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Ochratoxin A producing Penicillium verrucosum isolates from cereals reveal large AFLP fingerprinting variability

Frisvad, J.C.; Lund, F. and Elmholt, S. (2005) Ochratoxin A producing Penicillium verrucosum isolates from cereals reveal large AFLP fingerprinting variability. [.] Journal of Applied Microbiology, 98, pp. 684-692.

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Aims: To examine if molecular amplified fragment length polymorphism (AFLP) fingerprinting of the only ochratoxin A-producing species in European cereals, Penicillium verrucosum, can be used as a method in hazard analysis using critical control points (HACCP).
Methods and Results: A total of 321 isolates of P. verrucosum were isolated from ochratoxin A contaminated cereals from Denmark (oats), UK (wheat and barley) and Sweden (wheat). Of these, 236 produced ochratoxin A as determined by thin layer chromatography; 185 ochratoxin A-producing isolates were selected for AFLP fingerprinting. A total of 138 isolates had unique AFLP patterns, whereas 52 isolates could be allocated to small groups containing from two to four isolates with similar AFLP patterns. A total of 155 clones were found among the 185 P. verrucosum isolates, thus 84 % of the isolates may represent different genets of P. verrucosum. As the few isolates that were grouped often came from the same farm, and those groups that contained AFLP-identical isolates from different countries were morphotypically different. On single farms up to 35 clones were found. The few groups of ramets from the same genet indicated that a HACCP approach based on clones may require a very large number of AFLP analysis to work in practice, we recommend basing the HACCP approach on the actual species P. verrucosum. A more detailed characterization should rather be based on the profile of species present at different control points, or analysis of the mycotoxins ochratoxin A and citrinin in the isolates. Examination of 86 isolates with HPLC and diode array detection of P. verrucosum showed that 66% produced ochratoxin A, 87% produced citrinin, 92% produced verrucin and 100% produced verrucolone.
Conclusions: Among 184 ochratoxin A-producing Penicillium verrucosum, 155 clonal lineages were indicated by AFLP fingerprinting, indicating a high genetical diversity, yet the species P. verrucosum is phenotypically distinct and valid.
Significance and Impact of the Study: AFLP fingerprinting of Penicillium verrucosum indicates that genetic recombination takes place in this fungus.

EPrint Type:Journal paper
Subjects: Crop husbandry > Post harvest management and techniques
Research affiliation: Denmark > DARCOF II (2000-2005) > I.12 (PREMYTOX) Preventing mycotoxin problems
Deposited By: Elmholt, Susanne
ID Code:7392
Deposited On:28 Feb 2006
Last Modified:12 Apr 2010 07:32
Document Language:English
Refereed:Peer-reviewed and accepted

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