title: Evaluating Fumonisin Gene Expression in Fusarium verticillioides.
abstract: Transcript levels of key genes in a biosynthetic pathway are often taken as a proxy for metabolite production. This is the case of FUM1, encoding the first dedicated enzyme in the metabolic pathway leading to the production of the mycotoxins Fumonisins by fungal species belonging to the genus Fusarium. FUM1 expression can be quantified by different methods; here, we detail a protocol based on quantitative reverse transcriptase polymerase chain reaction (RT-qPCR), by which relative or absolute transcript abundance can be estimated in Fusaria grown in vitro or in planta. As very seldom commercial kits for RNA extraction and cDNA synthesis are optimized for fungal samples, we developed a protocol tailored for these organisms, which stands alone but can be also easily integrated with specific reagents and kits commercially available.
authors.affiliation: Department of Environmental Biology, University of Rome "Sapienza", Largo Cristina di Svezia, 24, 00165, Rome, Italy.
authors.forename: Valeria
authors.initials: V
authors.lastname: Scala
authors.affiliation: Department of Agricultural, Forest and Food Sciences, University of Turin, Largo Paolo Braccini, 2, 10095, Grugliasco (TO), Italy.
authors.forename: Ivan
authors.initials: I
authors.lastname: Visentin
authors.affiliation: Department of Agricultural, Forest and Food Sciences, University of Turin, Largo Paolo Braccini, 2, 10095, Grugliasco (TO), Italy. francesca.cardinale@unito.it.
authors.forename: Francesca
authors.initials: F
authors.lastname: Cardinale
mesh_terms: D053898:Biosynthetic Pathways / Q000235:genetics*; D037341:Fumonisins / Q000378:metabolism*; D005670:Fusarium / Q000235:genetics* / Q000378:metabolism*; D015966:Gene Expression Regulation, Fungal; D060888:Real-Time Polymerase Chain Reaction; D003313:Zea mays / Q000382:microbiology
chemical_list: D037341:Fumonisins
keywords: F. verticillioides; FUM pathway; FUM1; RT-qPCR; mRNA