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Next-generation Sequencing of 16S Ribosomal RNA Gene Amplicons
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High-Throughput DNA Sequence-Based Analysis of AMF Communities.

Íris Marisa Maxaieie Victorino1,2,3, Andrea Berruti4, Alberto Orgiazzi5

  • 1Institute for Sustainable Plant Protection, National Research Council (CNR), Torino, Italy. rvictori@unito.it.

Methods in Molecular Biology (Clifton, N.J.)
|May 17, 2020
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Summary

Arbuscular mycorrhizal fungi (AMF) are crucial plant symbionts. This study presents a standardized high-throughput sequencing protocol for identifying AMF in soil and root communities using DNA targets.

Keywords:
Arbuscular mycorrhizal fungi (AMF)DNA-based species characterizationHigh-throughput sequenceITS primersIllumina MiSeqSSU primers

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Area of Science:

  • Mycology
  • Plant Science
  • Ecology

Background:

  • Arbuscular mycorrhizal fungi (AMF) form essential symbiotic relationships with most terrestrial plants.
  • These fungi play vital roles in plant nutrition, water uptake, soil health, and pathogen resistance.
  • Understanding AMF distribution is critical for ecological and agricultural applications.

Purpose of the Study:

  • To establish a standardized, high-throughput sequencing workflow for AMF identification.
  • To enable accurate assessment of AMF communities in both soil and plant roots.
  • To facilitate large-scale ecological studies on AMF.

Main Methods:

  • Utilized high-throughput sequencing on the Illumina MiSeq platform.
  • Targeted two key DNA regions: the small subunit (SSU) and internal transcribed spacer (ITS).
  • Developed a protocol applicable to DNA extracted from both soil and root samples.

Main Results:

  • The described workflow allows for efficient and reliable identification of AMF taxa.
  • The protocol is adaptable for analyzing diverse AMF communities from different environmental matrices.
  • Provides a foundation for comparative studies on AMF diversity and abundance.

Conclusions:

  • The presented protocol offers a standardized approach for AMF community analysis.
  • This method supports ecological research by enabling robust AMF identification.
  • Facilitates a deeper understanding of AMF roles in various ecosystems.