Abstract
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.
Similar content being viewed by others
References
Abdallah MF, Girgin G, Baydar T, Krska R, Sulyok M (2017) Occurrence of multiple mycotoxins and other fungal metabolites in animal feed and maize samples from Egypt using LC-MS/MS. J Sci Food Agric 97:4419–4428. https://doi.org/10.1002/jsfa.8293
Adatia R, Heaton JM, Macgeorge KM, Mantle PG (1991) Acute histopathological changes produced by Penicillium aurantiogriseum nephrotoxin in the rat. Int J Exp Pathol 72:47–53
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
Axberg K, Jansson G, Svensson G, Hult K (1997) Varietal differences in accumulation of ochratoxin A in barley and wheat cultivars after inoculation of Penicillium verrucosum. Acta Agric Scand Sect B Soil Plant Sci 47:229–237. https://doi.org/10.1080/09064719709362465
Bennett JW, Klich M (2003) Mycotoxins. Clin Microbiol Rev 16:497–516. https://doi.org/10.1128/CMR.16.3.497
Ciegler A, Fennell DI, Sansing GA, Detroy RW, Bennett GA (1973) Mycotoxin-producing strains of Penicillium viridicatum: classification into subgroups. Appl Environ Microbiol 26:271–278
Coufal-Majewski S, Stanford K, McAllister T, Blakley B, McKinnon J, Chaves AV, Wang Y (2016) Impacts of cereal ergot in food animal production. Front Vet Sci 3:15. https://doi.org/10.3389/fvets.2016.00015
El-Neketi M, Ebrahim W, Lin W, Gedara S, Badria F, HE a S, Lai D, Proksch P (2013) Alkaloids and polyketides from Penicillium citrinum, an endophyte isolated from the Moroccan plant Ceratonia siliqua. J Nat Prod 76:1099–1104. https://doi.org/10.1021/np4001366
Fox EM, Howlett BJ (2008) Secondary metabolism: regulation and role in fungal biology. Curr Opin Microbiol 11:481–487. https://doi.org/10.1016/j.mib.2008.10.007
Frisvad JC, Filtenborg O (1983) Classification of terverticillate penicillia based on profiles of mycotoxins and other secondary metabolites. Appl Environ Microbiol 46:1301–1310
Frisvad JC, Filtenborg O (1989) Terverticillate penicillia: chemotaxonomy and mycotoxin production. Mycologia 81:837–861. https://doi.org/10.2307/3760103
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 441–476
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 49:1–174
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
Frisvad JC, Filtenborg O, Wicklow DT (1987) Terverticillate penicillia isolated from underground seed caches and cheek pouches of banner-tailed kangaroo rats (Dipodomys spectabilis). Can J Bot 65:765–773. https://doi.org/10.1139/b87-102
Frisvad JC, Seifert KA, Samson A, Mills JT (1994) Penicillium tricolor, a new mould species from Canadian wheat. Can J Bot 72:933–939. https://doi.org/10.1139/b94-118
Frisvad JC, Smedsgaard J, Larsen TO, Samson RA (2004) Mycotoxins , drugs and other extrolites produced by species in Penicillium subgenus Penicillium. Stud Mycol 49:201–241
Gerhards N, Neubauer L, Tudzynski P, Li S-M (2014) Biosynthetic pathways of ergot alkaloids. Toxins 6:3281–3295. https://doi.org/10.3390/toxins6123281
Giubergia S, Phippen C, Gotfredsen CH, Nielsen KF, Gram L (2016) Influence of niche-specific nutrients on secondary metabolism in Vibrionaceae. Appl Environ Microbiol 82:4035–4044. https://doi.org/10.1128/AEM.00730-16
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
