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.
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Acknowledgement
This work was supported by the Ministry of Research and Education through the project FIRB2008 “Futuro in Ricerca,” grant N. FIRB-RBFR08JKHI to V.S.
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Scala, V., Visentin, I., Cardinale, F. (2017). Evaluating Fumonisin Gene Expression in Fusarium verticillioides . In: Moretti, A., Susca, A. (eds) Mycotoxigenic Fungi. Methods in Molecular Biology, vol 1542. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6707-0_16
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DOI: https://doi.org/10.1007/978-1-4939-6707-0_16
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Publisher Name: Humana Press, New York, NY
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Online ISBN: 978-1-4939-6707-0
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