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Characterizing Mycoviruses.

Mahtab Peyambari1, Marilyn J Roossinck2

  • 1Center for Infectious Disease Dynamics, Millennium Science Complex, Pennsylvania State University, University Park, PA, USA.

Methods in Molecular Biology (Clifton, N.J.)
|September 6, 2018
PubMed
Summary

This study presents a protocol for detecting fungal viruses by isolating double-stranded (ds) RNA from infected fungi. Molecular methods like sequencing are then used for virus characterization.

Keywords:
DNaseNucleic acidPhenolRACERNase

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Area of Science:

  • Mycology
  • Virology
  • Molecular Biology

Background:

  • Traditional methods for fungal virus detection include serological and biological assays.
  • Modern molecular techniques like polymerase chain reaction (PCR) and sequencing offer enhanced sensitivity for virus study.
  • Double-stranded RNA (dsRNA) isolation is a key strategy for detecting mycoviruses, as it accumulates in infected cells.

Purpose of the Study:

  • To present a basic protocol for the isolation and molecular characterization of fungal viruses.
  • To detail a method for extracting dsRNA from fungal tissues for mycovirus detection.
  • To outline subsequent molecular diagnostic steps for virus identification and analysis.

Main Methods:

  • Fungal strains were cultured using a basic protocol.
  • Double-stranded RNA (dsRNA) was isolated from fungal tissues utilizing cellulose chromatography.
  • Molecular diagnostics included complementary DNA (cDNA) synthesis, sequencing, and 5' end determination via primer ligation.

Main Results:

  • A reproducible protocol for dsRNA isolation from fungal samples was established.
  • The protocol facilitates the detection and characterization of mycoviruses.
  • Subsequent molecular analyses enabled virus genome sequencing and identification.

Conclusions:

  • The described protocol provides an effective method for fungal virus detection and characterization.
  • dsRNA isolation coupled with molecular diagnostics is a powerful approach for studying mycoviruses.
  • This method contributes to the understanding of fungal virology and disease mechanisms.