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Papers by Antonella Susca

Research paper thumbnail of Fungal genomics: Organic acid production and secondary metabolism

Research paper thumbnail of Polymerase chain reaction (PCR) identification ofAspergillus nigerandAspergillus tubingensisbased on the calmodulin gene

Food Additives and Contaminants, 2007

HAL is a multidisciplinary open access archive for the deposit and dissemination of scientific re... more HAL is a multidisciplinary open access archive for the deposit and dissemination of scientific research documents, whether they are published or not. The documents may come from teaching and research institutions in France or abroad, or from public or private research centers. L'archive ouverte pluridisciplinaire HAL, est destinée au dépôt et à la diffusion de documents scientifiques de niveau recherche, publiés ou non, émanant des établissements d'enseignement et de recherche français ou étrangers, des laboratoires publics ou privés.

Research paper thumbnail of PCR Assay for Identification of Aspergillus Carbonarius and Aspergillus Japonicus

European Journal of Plant Pathology, 2004

Black Aspergilli, and in particular Aspergillus carbonarius, are the main causes of contamination... more 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.

Research paper thumbnail of Toxin Profile, Fertility and AFLP Analysis of Fusarium verticillioides from Banana Fruits

European Journal of Plant Pathology, 2004

Gibberella fujikuroi is composed of at least nine mating populations (MPs), corresponding to biol... more Gibberella fujikuroi is composed of at least nine mating populations (MPs), corresponding to biological species and assigned letters (from A to I). Each MP possesses a specific toxicological profile and a preferential host. Members of Fusarium verticillioides and F. thapsinum, anamorphs respectively of MPs A (G. moniliformis) and F (G. thapsina), share identical morphological traits, but they have a different preferential hosts (maize and sorghum, respectively) and toxin profiles, being able the only member of MP A to produce fumonisins and the only member of MP F to produce moniliformin. Isolates from banana fruits were identified morphologically as F. verticillioides. The isolates were analyzed for fumonisin and moniliformin production. While none of the isolates produced fumonisin, all the isolates produced moniliformin. The isolates were crossed with tester strains of MPs A and F, showing ability to produce fertile perithecia only when crossed by MP A tester strains isolated from maize. However, the time required for the formation of fertile perithecia and their size differed significantly from the usual fertile crosses of strains belonging to MP A. Pathogenicity tests using such isolates of F. verticillioides isolated from banana and a set of F. verticillioides isolates isolated from maize were also performed on banana fruits. The data showed that the isolates from banana were more pathogenic. Finally, isolates from banana and maize were compared using AFLP. The results revealed that isolates from banana and maize produced two distinctly different clusters. In conclusion, isolates of F. verticillioides from banana showed specific traits (toxin production, in vitro fertility, pathogenicity and molecular profiles), that were different to those of the same species from maize. This could reflect important differences in the ecology and natural history of the population from banana and should encourage further investigations into the mechanisms of toxin production and pathogenicity within the same MP.

Research paper thumbnail of In vitro study of fum gene expression and fumonisins production in Fusarium verticillioides under different ecological conditions

F. verticillioides is a fungus associated to maize ears all over the world. It can cause Fusarium... more F. verticillioides is a fungus associated to maize ears all over the world. It can cause Fusarium pink ear rot and produces fumonisins (FBs), mycotoxins found in both maize kernels and its derivatives. They are toxic to humans, in particular FB1 is classified as possibly carcinogenic. Fumonisins biosynthetic pathway is regulated by several genes belonging to FUM cluster whose behaviour in different ecological conditions was poorly studied. The aim of this work was to investigate the behaviour of 2 F. verticillioides strains grown in vitro both on a FB-inducing (Malt extract agar-MEA) and FB-inhibiting (Czapeck yeast agar-CYA) media. Fungal growth, FBs production and gene expression were studied in different temperature (T; 20-30) and water activity (aw; 0.90-0.99) regimes, in liquid cultures incubated for 7-21 days. The expression of two genes: FUM21 and FUM2 was considered; relative quantitation of gene expression was performed by RealTime (RT)-PCR. The results showed that, with aw fixed at 0.99, maximum FUM21 and FUM2 expression was observed at 30\ub0C after 14 days of incubation, while FBs production increased from 7 to 21 days. At fixed T (25\ub0C), fungal growth at 0.90 aw was not observed after 21 days of incubation. Gene expression increased with incubation time both at 0.95 and 0.99 aw, FB production increased till to 21 and 14 days respectively at 0.95 and 0.99 aw. In all the conditions studied the expression of the 2 genes considered followed a very similar trend, but FUM2 was much more expressed than FUM21 and more influenced by aw conditions

