Growth of an Aspergillus flavus Transformant Expressing Escherichia coli β-Glucuronidase in Maize Kernels Resistant to Aflatoxin Production

Kernels of a maize inbred that demonstrated resistance to aflatoxin production in previous studies were inoculated with an Aspergillus flavus strain containing the Escherichia coli β-d-glucuronidase reporter gene linked to a β-tubulin gene promoter and assessed for both fungal growth and aflatoxin accumulation. Prior to inoculation, kernels were pin-wounded through the pericarp to the endosperm, pin-wounded in the embryo region, or left unwounded. After 7 days incubation with the fungus, β-glucuronidase activity (fungal growth) in the kernels was quantified using a fluorogenic assay and aflatoxin B₁ content of the same kernels was analyzed. Kernels of a susceptible inbred, similarly treated, served as controls. Results indicate a positive relationship between aflatoxin levels and the amount of fungal growth. However, resistant kernels wounded through the pericarp to the endosperm before inoculation supported an increase in aflatoxin B₁ over levels observed in nonwounded kernels, without an increase in fungal growth. Wounding kernels of the resistant inbred through the embryo resulted in both the greatest fungal growth and the highest levels of aflatoxin B₁ for this genotype. Maintenance of resistance to aflatoxin B₁ in endosperm-wounded kernels may be due to the action of a mechanism which limits fungal access to the kernel embryo.