DNA-based species level detection of Glomeromycota: one PCR primer set for all arbuscular mycorrhizal fungi
DNA-based species level detection of Glomeromycota: one PCR primer set for all arbuscular mycorrhizal fungi
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DOI：
10.1111/j.1469-8137.2009.02835.x
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发表时间：
2009-01-01
期刊：
影响因子：
9.4
通讯作者：
Schuessler, Arthur
中科院分区：
文献类型：
--
作者：
Krueger, Manuela;Stockinger, Herbert;Schuessler, Arthur
目前，丛枝菌根真菌（AMF）的分子生态研究只有在针对整个群落时才能在物种水平以上进行。为了改善分子物种表征并允许在野外进行物种水平的群落分析，成功测试了一套新设计的 AMF 特异性 PCR 引物。对来自不同系统发育 AMF 谱系的核 rDNA 片段进行了测序和分析，设计了四种引物混合物，每种引物混合物针对小亚基 (SSU) 或大亚基 (LSU) rDNA 中的一个结合位点。为了实现物种分辨率，它们跨越了覆盖部分 SSU、整个内转录间隔区 (ITS) rDNA 区域和部分 LSU 的片段。新引物适用于从可能被其他生物体污染的材料（例如盆栽或田间样品）中特异性扩增 AMF rDNA，表征田间样品中 AMF 物种的多样性，并扩增 SSU-ITS-LSU 片段，以进行物种水平分辨率的系统发育分析。用于基于单个 rDNA 标记区域监测整个 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。New Phytological (2009) 183:212-223doi:10.1111/j.1469-8137.2009.02835.x。
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.New Phytologist (2009) 183: 212-223doi: 10.1111/j.1469-8137.2009.02835.x.