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Fusarium asiaticum, a dominant pathogen of Fusarium head blight (FHB) in East Asia, causes huge economic losses.Phenamacril, a novel cyanoacrylate fungicide, has been increasingly applying to control FHB in China, especially where resistance of F.asiaticum against carbendazim was severe.It is important to clarify the resistance related mechanisms of F.asiaticum to phenamacril so as to avoid control failures and to sustain the usefulness of the new product.In this study, a novel phenamacril-resistance related gene Famfs1 was obtained by employing cDNA random amplified polymorphic DNA (cDNA-RAPD) technique and was validated by genetic and biochemical assays.Compared with the corresponding progenitors, deletion of Famfs1 in the phenamacril-sensitive or-highly resistant strain caused significant decrease in effective concentrations inhibiting radial growth by 50% (EC50 value).Additionally, the biological fitness parameters (including mycelial growth under different stresses, conidiation, perithecia formation and virulence) of the deletion mutants attenuated significantly.Famfs1 was not only involved in the resistance of F.asiaticum to phenamacril, but also played important roles in adaptation of F.asiaticum to environment.Moreover, our data appear that cDNA-RAPD method can be a candidate technique to clone resistance related genes in fungi.