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 (qRT-PCR), 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.

Evaluating fumonisin gene expression in Fusarium verticilliioides

VISENTIN, IVAN;CARDINALE, Francesca
2017-01-01

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 (qRT-PCR), 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.
2017
Mycotoxigenic Fungi. Methods and Protocols
Humana Press Inc (Springer)
Methods in Molecular Biology
249
257
978-1-4939-6705-6
F. verticillioides, FUM pathway, FUM1, mRNA, RT-qPCR
Valeria Scala; Ivan Visentin; Francesca Cardinale
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2318/1634042
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