Hallas-Møller M, Nielsen KF, Frisvad JC (2016) Production of the Fusarium mycotoxin moniliformin by Penicillium melanoconidium. J Agric Food Chem 64:4505–4510. https://doi.org/10.1021/acs.jafc.6b00298
Hammerschmidt L, Wray V, Lin W, Kamilova E, Proksch P, Aly AH (2012) New styrylpyrones from the fungal endophyte Penicillium glabrum isolated from Punica granatum. Phytochem Lett 5:600–603. https://doi.org/10.1016/j.phytol.2012.06.003
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
Jakubczyk D, Cheng JZ, O’Connor SE (2014) Biosynthesis of the ergot alkaloids. Nat Prod Rep 31:1328–1338. https://doi.org/10.1039/C4NP00062E
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
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
Lai D, Brötz-Oesterhelt H, Müller WEG, Wray V, Proksch P (2013) Bioactive polyketides and alkaloids from Penicillium citrinum, a fungal endophyte isolated from Ocimum tenuiflorum. Fitoterapia 91:100–106. https://doi.org/10.1016/j.fitote.2013.08.017
Lam YKT, Dai P, Borris R, Dombrowski A, Ransom R, Young G, Beer M, Middlemiss D, Smith J (1994) A new indole from Penicillium daleae. J Antibiot (Tokyo) 47:724–726. https://doi.org/10.7164/antibiotics.47.724
Larsen TO, Frisvad JC, Jensen SR (1992) Aurantiamine, a diketopiperazine from two varieties of Penicillium aurantiogriseum. Phytochemistry 31:1613–1615. https://doi.org/10.1016/0031-9422(92)83116-G
Lê S, Josse J, Husson F (2008) FactoMineR: an R package for multivariate analysis. J Stat Softw 25:1–18. https://doi.org/10.1016/j.envint.2008.06.007
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
Lund F, Frisvad JC (1994) Chemotaxonomy of Penicillium aurantiogriseum and related species. Mycol Res 98:481–492. https://doi.org/10.1016/S0953-7562(09)80466-8
Macgeorge KM, Mantle PG (1990) Nephrotoxicity of Penicillium aurantiogriseum and P. commune from an endemic nephropathy area of Yugoslavia. Mycopathologia 112:139–145. https://doi.org/10.1007/BF00436643
Maiya S, Grundmann A, Li SM, Turner G (2006) The fumitremorgin gene cluster of Aspergillus fumigatus: identification of a gene encoding brevianamide F synthetase. ChemBioChem 7:1062–1069. https://doi.org/10.1002/cbic.200600003
Malir F, Ostry V, Pfohl-Leszkowicz A, Malir J, Toman J (2016) Ochratoxin a: 50 years of research. Toxins 8:12–15. https://doi.org/10.3390/toxins8070191
Mantle PG (1994) Renal histopathological responses to nephrotoxic Penicillium aurantiogriseum in the rat during pregnancy, lactation and after weaning. Nephron 66:93–98. https://doi.org/10.1159/000187773
Mantle PG, Miljkovic A, Udupa V, Dobrota M (1998) Does apoptosis cause renal atrophy in Balkan endemic nephropathy? Lancet 352:1118–1119. https://doi.org/10.1016/s0140-6736(05)79758-0
Mantle PG, McHugh KM, Fincham JE (2010) Contrasting nephropathic responses to oral administration of extract of cultured Penicillium polonicum in rat and primate. Toxins 2:2083–2097. https://doi.org/10.3390/toxins2082083
Miljkovic A, Pfohl-Leszkowicz A, Dobrota M, Mantle PG (2003) Comparative responses to mode of oral administration and dose of ochratoxin A or nephrotoxic extract of Penicillium polonicum in rats. Exp Toxicol Pathol 54:305–312. https://doi.org/10.1078/0940-2993-00262
Miyao K (1960) The structure of fungisporin (studies on fungisporin III). Bull Agric Chem Soc Japan 24:23–30. https://doi.org/10.1080/03758397.1960.10857626
