title: The application of laser microdissection to profiling fungal pathogen gene expression in planta.
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.
authors.affiliation: National Key Laboratory of Plant Molecular Genetics, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China.
authors.forename: Wei-Hua
authors.initials: WH
authors.lastname: Tang
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhang
authors.forename: Jon
authors.initials: J
authors.lastname: Duvick
mesh_terms: D004867:Equipment Design; D005658:Fungi / Q000235:genetics* / Q000302:isolation & purification; D020869:Gene Expression Profiling / Q000379:methods*; D007834:Lasers; D042282:Microdissection / Q000295:instrumentation / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D010944:Plants / Q000382:microbiology* / Q000648:ultrastructure; D012331:RNA, Fungal / Q000235:genetics* / Q000302:isolation & purification; D016610:Tissue Embedding / Q000379:methods; D016707:Tissue Fixation / Q000379:methods
chemical_list: D012331:RNA, Fungal