Trichothecene genotypes and production profiles of Fusarium graminearum isolates obtained from barley cultivated in Argentina.

Fusarium graminearum is one of the most important pathogens isolated from small cereal grains with Fusarium Head Blight symptoms. The presence of this fungus is often linked to the occurrence of several mycotoxins in barley and wheat. The aim of our study was to characterize trichothecene genotypes and production profiles of F. graminearum sensu stricto isolates obtained from barley grains in Argentina. A total of 110 F. graminearum s.s. isolates were analyzed by PCR assays to predict deoxynivalenol (DON), 15-acetyldeoxynivalenol (15-ADON), 3-acetyldeoxynivalenol (3-ADON) and nivalenol (NIV) production, and all isolates were found to belong to the same molecular 15-ADON genotype. Trichothecene production in autoclaved rice was analyzed by using gas chromatography (GC) and confirmed by GC-MS. Of the 110 isolates, 95% were able to produce DON, 71% produced 15-ADON, 63% 3-ADON and 52% NIV. With the exception of a single isolate, all isolates that produced NIV, also produced DON. However, the NIV production was very low, ranging from 0.13 to 0.30 µg/g. Six different production profiles of DON and its acetyl-derivatives were detected, the predominant being simultaneous production of DON, 3-ADON and 15-ADON, followed by DON production, and DON and 15-ADON co-production. This work is the first attempt to characterize the trichothecene genotypes and production profiles of F. graminearum s.s. isolates from Argentinean barley.

Genetic diversity in Fusarium graminearum from a major wheat-producing region of Argentina.

The Fusarium graminearum species complex (FGSC) is a group of mycotoxigenic fungi that are the primary cause of Fusarium head blight (FHB) of wheat worldwide. The distribution, frequency of occurrence, and genetic diversity of FGSC species in cereal crops in South America is not well understood compared to some regions of Asia, Europe and North America. Therefore, we examined the frequency and genetic diversity of a collection of 183 FGSC isolates recovered from wheat grown during multiple growing seasons and across a large area of eastern Argentina, a major wheat producing region in South America. Sequence analysis of the translation elongation factor 1-α and ß-tubulin genes as well as Amplified Fragment Length Polymorphism (AFLP) analyses indicated that all isolates were the FGSC species F. graminearum sensu stricto. AFLP analysis resolved at least 11 subgroups, and all the isolates represented different AFLP haplotypes. AFLP profile and geographic origin were not correlated. Previously obtained trichothecene production profiles of the isolates revealed that the 15-acetyldeoxynivalenol chemotype was slightly more frequent than the 3-acetyldeoxynivalenol chemotype among the isolates. These data extend the current understanding of FGSC diversity and provide further evidence that F. graminearum sensu stricto is the predominant cause of FHB in the temperate main wheat-growing area of Argentina. Moreover, two isolates of F. crookwellense and four of F. pseudograminearum were also recovered from wheat samples and sequenced. The results also suggest that, although F. graminearum sensu stricto was the only FGSC species recovered in this study, the high level of genetic diversity within this species should be considered in plant breeding efforts and development of other disease management strategies aimed at reducing FHB.

AFLP variability, toxin production, and pathogenicity of Alternaria species from Argentinean tomato fruits and puree.

Large amounts of tomato fruits and derived products are produced in Argentina and may be contaminated by Alternaria toxins. Limited information is available on the genetic variability, toxigenicity, and pathogenicity of Alternaria strains occurring on tomato. We analyzed 65 Alternaria strains isolated in Argentina from tomato fruits affected by black mould and from tomato puree, using amplified fragment length polymorphisms (AFLPs) technique. AFLP analysis resolved the set of strains in 3 main clusters (DICE similarity values of 58 and 60%) corresponding to A. alternata/tenuissima (44 strains), A. arborescens (15 strains) and to an unknown group (6 strains). Most of the representative strains, belonging to each AFLP cluster, when cultured on rice, produced tenuazonic acid (up to 46,760 mg/kg), alternariol monomethyl ether (AME, up to 1860 mg/kg), and alternariol (up to 70 mg/kg). The toxin profile related to the strains was not related to any AFLP cluster, except for AME which was produced at lower level by A. arborescens. Most of strains were pathogenic on two types of commonly cultivated tomato fruits. These findings provide new information on the variability within the Alternaria species complex associated with tomato disease.

Ochratoxin A production by a mixed inoculum of Aspergillus carbonarius at different conditions of water activity and temperature.

Growth rate, lag phase and OTA production of a mixed inoculum of four Aspergillus carbonarius strains were compared to the behaviour of the single strains at 30 degrees C on Czapek Yeast Extract Agar. Significant differences between radial growth rates and lag phases of the different isolates were observed; however, no significant differences between growth rates of each strain and the mixed inoculum were detected. When the four strains were inoculated simultaneously, the lag phase was the same than the higher value obtained for individual strains, suggesting mycelial interactions between the A. carbonarius isolates. The four strains differed in maximum OTA yield, and the toxin accumulation by the mixed inoculum showed intermediate levels at each time point. The effects of water activity (0.83, 0.85, 0.87, 0.89, 0.90, 0.93 and 0.95) and temperature (15, 20, 25, 30, and 35 degrees C) on OTA production by the mixed inoculum were studied at 7, 14, 21 and 28days of incubation. The limiting water activity for OTA production by the mixed inoculum was 0.87, showing xerotolerant behaviour of the strains isolated from dried vine fruits. Results obtained were similar to those reported for single A. carbonarius strains from European countries, Israel, Australia and South America. The similar trend in the response of the different isolates to the variation of environmental parameters may be of interest for the building of predictive models.

Toxigenic potential of Fusarium graminearum sensu stricto isolates from wheat in Argentina.

The toxigenic potential of 144 Fusarium graminearum sensu stricto strains isolated from wheat in different subregions of the main production area in Argentina during 2001, 2003 and 2004 harvests was studied. The mycotoxin producing ability of the isolates was examined by Gas Chromatography with Electron Capture Detection and confirmed by Mass Spectrometry. Analysis of the trichothecene chemotype distribution across the Argentinean wheat cropping area revealed that 15-acetyldeoxynivalenol was the most common chemotype. Two other chemotypes, the 3-acetyldeoxynivalenol and 3- and 15-acetyldeoxynivalenol, were always present. The number of isolates with simultaneous production of both acetylderivatives increased along the years. The frequency of isolates with different toxigenic profile did not shown significant differences by subregions. An increased vigilance over crop seasons and geographical areas should be carried out to detect changes in the chemotypes distribution.