Patulin risk associated with blue mould of pome fruit marketed in southern Italy

Main Article Content

S.M. Sanzani
A. Susca
S. Mastrorosa
M. Solfrizzo


Penicillium, mycotoxin, isoepoxydon dehydrogenase, apple, pear


Blue mould is one of the most important postharvest diseases of pome fruit in all producing countries. It is mainly associated to Penicillium expansum that produces the mycotoxin patulin, although other species might be involved. The aim of the present study was to characterise Penicillium isolates associated with blue mould decay of pome fruit marketed in Apulia region (southern Italy), and verify their ability to produce patulin in vitro. Twenty-nine isolates of Penicillium spp. were recovered from pome fruit showing visible blue mould symptoms, and analysed for patulin production. After fungal isolation, the fruits were singularly analysed for patulin content. In general, the isolates proved to produce patulin and most of the pome fruit contained significant amounts of patulin, but there was no quantitative correspondence between in vitro and in vivo toxin accumulation. Isolate identification at species level was based on DNA analysis by P. expansum species-specific primers and sequencing of ?-tubulin gene. Furthermore, fungal isolates were tested for the occurrence of the patN gene coding the enzyme isoepoxydon dehydrogenase (IDH), involved in patulin metabolic pathway and considered a useful indicator of critical control points for patulin contamination. All 26 isolates identified as P. expansum were positive for patN and produced patulin. Moreover, three pear isolates belonging to other Penicillium species were found. They were positive for patN, but only two actually produced patulin. It can be concluded that toxigenic P. expansum isolates are associated with blue mould of pome fruit marketed in Apulia, thus a rapid detection is important to avoid patulin contamination beyond regulatory limits. Nevertheless, the presence of patN gene alone cannot be considered a predictive assay for patulin production. An evaluation of its expression level should be carried out.

