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Comprehensive molecular system to study the presence, growth and ochratoxin A biosynthesis of Penicillium verrucosum in wheat

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Based on the sequence of the ochratoxin A polyketide synthase gene (otapksPV), a polymerase chain reaction (PCR) system for the specific detection of Penicillium verrucosum in wheat has been developed. In a further approach, a real-time PCR system has been applied to determine the growth kinetics of P. verrucosum in wheat at cell numbers above 103 colony-forming units (cfu) ml-1. The data obtained by real-time PCR correlated well with the data obtained by the plate count technique. For this purpose, the DNA was isolated directly from contaminated wheat without any further enrichment step. In a reverse transcriptase real-time PCR, the expression of the otapksPV gene in wheat was detected 22 days after inoculation and storage at ambient temperature. Reasonable amounts of ochratoxin A, however, could not be detected before day 30. This early activation of ochratoxin A related genes was confirmed by microarray analysis.
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Keywords: cereals; molecular biology; mycotoxins; ochratoxin A; polymerase chain reaction (PCR)

Document Type: Research Article

Affiliations: 1: Max Rubner Institute-Federal Research Centre of Nutrition and Food, Karlsruhe, Germany 2: Bavarian State Research Centre for Agriculture, Institute of Animal Nutrition and Forage, Poing, Germany 3: European Commission, Joint Research Centre, Institute for Reference Materials and Measurements, Geel, Belgium

Publication date: August 1, 2008

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