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Penicillium roqueforti PR toxin gene cluster characterization

  • Applied genetics and molecular biotechnology
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Abstract

PR toxin is a well-known isoprenoid mycotoxin almost solely produced by Penicillium roqueforti after growth on food or animal feed. This mycotoxin has been described as the most toxic produced by this species. In this study, an in silico analysis allowed identifying for the first time a 22.4-kb biosynthetic gene cluster involved in PR toxin biosynthesis in P. roqueforti. The pathway contains 11 open reading frames encoding for ten putative proteins including the major fungal terpene cyclase, aristolochene synthase, involved in the first farnesyl-diphosphate cyclization step as well as an oxidoreductase, an oxidase, two P450 monooxygenases, a transferase, and two dehydrogenase enzymes. Gene silencing was used to study three genes (ORF5, ORF6, and ORF8 encoding for an acetyltransferase and two P450 monooxygenases, respectively) and resulted in 20 to 40% PR toxin production reductions in all transformants proving the involvement of these genes and the corresponding enzyme activities in PR toxin biosynthesis. According to the considered silenced gene target, eremofortin A and B productions were also affected suggesting their involvement as biosynthetic intermediates in this pathway. A PR toxin biosynthesis pathway is proposed based on the most recent and available data.

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Acknowledgements

The authors would like to thank Dr. Olivier Puel from INRA Toulouse for useful discussion on PR toxin identification and for kindly providing a PR toxin sample. We are also grateful to Pr. Joëlle Dupont from the MNHN for providing the P. roqueforti FM164 genome sequence, Nolwenn Hymery for RT-qPCR statistical analyses, and Riccardo Baroncelli for bioinformatics assistance for PR toxin clusters.

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Authors and Affiliations

  1. Université de Brest, EA 3882 Laboratoire Universitaire de Biodiversité et d’Ecologie Microbienne, IBSAM, ESIAB, Technopôle Brest-Iroise, Plouzané, 29280, Brest, France

    Pedro I. Hidalgo, Elisabeth Poirier, Justine Piqueras, Laurence Meslet-Cladière, Emmanuel Coton & Monika Coton

  2. mAbxience, Upstream Production, Parque Tecnológico de León, Julia Morros s/n, Armunia, 24009, León, Spain

    Ricardo V. Ullán

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  1. Pedro I. Hidalgo
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  2. Elisabeth Poirier
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  5. Laurence Meslet-Cladière
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  6. Emmanuel Coton
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  7. Monika Coton
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Correspondence to Monika Coton.

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This study was supported by funding from the Région Bretagne, France (PENIMET 7923, Stratégie d’Attractivité Durable), to Monika Coton for a post-doctoral fellowship for Pedro Hidalgo.

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All authors declare that they have no conflict of interest.

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This article does not contain any studies with human participants or animals performed by any of the authors.

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Hidalgo, P.I., Poirier, E., Ullán, R.V. et al. Penicillium roqueforti PR toxin gene cluster characterization. Appl Microbiol Biotechnol 101, 2043–2056 (2017). https://doi.org/10.1007/s00253-016-7995-5

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