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采用未培养技术对荷斯坦奶牛瘤胃细菌多样性进行初步分析
引用本文:王远亮,杨瑞红,毛爱军,王加启,董志扬.采用未培养技术对荷斯坦奶牛瘤胃细菌多样性进行初步分析[J].微生物学报,2005,45(6):915-919.
作者姓名:王远亮  杨瑞红  毛爱军  王加启  董志扬
作者单位:1. 中国科学院微生物研究所,北京,100080
2. 中国农业科学院畜牧研究所,北京,10093;新疆农业大学农学院,乌鲁木齐,830052
3. 中国农业科学院畜牧研究所,北京,10093
基金项目:国家“863计划”(2004AA214080)~~
摘    要:采用未培养(Culture independent)技术直接从荷斯坦奶牛瘤胃液中提取瘤胃细菌微生物混合DNA(也叫元基因组DNA),利用细菌16SrDNA通用引物27F与1492R,扩增瘤胃混合微生物的16SrDNA,根据16SrDNA序列对瘤胃细菌多样性进行初步分析。通过16SrDNA序列同源性分析,发现有多于一半以上的序列与可培养的菌株的同源性小于90%,属于不可培养的菌株。选用45条测得序列与已知序列构建系统发育树,分析结果表明,它们分属于两大类LGCGPB(the lowG CGram positivebac teria)和CFB(Cytophaga_Flexibacter $CBacteroides group),剩下的克隆尚难确定其分类地位,可能是代表新属和种的序列,这些序列已向GenBank提交并得到序列号(AY986777_AY986791)。

关 键 词:瘤胃  微生物多样性  系统发育分析
文章编号:0001-6209(2005)06-0915-05
收稿时间:2005-04-15
修稿时间:2005-09-11

Phylogenetic diversity analyse of Rumen Bacteria using culture independent method
WANG Yuan-liang,YANG Rui-hong,MAO Ai-jun,WANG Jia-qi,DONG Zhi-yang.Phylogenetic diversity analyse of Rumen Bacteria using culture independent method[J].Acta Microbiologica Sinica,2005,45(6):915-919.
Authors:WANG Yuan-liang  YANG Rui-hong  MAO Ai-jun  WANG Jia-qi  DONG Zhi-yang
Institution:1 Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, China ; 2 Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 10093, China ; 3 Agriculture College, Xinjiang Agriculture University, Urimqi 830052, China
Abstract:Culture independent method was used to study the diversity of rumen bacteria. Molecular diversity of rumen bacteria was analyzed by PCR amplification and sequencing of 16S rDNA clone libraries prepared from the rumen content of Holstein cows. The total DNA directly extracted from rumen fluid was used as PCR template. Bacteria universal primer 27F and 1492R was used as primer. Random clones, containing almost full size 16S rDNA sequences (about 1.5 kb long), were sequenced and subjected to an on line similarity search. The 16S rDNA sequence analysis indicated that more than half of the sequences belonged to the not-yet-cultured groups. The 16S rDNA similarity levels with cultured species was less than 90%. The bacterial community structure was also revealed by phylogenetic tree of known sequences and selected sequence. In the library from the rumen fluid, the sequences were mainly affiliated with the following major phyla: low G C Gram-positive bacteria, Cytophaga-Flexibacter-Bacteroides, and the remaining sequences were placed within not-yet-uncultured groups that had an uncertain affiliation. These several sequences are likely to represent novel taxonomic groupings. The nucleotide sequences have been submitted to the GenBank/EMBL /DDBJ databases under accession numbers AY986777-AY986791.
Keywords:Rumen  Microbiology diversity  Phylogenetic analysis  
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