Journal Article

High-resolution community profiling of arbuscular mycorrhizal fungi

Marcel G. A. van der Heijden, Matthias C. Rillig, S. Franz Bender, Fabio Mascher, Stefan Hempel, Giancarlo Russo, Klaus Schlaeppi, Tessa Camenzind, Andrea Patrignani

2016

Overview

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.

• Publisher: New Phytologist Trust,Wiley Subscription Services, Inc
• Subject: AMF community analyses / AMF inoculation / arbuscular mycorrhizal fungi (AMF) / DNA / DNA primers / DNA, Fungal - genetics / Glomeromycota - physiology / Glomeromycota diversity / inoculum / internal transcribed spacers / Methods / microbiome / Mycorrhizae - physiology / mycorrhizal fungi / Operon - genetics / polymerase chain reaction / protein subunits / ribosomal proteins / RNA, Ribosomal - genetics / Sequence Analysis, DNA / single molecule real‐time (SMRT) sequencing / soil / Soil Microbiology