Abstract
The determination of aflatoxin production ability and differentiation of aflatoxigenic strains can be assessed by monitoring the expression of one or several key genes using reverse transcription polymerase chain reaction (RT-PCR). We herein describe the methods for RNA induction, extraction, and quality determination, and the RT-PCR conditions used to evaluate the ability of a given Aspergillus strain to produce aflatoxins.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Yu J, Bhatnagar D, Cleveland TE (2004) Completed sequence of aflatoxin pathway gene cluster in Aspergillus parasiticus. FEBS Lett 564:126–130
Rodrigues P, Venâncio A, Kozakiewicz Z, Lima N (2009) A polyphasic approach to the identification of aflatoxigenic and non-aflatoxigenic strains of Aspergillus Section Flavi isolated from Portuguese almonds. Int J Food Microbiol 129:187–193
Paterson RPM (2006) Identification and quantification of mycotoxigenic fungi by PCR. Process Biochem 41:1467–1474
Sweeney MJ, Pàmies P, Dobson ADW (2000) The use of reverse transcription-polymerase chain reaction (RT-PCR) for monitoring aflatoxin production in Aspergillus parasiticus 439. Int J Food Microbiol 56:97–103
Scherm B, Palomba M, Serra D et al (2005) Detection of transcripts of the aflatoxin genes aflD, aflO, and aflP by reverse-transcription-polymerase chain reaction allows differentiation of aflatoxin-producing isolates of Aspergillus flavus and Aspergillus parasiticus. Int J Food Microbiol 98:201–210
Degola F, Berni E, Dall’Asta C et al (2007) A multiplex RT-PCR approach to detect aflatoxigenic strains of Aspergillus flavus. J Appl Microbiol 103:409–417
Jamali M, Karimipour M, Shams-Ghahfarokhi M et al (2013) Expression of aflatoxin genes aflO (omtB) and aflQ (ordA) differentiates levels of aflatoxin production by Aspergillus flavus strains from soils of pistachio orchards. Res Microbiol 164:293–299
Heptinstall J, Rapley R (2000) Spectrophotometric analysis of nucleic acids. In: Rapley R (ed) The nucleic acid protocols handbook. Humana Press, USA
Rio DC, Ares M Jr, Hannon GJ, Nilsen TW (2010) Purification of RNA using TRIzol (TRI reagent). Cold Spring Harb Protoc (6):pdb.prot5439
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media LLC
About this protocol
Cite this protocol
Rodrigues, P.C.A., Gil-Serna, J., González-Jaén, M.T. (2017). Evaluating Aflatoxin Gene Expression in Aspergillus Section Flavi. 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_15
Download citation
DOI: https://doi.org/10.1007/978-1-4939-6707-0_15
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-6705-6
Online ISBN: 978-1-4939-6707-0
eBook Packages: Springer Protocols