High-resolution community profiling of arbuscular mycorrhizal fungi.
High-resolution community profiling of arbuscular mycorrhizal fungi.
复制标题
丛枝菌根真菌的高分辨率群落分析。
作者：
Klaus Schlaeppi;S. Bender;F. Mascher;G. Russo;A. Patrignani;Tessa Camenzind;Stefan Hempel;M. Rillig;M. G. Heijden;M. G. Heijden
利用核糖体小亚基(SSU)或内转录间隔区(ITS)DNA序列对丛枝菌根真菌(AMF)进行群落分析通常存在分辨率或覆盖率低的问题。我们开发了一种新的基于测序的方法，用于AMF群落的高分辨率和特异性分析。我们利用先前建立的AMF特异的PCR引物，扩增出覆盖SSU、ITS和部分大核糖体亚单位(LSU)部分的C.1.5kb长片段，并用单分子实时(SMRT)测序对得到的扩增产物进行了测序。该方法适用于土壤和根系样品，检测到了所有主要的AMF家族，并成功地区分了亲缘关系密切的AMF物种，而SSU序列无法区分这些物种。在接种试验中，我们可以在分子水平上对引入的AMF接种物进行追踪。其中一个引进的菌株几乎取代了当地的菌株(S)，这表明接种丛枝菌根真菌可以对本土群落产生深刻的影响。该方法为研究人员提供了一种强大的新工具，用于AMF群落分析，因为它结合了改进的特异性和增强的分辨率，而中等测序吞吐量的缺点对于AMF等低多样性群体似乎不那么重要。
Community analyses of arbuscular mycorrhizal fungi (AMF) using ribosomal small subunit (SSU) or internal transcribed spacer (ITS) DNA sequences often suffer from low resolution or coverage. We developed a novel sequencing based approach for a highly resolving and specific profiling of AMF communities. We took advantage of previously established AMF-specific PCR primers that amplify a c. 1.5-kb long fragment covering parts of SSU, ITS and parts of the large ribosomal subunit (LSU), and we sequenced the resulting amplicons with single molecule real-time (SMRT) sequencing. The method was applicable to soil and root samples, detected all major AMF families and successfully discriminated closely related AMF species, which would not be discernible using SSU sequences. In inoculation tests we could trace the introduced AMF inoculum at the molecular level. One of the introduced strains almost replaced the local strain(s), revealing that AMF inoculation can have a profound impact on the native community. The methodology presented offers researchers a powerful new tool for AMF community analysis because it unifies improved specificity and enhanced resolution, whereas the drawback of medium sequencing throughput appears of lesser importance for low-diversity groups such as AMF.
影响因子：
2.2
作者：
Mosher, Jennifer J.;Bernberg, Erin L.;Kaplan, Louis A.
通讯作者：
Kaplan, Louis A.
影响因子：
9.4
作者：
Dumbrell, Alex J.;Ashton, Peter D.;Helgason, Thorunn
通讯作者：
Helgason, Thorunn
DOI：
10.1073/pnas.1117018109
发表时间：
2012-04-17
影响因子：
11.1
作者：
Schoch, Conrad L.;Seifert, Keith A.;Schindel, David
通讯作者：
Schindel, David