Fusarium head blight (FHB) results in yield loss and damaging contamination of cereal grains and ... more Fusarium head blight (FHB) results in yield loss and damaging contamination of cereal grains and can be caused by several Fusarium species. The objective of the present study was to determine, in a greenhouse experiment on winter wheat, how FHB was affected by timing of infection (0, 3, 6 or 9 days after anthesis, daa) by the aggressive species Fusarium graminearum compared to the relatively weak species Fusarium avenaceum, Fusarium poae and Fusarium acuminatum. Measures of FHB development were: symptoms in spikes (visually assessed), fungal biomass (quantified by real time quantitative PCR) and accumulation of fungal secondary metabolites (quantified by liquid chromatography-tandem mass spectrometry) in kernels. With regard to symptoms, F. graminearum was unaffected by inoculation timing, while the weaker pathogens caused greater disease severity at later timings. In contrast, the accumulation of F. graminearum biomass was strongly affected by inoculation timing (3 daa ≥ 6 daa ≥ 0 daa = 9 daa), while colonization by the weaker pathogens was less influenced. Similarly, F. graminearum secondary metabolite accumulation was affected by inoculation timing (3 daa ≥ 6 daa ≥ 0 daa = 9 daa), while that of the weaker species was less affected. However, secondary me-tabolites produced by these weaker species tended to be higher from intermediate-late inoculations (6 daa). Overall, infection timing appeared to play a role particularly in F. graminearum colonization and secondary metabolite accumulation. However, secondary metabolites of weaker Fusarium species may be relatively more abundant when environmental conditions promote spore dispersal later in anthesis, while secondary metabolites produced by F. graminearum are relatively favored by earlier conducive conditions.
Fusarium crown rot (FCR), an important disease of wheat and barley, is mainly caused by Fusarium ... more Fusarium crown rot (FCR), an important disease of wheat and barley, is mainly caused by Fusarium graminearum, F. culmorum and F. pseudograminearum, which are also responsible for mycotoxin production. This is the first comparative investigation of their colonization on barley plants after stem base inoculation. At plant maturity, FCR symptoms were visually evaluated, fungal biomass was quantified by Real-Time quantitative PCR and deoxyniva-lenol (DON) was detected by enzyme-linked immunosorbent assay (ELISA). All the inoculated strains caused the typical FCR necrotic symptoms. Real-Time PCR analysis showed that F. graminearum and F. culmorum were present in the head tissues, while F. pseudograminearum colonized only up to the area including the second node of the stem. Conversely, DON was detected up to the head for all the three species. This study shows that, as already demonstrated in previous research for wheat, DON may be detected up to the head as a consequence of stem base infection by the three FCR agents.
In recent years, due to the negative impact of toxigenic mycobiota and of the accumulation of the... more In recent years, due to the negative impact of toxigenic mycobiota and of the accumulation of their secondary metabolites in malting barley grains, monitoring the evolution of fungal communities in a certain cultivation area as well as detecting the different mycotoxins present in the raw material prior to malting and brewing processes have become increasingly important. In this study, a survey was carried out on malting barley samples collected after their harvest in the Umbria region (central Italy). Samples were analyzed to determine the composition of the fungal community, to identify the isolated Fusarium species, to quantify fungal secondary metabolites in the grains and to characterize the in vitro mycotoxigenic profile of a subset of the isolated Fusarium strains. The fungal community of barley grains was mainly composed of microorganisms belonging to the genus Alternaria (77%), followed by those belonging to the genus Fusarium (27%). The Fusarium head blight (FHB) complex was represented by nine species with the predominance of Fusarium poae (37%), followed by Fusarium avenaceum (23%), Fusarium graminearum (22%) and Fusarium tricinctum (7%). Secondary metabolites bio-synthesized by Alternaria and Fusarium species were present in the analyzed grains. Among those biosynthesized by Fusarium species, nivalenol and enniatins were the most prevalent ones. Type A trichothecenes (T-2 and HT-2 toxins) as well as beauvericin were also present with a high incidence. Conversely, the number of samples contaminated with deoxynivalenol was low. Conjugated forms, such as deoxynivalenol-3-glucoside and HT-2-glucoside, were detected for the first time in malting barley grains cultivated in the surveyed area. In addition, strains of F. avenaceum and F. tricinctum showed the ability to biosynthesize in vitro high concentrations of enniatins. The analysis of fungal secondary metabolites, both in the grains and in vitro, revealed also the presence of other compounds, for which further investigations will be required. The combination of microbiological analyses, of molecular biology assays and of multi-mycotoxin screening shed light on the complexity of the fungal community and its secondary metabolites released in malting barley.