Research paper thumbnail of Evidence for birth-and-death evolution and horizontal transfer of the fumonisin mycotoxin biosynthetic gene cluster in Fusarium

In fungi, genes required for synthesis of secondary metabolites are often clustered. The FUM gene... more In fungi, genes required for synthesis of secondary metabolites are often clustered. The FUM gene cluster is required for synthesis of fumonisins, a family of toxic secondary metabolites produced by species in the Fusarium (Gibberella) fujikuroi species complex (FFSC). Fumonisins are a health and agricultural concern because their consumption is epidemiologically associated with cancer and neural tube defects in humans and other animals. Among FFSC species, the FUM cluster is uniform in gene order and orientation, but located in different genomic positions. Phylogenetic analyses indicated discord between species phylogenies and FUM gene-based phylogenies. Subsequent constraint analyses confirmed the discord, and analyses of variation in synonymous sites indicated that cluster divergence predated, in some cases, and postdated, in one case, divergence of lineages of Fusarium in which the cluster occurs. The results are not consistent with the discord resulting from transspecies evolution of ancestral cluster alleles, or with interspecies hybridization, but are consistent with duplication of the cluster within an FFSC ancestor and subsequent loss and sorting of paralogous clusters in a manner consistent with the birth-and-death evolution seen in several multigene families. Although the results are also consistent with horizontal transfer of the cluster, such a model is less parsimonious because it requires multiple transfer events from unknown but related donors to multiple FFSC recipients. However, the analyses do provide strong support for horizontal transfer of the cluster from FFSC to another Fusarium lineage. Thus, despite conservation of gene organization within it, the FusariumFUM cluster has had a complex evolutionary history.

Research paper thumbnail of Evidence for birth-and-death evolution of a secondary metabolite biosynthetic gene cluster and its relocation within and between genomes of the filamentous fungus Fusarium

Research paper thumbnail of Insights into the evolution of toxin biosynthesis in the fungus Fusarium

Research paper thumbnail of A Species-Specific PCR Assay Based on the Calmodulin Partial Gene for Identification of Fusarium Verticillioides, F. Proliferatum and F. Subglutinans

European Journal of Plant Pathology, 2004

Fusarium proliferatum, F. subglutinans and F. verticillioides are the most important Fusarium spe... more Fusarium proliferatum, F. subglutinans and F. verticillioides are the most important Fusarium species occurring on maize worldwide , capable of producing a wide range of mycotoxins which are a potential health hazard for animals and humans. The ribosomal internal transcribed spacer and a portion of the calmodulin gene were sequenced and analysed in order to design species-specific primers useful for diagnosis. The primer pairs were based on a partial calmodulin gene sequence. Three pairs of primers (PRO1/2, SUB1/2 and VER 1/2) produced PCR products of 585, 631 and 578 bp for F. proliferatum, F. subglutinans and F. verticillioides, respectively. Primer specificity was confirmed by analyzing DNA of 150 strains of these species, mostly isolated from maize in Europe and USA. The sensitivity of primers was 12.5 pg when the pure total genomic DNA of each species was analyzed. The developed PCR assay should provide a powerful tool for the detection of toxigenic fungi in maize kernels.

Research paper thumbnail of Convergent and divergent evolution of the trichothecene mycotoxin biosynthetic gene cluster in the Fusarium

Research paper thumbnail of Toxigenic fungi and mycotoxins. Diagnosis and molecular kits

Research paper thumbnail of Biodiversity of important toxigenic fungi that threaten food safety

Research paper thumbnail of Characterization of the fumonisin B2 biosynthetic gene cluster in Aspergillus niger and A. awamori

Research paper thumbnail of Fumonisin production by black Aspergillus species in maize

Research paper thumbnail of Funghi tossigeni e micotossine: diagnosi e kit molecolari