Morren JA, Galvez-Jimenez N (2010) Where is dihydroergotamine mesylate in the changing landscape of migraine therapy? Expert Opin Pharmacother 11:3085–3093. https://doi.org/10.1517/14656566.2010.533839
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
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
Muñoz K, Vega M, Rios G, Geisen R, Degen GH (2011) Mycotoxin production by different ochratoxigenic Aspergillus and Penicillium species on coffee- and wheat-based media. Mycotoxin Res 27:239–247. https://doi.org/10.1007/s12550-011-0100-0
Nielsen KF, Månsson M, Rank C, Frisvad JC, Larsen TO (2011) Dereplication of microbial natural products by LC-DAD-TOFMS. J Nat Prod 74:2338–2348. https://doi.org/10.1021/np200254t
Oliveira MS, Rocha A, Sulyok M, Krska R, Mallmann CA (2017) Natural mycotoxin contamination of maize (Zea mays L.) in the south region of Brazil. Food Control 73:127–132. https://doi.org/10.1016/j.foodcont.2016.07.033
Perez-Lloret S, Rascol O (2010) Dopamine receptor agonists for the treatment of early or advanced Parkinsons disease. CNS Drugs 24:941–968. https://doi.org/10.2165/11537810-000000000-00000
Pitt JI (1979) The genus Penicillium and its teleomorphic states Eupenicillium and Talaromyces. Academic Press, London
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, Ames
Roberts A, Beaumont C, Manzarpour A, Mantle P (2016) Purpurolic acid: a new natural alkaloid from Claviceps purpurea (Fr.) Tul. Fungal Biol 120:104–110. https://doi.org/10.1016/j.funbio.2015.10.011
Samson RA, Houbraken J, Thrane U, Frisvad JC, Andersen B (2010) Food and indoor fungi, CBS Labora. CBS-KNAW, Utrecht
Sansing GA, Lillehoj EB, Detroy RW, Miller MA (1976) Synergistic toxic effects of citrinin, ochratoxin A and penicillic acid in mice. Toxicon 14:213–220. https://doi.org/10.1016/0041-0101(76)90009-X
Savard ME, Miller JD, Blais LA, Seifert KA, Samson RA (1994) Secondary metabolites of Penicillium bilaii strain PB-50. Mycopathologia 127:19–27. https://doi.org/10.1007/BF01104007
Scott DB (1964) Toxigenic fungi isolated from cereal and legume products. Mycopathol Mycol Appl 25:213–222. https://doi.org/10.1007/bf02049914
Šegvić Klarić M, Rašić D, Peraica M (2013) Deleterious effects of mycotoxin combinations involving ochratoxin A. Toxins 5:1965–1987. https://doi.org/10.3390/toxins5111965
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
Smedsgaard J (1997) Micro-scale extraction procedure for standardized screening of fungal metabolite production in cultures. J Chromatogr A 760:264–270. https://doi.org/10.1016/S0021-9673(96)00803-5
Smith M, Madec S, Coton E, Hymery N (2016) Natural co-occurrence of mycotoxins in foods and feeds and their in vitro combined toxicological effects. Toxins 94:1–36. https://doi.org/10.3390/toxins8040094
Speijers GJA, Speijers MHM (2004) Combined toxic effects of mycotoxins. Toxicol Lett 153:91–98. https://doi.org/10.1016/j.toxlet.2004.04.046
Stoev SD, Vitanov S, Anguelov G, Creppy EE (2001) Experimental mycotoxic nephropathy in pigs provoked by a diet containing ochratoxin A and penicillic acid. Vet Res Commun 25:205–223. https://doi.org/10.1023/A:1006433709685
Streit E, Schwab C, Sulyok M, Naehrer K, Krska R, Schatzmayr G (2013) Multi-mycotoxin screening reveals the occurrence of 139 different secondary metabolites in feed and feed ingredients. Toxins 5:504–523. https://doi.org/10.3390/toxins5030504
