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Heritability study of eGFP-transformedAspergillus flavus strains

in World Mycotoxin Journal
Autor:in:
G.G. Moore Southern Regional Research Center, Agricultural Research Service, SDA, 1100 Robert E. Lee Blvd, New Orleans, LA 70124, USA

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Field inoculation with non-aflatoxigenicAspergillus flavus is a preferred method for pre-harvest biocontrol of aflatoxin contamination of maize, cottonseed, and groundnut. Rationale for using theseA. flavus strains is that they (1) maintain persistent control of aflatoxigenic fungi in the field, and (2) are incapable of out-crossing. Trackable field-released biocontrol strains will be beneficial to study the movement and longevity of non-aflatoxigenicA. flavus strains. Incorporating a naturally-occurring compound such as enhanced green fluorescent protein (eGFP) into a biocontrol strain might allow observation of its behaviour in field settings. The success of long-term field testing of eGFP-expressingA. flavus strains depends on their ability to maintain fluorescence throughout growth. Additionally, to ensure accurate tracking of the fluorescent atoxigenic strain, the likelihood of their out-crossing with individuals from the native population must be determined.In vitro mating experiments paired each of six different eGFP-transformed atoxigenic strains with a highly fertile toxigenicA. flavus isolate. Findings indicate that the eGFP gene, and possibly the aflatoxin cluster, is heritable by the F1 progeny. Not all cultured ascospores were fluorescent, but subsequent growth arising from a single fluorescent ascospore exhibited fluorescence similar to the eGFP parent. Observed mixed-fluorescence among conidia in a single chain suggests heterokaryosis at the moment of conidiogenesis. Mycotoxin assays showed that some fluorescent F1 individuals produce aflatoxin and/or cyclopiazonic acid which would indicate they are recombinant offspring. The findings in this laboratory study lend support to concern that atoxigenic strains are not impervious to genetic recombination and for which, if possible in a natural environment, repeated use could pose a risk of increasing the occurrence of aflatoxigenic individuals in treated fields.

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