Research paper thumbnail of Good Food Project: Detection of Aspergillus carbonarius and other black Aspergilli on grapes by direct multi-detection DNA microarray

Research paper thumbnail of MBLabDB: a social database for molecular biodiversity data

Research paper thumbnail of Aspergillus uvarum sp. nov., an uniseriate black Aspergillus species isolated from grapes in Europe

INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY, 2008

Research paper thumbnail of Molecular biodiversity of mycotoxigenic fungi that threaten food safety

International Journal of Food Microbiology, 2013

Fungal biodiversity is one of the most important contributors to the occurrence and severity of m... more Fungal biodiversity is one of the most important contributors to the occurrence and severity of mycotoxin contamination of crop plants. Phenotypic and metabolic plasticity has enabled mycotoxigenic fungi to colonize a broad range of agriculturally important crops and to adapt to a range of environmental conditions. New mycotoxin-commodity combinations provide evidence for the ability of fungi to adapt to changing conditions and the emergence of genotypes that confer enhanced aggressiveness toward plants and/or altered mycotoxin production profiles. Perhaps the most important contributor to qualitative differences in mycotoxin production among fungi is variation in mycotoxin biosynthetic genes. Molecular genetic and biochemical analyses of toxigenic fungi have elucidated specific differences in biosynthetic genes that are responsible for intra- and inter-specific differences in mycotoxin production. For Aspergillus and Fusarium, the mycotoxigenic genera of greatest concern, variation in biosynthetic genes responsible for production of individual families of mycotoxins appears to be the result of evolutionary adaptation. Examples of such variation have been reported for: a) aflatoxin biosynthetic genes in Aspergillus flavus and Aspergillus parasiticus; b) trichothecene biosynthetic genes within and among Fusarium species; and c) fumonisin biosynthetic genes in Aspergillus and Fusarium species. Understanding the variation in these biosynthetic genes and the basis for variation in mycotoxin production is important for accurate assessment of the risks that fungi pose to food safety and for prevention of mycotoxin contamination of crops in the field and in storage.

Research paper thumbnail of AFLP characterization of Southern Europe population of Aspergillus Section Nigri from grapes

International Journal of Food Microbiology, 2006

Research paper thumbnail of Fungal genomics: Organic acid production and secondary metabolism

Research paper thumbnail of Polymerase chain reaction (PCR) identification ofAspergillus nigerandAspergillus tubingensisbased on the calmodulin gene

Food Additives and Contaminants, 2007

HAL is a multidisciplinary open access archive for the deposit and dissemination of scientific re... more HAL is a multidisciplinary open access archive for the deposit and dissemination of scientific research documents, whether they are published or not. The documents may come from teaching and research institutions in France or abroad, or from public or private research centers. L'archive ouverte pluridisciplinaire HAL, est destinée au dépôt et à la diffusion de documents scientifiques de niveau recherche, publiés ou non, émanant des établissements d'enseignement et de recherche français ou étrangers, des laboratoires publics ou privés.

Research paper thumbnail of PCR Assay for Identification of Aspergillus Carbonarius and Aspergillus Japonicus

European Journal of Plant Pathology, 2004

Black Aspergilli, and in particular Aspergillus carbonarius, are the main causes of contamination... more 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.

Research paper thumbnail of Toxin Profile, Fertility and AFLP Analysis of Fusarium verticillioides from Banana Fruits

European Journal of Plant Pathology, 2004

Gibberella fujikuroi is composed of at least nine mating populations (MPs), corresponding to biol... more Gibberella fujikuroi is composed of at least nine mating populations (MPs), corresponding to biological species and assigned letters (from A to I). Each MP possesses a specific toxicological profile and a preferential host. Members of Fusarium verticillioides and F. thapsinum, anamorphs respectively of MPs A (G. moniliformis) and F (G. thapsina), share identical morphological traits, but they have a different preferential hosts (maize and sorghum, respectively) and toxin profiles, being able the only member of MP A to produce fumonisins and the only member of MP F to produce moniliformin. Isolates from banana fruits were identified morphologically as F. verticillioides. The isolates were analyzed for fumonisin and moniliformin production. While none of the isolates produced fumonisin, all the isolates produced moniliformin. The isolates were crossed with tester strains of MPs A and F, showing ability to produce fertile perithecia only when crossed by MP A tester strains isolated from maize. However, the time required for the formation of fertile perithecia and their size differed significantly from the usual fertile crosses of strains belonging to MP A. Pathogenicity tests using such isolates of F. verticillioides isolated from banana and a set of F. verticillioides isolates isolated from maize were also performed on banana fruits. The data showed that the isolates from banana were more pathogenic. Finally, isolates from banana and maize were compared using AFLP. The results revealed that isolates from banana and maize produced two distinctly different clusters. In conclusion, isolates of F. verticillioides from banana showed specific traits (toxin production, in vitro fertility, pathogenicity and molecular profiles), that were different to those of the same species from maize. This could reflect important differences in the ecology and natural history of the population from banana and should encourage further investigations into the mechanisms of toxin production and pathogenicity within the same MP.

