Abstract
Black Aspergilli, and in particular Aspergillus carbonarius, are the main causes of contamination of grapes and their by-products by ochratoxin A. A PCR-based method was developed to detect DNA of A. carbonarius and A. japonicus. Two pairs of primers (CARBO1/2 and JAPO1/2) designed from the calmodulin gene, produced PCR products of 371 and 583 bp for A. carbonarius and A. japonicus, respectively. Primer specificity was tested with DNA of 107 strains belonging to Aspergillus section Nigri isolated mostly from grapes in Europe. The sensitivity of primers CARBO1/2 and JAPO1/2 was 12.5 pg when using pure total genomic DNA of the two species. The developed primers provide a powerful tool for detection of the main ochratoxigenic producing Aspergillus species in grapes.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Abarca ML, Bragulat MR, Castella G and Cabanes FJ (1994) Ochratoxin A production by strains of Aspergillus niger var. niger. Applied and Environmental Microbiology 60: 2650-2652
Accensi F, Cano J, Figuera L, Abarca ML and Cabanes FJ (1999) New PCR method to differentiate species in the Aspergillus niger aggregate. FEMS Microbiology Letters 180: 191-196
Al Musallam A (1980) Revision of the black Aspergillus species. Ph.D. thesis. University of Utrecht, Utrecht, The Netherlands
Battilani P and Pietri A (2002) Ochratoxin A in grapes and wine. European Journal of Plant Pathology 108: 639-643
Bennet JW and Klich MA (1992) Aspergillus: Biology and Industrial Applications. Butterworths-Heinemann, Boston, USA
Gams W, Christensen M, Onions AHS, Pitt JI and Samson RA (1985) Infrageneric taxa of Aspergillus. In: Samson RA and Pitt JI (ed) Advances in Penicillium and Aspergillus Systematics (pp. 55–61). Plenum Press, New York, USA
Geiser DM, Harbinski FM and Taylor JW (2000) Molecular and analytical tools for characterizing Aspergillus and Penicillium species at the intra-and interspecific levels. In: Samson RA and Pitt JI (eds) Integration of Modern Taxonomic Methods of Penicillium and Aspergillus Classification (pp 381–394). Harwood Academic Publishers, Amsterdam, The Netherlands
Hamari Z, Kevei F, Kovács É, Varga J, Kozakiewicz Z and Croft JH (1997) Molecular and phenotypic characterization of Aspergillus japonicus and Aspergillus aculeatus strains with special regard to their mitochondrial DNA polymorphisms. Antonie Van Leeuwenhoek 72: 337-347
IARC (1993) Some Naturally Occurring Substances: Food Items and Constituents, Heterocyclic Aromatic Amines and Mycotoxins, IARC Monographs on the Evaluation of Carcinogenic Risks to Humans. Vol. 56. (pp 397–444; 445-466; 467-488). International Agency for Research on Cancer, Lyon, France
Klich MA and Pitt JI (1988) A Laboratory Guide to Common Aspergillus Species and their Teleomorphs. CSIRO Division of Food Processing, North Ryde, Australia
Kozakiewicz Z (1989) Aspergillus species on stored products. Mycological Papers 161, CAB International Mycological Institute, Oxon, UK
O'Donnell K, Nirenberg HI, Aoki T and Cigelnik E (2000) A multigene phylogeny of the Gibberella fujikuroi species complex: detection of additional phylogenetically distinct species. Mycoscience 41: 61-78
Otteneder H and Majerus P (2000) Occurrence of ochratoxin A (OTA) in wines: influence of the type of wine and its geographical origin. Food Additives and Contaminants 17: 793-798
Parenicova L, Skouboe P, Samson RA, Rossen L and Visser J (2000) Molecular tools for the classification of black Aspergilli. In: Samson RA and Pitt JI (eds) Integration of Modern Taxonomic Methods of Penicillium and Aspergillus Classification (pp. 413–424) Harwood Academic Publishers, Amsterdam, The Netherlands
Parenicova L, Skouboe P, Frisvad J, Samson RA, Rossen L, Ten Hoor-Suykerbuyk M and Visser J (2001) Combined molecular and biochemical approach identifies Aspergillus japonicus and Aspergillus aculeatus as two species. Applied and Environmental Microbiology 7: 521-527
Raper KB and Fennell DI (1965) The Genus Aspergillus. Robert E Krieger Publishing Company, Krieger Drive, Malabar, FL, USA, 686 pp
Samson RA (1992) Current taxonomic schemes of the genus Aspergillus and teleomorphs. In: Bennet JW and Klich MA (ed) Aspergillus: Biology and Industrial Applications (pp 355–3909) Butterworths-Heinemann, Boston, USA
Taylor JW, Geiser DM, Burt A and Koufopanou V (1999) The Evolutionary Biology and Population Genetics Underlying Fungal Strain Typing. Clinical Microbiology Reviews 12: 126-146
Téren J, Varga J, Hamari Z, Rinyu Éand Kevei F (1996) Immunochemical detection of ochratoxin A in black Aspergillus strains. Mycopathologia 134: 171-176
van Peij H, Pel H, van den Hombergh M, van den Berg R, van Rooijen A, van Ooyen and van Dijk A (2002) the DSM Aspergillus niger genome project. In 6th European Conference on Fungal Genetics, Pisa, Italy
Varga J, Kevei F, Rinyu É, Téren J and Kozakiewicz Z (1996) Ochratoxin production by Aspergillus species. Applied and Environmental Microbiology 62: 4461-4464
Zimmerli B and R Dick (1996) Ochratoxin A in table wine and grape-juice: Occurrence and risk assessment. Food Additives and Contaminants 13: 655-668
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Perrone, G., Susca, A., Stea, G. et al. PCR Assay for Identification of Aspergillus Carbonarius and Aspergillus Japonicus . European Journal of Plant Pathology 110, 641–649 (2004). https://doi.org/10.1023/B:EJPP.0000032403.08921.49
Issue Date:
DOI: https://doi.org/10.1023/B:EJPP.0000032403.08921.49