Morphological and chemical characterization of Alternaria populations from apple fruit.

Alternaria is a frequent contaminant of apple fruit, causing severe economic losses. It can produce external lesions and mouldy core, characterised by a rotten area in the apple core. In the present study, morphological and chemical characterization of Alternaria from apples was performed, evaluating differences related to agricultural practices and type of disease. A low morphological diversity was observed; most of the isolates were identified as A. tenuissima sp.-grp. (95 %). A. arborescens sp.-grp. and A. gaisen sp.-grp. were present in a proportion of 1 %, and 3 % of the isolates showed intermediate characteristics between these sp.-grps. and were identified as Alternaria sp. The chemical diversity was greater; 27 secondary metabolites were produced by the apple isolates. The most frequents were altertoxin-I (85 %), altechromone A (76 %), tentoxin (69 %), and tenuazonic acid (68 %). The alternariols were produced in a lower frequency when comparing with isolates from other crops; alternariol, 58 % and alternariol monomethyl ether, 57 %. The predominant secondary metabolite profile included compounds from different chemical families, such as dibenzopyrones, tetramic acids, perylene quinones, and cyclic tetrapeptides. A wider metabolomic capacity was observed in isolates from conventional apples when compared to those from organic fruit, with the predominance of strong producers of altertoxins and alternariols. The isolates from mouldy core showed higher ability to produce metabolites from different chemical families than those from external lesions. The wide chemical diversity of the Alternaria apple population should be considered to assess the health risk associated with apple by-products.

From field to process: How storage selects toxigenic Alternaria spp. causing mouldy core in Red Delicious apples.

Apple is a major crop in Argentina where 50% of the production is derived to by-products. Industries process either recently harvested apples or fruit stored for up to 9 months. This crop is susceptible to fungal diseases both external and internal, such as mouldy core (MC). The incidence of fungal pathogens changes during storage, as well as the risk associated with their presence since some contaminants belong to mycotoxigenic genera. The objective of this study was to characterize the fungal contaminants of Red Delicious apple fruit in Argentina evaluating their evolvement from field to process, with main interest on MC causal agents and mycotoxigenic species. A total of 240 apples were analysed; 140, recently harvested and intended for fresh consumption (C), and 100 stored for 9 months in a refrigerated chamber (0-3 °C) and destined to industrialization (I). The 86% of fresh consumption apples showed external fungal lesions, and only 14% were undamaged; MC incidence was 34%. High biodiversity was observed; Penicillium was the predominant genus (54%), followed by Alternaria spp. (41%). Only 3% of industrialization fruit were undamaged, 48% had external lesions and 51% MC. However, biodiversity was lower in these apples. Alternaria spp. was recovered from 60% of apples, mainly causing MC, while Penicillium spp. took second place (34%). All the Alternaria isolates belonged to Section Alternaria with A. tenuissima as the predominant species-group. Alternariol was synthesised by 75% of the isolates, while both alternariol monomethyl-ether and tenuazonic acid by 76%. From the 100 I apples, 93 were contaminated with at least one of these mycotoxins. Alternaria was the main causal agent of MC in Argentinean Red Delicious apples, and fruit affected by this disease might be incorporated into the process line, with a consequent risk of mycotoxin contamination in apple by-products.

Secondary metabolite profiles of small-spored Alternaria support the new phylogenetic organization of the genus.

The group of the small-spored Alternaria species is particularly relevant in foods due to its high frequency and wide distribution in different crops. These species are responsible for the accumulation of mycotoxins and bioactive secondary metabolites in food. The taxonomy of the genus has been recently revised with particular attention on them; several morphospecies within this group cannot be segregated by phylogenetic methods, and the most recent classifications proposed to elevate several phylogenetic species-groups to the taxonomic status of section. The purpose of the present study was to compare the new taxonomic revisions in Alternaria with secondary metabolite profiles with special focus on sections Alternaria and Infectoriae and food safety. A total of 360 small-spored Alternaria isolates from Argentinean food crops (tomato fruit, pepper fruit, blueberry, apple, wheat grain, walnut, pear, and plum) was morphologically identified to species-group according to Simmons (2007), and their secondary metabolite profile was determined. The isolates belonged to A. infectoria sp.-grp. (19), A. tenuissima sp.-grp. (262), A. arborescens sp.-grp. (40), and A. alternata sp.-grp. (7); 32 isolates, presenting characteristics overlapping between the last three groups, were classified as Alternaria sp. A high chemical diversity was observed; 78 different metabolites were detected, 31 of them of known chemical structure. The isolates from A. infectoria sp.-grp. (=Alternaria section Infectoriae) presented a specific secondary metabolite profile, different from the other species-groups. Infectopyrones, novae-zelandins and phomapyrones were the most frequent metabolites produced by section Infectoriae. Altertoxin-I and alterperylenol were the only compounds that these isolates produced in common with members of section Alternaria. None of the well-known Alternaria toxins, considered relevant in foods, namely alternariol (AOH), alternariol monomethyl ether (AME), tenuazonic acid (TeA), tentoxin (TEN) or altenuene (ALT), were produced by isolates of this section. On the other hand, strains from section Alternaria (A. tenuissima, A. arborescens, and A. alternata sp.-grps.) shared a common metabolite profile, indistinguishable from each other. AOH, AME, ALT, TEN, and TeA were the most frequently mycotoxins produced, together with pyrenochaetic acid A and altechromone A. Alternaria section Alternaria represents a substantial risk in food, since their members in all types of crops are able to produce the toxic metabolites.