Secondary metabolite production by cereal-associated penicillia during cultivation on cereal grains
Cereals are vulnerable substrates for fungal growth and subsequent mycotoxin contamination. One of the major fungal genera to colonize the ecosystem of stored grain is Penicillium, especially species in the series of Viridicata and Verrucosa. Culturing these species on grains, we hoped to induce the production of relevant secondary metabolites produced by these fungi in the early stage of cereal breakdown. In a multivariate setup six different cereal grains (wheat, rye, barley, oat, rice, and maize), one kind of white beans, and two standard fungal media, Yeast Extract Sucrose agar (YES agar) and Czapek Yeast Autolysate agar (CYA agar), were inoculated with the ten most important cereal-associated species from Penicillium (P. aurantiogriseum, P. cyclopium, P. freii, P. melanoconidium, P. neoechinulatum, P. polonicum, P. tricolor, P. viridicatum, P. hordei, and P. verrucosum). P. nordicum is a meat-associated species, which was included due to its chemical association with P. verrucosum, in addition to see if a substrate change would alter the profile of known chemistry. We found that cereals function very well as substrates for secondary metabolite production, but did not present significantly different secondary metabolite profiles, concerning known chemistry, as compared to standard laboratory agar media. However, white beans altered the semi-quantitative secondary metabolite profiles for several species. Correlations between substrates and certain metabolites were observed, as illuminated by principal component analysis. Many bioactive secondary metabolites were observed for the first time in the analyzed fungal species, including ergot type alkaloids in P. hordei.
KeywordsFilamentous fungi Cereals Mycotoxins Penicillium Viridicata Chemotaxonomy Secondary metabolites
We are grateful to Agilent Technologies for the Thought Leader Donation of the UHPLC-QTOFMS system.
We would like to thank the Novo Nordisk Foundation grant #NNF 130 C0005201 for the funding of this study.
Compliance with ethical standards
The authors declare that they have no conflict of interest.
Human and animal rights and informed consent
This article does not contain any studies with human participants or animals performed by any of the authors.
- Antia BS, Aree T, Kasettrathat C, Wiyakrutta S, Ekpa OD, Ekpe UJ, Mahidol C, Ruchirawat S, Kittakoop P (2011) Itaconic acid derivatives and diketopiperazine from the marine-derived fungus Aspergillus aculeatus CRI322-03. Phytochemistry 72:816–820. https://doi.org/10.1016/j.phytochem.2011.02.013 CrossRefPubMedGoogle Scholar
- Ciegler A, Fennell DI, Sansing GA, Detroy RW, Bennett GA (1973) Mycotoxin-producing strains of Penicillium viridicatum: classification into subgroups. Appl Environ Microbiol 26:271–278Google Scholar
- Frisvad JC, Samson RA (1991) Mycotoxin produced by species of Penicillium and Aspergillus occurring in cereals. In: Chelkowski J (ed) Cereal grain: mycotoxins, fungi and quality in drying and storage. Elsevier Inc., Amsterdam, pp 441–476Google Scholar
- Frisvad JC, Samson RA (2004) Polyphasic taxonomy of Penicillium subgenus Penicillium: a guide to identification of food and air-borne terverticillate Penicillia and their mycotoxins. Stud Mycol 49:1–174Google Scholar
- Frisvad JC, Thrane U (1987) Standardized high-performance liquid chromatography of 182 mycotoxins and other fungal metabolites based on alkylphenone retention indices and UV-VIS spectra (diodearray detection). J Chromatogr A 404:195–214. https://doi.org/10.1016/S0021-9673(01)86850-3 CrossRefGoogle Scholar
- Frisvad JC, Smedsgaard J, Larsen TO, Samson RA (2004) Mycotoxins , drugs and other extrolites produced by species in Penicillium subgenus Penicillium. Stud Mycol 49:201–241Google Scholar