Fusarium pseudograminearum, F. culmorum and F. graminearum are the most important fusarium crown ... more Fusarium pseudograminearum, F. culmorum and F. graminearum are the most important fusarium crown rot (FCR) causal agents. They have the common ability to biosynthesize deoxynivalenol (DON). To elucidate the behaviour of each of the three species, a comparative study was carried out to investigate symptom progression, fungal systemic growth and translocation of DON following stem base inoculation of soft wheat. FCR symptoms were mainly localized in the inoculated area, which extended up to the second node for all inoculated species. Only the most aggressive strains caused symptoms up to the third node. Real-time quantitative PCR showed that fungal colonization reached the third node for all the tested species, but a low percentage of plants showed colonization above the third node following inoculation with the most aggressive strains. Fungal growth was detected in symptomless tissues but none of the three species was able to colonize as far as the head tissues. However, even if the pathogens were not detected in the heads, DON was detected in head tissues of the plants inoculated with the most aggressive strains. These results demonstrate that F. pseudograminearum, F. culmorum and F. graminearum, under the same experimental conditions, follow a similar pattern of symptom progression, fungal colonization and DON translocation after stem base infection. Differences in the extent of symptoms, fungal colonization and mycotoxin distribution were mainly attributable to strain aggressiveness. These findings provide comparative information on the events following infection of the stem base of wheat by three of the most important FCR casual agents.
Durum wheat samples harvested in central Italy (Umbria) were analyzed to: evaluate the occurrence... more Durum wheat samples harvested in central Italy (Umbria) were analyzed to: evaluate the occurrence of the fungal community in the grains, molecularly identify the Fusarium spp. which are part of the Fusarium head blight (FHB) complex and characterize the in vitro secondary metabolite profiles of a subset of Fusarium strains. The Fusarium genus was one of the main components of the durum wheat fungal community. The FHB complex was composed of eight species: Fusarium avenaceum (61%), F. graminearum (22%), F. poae (9%), F. culmorum (4%), F. proliferatum (2%), F. sporotrichioides (1%), F. sambucinum (0.5%) and F. langsethiae (0.5%). F. graminearum population was mainly composed of the 15-acetyldeoxynivalenol chemotype, while, F. culmorum population was composed of the 3-acetyldeoxynivalenol chemotype. In vitro characterization of secondary metabolite biosynthesis was conducted for a wide spectrum of substances, showing the mycotoxigenic potential of the species complex. F. avenaceum strains were characterized by high enniantin and moniliformin production. F. graminearum strains were in prevalence deoxynivalenol producers. F. poae strains were characterized by a high biosynthesis of beauvericin like the F. sporotrichioides strain which was also found to be a high T-2/HT-2 toxins producer. Production of aurofusarin, butenolide, gibepyrone D, fusarin C, apicidin was also reported for the analyzed strains.
In this study, conducted for three years on eleven malting barley varieties cultivated in central... more In this study, conducted for three years on eleven malting barley varieties cultivated in central Italy, the incidence of different mycotoxigenic fungal genera, the identification of the Fusarium species associated with the Fusarium Head Blight (FHB) complex, and kernels contamination with deoxynivalenol (DON) and T-2 mycotoxins were determined. The influence of climatic conditions on Fusarium infections and FHB complex composition was also investigated. Fusarium species were always present in the three years and the high average and maximum temperatures during anthesis mainly favored their occurrence. The FHB complex was subject to changes during the three years and the main causal agents were F. poae, F. avenaceum, F. tricinctum and F. graminearum, which, even if constantly present, never represented the principal FHB agent. The relative incidence of Fusarium species changed because of climatic conditions occurring during the seasons. The FHB complex was composed of many different Fusarium species and some of them were associated with a specific variety and/or with specific weather parameters, indicating that the interaction between a certain plant genotype and climatic conditions may influence the presence of Fusarium spp. causing infections. With regard to mycotoxin contamination, T-2 toxin, in some cases, was found in kernels at levels that exceeded EU recommended values.
A study was carried out on 43 malting barley samples collected in 2013 across the Umbria region (... more A study was carried out on 43 malting barley samples collected in 2013 across the Umbria region (central Italy) to determine the incidence of the principal mycotoxigenic fungal genera, to identify the Fusarium species isolated from the grains, and to detect the presence of 34 fungal secondary metabolites by liquid chromatography−high-resolution mass spectrometry. The multimycotoxin-method development involved the evaluation of both a two-step solvent and QuEChERS protocol for metabolite extraction. The former protocol was selected because of better accuracy, which was evaluated on the basis of spike-recovery experiments. The most frequently isolated fungal species belonged to the genera Alternaria and Fusarium. The predominant Fusarium species was F. avenaceum, followed by F. graminearum. HT-2 toxin was the most frequently detected mycotoxin, followed by enniatin B, enniatin B1, T-2 toxin, and nivalenol. As a consequence of the observed mixed fungal infections, mycotoxin co-occurrence was also detected. A combination of mycological and mycotoxin analyses allowed the ability to obtain comprehensive information about the presence of mycotoxigenic fungi and their contaminants in malting barley cultivated in a specific geographic area.
