title: Membrane-assisted culture of fungal mycelium on agar plates for RNA extraction and pharmacological analyses.
abstract: Fungal mycelium grown in liquid culture is easy to harvest for RNA extraction and gene expression analyses, but liquid cultures often develop rather heterogeneously. In contrast, growth of fungal mycelium on agar plates is highly reproducible. However, this biological material cannot be harvested easily for downstream analyses. This article describes a PVDF (polyvinylidene difluoride) membrane-assisted agar plate culture method that enables the harvest of mycelium grown on agar plates. This culture method leads to a strongly reduced variation in gene expression between biological replicates and requires less growth space as compared with liquid cultures.
authors.affiliation: Plant Nutrition Laboratory, Institute of Agricultural and Nutritional Sciences (IAEW), Faculty of Natural Sciences III, Martin Luther University of Halle-Wittenberg, 06099 Halle (Saale), Germany.
authors.forename: Mario
authors.initials: M
authors.lastname: Lange
authors.affiliation: Plant Nutrition Laboratory, Institute of Agricultural and Nutritional Sciences (IAEW), Faculty of Natural Sciences III, Martin Luther University of Halle-Wittenberg, 06099 Halle (Saale), Germany.
authors.forename: Carolin
authors.initials: C
authors.lastname: Müller
authors.affiliation: Plant Nutrition Laboratory, Institute of Agricultural and Nutritional Sciences (IAEW), Faculty of Natural Sciences III, Martin Luther University of Halle-Wittenberg, 06099 Halle (Saale), Germany. Electronic address: edgar.peiter@landw.uni-halle.de.
authors.forename: Edgar
authors.initials: E
authors.lastname: Peiter
mesh_terms: D000362:Agar; D020231:Colletotrichum / Q000187:drug effects / Q000235:genetics / Q000254:growth & development*; D015169:Colony Count, Microbial; D015966:Gene Expression Regulation, Fungal; D008567:Membranes, Artificial; D025282:Mycelium / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development*; D011145:Polyvinyls; D012331:RNA, Fungal / Q000235:genetics / Q000302:isolation & purification*; D013499:Surface Properties; D014230:Triazoles / Q000494:pharmacology*; D005465:Fluorocarbon Polymers
chemical_list: D000362:Agar; D008567:Membranes, Artificial; D011145:Polyvinyls; D012331:RNA, Fungal; D014230:Triazoles; C024865:polyvinylidene fluoride; D005465:Fluorocarbon Polymers; C087114:tebuconazole
keywords: Colletotrichum graminicola; Fungal growth; Gene expression; PVDF membrane; RNA extraction