Jul 7, 2016

High-resolution community profiling of arbuscular mycorrhizal fungi

The New Phytologist
Klaus SchlaeppiMarcel G A van der Heijden

Abstract

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 importan...Continue Reading

  • References42
  • Citations15

References

  • References42
  • Citations15

Citations

Mentioned in this Paper

Glomeromycota
NCOR2 wt Allele
Ribosome Subunits, Large
Research Personnel
SSU-1 protein, C elegans
Vaccination
SSU rRNA Binding
Sequencing
High Throughput Analysis
Oligonucleotide Primers

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