A multi-mycotoxin analysis method based on liquid chromatography coupled to triple quadrupole mas... more A multi-mycotoxin analysis method based on liquid chromatography coupled to triple quadrupole mass spectrometry was validated and applied for the determination of twenty-six mycotoxins, including eight trichothecenes, three fumonisins, sterigmatocystin, ochratoxin A, four aflatoxins, zearalenone, five “emerging” mycotoxins and three Alternaria mycotoxins in 74 durum wheat samples from central Italy. Eightyeight percent of the analysed samples contained one or more mycotoxins, with enniatin B showing the highest contamination levels (78% of samples with levels ranging from 23 to 1826 ng/g), followed by enniatin B1 (10–1384 ng/g) and deoxynivalenol (48–2267 ng/g). This work gives an important contribution in terms of mycotoxin analysis and occurrence in durum wheat and in assessing the mycotoxicological risk posed by both traditional and emerging mycotoxins for the final consumers of durum wheat derived products.
Fusarium head blight (FHB) results in yield loss and damaging contamination of cereal grains and ... more Fusarium head blight (FHB) results in yield loss and damaging contamination of cereal grains and can be caused by several Fusarium species. The objective of the present study was to determine, in a greenhouse experiment on winter wheat, how FHB was affected by timing of infection (0, 3, 6 or 9 days after anthesis, daa) by the aggressive species Fusarium graminearum compared to the relatively weak species Fusarium avenaceum, Fusarium poae and Fusarium acuminatum. Measures of FHB development were: symptoms in spikes (visually assessed), fungal biomass (quantified by real time quantitative PCR) and accumulation of fungal secondary metabolites (quantified by liquid chromatography-tandem mass spectrometry) in kernels. With regard to symptoms, F. graminearum was unaffected by inoculation timing, while the weaker pathogens caused greater disease severity at later timings. In contrast, the accumulation of F. graminearum biomass was strongly affected by inoculation timing (3 daa ≥ 6 daa ≥ 0 daa = 9 daa), while colonization by the weaker pathogens was less influenced. Similarly, F. graminearum secondary metabolite accumulation was affected by inoculation timing (3 daa ≥ 6 daa ≥ 0 daa = 9 daa), while that of the weaker species was less affected. However, secondary me-tabolites produced by these weaker species tended to be higher from intermediate-late inoculations (6 daa). Overall, infection timing appeared to play a role particularly in F. graminearum colonization and secondary metabolite accumulation. However, secondary metabolites of weaker Fusarium species may be relatively more abundant when environmental conditions promote spore dispersal later in anthesis, while secondary metabolites produced by F. graminearum are relatively favored by earlier conducive conditions.
Fusarium crown rot (FCR), an important disease of wheat and barley, is mainly caused by Fusarium ... more Fusarium crown rot (FCR), an important disease of wheat and barley, is mainly caused by Fusarium graminearum, F. culmorum and F. pseudograminearum, which are also responsible for mycotoxin production. This is the first comparative investigation of their colonization on barley plants after stem base inoculation. At plant maturity, FCR symptoms were visually evaluated, fungal biomass was quantified by Real-Time quantitative PCR and deoxyniva-lenol (DON) was detected by enzyme-linked immunosorbent assay (ELISA). All the inoculated strains caused the typical FCR necrotic symptoms. Real-Time PCR analysis showed that F. graminearum and F. culmorum were present in the head tissues, while F. pseudograminearum colonized only up to the area including the second node of the stem. Conversely, DON was detected up to the head for all the three species. This study shows that, as already demonstrated in previous research for wheat, DON may be detected up to the head as a consequence of stem base infection by the three FCR agents.
