393 The Potential of Saprophytic Yeasts Antagonistic to Aspergillus flavus in Reducing Aflatoxin Contamination of Tree Nuts
California is the major state for producing almonds, pistachios, and walnuts, with a total market value of $1.6 billion. Both domestic and export markets of these nuts presently allow a maximum level of aflatoxin B1 contamination in the edible nuts to be 20 ppb. Even very low degrees of infection of the nuts by A. flavus can result in aflatoxin levels above the mandatory standards. Biological control to reduce the population of and to inhibit the biosynthesis of A. flavus in orchards may be useful to decrease infection and thus aflatoxin content in the edible nuts. Certain saprophytic yeasts were shown to effectively compete with postharvest fungal pathogens such as Penicillium expansum and Botrytis cinerea. The potential of saprophytic yeasts to reduce aflatoxin contamination in tree nuts has not been hitherto extensively explored. A safe visual bioassay for screening yeasts antagonistic to A. flavus has been developed. The nor mutant of A. flavus has a defective norsolorinic acid reductase and blocks the aflatoxin biosynthetic pathway, resulting in the accumulation of norsolorinic acid, a bright red-orange pigment. We used the nor mutant in the assay to screen yeasts strains for their ability to inhibit aflatoxin production by visually scoring the accumulation of this pigment as well as the growth and sporulation of the fungus. Yeast strains that reduced the red-orange pigment accumulation in the nor mutant were identified and shown to inhibit aflatoxin biosynthesis of several toxigenic strains of A. flavus.