Sanger and next generation sequencing in the characterisation of arbuscular mycorrhizal fungi (AMF) in Pancratium maritimum L. (Amaryllidaceae), a representative plant species of Mediterranean sand dunes

Planta. 2018 Dec;248(6):1443-1453. doi: 10.1007/s00425-018-2981-z. Epub 2018 Aug 20.

Abstract

An interesting AMF colonization microcosm has been detected in the roots of Pancratium maritimum (sea daffodil). Both sequencing techniques (Sanger and NGS) have been used for AMF characterisation, showing a balanced trade-off between pros and cons. By Sanger and next generation sequencing of rRNA nuclear molecular markers (SSU-ITS-LSU and ITS2, respectively), the presence of AMF communities in the roots of P. maritimum was evaluated. Our results shed light on the presence of AMF in sea daffodil and the diversity of assemblages of AMF detected after Sanger sequencing of the SSU-ITS-LSU marker is much higher than that determined following NGS sequencing of ITS2 alone.

Keywords: DNA barcoding; ITS2; Illumina MiSeq; Psammophilic plant; Root; SSU–ITS–LSU; Sea daffodil.

MeSH terms

  • Amaryllidaceae / microbiology*
  • DNA Barcoding, Taxonomic
  • DNA, Fungal / chemistry
  • DNA, Fungal / genetics
  • DNA, Ribosomal / chemistry
  • DNA, Ribosomal / genetics
  • Fungi / genetics*
  • Fungi / isolation & purification
  • High-Throughput Nucleotide Sequencing
  • Mycorrhizae / genetics*
  • Mycorrhizae / isolation & purification
  • Plant Roots / microbiology
  • Sequence Analysis, DNA

Substances

  • DNA, Fungal
  • DNA, Ribosomal