In recent years, due to the negative impact of toxigenic mycobiota and of the accumulation of the... more In recent years, due to the negative impact of toxigenic mycobiota and of the accumulation of their secondary metabolites in malting barley grains, monitoring the evolution of fungal communities in a certain cultivation area as well as detecting the different mycotoxins present in the raw material prior to malting and brewing processes have become increasingly important. In this study, a survey was carried out on malting barley samples collected after their harvest in the Umbria region (central Italy). Samples were analyzed to determine the composition of the fungal community, to identify the isolated Fusarium species, to quantify fungal secondary metabolites in the grains and to characterize the in vitro mycotoxigenic profile of a subset of the isolated Fusarium strains. The fungal community of barley grains was mainly composed of microorganisms belonging to the genus Alternaria (77%), followed by those belonging to the genus Fusarium (27%). The Fusarium head blight (FHB) complex was represented by nine species with the predominance of Fusarium poae (37%), followed by Fusarium avenaceum (23%), Fusarium graminearum (22%) and Fusarium tricinctum (7%). Secondary metabolites bio-synthesized by Alternaria and Fusarium species were present in the analyzed grains. Among those biosynthesized by Fusarium species, nivalenol and enniatins were the most prevalent ones. Type A trichothecenes (T-2 and HT-2 toxins) as well as beauvericin were also present with a high incidence. Conversely, the number of samples contaminated with deoxynivalenol was low. Conjugated forms, such as deoxynivalenol-3-glucoside and HT-2-glucoside, were detected for the first time in malting barley grains cultivated in the surveyed area. In addition, strains of F. avenaceum and F. tricinctum showed the ability to biosynthesize in vitro high concentrations of enniatins. The analysis of fungal secondary metabolites, both in the grains and in vitro, revealed also the presence of other compounds, for which further investigations will be required. The combination of microbiological analyses, of molecular biology assays and of multi-mycotoxin screening shed light on the complexity of the fungal community and its secondary metabolites released in malting barley.
Fusarium pseudograminearum, F. culmorum and F. graminearum are the most important fusarium crown ... more Fusarium pseudograminearum, F. culmorum and F. graminearum are the most important fusarium crown rot (FCR) causal agents. They have the common ability to biosynthesize deoxynivalenol (DON). To elucidate the behaviour of each of the three species, a comparative study was carried out to investigate symptom progression, fungal systemic growth and translocation of DON following stem base inoculation of soft wheat. FCR symptoms were mainly localized in the inoculated area, which extended up to the second node for all inoculated species. Only the most aggressive strains caused symptoms up to the third node. Real-time quantitative PCR showed that fungal colonization reached the third node for all the tested species, but a low percentage of plants showed colonization above the third node following inoculation with the most aggressive strains. Fungal growth was detected in symptomless tissues but none of the three species was able to colonize as far as the head tissues. However, even if the pathogens were not detected in the heads, DON was detected in head tissues of the plants inoculated with the most aggressive strains. These results demonstrate that F. pseudograminearum, F. culmorum and F. graminearum, under the same experimental conditions, follow a similar pattern of symptom progression, fungal colonization and DON translocation after stem base infection. Differences in the extent of symptoms, fungal colonization and mycotoxin distribution were mainly attributable to strain aggressiveness. These findings provide comparative information on the events following infection of the stem base of wheat by three of the most important FCR casual agents.
Durum wheat samples harvested in central Italy (Umbria) were analyzed to: evaluate the occurrence... more Durum wheat samples harvested in central Italy (Umbria) were analyzed to: evaluate the occurrence of the fungal community in the grains, molecularly identify the Fusarium spp. which are part of the Fusarium head blight (FHB) complex and characterize the in vitro secondary metabolite profiles of a subset of Fusarium strains. The Fusarium genus was one of the main components of the durum wheat fungal community. The FHB complex was composed of eight species: Fusarium avenaceum (61%), F. graminearum (22%), F. poae (9%), F. culmorum (4%), F. proliferatum (2%), F. sporotrichioides (1%), F. sambucinum (0.5%) and F. langsethiae (0.5%). F. graminearum population was mainly composed of the 15-acetyldeoxynivalenol chemotype, while, F. culmorum population was composed of the 3-acetyldeoxynivalenol chemotype. In vitro characterization of secondary metabolite biosynthesis was conducted for a wide spectrum of substances, showing the mycotoxigenic potential of the species complex. F. avenaceum strains were characterized by high enniantin and moniliformin production. F. graminearum strains were in prevalence deoxynivalenol producers. F. poae strains were characterized by a high biosynthesis of beauvericin like the F. sporotrichioides strain which was also found to be a high T-2/HT-2 toxins producer. Production of aurofusarin, butenolide, gibepyrone D, fusarin C, apicidin was also reported for the analyzed strains.
