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新疆沙湾冷泉沉积物中免培养古菌多样性初步研究
引用本文:曾军,杨红梅,吴江超,徐建华,张涛,孙建,娄恺. 新疆沙湾冷泉沉积物中免培养古菌多样性初步研究[J]. 微生物学报, 2010, 50(5): 574-579
作者姓名:曾军  杨红梅  吴江超  徐建华  张涛  孙建  娄恺
作者单位:1. 新疆大学生命科学与技术学院,乌鲁木齐,830046;新疆农业科学院微生物研究所,乌鲁木齐,830091
2. 新疆农业科学院微生物研究所,乌鲁木齐,830091
基金项目:973计划前期研究专项(2008CB417214);新疆特殊环境微生物实验室开放课题(XJYS0203-2009-02)
摘    要:【目的】了解新疆沙湾冷泉沉积物的古菌组成及多样性。【方法】采用免培养法,液氮研磨提取冷泉沉积物总DNA,使用古菌通用引物进行16S rRNA基因扩增,构建16S rRNA基因文库。对阳性克隆进行HhaI限制性酶切分型,选出具有不同酶切图谱的序列进行测序,将所得序列与GenBank数据库中序列比对并构建16S rRNA基因系统发育树。【结果】从冷泉沉积物古菌16S rRNA基因文库中随机挑选了121个阳性克隆,共得到22个不同的可操作分类单元,BLAST结果表明全部克隆子归属于泉古菌门(Crenarchaeote)中免培养类群。系统发育分析归类为Soil-Freshwater-subsurface group和MarinegroupI,2个亚群并且各占整个文库的50%。其中40%左右的克隆子与具有无机碳和硝酸盐同化能力的泉古菌有高的相似性。此外还发现40%的克隆子与低温泉古菌类群具有很高的相似性。【结论】新疆沙湾冷泉沉积物中古菌类群多样性较低,但存有大量高度适应此低温、贫营养环境的泉古菌类群。

关 键 词:关键词:冷泉;地下水;古菌多样性;16S rRNA同源性
收稿时间:2009-11-07
修稿时间:2010-01-11

Uncultured archaeal diversity in cold spring sediment of Shawan, Xinjiang, China
Jun Zeng,Hongmei Yang,Jiangchao Wu,Jianhua Xu,Tao Zhang,Jian Sun and Kai Lou. Uncultured archaeal diversity in cold spring sediment of Shawan, Xinjiang, China[J]. Acta microbiologica Sinica, 2010, 50(5): 574-579
Authors:Jun Zeng  Hongmei Yang  Jiangchao Wu  Jianhua Xu  Tao Zhang  Jian Sun  Kai Lou
Abstract:Abstract: [Objective] We surveyed the composition and diversity of archaea in Xinjiang Shawan cold spring sediment. [Methods] We studied the archaeal diversity in the cold spring sediment by direct extracting environmental DNA with liquid nitrogen grinding method and constructing clone libraries of 16S rRNA gene amplified with archaeal-speci?c primers. Amplified 16S rRNA gene fragments were analyzed by Restriction Fragment Length Polymorphism (RFLP), and the unique RFLP patterns were selected for sequencing, alignment and constructing 16S rRNA gene phylogenetic tree. [Results] A total of 121 positive clones were randomly screened from the library and 22 Operational Taxonomic Units (OTUs) were determined. BLAST analysis indicated that all OTUs were affiliated with the phylum Crenarchaeota. Phylogenetic analysis classified them into two subgroups of Soil-Freshwater-subsurface group, and Marine group I and represented 50% of total clones, respectively. Of them, clones with the potential to assimilate nitrate accounted for 40% of the total archaeal clones. In addition, 40% of clones were related to the cold-loving Crenarchaeota. [Conclusion] These results suggested that the spring sediment harbors a low diversity of archaea, however, a large fraction of Crenarchaeota indigenous species might exist and well adapted to the cold and oligotraphic environment.
Keywords:Keywords: cold spring   groundwater   achaea   16S rRNA homogeny
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