Antifungal mechanism of isothiocyanates against Cochliobolus heterostrophus

Background: Isothiocyanates (ITCs) generated from the 'glucosinolates-myrosinase' defense system in the Brassicaceae exhibit broad antagonistic activity to various fungal pathogens. Nevertheless, the antifungal activity of ITCs to non-adapted fungi of Brassicaceae plants were seldom determined. The inhibitory effects of ITCs on Cochliobolus heterostrophus were evaluated and the antagonistic mechanism was explored.

Results: The mycelium growth of C. heterostrophus was hindered significantly by allyl, 4-(methylthio)-butyl, and phenyethyl ITCs, 4MTB-ITC exhibited the highest inhibitory effect on mycelium growth with an IC₅₀ value of 53.4 μmol L^-1 . In addition, ITCs exhibited obvious inhibitory effect on conidia germination and pathogenicity of C. heterostrophus. Proteomic analysis indicated that the inhibition of C. heterostrophus by A-ITC downregulated the expression of genes related to energy metabolism, oxidoreductase activity, melanin biosynthesis, and cell wall-degrading enzymes. Furthermore, mutants ΔChtrx2 and ΔChnox1 showed increased sensitivity to ITCs, and melanin biosynthesis was inhibited significantly in C. heterostrophus in response to A-ITC. Interestingly, unlike other pathogens that infected Brassicaceae plants, the SaxA in C. heterostrophus displayed no function in ITC degradation. In addition, the ITCs also exhibited obvious inhibitory effect on mycelium growth of Setosphaeria turcica, Fusarium graminearum, and Magnaporthe oryzae.

Conclusion: This study indicated that non-Brassicaceae-adapted pathogens are more sensitive to ITCs, and ITCs could have applications in protecting non-Brassicaceae crops in future. In addition, loss of ChNOX1 and ChTRX2 increased the sensitivity of C. heterostrophus to ITCs. Our results provided potential utilization of ITCs to control diseases caused by non-Brassicaceae pathogenic fungi. © 2022 Society of Chemical Industry.