通过提取环境总16S rDNA和总16S rRNA分别构建DNA克隆文库和反转录cDNA文库,并进行克隆测序和序列分析,探讨湖光岩玛珥湖浮游细菌和浮游活性细菌的遗传多样性。结果表明：构建两个水层的4个克隆文库,即1 m和10 m水层总菌的16S rDNA文库和总活性菌的16S rcDNA文库,共获得413条序列,分属15门、32目、52科;1 m水层的rDNA文库与rcDNA文库中Verrucomicrobia类群所占比例最大（36.8%和32.1%）,其次为Alphaproteobacteri（a 15.1%和16%）和Betaproteobacteri（a 17.9%和16%）;在10 m水层的2个文库中,Alphaproteobacteria类群均占绝对优势,分别为63.5%（rDNA）和43.8%（rcDNA）,其次为Verrucomicrobia（11.5%和14.3%）,其他细菌类群包括Acidobacteria、Chloroflexi、Firmicutes、待定门OD1、Planctomycetes、Spirochetes和Deltaproteobacteria,比例在0.8%~3.8%之间;在科的水平上,SAR11为湖光岩玛珥湖最为丰富的类群,占总克隆数的26.9%,其中在10 m水层的比例高达51.5%。序列同源性分析表明,绝大多数序列（89.8%）来源于新种,其中19.6%的序列可能来自于新属,69%的序列可能来自于全新的科。湖光岩玛珥湖浮游细菌群落结构较为独特,有大量的浮游细菌种类尚未获得相应的纯培养物,蕴含着丰富而新颖的微生物资源。

Huguangyan Maar Lake belongs to a unique type of volcanic lake. Little is known about the microbial diversity in Maar Lake. So it is important to study the bacterial diversity in the water body for protecting and developing Maar Lake microbial resources. Total DNA and RNA were extracted from planktonic microbial communities in the Huguangyan Maar Lake, and 16S rDNA and 16S rcDNA clone libraries were constructed, and a number of clones were sequenced with the aim to analyze the genetic diversity of total bacterioplankton and active planktonic bacteria. Four clone libraries were constructed from the community DNA and cDNA of the 1 m and 10 m layers of the Huguangyan Maar Lake with 413 sequences reported, which were further classified into 15 phyla, 32 orders, and 52 families. Within the 16S rDNA library and 16S rcDNA library of the 1m layer, Verrucomicrobia occupied the largest proportion (36.8% and 32.1%, respectively), followed by Alphaproteobacteria (15.1% and 16%, respectively) and B

目的应用T-RFLP分析胆固醇结石中微生物群落结构。方法应用末端标记限制性片段长度多态性（Terminal Restriction Fragment Length Polymorphism，T-RFLP）和克隆文库分析，以微生物群落16S rRNA基因（16S rDNA）为目标，对8例常规细菌培养阴性的纯胆固醇患者胆囊结石和胆汁中的微生物群落结构进行解析和比较。结果发现8例纯胆固醇结石患者胆汁中未找到细菌存在的证据，结石中细菌16S rDNA阳性率为37.5%（3/8），细菌16S rDNA片段测序表明细菌群落与肠杆菌科和微球菌科的微生物有较高的相似性，同时细菌群落中检出了大量的未培养微生物（Uncultured bacterium clone）。结论运用T-RFLP方法分析16S rDNA克隆片段能够有效评估纯胆固醇结石中的细菌群落结构的多样性。

Objective To analysis of microbial community structure in cholesterol calculus by T-RFLP (Terminal Restriction Fragment Length Polymorphism). Methods Small subunit rRNA gene (16S rDNA) was retrieved from 8 patients'' gallstone and bile with cholesterol calculus. The microbial community structure of these cholesterol calculus patients'' gallstone and bile were investigated by T-RFLP and clone libraries approaches. Results There was no bacterium found in bile from 8 patients with cholesterol calculus. The positive rate of bacterial 16S rDNA in stone was 37.5%(3/8). Bacterial 16S rDNA sequencing fragments showed that microbial community and Enterobacteriaceae and Micrococcaceae microorganisms had high similarity. Meanwhile, there were detected a number of uncultured bacterium from microbial community. Conclusion Analysis of 16S rDNA cloned fragment could effectively evaluate the diversity of bacterial community structure by T-RFLP method in cholesterol stone.

采用16S rDNA克隆文库法对某刺参养殖场患病刺参Apostichopus japonicus幼苗(体长为3~5 cm)表皮的菌群结构进行分析。结果表明：化皮参苗病灶组织和未明显化皮参苗表皮菌群均由γ-变形菌纲、β-变形菌纲和α-变形菌纲组成，优势菌为γ-变形菌纲，主要包括志贺氏菌Shigella sp.、不动杆菌Acineto-bacter sp.和假单胞菌Pseudomonas sp.；化皮参苗病灶组织中志贺氏菌、不动杆菌和假单胞菌比例分别为49%、17%和5%，未明显化皮参苗表皮中3类菌的比例分别为20%、26%、9%。将该养殖场育苗池海水作为对照并进行菌群结构分析，结果显示，水中菌群同样由γ-变形菌纲、β-变形菌纲和α-变形菌纲组成，优势菌也为γ-变形菌纲，主要包含弧菌Vibrio sp.(75%)与志贺氏菌(12%)。此外，对化皮参苗病灶处进行细菌分离培养，得到一株优势菌，经16S rDNA初步鉴定为弧菌。研究表明，采用16S rDNA克隆文库法所得结果与传统的病原分离培养法存在差异。

