Enriching plant microbiota for a metagenomic library construction
Wang, Hao-Xin1,2; Geng, Zhao-Liang1,2; Zeng, Ying1; Shen, Yue-Mao1
2008-10-01
发表期刊ENVIRONMENTAL MICROBIOLOGY
ISSN1462-2912
卷号10期号:10页码:2684-2691
摘要Plant microbiota (the microorganisms that live in any associations with plant tissues) represents a rather unexplored area of metagenomic research compared with soils and oceans. Constructing a metagenomic library for plant microbiota is technically challenging. Using all the biomass without pre-enrichment could lead to vast proportions of the host plant DNA in the metagenomic library, doubtless obliterating the microbial contribution. Therefore, the first and essential step is to enrich for the constituent microorganisms from plant tissues. Here, a strong enrichment for plant microbiota was achieved by coupling SDS (sodium dodecyl sulfate) with NaCl, creating a predominantly microbial metagenomic library that contains 88% bacterial inserts. 16S rDNA sequence analysis revealed that the metagenomic DNA of enrichments originates from very diverse microorganisms. At least 74 distinct ribotypes (at a 97% threshold) from seven different bacterial phyla were identified and mainly distributed among Actinobacteria and Proteobacteria. Additionally, a simplified version of Amplified Ribosomal DNA Restriction Analysis (ARDRA) was developed for a quick and efficient assessment of the enriching procedures. This work opens further insight into the great biotechnical potential of plant microbiota, holding more potential for drug discovery through a metagenomic strategy, and paving the way for recovery and biochemical characterization of functional gene repertoire from plant microbiota.
关键词Ribosomal-rna Sequences Uncultured Microorganisms Diversity Database Project Genomes Dna
资助信息National Natural Science Foundation of China [30430020]
学科领域Microbiology
DOI10.1111/j.1462-2920.2008.01689.x
收录类别SCI
语种英语
WOS研究方向Microbiology
WOS类目Microbiology
WOS记录号WOS:000259147900019
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被引频次:25[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.kib.ac.cn/handle/151853/1715
专题植物化学与西部植物资源持续利用国家重点实验室
作者单位1.Chinese Acad Sci, State Key Lab Phytochem & Plant Resources W China, Kunming Inst Bot, Kunming 650204, Yunnan, Peoples R China
2.Chinese Acad Sci, Grad Sch, Beijing 100039, Peoples R China
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Wang, Hao-Xin,Geng, Zhao-Liang,Zeng, Ying,et al. Enriching plant microbiota for a metagenomic library construction[J]. ENVIRONMENTAL MICROBIOLOGY,2008,10(10):2684-2691.
APA Wang, Hao-Xin,Geng, Zhao-Liang,Zeng, Ying,&Shen, Yue-Mao.(2008).Enriching plant microbiota for a metagenomic library construction.ENVIRONMENTAL MICROBIOLOGY,10(10),2684-2691.
MLA Wang, Hao-Xin,et al."Enriching plant microbiota for a metagenomic library construction".ENVIRONMENTAL MICROBIOLOGY 10.10(2008):2684-2691.
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