Population genetic analysis and trichothecene profiling of Fusarium graminearum from wheat in Uruguay.

Fusarium graminearum sensu stricto (F. graminearum s.s.) is the major causal agent of Fusarium head blight of wheat worldwide, and contaminates grains with trichothecene mycotoxins that cause serious threats to food safety and animal health. An important aspect of managing this pathogen and reducing mycotoxin contamination of wheat is knowledge regarding its population genetics. Therefore, isolates of F. graminearum s.s. from the major wheat-growing region of Uruguay were analyzed by amplified fragment length polymorphism assays, PCR genotyping, and chemical analysis of trichothecene production. Of the 102 isolates identified as having the 15-ADON genotype via PCR genotyping, all were DON producers, but only 41 strains were also 15-ADON producers, as determined by chemical analysis. The populations were genotypically diverse but genetically similar, with significant genetic exchange occurring between them. Analysis of molecular variance indicated that most of the genetic variability resulted from differences between isolates within populations. Multilocus linkage disequilibrium analysis suggested that the isolates had a panmictic population genetic structure and that there is significant recombination occurs in F. graminearum s.s. In conclusion, tour findings provide the first detailed description of the genetic structure and trichothecene production of populations of F. graminearum s.s. from Uruguay, and expands our understanding of the agroecology of F. graminearum and of the correlation between genotypes and trichothecene chemotypes.

Endophytic bacteria from wheat grain as biocontrol agents of Fusarium graminearum and deoxynivalenol production in wheat.

In Uruguay, Fusarium graminearum is the most common species that infects wheat and is responsible for Fusarium head blight (FHB) and contamination of grain with deoxynivalenol (DON). The aim of this work was to select bacterial endophytes isolated from wheat grain to evaluate their antagonistic ability against F. graminearum and DON production in vitro and under field conditions. Four strains identified as Bacillus megaterium (BM1) and Bacillus subtilis (BS43, BSM0 y BSM2) significantly reduced fungal growth and spore germination of F. graminearum. This antagonist activity remained unchanged after the bacterial cultures were heat treated. Under field conditions, treatments with antagonist BM1 was the most effective, reducing the FHB incidence and severity by 93 and 54 %, respectively, and the production of DON by 89.3 %.

Resistance to Chalkbrood disease in Apis mellifera L. (Hymenoptera: Apidae) colonies with different hygienic behaviour.

Chalkbrood disease affects the larvae of honeybees Apis mellifera L. and is caused by the fungus Ascosphaera apis. Infected larvae die when they are stretched in the cap cell and suffer a gradual hardening that ends in a very hard structure (mummie). Several studies have demonstrated that colonies that express an efficient hygienic behaviour (uncapping of cell and subsequent removal of dead brood) exhibit a higher resistance to the disease. However, it remains unclear whether the advantage of hygienic colonies over less hygienic ones lies in the ability to remove mummies or in the early detection of infected larvae and its cannibalization before they harden. To elucidate this aspect, the hygienic behaviour of 24 colonies, which were subsequently provided with pollen cakes containig A. apis, was evaluated. The number of mummies and the number of partially cannibalized and whole larvae in uncapped cells were recorded. The most hygienic colonies controlled the disease better. These colonies also had a higher tendency to uncap cells that contained infected larvae and cannibalize them. The presence of A. apis in partially cannibalized and whole larvae in uncapped cells indicate that the advantage of hygienic colonies over less hygienic ones lies in the early detection of infected larvae death and their quick removal from the cell before they become mummies.

Resistance to Chalkbrood Disease in Apis mellifera L. (Hymenoptera: Apidae) Colonies with Different Hygienic Behaviour

Chalkbrood disease affects the larvae of honeybees Apis mellifera L. and is caused by the fungus Ascosphaera apis. Infected larvae die when they are stretched in the cap cell and suffer a gradual hardening that ends in a very hard structure (mummie). Several studies have demonstrated that colonies that express an efficient hygienic behaviour (uncapping of cell and subsequent removal of dead brood) exhibit a higher resistance to the disease. However, it remains unclear whether the advantage of hygienic colonies over less hygienic ones lies in the ability to remove mummies or in the early detection of infected larvae and its cannibalization before they harden. To elucidate this aspect, the hygienic behaviour of 24 colonies, which were subsequently provided with pollen cakes containig A. apis, was evaluated. The number of mummies and the number of partially cannibalized and whole larvae in uncapped cells were recorded. The most hygienic colonies controlled the disease better. These colonies also had a higher tendency to uncap cells that contained infected larvae and cannibalize them. The presence of A. apis in partially cannibalized and whole larvae in uncapped cells indicate that the advantage of hygienic colonies over less hygienic ones lies in the early detection of infected larvae death and their quick removal from the cell before they become mummies.

Correlation of rainfall and levels of deoxynivalenol in wheat from Uruguay, 1997-2003.

A total of 286 wheat samples for human consumption collected during 1997-2003 from four wheat-producing localities of south-western Uruguay were screened for deoxynivalenol (DON). Quantification was carried on by an immunochemical method using immunoaffinity columns and fluorimetric detection. The incidence of DON was high during the whole survey (58.5-100%), except in 1998 and 1999 in which no contamination occurred. During 2001 and 2002, 100% of samples contained detectable levels of DON, being the mean DON contents 6593 and 5880 µg kg(-1), respectively. The annual maximum levels ranged from 8800 to 11,400 µg kg(-1). A positive correlation between DON levels and precipitation was seen. The 70% of wheat samples destined for human consumption were contaminated with DON. To avoid the introduction of contaminated materials into the food chain process, the adoption of regular screening of the DON level in wheat is recommended, particularly in years with heavy rainfall during the flowering-to-early stages of grain maturity months.

Endophytic fungi from flowers, capsules and seeds of Eucalyptus globulus.

The main goal of this work was to detect whether Cytospora chrysosperma and Fusicoccum eucalypti are present as endophytes of symptomless hypocotyls, cotyledons, flowers, capsules, peduncles of flowers in order to interpret an earlier finding of their presence in seeds, seedling stems and twigs of E. globulus. Segments from these organs as well as from bark and the xylem from flower peduncles were surface-sterilized and plated on 2% malt-agar. All plates were incubated at 24 degrees C for six weeks or more depending on the growth rate of fungi. C. chrysosperma was asymptomatically present in flowers, capsules, hypocotyls, cotyledons and peduncles. F. eucalypti was isolated from asymptomatic flowers and capsules. It is probable that C. chrysosperma spreads during seed germination colonizing seedling stems through hypocotyl and cotyledon.

The effect of fungal competition on colonization of maize grain by Fusarium moniliforme, F. proliferatum and F. graminearum and on fumonisin B1 and zearalenone formation.

The effect of water activity (0.98, 0.95, 0.93) and temperature (15, 25 degrees C) on fungal growth and toxin production from interactions between isolates of Fusarium moniliforme and F. proliferatum producing fumonisin, and an isolate of F. graminearum producing zearalenone, incubated at the same time on irradiated maize grains were determined in vitro. Populations (CFUs) of F. moniliforme and F. proliferatum were reduced to a greater or lesser extent by the presence of F. graminearum under all conditions tested, while that the presence of F. moniliforme or F. proliferatum had a minor inhibitory effect on fungal populations of F. graminearum. Fumonisin B, production by F. proliferatum was inhibited under all conditions tested, while fumonisin B1 production by F. moniliforme was inhibited at 15 degrees C and enhanced at 25 degrees C in the presence of F. graminearum. The level of zearalenone was not significantly modified in the presence of F. moniliforme and F. proliferatum under the conditions tested.