Studer RO (1969) Synthesis and structure of fungisporin. Experientia 25:899. https://doi.org/10.1007/BF01898048
Studer-Rohr I, Dietrich DR, Schlatter J, Schlatter C (1995) The occurrence of ochratoxin-A in coffee. Food Chem Toxicol 33:341–355
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
Tanenbaum SW, Nakajima S, Marx G (1969) The role of polyketides in secondary metabolite production by penicillia. Biotechnol Bioeng 11:1135–1156. https://doi.org/10.1002/bit.260110610
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
Tudzynski B (2014) Nitrogen regulation of fungal secondary metabolism in fungi. Front Microbiol 5:1–15. https://doi.org/10.3389/fmicb.2014.00656
Urbano GR, Taniwaki MH, Vicentini MC (2001) Occurrence of ochratoxin A—producing fungi in raw Brazilian coffee. J Food Prot 64:1226–1230. https://doi.org/10.4315/0362-028x-64.8.1226
Vinokurova NG, Boichenko LV, Arinbasarov MU (2003) Production of alkaloids by fungi of the genus Penicillium grown on wheat grain. Appl Biochem Microbiol 39:457–460. https://doi.org/10.1023/A:1024576703367
Vrabcheva T, Usleber E, Dietrich R, Märtlbauer E (2000) Co-occurrence of ochratoxin A and citrinin in cereals from bulgarian villages with a history of Balkan endemic nephropathy. J Agric Food Chem 48:2483–2488. https://doi.org/10.1021/jf990891y
Wawrzyniak J, Waśkiewicz A (2014) Ochratoxin A and citrinin production by Penicillium verrucosum on cereal solid substrates. Food Addit Contam Part A Chem Anal Control Expo Risk Assess 31:139–148. https://doi.org/10.1080/19440049.2013.861933
Wickham H (2016) ggplot2: elegant graphics for data analysis, 2nd edn. Springer, New York
Winblad B, Fioravanti M, Dolezal T, Logina I, Milanov IG, Popescu DC, Solomon A (2008) Therapeutic use of nicergoline. Clin Drug Investig 28:533–552. https://doi.org/10.1111/j.1365-2354.2008.00917.x
Wontner-smith TJ, Bruce DM, Cardwell SK, Armitage DM, Jennings P (2014) Ochratoxin A production during ambient-air drying. J Stored Prod Res 56:1–7. https://doi.org/10.1016/j.jspr.2013.11.007
Xu W, Gavia DJ, Tang Y (2014) Biosynthesis of fungal indole alkaloids. Nat Prod Rep 31:1474–1487. https://doi.org/10.1039/c4np00073k
Yeulet SE, Mantl PG, Rudge MS, Greig JB (1988) Nephrotoxicity of Penicillium aurantiogriseum, a possible factor in the aetiology of Balkan Endemic Nephropathy. Mycopathologia 120:21–30. https://doi.org/10.1007/bf00436248
Zhi HX, Fang Y, Du L, Zhu T, Duan L, Chen J, Gu QQ, Zhu WM (2007) Aurantiomides A-C, quinazoline alkaloids from the sponge-derived fungus Penicillium aurantiogriseum SP0-19. J Nat Prod 70:853–855. https://doi.org/10.1021/np060516h
Acknowledgments
We are grateful to Agilent Technologies for the Thought Leader Donation of the UHPLC-QTOFMS system.
Funding
We would like to thank the Novo Nordisk Foundation grant #NNF 130 C0005201 for the funding of this study.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
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.
Electronic supplementary material
ESM 1
(PDF 20327 kb)
Rights and permissions
About this article
Cite this article
Hallas-Møller, M., Nielsen, K.F. & Frisvad, J.C. Secondary metabolite production by cereal-associated penicillia during cultivation on cereal grains. Appl Microbiol Biotechnol 102, 8477–8491 (2018). https://doi.org/10.1007/s00253-018-9213-0
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00253-018-9213-0