The application of laser microdissection to profiling fungal pathogen gene expression in planta

Methods Mol Biol. 2012:835:219-36. doi: 10.1007/978-1-61779-501-5_14.

Abstract

Laser microdissection (LM) of plant tissues infected with a fluorescent protein-tagged fungus is a useful method for obtaining samples highly enriched in fungal RNA for downstream analysis such as hybridization to a microarray. This paper outlines the requirements for successful LM of infected tissues and details a set of protocols for (1) preparing and sectioning infected tissue samples under conditions that preserve both RNA integrity and cytological features; (2) capturing fungal structures via LM; and (3) extraction and amplification of transcripts for further analysis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Equipment Design
  • Fungi / genetics*
  • Fungi / isolation & purification
  • Gene Expression Profiling / methods*
  • Lasers
  • Microdissection / instrumentation
  • Microdissection / methods*
  • Oligonucleotide Array Sequence Analysis / methods
  • Plants / microbiology*
  • Plants / ultrastructure
  • RNA, Fungal / genetics*
  • RNA, Fungal / isolation & purification
  • Tissue Embedding / methods
  • Tissue Fixation / methods

Substances

  • RNA, Fungal