psychro-tolerant bacteria were cultured and used for screening the cold-active enzymes. Some characteristics of cold-active protease and chitinase were analyzed then.Microbial 16S rDNA (ribosomal DNA) clone libraries of deep sea sediments were constructed ...更多and studied by PCR-RFLP (Restriction Fragment Length Polymorphism) and phylogenetic analysis based on 16S rDNA sequences. The microbial diversity and community structures of deep sea sediments collected from two different sea area including the west Pacific "Warm Pool" and the east Pacific "Manganese nodule" area, as well as the interaction between microbial community and environment, were analyzed based on these studies. Members of the γ-proteobctcteria and α-proteobacteria were found to be most abundant in both areas, and members of β-proteobacteria were seldom detected. The difference of microbial group between different sea areas was the 8- and e- subdivision of proteobacteria, which was abundant at upper sediment of "Warm Pool" area while they were not detected at the sediments of "Manganese nodule" area. At the core sediment of "Warm Pool" area, members of δ-proteobacteria reduced from upper to lower sediments. Members of the CFB (Cytophaga-Flexibacter-Bacteroides) were found to be rich in upper sediment of "Warm Pool" while they were not detected in sediment from "Manganese nodule" area, which indicated that there were more organic substances in the "Warm Pool" area. The results of archaeal diversity analysis showed that most of the members belonged to Crenarchaeota, and included many unidentified archaea species.Abundant bacteria related to the metabolism of sulfur and methane were found in sediment collected from "Warm Pool" area, which indicated that the metabolism of sulfur and methane played an important role in the substance and energy conversion of this area. The distribution and amount analysis of these bacteria in different layers of core sediment indicated that there was an intact cycle that coupled sulfur metabolism with methane metabolism existed in this area, which may be the microbial response to the environment because there was seldom similar bacteria detected from "Manganese nodule" area sediment by DNA-DNA hybridization with specific oligonucleotide probe and 16S rDNA clone library analysis. It could play an important role in the evolution of "Warm Pool" area.More than 300 pure psychrophilic and psychro-tolerant bacteria were isolated from environmental samples. The analysis of growth indicated that all strains could grow well at 4℃15℃ while they could grow poorly at 25℃30℃, and few of them could grow constrainedly at 37℃. More psychrophiles isolated from antarctic samples than that from deep sea sediments. The molecular classification and identification of psychrophiles based on 16S rDNA sequences indicated that Pseudoalteromonas sp. and Pseudomonas sp. were common in strains isolated.The unique 16S rDNA fragment of marine psychrophiles was studied based on the comparison of 16S rDNA sequences. The results indicated that the PCR primers designed could distinguish between marine bacteria and terrestrial bacteria, and could be applied in distinguishing the psychrophiles.Sixteen strains which could produce cold-active protease and chitinase were screened by selective medium and  还原水利论文8?/\n5bgU ^&F

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【中文关键词】 深海; 微生物分子生态; 西太平洋暖池; 环境响应; 低温酶
_|zgoY!gL+d1l"K0【英文关键词】 deep sea; microbial molecular ecology; the west Pacific Ocean "Warm Pool"; environmental response; cold-active enzyme水利论文W(S$~5~j(L,}

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TAG: 沉积物 生态学 太平洋 微生物 徐洵
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