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Deletion of Msg5 and Yvh1 led to significant defects in conidiation, sclerotia formation, aflatoxin production and crop illness. Also, dual knockout mutants (ΔMsg5/ΔPtp1, ΔMsg5/ΔPtp2 and ΔMsg5/ΔOca2) displayed comparable defects to those observed in the ΔMsg5 single mutant, indicating that Msg5 plays a significant part within the legislation of development and pathogenicity in A. flavus. Notably, we found t