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基于DNA的球囊菌门物种水平检测:一套适用于所有丛枝菌根真菌的PCR引物。

DNA-based species level detection of Glomeromycota: one PCR primer set for all arbuscular mycorrhizal fungi.

作者信息

Krüger Manuela, Stockinger Herbert, Krüger Claudia, Schüßler Arthur

机构信息

Ludwig-Maximilians-University Munich, Dept Biology I, Genetics, Großhaderner Strasse 4, D-82152 Planegg-Martinsried, Germany.

出版信息

New Phytol. 2009;183(1):212-223. doi: 10.1111/j.1469-8137.2009.02835.x. Epub 2009 Apr 8.

DOI:10.1111/j.1469-8137.2009.02835.x
PMID:19368665
Abstract
  • At present, molecular ecological studies of arbuscular mycorrhizal fungi (AMF) are only possible above species level when targeting entire communities. To improve molecular species characterization and to allow species level community analyses in the field, a set of newly designed AMF specific PCR primers was successfully tested. * Nuclear rDNA fragments from diverse phylogenetic AMF lineages were sequenced and analysed to design four primer mixtures, each targeting one binding site in the small subunit (SSU) or large subunit (LSU) rDNA. To allow species resolution, they span a fragment covering the partial SSU, whole internal transcribed spacer (ITS) rDNA region and partial LSU. * The new primers are suitable for specifically amplifying AMF rDNA from material that may be contaminated by other organisms (e.g., samples from pot cultures or the field), characterizing the diversity of AMF species from field samples, and amplifying a SSU-ITS-LSU fragment that allows phylogenetic analyses with species level resolution. * The PCR primers can be used to monitor entire AMF field communities, based on a single rDNA marker region. Their application will improve the base for deep sequencing approaches; moreover, they can be efficiently used as DNA barcoding primers.
摘要

目前,在针对整个群落时,丛枝菌根真菌(AMF)的分子生态学研究仅在物种水平以上才可行。为了改进分子物种表征并实现田间物种水平的群落分析,一组新设计的AMF特异性PCR引物已成功测试。对来自不同系统发育AMF谱系的核rDNA片段进行测序和分析,以设计四种引物混合物,每种引物混合物靶向小亚基(SSU)或大亚基(LSU)rDNA中的一个结合位点。为了实现物种分辨率,它们跨越一个片段,覆盖部分SSU、整个内部转录间隔区(ITS)rDNA区域和部分LSU。新引物适用于从可能被其他生物污染的材料(如盆栽培养物或田间样本)中特异性扩增AMF rDNA,表征田间样本中AMF物种的多样性,并扩增一个SSU-ITS-LSU片段,以便进行具有物种水平分辨率的系统发育分析。基于单个rDNA标记区域,PCR引物可用于监测整个AMF田间群落。它们的应用将改善深度测序方法的基础;此外,它们可以有效地用作DNA条形码引物。

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