Molecular biomass and MetaTaxogenomic assessment of soil microbial communities as influenced by soil DNA extraction procedure

焦测序 放线菌门 厚壁菌 生物 稀薄(生态学) DNA提取 土壤水分 克雷纳恰奥塔 土壤微生物学 16S核糖体RNA 生物量(生态学) 蛋白质细菌 萃取(化学) 生物多样性 生态学 聚合酶链反应 遗传学 细菌 基因 色谱法 化学
作者
Sébastien Terrat,Richard Christen,Samuel Dequiedt,Mélanie Lelièvre,Virginie Nowak,Tiffanie Régnier,Dipankar Bachar,Pierre Plassart,Patrick Wincker,Claudy Jolivet,Antonio Bispo,Philippe Lemanceau,Pierre‐Alain Maron,Christophe Mougel,Lionel Ranjard
出处
期刊:Microbial biotechnology [Wiley]
卷期号:5 (1): 135-141 被引量:163
标识
DOI:10.1111/j.1751-7915.2011.00307.x
摘要

Summary Three soil DNA extraction procedures (homemade protocols and commercial kit) varying in their practicability were applied to contrasting soils to evaluate their efficiency in recovering: (i) soil DNA and (ii) bacterial diversity estimated by 16S rDNA pyrosequencing. Significant differences in DNA yield were systematically observed between tested procedures. For certain soils, 10 times more DNA was recovered with one protocol than with the others. About 15 000 sequences of 16S rDNA were obtained for each sample which were clustered to draw rarefaction curves. These curves, as well as the PCA ordination of community composition based on OTU clustering, did not reveal any significant difference between procedures. Nevertheless, significant differences between procedures were highlighted by the taxonomic identification of sequences obtained at the phylum to genus levels. Depending on the soil, differences in the number of genera detected ranged from 1% to 26% between the most and least efficient procedures, mainly due to a poorer capacity to recover populations belonging to Actinobacteria , Firmicutes or Crenarchaeota . This study enabled us to rank the relative efficiencies of protocols for their recovery of soil molecular microbial biomass and bacterial diversity and to help choosing an appropriate soil DNA extraction procedure adapted to novel sequencing technologies.

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