Abstract 68 | PDF Downloads 57


Beretta, B., Gaiaschi, A., Galli, C.L. and Restani, P., 2000. Patulin in apple-based foods: occurrence and safety evaluation. Food Additives and Contaminants 17: 399-406.
Castoria, R., Mannina, L., Durán-Patrón, R., Maffei, F., Sobolev, A.P., De Felice, D.V., Pinedo-Rivilla, C., Ritieni, A., Ferracane, R. and Wright, S.A.I., 2012. Conversion of the mycotoxin patulin to the less toxic desoxypatulinic acid by the biocontrol yeast Rhodosporidium kratochvilovae strain LS11. Journal of Agricultural and Food Chemistry 59: 11571-11578.
Chang, P.K., Horn, B.W. and Dorner, J.W., 2005. Sequence breakpoints in the aflatoxin biosynthesis gene cluster and flanking regions in nonaflatoxigenic Aspergillus flavus isolates. Fungal Genetics and Biology 42: 914-923.
Codex Alimentarius Commission (CAC), 2003. Code of practice for the prevention and reduction of patulin contamination in apple juice and apple juice ingredients in other beverages. CAC/RCP 50: 1-6.
Eisele, T.A. and Drakeb, R.S., 2005. The partial compositional characteristics of apple juice from 175 apple varieties. Journal of Food Composition and Analysis 18: 3213-3221.
European Commission (EC), 2006. Commission Regulation No. 1881/2006 of setting maximum levels of certain contaminants in foodstuffs. Official Journal of the European Community L364: 7-16.
FAOSTAT, 2012. Database. Available at:
Frisvad, J.C., Thrane, U.L.F. and Samson, R.A., 2007. Mycotoxin producers. Mycology Series 25: 135.
Funes, G.J. and Resnik, S.L., 2009. Determination of patulin in solid and semisolid apple and pear products marketed in Argentina. Food Control 20: 277-280.
Glass, N.L. and Donaldson, G.C., 1995. Development of primer sets designed for use with the PCR to amplify conserved genes from filamentous ascomycetes. Applied and Environmental Microbiology 61: 1323-1330.
I.Stat, 2011. Database. Available at:
Iwahashi, Y., Hosoda, H., Park, J., Lee, J., Suzuki, Y., Kitagawa, E., Murata, S., Jwa, N., Gu, M. and Iwahashi, H., 2006. Mechanisms of patulin toxicity under the condition that cause growth inhibition to yeast cells. Journal of Agriculture and Food Chemistry 54: 1936-1942.
MacDonald, S., Long, M., Gilbert, J. and Felgueiras, I., 2000. Liquid chromatographic method for determination of patulin in clear and cloudy apple juices and apple puree: collaborative study. Journal of AOAC International 83: 1387-1394.
Marek, P., Annamalai, T. and Venkitanarayanan, K., 2003. Detection of Penicilliumexpansum by polymerase chain reaction. International Journal of Food Microbiology 89: 139-144.
Moake, M.M., Padilla-Zakour, O.I. and Worobo, R.W., 2005. Comprehensive review of patulin control methods in foods. Comprehensive Reviews of Food Science and Food Safety 1: 8-21.
Morales, H., Barros, G., Marín, S., Chulze, S., Ramos, A.J. and Sanchis, V., 2008b. Effects of apple and pear varieties and pH on patulin accumulation by Penicillium expansum. Journal of the Science of Food and Agriculture 88: 2738-2743.
Morales, H., Sanchis, V., Coromines, J., Ramos, A.J. and Marin, S., 2008a. Inoculum size and intraspecific interactions affects Penicillium expansum growth and patulin accumulation in apples. Food Microbiology 25: 378-385.
Napolitano, A., Cascone, A., Graziani, G., Ferracane, R., Scalfi, L., Di Vaio, C., Ritieni, A. and Fogliano, V., 2004. Influence of variety and storage on the polyphenol composition of apple flesh. Journal of Agriculture and Food Chemistry 52: 6526-6531.
Paterson, R.R.M., 2007. The isoepoxydon dehydrogenase gene PCR profile is useful in fungal taxonomy. Revista Iberoamericana de Micología 24: 289-293.
Paterson, R.R.M., Archer, S., Kozakiewicz, Z., Lea, A., Locke, T. and O’Grady, E., 2000. A gene probe for the patulin metabolic pathway with potential for use in patulin and novel disease control. Biocontrol Science and Technology 10: 509-512.
Paterson, R.R.M., Kozakiewicz, Z., Locke, T., Brayford, D. and Jones, S.C.B., 2003. Novel use of the isoepoxydon dehydrogenase gene probe of the patulin metabolic pathway and chromatography to test penicillia isolated from apple production systems for the potential to contaminate apple juice with patulin. Food Microbiology 20: 359-364.
Pianzzola, M.J., Moscatelli, M. and Vero, S., 2004. Characterization of Penicillium isolates associated with blue mold on apple in Uruguay. Plant Disease 88: 23-28.
Piemontese, L., Solfrizzo, M. and Visconti, A., 2005. Occurrence of patulin in conventional and organic fruit products in Italy and subsequent exposure assessment. Food Additives and Contaminants 22: 437-442.
Ricelli, A., Baruzzi, F., Solfrizzo, M., Morea, M. and Fanizzi, F.P., 2007. Biotransformation of Patulin by Gluconobacter oxydans. Applied and Environmental Microbiology 73(3): 3785-3792.
Ritieni, A., 2003. Patulin in Italian commercial apple products. Journal of Agriculture and Food Chemistry 51: 6086-6090.
Samson, R.A. and Pitt, J.I., 2000. Integration of modern taxonomic methods for Penicillium and Aspergillus classification. Harwood Academic Publishers, London, UK, 524 pp.
Sanderson, P.G. and Spotts, R.A., 1995. Postharvest decay of winter pear and apple fruit caused by species of Penicillium. Phytopathology 85: 103-110.
Sanzani, S.M., De Girolamo, A., Schena, L., Solfrizzo, M., Ippolito, A. and Visconti, A., 2009a. Control of Penicillium expansum and patulin accumulation on apples by quercetin and umbelliferone. European Food Research and Technology 228: 381-389.
Sanzani, S.M., Li Destri Nicosia, M.G., Faedda, R., Cacciola, S.O. and Schena, L., 2014. Use of quantitative PCR detection methods to study biocontrol agents and phytopathogenic fungi and oomycetes in environmental samples. Journal of Phytopathology 162: 1-13.
Sanzani, S.M., Montemurro, C., Di Rienzo, V., Solfrizzo, M. and Ippolito, A., 2013. Genetic structure and natural variation associated with host of origin in Penicillium expansum strains causing blue mould. International Journal of Food Microbiology 165: 111-120.
Sanzani, S.M., Schena, L., Nigro, F., De Girolamo, A. and Ippolito, A., 2009b. Effect of quercetin and umbelliferone on the transcript level of Penicillium expansum genes involved in patulin biosynthesis. European Journal of Plant Pathology 125: 223-233.
Sarubbi, F., Formisano, G., Auriemma, G., Arrichiello, A. and Palomba, R., 2016. Patulin in homogenized fruit’s and tomato products. Food Control 59: 420-423.
Schena, L., Li Destri Nicosia, M.G., Sanzani, S.M., Faedda, R., Ippolito, A. and Cacciola, S.O., 2013. Development of quantitative PCR detection methods for phytopathogenic fungi and oomycetes. Journal of Plant Pathology 95: 7-24.
Sommer, N.F., Buchanan, J.R. and Fortlage, R.J., 1974. Production of patulin by Penicilliumexpansum. Applied Environmental Microbiology 28: 589-593.
Spadaro, D., Ciavorella, A., Frati, S., Garibaldi, A. and Gullino, M.L., 2007. Incidence and level of patulin contamination in pure and mixed apple juices marketed in Italy. Food Control 18: 1098-1102.
Spadaro, D., Garibaldi, A. and Gullino, M.L., 2008. Efficacy of biocontrol yeasts against Penicillium expansum and patulin on different cultivars of apple in postharvest. Acta Horticulturae 873: 191-196.
Susca, A., Proctor, R.H., Butchko, R.A., Haidukowski, M., Stea, G., Logrieco, A. and Moretti, A., 2014. Variation in the fumonisin biosynthetic gene cluster in fumonisin-producing and nonproducing black aspergilli. Fungal Genetics and Biology 73: 39-52.
Ward, T.J., Bielawski, J.P., Kistler, H.C., Sullivan, E. and O’Donnell, K., 2002. Ancestral polymorphism and adaptive evolution in the trichothecene mycotoxin gene cluster of phytopathogenic Fusarium. Proceedings of the National Academy of Sciences of the USA 99: 9278-9283.
Weidenbörner, M., 2001. Encyclopaedia of food mycotoxins. Springer-Verlag, Berlin, Germany.