- Grijseels S, Nielsen JC, Randelovic M, Nielsen J, Nielsen KF, Workman M, Frisvad JC (2016) Penicillium arizonense, a new, genome sequenced fungal species, reveals a high chemical diversity in secreted metabolites. Sci Rep 6:35112. https://doi.org/10.1038/srep35112 CrossRefPubMedPubMedCentralGoogle Scholar
- Hautbergue T, Puel O, Tadrist S, Meneghetti L, Péan M, Delaforge M, Debrauwer L, Oswald IP, Jamin EL (2017) Evidencing 98 secondary metabolites of Penicillium verrucosum using substrate isotopic labeling and high-resolution mass spectrometry. J Chromatogr B 1071:29–43. https://doi.org/10.1016/j.jchromb.2017.03.011 CrossRefGoogle Scholar
- Kildgaard S, Mansson M, Dosen I, Klitgaard A, Frisvad JC, Larsen TO, Nielsen KF (2014) Accurate dereplication of bioactive secondary metabolites from marine-derived fungi by UHPLC-DAD-QTOFMS and a MS/HRMS library. Mar Drugs 12:3681–3705. https://doi.org/10.3390/md12063681 CrossRefPubMedPubMedCentralGoogle Scholar
- Klitgaard A, Iversen A, Andersen MR, Larsen TO, Frisvad JC, Nielsen KF (2014) Aggressive dereplication using UHPLC-DAD-QTOF: screening extracts for up to 3000 fungal secondary metabolites. Anal Bioanal Chem 406:1933–1943. https://doi.org/10.1007/s00216-013-7582-x CrossRefPubMedPubMedCentralGoogle Scholar
- Li J, Wang J, Jiang C-S, Li G, Guo Y-W (2014) (+)-Cyclopenol, a new naturally occurring 7-membered 2,5-dioxopiperazine alkaloid from the fungus Penicillium sclerotiorum endogenous with the Chinese mangrove Bruguiera gymnorrhiza. J Asian Nat Prod Res 16(5):542–548. https://doi.org/10.1080/10286020.2014.911290 CrossRefPubMedGoogle Scholar
- Mulinge SK, Chesters CGC (1970) Ecology of fungi associated with moist stored barley grain. Ann Appl Biol 65:277–284. https://doi.org/10.1111/j.1744-7348.1970.tb04588.x CrossRefGoogle Scholar
- Muller HM, Boley A (1993) Studies on the refrigerated storage of wheat (Triticum aestivum). 2. Ergosterol, xanthomegnin, viomellein and brevianamide A after inoculation with Penicillium viridicatum. Zentralbl Mikrobiol 148:419–431. https://doi.org/10.1016/S0232-4393(11)80307-0 CrossRefPubMedGoogle Scholar
- Pitt JI (1979) The genus Penicillium and its teleomorphic states Eupenicillium and Talaromyces. Academic Press, LondonGoogle Scholar
- R Core Team (2015) R: language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. http://www.r-project.org/
- Richard JL, Payne GA (2003) Mycotoxins: risk in plant, animal, and human systems. Council for Agricultural Science and Technology (CAST) Report, AmesGoogle Scholar
- Samson RA, Houbraken J, Thrane U, Frisvad JC, Andersen B (2010) Food and indoor fungi, CBS Labora. CBS-KNAW, UtrechtGoogle Scholar
- Shimada A, Kusano M, Takeuchi S, Inokuchi T, Fujioka S, Kimura Y (2002) Aspterric acid and 6-hydroxymellein, inhibitors of pollen development in Arabidopsis thaliana, produced by Aspergillus terreus. Zeitschrift fur Naturforsch - sect C. J Biosci 57:459–464. https://doi.org/10.1515/znc-2002-5-610 CrossRefGoogle Scholar
- Sumner LW, Amberg A, Barrett D, Beale MH, Beger R, Daykin CA, Fan TWM, Fiehn O, Goodacre R, Griffin JL, Hankemeier T, Hardy N, Harnly J, Higashi R, Kopka J, Lane AN, Lindon JC, Marriott P, Nicholls AW, Reily MD, Thaden JJ, Viant MR (2007) Proposed minimum reporting standards for chemical analysis: Chemical Analysis Working Group (CAWG) Metabolomics Standards Initiative (MSI). Metabolomics 3:211–221. https://doi.org/10.1007/s11306-007-0082-2 CrossRefPubMedPubMedCentralGoogle Scholar
- Tsuda Y, Kaneda M, Tada A, Nitta K, Yamamoto Y, Iitaka Y (1978) Aspterric acid, a new sesquiterpenoid of the carotane group, a metabolite from Aspergillus terreus IFO-6123. X-ray crystal and molecular structure of its p-bromobenzoate. J C S Chem Commun 160–161. https://doi.org/10.1039/c39780000160