Determination of the Mycovirome of a Necrotrophic Fungus.

Methods Mol Biol

Centro de Biotecnología y Genómica de Plantas, Universidad Politécnica de Madrid (UPM)-Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Madrid, Spain.

Published: December 2023

AI Article Synopsis

  • Next-generation sequencing (NGS) of total RNA has enhanced our understanding of viruses affecting various hosts, including fungi, by revealing new viruses and their evolutionary patterns.
  • The text outlines specific procedures for isolating the plant pathogenic fungus Botrytis cinerea from grapevine plants with gray mold, as well as methods for culturing and maintaining this fungus.
  • It details the process of RNA extraction for NGS, a bioinformatics pipeline for identifying mycoviruses in the samples, and techniques for validating the detected mycoviruses.

Article Abstract

Next-generation sequencing (NGS) of total RNA has allowed the detection of novel viruses infecting different hosts, such as fungi, increasing our knowledge on virus horizontal transfer events among different hosts, virus diversity, and virus evolution. Here, we describe the detailed protocols for the isolation of the plant pathogenic fungus Botrytis cinerea, from grapevine plants showing symptoms of the mold gray disease, the culture and maintenance of the isolated B. cinerea strains, the extraction of total RNA from B. cinerea strains for NGS, the bioinformatics pipeline designed and followed to detect mycoviruses in the sequenced samples, and the validation of the in silico detected mycoviruses by different approaches.

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http://dx.doi.org/10.1007/978-1-0716-3515-5_6DOI Listing

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