Study of the phenotypic and genotypic biodiversity of potentially ochratoxigenic black aspergilli isolated from grapes

Ochratoxin A (OTA) is a mycotoxin with nephrotoxic, carcinogenic, teratogenic and immunotoxic effects, naturally found in agricultural products including grapes and wine. Black Aspergillus species (Section Nigri) are mainly responsible for OTA accumulation in wine grapes and in particular Aspergillus carbonarius and Aspergillus niger aggregate. The biodiversity of potentially ochratoxigenic strains of black aspergilli from different French vineyards in the southern Mediterranean region of Languedoc-Roussillon was studied. One hundred and eighty nine black strains were isolated from grapes and studied according to harvest year, production zone, grape variety and pre-harvest treatment of grapevines. The strains were identified and classified in two groups according to macroscopic and microscopic characters; these were called the A. carbonarius representative group and the A. niger aggregate representative group. Members of each group were classified in subgroups based on macroscopic morphological colony characters. Strain biodiversity was studied according to phenotypic and genotypic characterization and to the OTA production of selected strains on PDA medium. After identification was confirmed by specific PCR using primer pair ITS1/CAR and ITS1/NIG, 24 potential ochratoxigenic strains belonging to A. carbonarius and A. niger aggregate were discriminated by RAPD-PCR using 8 different OPC primers. The use of specific primers supported the identification based on phenotypic and morphological characters. RAPD-PCR patterns demonstrated a considerable diversity among the strains. Clustering among A. niger aggregate strains was associated with production zone and harvest year, but not grape variety or pre-harvest treatment. Clustering among A. carbonarius strains was not associated with any of the above parameters. OTA production of strains on culture medium seemed to correlate better with morphological characters than with genotypic profiles. No clear relation could be established between phenotypic and genotypic characters of the studied black aspergilli.