Research paper thumbnail of In vitro study of fum gene expression and fumonisins production in Fusarium verticillioides under different ecological conditions

F. verticillioides is a fungus associated to maize ears all over the world. It can cause Fusarium... more F. verticillioides is a fungus associated to maize ears all over the world. It can cause Fusarium pink ear rot and produces fumonisins (FBs), mycotoxins found in both maize kernels and its derivatives. They are toxic to humans, in particular FB1 is classified as possibly carcinogenic. Fumonisins biosynthetic pathway is regulated by several genes belonging to FUM cluster whose behaviour in different ecological conditions was poorly studied. The aim of this work was to investigate the behaviour of 2 F. verticillioides strains grown in vitro both on a FB-inducing (Malt extract agar-MEA) and FB-inhibiting (Czapeck yeast agar-CYA) media. Fungal growth, FBs production and gene expression were studied in different temperature (T; 20-30) and water activity (aw; 0.90-0.99) regimes, in liquid cultures incubated for 7-21 days. The expression of two genes: FUM21 and FUM2 was considered; relative quantitation of gene expression was performed by RealTime (RT)-PCR. The results showed that, with aw fixed at 0.99, maximum FUM21 and FUM2 expression was observed at 30\ub0C after 14 days of incubation, while FBs production increased from 7 to 21 days. At fixed T (25\ub0C), fungal growth at 0.90 aw was not observed after 21 days of incubation. Gene expression increased with incubation time both at 0.95 and 0.99 aw, FB production increased till to 21 and 14 days respectively at 0.95 and 0.99 aw. In all the conditions studied the expression of the 2 genes considered followed a very similar trend, but FUM2 was much more expressed than FUM21 and more influenced by aw conditions

Research paper thumbnail of Evidence for birth-and-death evolution and horizontal transfer of the fumonisin mycotoxin biosynthetic gene cluster in Fusarium

In fungi, genes required for synthesis of secondary metabolites are often clustered. The FUM gene... more In fungi, genes required for synthesis of secondary metabolites are often clustered. The FUM gene cluster is required for synthesis of fumonisins, a family of toxic secondary metabolites produced by species in the Fusarium (Gibberella) fujikuroi species complex (FFSC). Fumonisins are a health and agricultural concern because their consumption is epidemiologically associated with cancer and neural tube defects in humans and other animals. Among FFSC species, the FUM cluster is uniform in gene order and orientation, but located in different genomic positions. Phylogenetic analyses indicated discord between species phylogenies and FUM gene-based phylogenies. Subsequent constraint analyses confirmed the discord, and analyses of variation in synonymous sites indicated that cluster divergence predated, in some cases, and postdated, in one case, divergence of lineages of Fusarium in which the cluster occurs. The results are not consistent with the discord resulting from transspecies evolution of ancestral cluster alleles, or with interspecies hybridization, but are consistent with duplication of the cluster within an FFSC ancestor and subsequent loss and sorting of paralogous clusters in a manner consistent with the birth-and-death evolution seen in several multigene families. Although the results are also consistent with horizontal transfer of the cluster, such a model is less parsimonious because it requires multiple transfer events from unknown but related donors to multiple FFSC recipients. However, the analyses do provide strong support for horizontal transfer of the cluster from FFSC to another Fusarium lineage. Thus, despite conservation of gene organization within it, the FusariumFUM cluster has had a complex evolutionary history.