Flora on eipthelia of diseased juvenile sea cucumber Apostichopus japonicus with body length of 3-5 cm showing body wall lesions was studied by a method of 16S rDNA cloning library. The results showed that the bacte-ria were composed of Alphaproteo bacteria, Betaproteo bacteria and Gammaproteo bacteria, with the predominant flora of Shigella sp. , Acinetobacter sp. and Pseudomonas sp. on the lesions of the diseased sea cucumber as well as sea cucumber without obvious symptom. There was 49% of Shigella sp. , 17% of Acinetobacter sp. and 5% of Pseudomonas sp. in the diseased sea cucumber, while there was 20% of Shigella sp. , 26% of Acinetobacter sp. and 9% of Pseudomonas sp. in those without lesions. The members of Alphaproteo bacteria, Betaproteo bacteria and Gammaproteo bacteria were found in the seawater from the culture ponds as a control group, with predominant phaproteobacteria including Vibrio sp. (75%) and Shigella sp. (12%). Also, a predominant bacterium strain was isolated fro

通过构建16S rDNA克隆文库对象山港南沙岛不同养殖模式（贝类养殖、藻类养殖及网箱养殖）表层沉积物微生物多样性和群落结构特征进行了比较和分析,共获取136个OUT。其中,贝类养殖区、藻类养殖区和网箱养殖区OTU分别为58、48和57个。各站位OTU分布差异明显,表现出高度的多样性。基于16S rDNA序列的生物多样性和丰富度分析表明,网箱养殖区丰富度指数ACE为739,香浓指数H？为3.8,均为最高值,丰富度指数Chao为245,略低于于贝类养殖区。贝类养殖区丰富度指数Chao为303,在各养殖区中最高。藻类养殖区丰富度指数ACE为174、Chao为89,香浓指数H？为3.6,均为最低值。系统发育分析表明,南沙岛各养殖区的优势种群均为变形菌门（Proteobacteria）,但是藻类养殖区微生物群落结构与其他养殖区域相比,16S rDNA克隆文库差异显著,其中根瘤菌属（Rhizobium）及其他光合细菌在藻类养殖区分布较多。网箱养殖区沉积物表层微生物群落中出现了与环境污染密切相关的菌群,如志贺氏菌属（Shigella）、埃希氏菌属（Escherichia）和ε-变形菌纲的微生物种群,揭示网箱养殖对底质沉积物环境的影响较大。

Xiangshan Bay, the biggest marine aquaculture base in Zhejiang Province, is a semi- enclosed bay with slow water exchange rate. In the center of Xiangshan Bay lies Nansha Bay where a variety of mariculture models are applied. To better understand the structures and diversity of sediment microbial communities in different mariculture models, we constructed the 16S rDNA clone library for the analysis of samples from the shellfish culture, the seaweed culture and the fish cage culture. We obtained 136 OTUs from three sampling models that included 58 OTUs from the shellfish culture, 48 from the seaweed culture, and 57 from the fish cage culture. The distribution patterns of OTUs were highly different between the three sampling models which indicated the distinct microbial community structures. The calculation of species richness (Chao), evenness (ACE), and diversity (Shannon) were 245/739/3.8 (fish cage culture), 313/330/3.6 (shellfish culture) and 89/174/3.6 (seaweed culture) respectively

在制浆造纸过程中,白水的循环回用加重了纸机微生物污染的问题,促进了纸机腐浆的形成,影响了纸机的正常运转。此次研究用滤膜抽滤富集细菌菌体,对白水样品直接提取DNA,通过16S rDNA序列分析,构建16S rDNA文库,对芬欧汇川(UPM)常熟纸厂PM1纸机白水中的细菌多样性进行了研究。获得的61个16S rDNA序列可分为28个可操作分类单元(OTUs),分属于6个不同门类,优势菌种顺序为变形菌门(Proteobacteria)(77.05%)、厚壁菌门(Firmicutes)(11.48%)、放线菌门(Actinobacteria)(4.92%)、异常球菌-栖热菌门(Deinococcus-Thermus)(3.28%)、拟杆菌门(Bacteroidetes)(1.64%)、蓝藻门(Cyanobacteria)(1.64%)。造纸白水具有较丰富的细菌多样性,以变形菌为优势菌群,占库容总量的77.05%,其中尤以β-变形菌群为主,占库容总量的45.90%,而首次在中国纸机鉴定出的水生施莱格尔氏菌(Schlegelella aquatica)为克隆文库的优势菌株,另外还含有2个菌种属于异常球菌-栖热菌门,该菌门为红色腐浆形成菌种。

Paper machines offer a suitable environment for microbial growth, which results in slime problems during the process of papermaking. Investigation of the bacterial diversity in whitewater which recycled in paper mill UPM was car-ried out in this paper. Total DNA were directly extracted from the whitewater sample. 16S rDNA were amplified directly from purified DNA by PCR with universally bacteria-specific rDNA primers and cloned. Sixty-one bacteria were identified by partial sequencing of their 16S rDNAs. Twenty-eight operational taxonomic units ( OTUs) were clustered in the follow-ing phyla including Proteobacteria ( 77. 05%) , Firmicutes ( 11. 48%) , Actinobacteria ( 4. 92%) , Deinococcus-Thermus (3.28%), Bacteroidetes(1.64%),Cyanobacteria(1.64%). Bacterial diversity was high in the whitewater sample. The results showed a high proportion of Schlegelella aquatica in the white water samples. It was the first time that Schlegelella aquatica strain had been identified from a paper machine