Metagenomics Approaches Based on Virion-Associated Nucleic Acids (VANA): An Innovative Tool for Assessing Without A Priori Viral Diversity of Plants

Methods Mol Biol. 2015:1302:249-57. doi: 10.1007/978-1-4939-2620-6_18.

Abstract

This chapter describes an efficient approach that combines quality and yield extraction of viral nucleic acids from plants containing high levels of secondary metabolites and a sequence-independent amplification procedure for both the inventory of known plant viruses and the discovery of unknown ones. This approach turns out to be a useful tool for assessing the virome (the genome of all the viruses that inhabit a particular organism) of plants of interest. We here show that this approach enables the identification of a novel Potyvirus member within a single plant already known to be infected by two other Potyvirus species.

MeSH terms

  • DNA, Viral / analysis*
  • DNA, Viral / genetics
  • Dioscorea / virology*
  • Genome, Viral
  • Metagenomics*
  • Plant Diseases / virology
  • Plant Viruses / classification*
  • Plant Viruses / genetics
  • Plant Viruses / isolation & purification
  • RNA, Viral / analysis*
  • RNA, Viral / genetics
  • Real-Time Polymerase Chain Reaction / methods*
  • Reverse Transcriptase Polymerase Chain Reaction / methods*
  • Species Specificity
  • Virion / genetics*

Substances

  • DNA, Viral
  • RNA, Viral