Research paper thumbnail of Evidence for birth-and-death evolution of a secondary metabolite biosynthetic gene cluster and its relocation within and between genomes of the filamentous fungus Fusarium

Research paper thumbnail of Insights into the evolution of toxin biosynthesis in the fungus Fusarium

Research paper thumbnail of A Species-Specific PCR Assay Based on the Calmodulin Partial Gene for Identification of Fusarium Verticillioides, F. Proliferatum and F. Subglutinans

European Journal of Plant Pathology, 2004

Fusarium proliferatum, F. subglutinans and F. verticillioides are the most important Fusarium spe... more Fusarium proliferatum, F. subglutinans and F. verticillioides are the most important Fusarium species occurring on maize worldwide , capable of producing a wide range of mycotoxins which are a potential health hazard for animals and humans. The ribosomal internal transcribed spacer and a portion of the calmodulin gene were sequenced and analysed in order to design species-specific primers useful for diagnosis. The primer pairs were based on a partial calmodulin gene sequence. Three pairs of primers (PRO1/2, SUB1/2 and VER 1/2) produced PCR products of 585, 631 and 578 bp for F. proliferatum, F. subglutinans and F. verticillioides, respectively. Primer specificity was confirmed by analyzing DNA of 150 strains of these species, mostly isolated from maize in Europe and USA. The sensitivity of primers was 12.5 pg when the pure total genomic DNA of each species was analyzed. The developed PCR assay should provide a powerful tool for the detection of toxigenic fungi in maize kernels.

Research paper thumbnail of Convergent and divergent evolution of the trichothecene mycotoxin biosynthetic gene cluster in the Fusarium

Research paper thumbnail of Toxigenic fungi and mycotoxins. Diagnosis and molecular kits

Research paper thumbnail of Biodiversity of important toxigenic fungi that threaten food safety

Research paper thumbnail of Characterization of the fumonisin B2 biosynthetic gene cluster in Aspergillus niger and A. awamori

Research paper thumbnail of Fumonisin production by black Aspergillus species in maize

Research paper thumbnail of Funghi tossigeni e micotossine: diagnosi e kit molecolari

Research paper thumbnail of Good Food Project: Detection of Aspergillus carbonarius and other black Aspergilli on grapes by direct multi-detection DNA microarray

Research paper thumbnail of MBLabDB: a social database for molecular biodiversity data

Research paper thumbnail of Aspergillus uvarum sp. nov., an uniseriate black Aspergillus species isolated from grapes in Europe

INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY, 2008

Research paper thumbnail of Molecular biodiversity of mycotoxigenic fungi that threaten food safety

International Journal of Food Microbiology, 2013

Fungal biodiversity is one of the most important contributors to the occurrence and severity of m... more Fungal biodiversity is one of the most important contributors to the occurrence and severity of mycotoxin contamination of crop plants. Phenotypic and metabolic plasticity has enabled mycotoxigenic fungi to colonize a broad range of agriculturally important crops and to adapt to a range of environmental conditions. New mycotoxin-commodity combinations provide evidence for the ability of fungi to adapt to changing conditions and the emergence of genotypes that confer enhanced aggressiveness toward plants and/or altered mycotoxin production profiles. Perhaps the most important contributor to qualitative differences in mycotoxin production among fungi is variation in mycotoxin biosynthetic genes. Molecular genetic and biochemical analyses of toxigenic fungi have elucidated specific differences in biosynthetic genes that are responsible for intra- and inter-specific differences in mycotoxin production. For Aspergillus and Fusarium, the mycotoxigenic genera of greatest concern, variation in biosynthetic genes responsible for production of individual families of mycotoxins appears to be the result of evolutionary adaptation. Examples of such variation have been reported for: a) aflatoxin biosynthetic genes in Aspergillus flavus and Aspergillus parasiticus; b) trichothecene biosynthetic genes within and among Fusarium species; and c) fumonisin biosynthetic genes in Aspergillus and Fusarium species. Understanding the variation in these biosynthetic genes and the basis for variation in mycotoxin production is important for accurate assessment of the risks that fungi pose to food safety and for prevention of mycotoxin contamination of crops in the field and in storage.

Research paper thumbnail of AFLP characterization of Southern Europe population of Aspergillus Section Nigri from grapes

International Journal of Food Microbiology, 2006