In this study, conducted for three years on eleven malting barley varieties cultivated in central... more In this study, conducted for three years on eleven malting barley varieties cultivated in central Italy, the incidence of different mycotoxigenic fungal genera, the identification of the Fusarium species associated with the Fusarium Head Blight (FHB) complex, and kernels contamination with deoxynivalenol (DON) and T-2 mycotoxins were determined. The influence of climatic conditions on Fusarium infections and FHB complex composition was also investigated. Fusarium species were always present in the three years and the high average and maximum temperatures during anthesis mainly favored their occurrence. The FHB complex was subject to changes during the three years and the main causal agents were F. poae, F. avenaceum, F. tricinctum and F. graminearum, which, even if constantly present, never represented the principal FHB agent. The relative incidence of Fusarium species changed because of climatic conditions occurring during the seasons. The FHB complex was composed of many different Fusarium species and some of them were associated with a specific variety and/or with specific weather parameters, indicating that the interaction between a certain plant genotype and climatic conditions may influence the presence of Fusarium spp. causing infections. With regard to mycotoxin contamination, T-2 toxin, in some cases, was found in kernels at levels that exceeded EU recommended values.
A study was carried out on 43 malting barley samples collected in 2013 across the Umbria region (... more A study was carried out on 43 malting barley samples collected in 2013 across the Umbria region (central Italy) to determine the incidence of the principal mycotoxigenic fungal genera, to identify the Fusarium species isolated from the grains, and to detect the presence of 34 fungal secondary metabolites by liquid chromatography−high-resolution mass spectrometry. The multimycotoxin-method development involved the evaluation of both a two-step solvent and QuEChERS protocol for metabolite extraction. The former protocol was selected because of better accuracy, which was evaluated on the basis of spike-recovery experiments. The most frequently isolated fungal species belonged to the genera Alternaria and Fusarium. The predominant Fusarium species was F. avenaceum, followed by F. graminearum. HT-2 toxin was the most frequently detected mycotoxin, followed by enniatin B, enniatin B1, T-2 toxin, and nivalenol. As a consequence of the observed mixed fungal infections, mycotoxin co-occurrence was also detected. A combination of mycological and mycotoxin analyses allowed the ability to obtain comprehensive information about the presence of mycotoxigenic fungi and their contaminants in malting barley cultivated in a specific geographic area.
A multi-mycotoxin analysis method based on liquid chromatography coupled to triple quadrupole mas... more A multi-mycotoxin analysis method based on liquid chromatography coupled to triple quadrupole mass spectrometry was validated and applied for the determination of twenty-six mycotoxins, including eight trichothecenes, three fumonisins, sterigmatocystin, ochratoxin A, four aflatoxins, zearalenone, five “emerging” mycotoxins and three Alternaria mycotoxins in 74 durum wheat samples from central Italy. Eightyeight percent of the analysed samples contained one or more mycotoxins, with enniatin B showing the highest contamination levels (78% of samples with levels ranging from 23 to 1826 ng/g), followed by enniatin B1 (10–1384 ng/g) and deoxynivalenol (48–2267 ng/g). This work gives an important contribution in terms of mycotoxin analysis and occurrence in durum wheat and in assessing the mycotoxicological risk posed by both traditional and emerging mycotoxins for the final consumers of durum wheat derived products.
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spectrometry was validated and applied for the determination of twenty-six mycotoxins, including eight
trichothecenes, three fumonisins, sterigmatocystin, ochratoxin A, four aflatoxins, zearalenone, five “emerging”
mycotoxins and three Alternaria mycotoxins in 74 durum wheat samples from central Italy. Eightyeight
percent of the analysed samples contained one or more mycotoxins, with enniatin B showing the
highest contamination levels (78% of samples with levels ranging from 23 to 1826 ng/g), followed by enniatin
B1 (10–1384 ng/g) and deoxynivalenol (48–2267 ng/g). This work gives an important contribution
in terms of mycotoxin analysis and occurrence in durum wheat and in assessing the mycotoxicological
risk posed by both traditional and emerging mycotoxins for the final consumers of durum wheat derived
products.
spectrometry was validated and applied for the determination of twenty-six mycotoxins, including eight
trichothecenes, three fumonisins, sterigmatocystin, ochratoxin A, four aflatoxins, zearalenone, five “emerging”
mycotoxins and three Alternaria mycotoxins in 74 durum wheat samples from central Italy. Eightyeight
percent of the analysed samples contained one or more mycotoxins, with enniatin B showing the
highest contamination levels (78% of samples with levels ranging from 23 to 1826 ng/g), followed by enniatin
B1 (10–1384 ng/g) and deoxynivalenol (48–2267 ng/g). This work gives an important contribution
in terms of mycotoxin analysis and occurrence in durum wheat and in assessing the mycotoxicological
risk posed by both traditional and emerging mycotoxins for the final consumers of durum wheat derived
products.