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Movement and Longevity of Aspergillus flavus Propagules and Factors that Contribute to and Influence their Colonization and Production

By Brandon Hassett

Abstract

Aflatoxin contamination accounts for millions of dollars worth of losses for corn and cotton in Texas. Two atoxigenic strains of Aspergillus flavus, AF36 and Afla-Guard, are labeled for its management. The purpose of this study was to measure differences in the ability of these strains to sporulate and to track movement of their conidia in corn and cotton fields. Sporulation was evaluated by incubating the two strains on their commercial formulations (inoculated on cereal grains) at six constant humidity levels ranging from 0-100%, using closed chambers with saturated salt solutions. Conidial production by Afla-Guard was 3-fold greater than that of AF36 at 100% humidity. Sporulation of the two strains was also evaluated on one substrate by inoculating their conidia on sterile, hulled barley. After 3 days, there was a 234-fold increase in conidia recovered from the barley inoculated with Afla-Guard, compared with a 21-fold increase in conidia recovered from the AF36-inoculated barley. These data suggest that the Afla-Guard strain sporulates better than the AF36 strain, which may be a factor in effectiveness for biological control. An in vitro de-Wit competition experiment showed that sporulation by the Afla-Guard strain was not affected by co-inoculation with either AF36 or the wildtype NRRL3357 toxin producing strain. To measure conidial movement, an Afla-Guard nitrate non-utilizing mutant colonizing autoclaved corn seed, was placed at one point in a field of cotton and corn. For detection, aliquots washed from leaf samples were plated onto a medium containing potassium chlorate. The mutant was recovered at a maximum distance of 6.4 m in corn fields along the same row and as far as 10.2 m across rows from the point source. In cotton fields, the mutant was recovered at 9.1 meters along the same row and 6.1 m across rows from the point source. There was no recovery at 24.3 m from the point source - the maximum distance evaluated. The experiment was repeated in a second year with similar results. These data suggest that plots in field trials may not need wide separation in order to avoid cross contamination. To assess the viability of a toxigenic and atoxigenic strain of A. flavus over time, polycarbonate packets containing conidia and sclerotia of both strains were buried in Ships Clay soil with the matric potential held constant at -24 kPa or -154 kPa. After 10 months, viable conidia were recovered in all treatments. After 14 months, viability of the atoxigenic strain incubated at -154 kPa ?m was lost, while other treatments remained viable. Ears of corn were inoculated via silk channel at different stages of silk senescence. Sclerotia were enumerated from the same plants following harvest of the crop. Sclerotial production by A. flavus was greatest from ears with silks inoculated at senescence, compared with inoculation when silks were green. The isolation frequency of Penicillium sp. from surface-sterilized kernels at harvest was the highest from ears that were inoculated with A. flavus when silks were fresh, as compared with A. flavus inoculation of ears with senescent silks. A Fusarium and Penicillium species was isolated from harvested kernels, and their sterile Czapek-Dox broth culture filtrates were tested for their effect on development of three strains of A. flavus on agar. The Penicillium broth filtrate greatly reduced sclerotial numbers relative to the control and the Fusarium filtrate (P<0.05). When A. flavus was grown in the presence of autoclaved Penicillium culture filtrate, there was no effect on sclerotial production. The Penicillium filtrate increased the rate of radial hyphal growth of the A. flavus isolates on agar compared to the control and the Fusarium culture filtrate.

Topics: aflatoxin, Aspergillus, atoxigenic, biocontrol, conidia, matric potential, sclerotia
Year: 2012
OAI identifier: oai:oaktrust.library.tamu.edu:1969.1/ETD-TAMU-2012-08-11710

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