Transcriptomic Analyses of Phytoplasmas

Methods Mol Biol. 2019:1875:239-251. doi: 10.1007/978-1-4939-8837-2_18.

Abstract

Transcriptomic analyses addressed to study phytoplasma gene expression may present few difficulties due to the uncultivable nature of these intracellular, obligate pathogens. While RNA extraction from insect vectors does not imply any particular adaptation of the protocols used in most commercial kits, RNA isolation from phytoplasma-infected plants can be a challenging task, given the high levels of polyphenol contents and accumulation of sucrose and starch in the different plant tissues. Here, we describe two different transcriptomic approaches, one focused on RNA phytoplasma sequencing and the other on phytoplasma quantitative gene expression in relation to pathogen load.

Keywords: Expression index; Gene expression; Phytoplasma load; Quantitative RT-PCR; RNA sequencing.

MeSH terms

  • Animals
  • Bacterial Load
  • Gene Expression Profiling / methods*
  • Gene Expression Regulation, Bacterial
  • Phytoplasma / genetics*
  • Phytoplasma / pathogenicity
  • Plant Leaves / chemistry
  • Plant Leaves / microbiology
  • Plants / chemistry
  • Plants / microbiology*
  • Polyphenols / chemistry
  • Reverse Transcriptase Polymerase Chain Reaction / methods
  • Sequence Analysis, RNA
  • Starch / chemistry
  • Sucrose / chemistry

Substances

  • Polyphenols
  • Sucrose
  • Starch