〔Abstract〕 Objective To explore the role of Argonaute(Ago) gene and RNA-Dependent RNA Polymerase(RDRP) gene of Aspergillus flavus in the growth and development about the RNAi mechanism. Methods A. flavus Ago1, Ago2, RDRP1, RDRP3 gene mutant strains were constructed by homologous recombination. The growth and development of the mutant strains were observed on potato dextrose agar(PDA)+ uracil uridine(UU) medium inoculated with 3 μl 106CFU/mL spores. 200,400 μg cell wall pressure agent conidored(CR), 0.8 mol/L,1.6 mol/L osmotic pressure agent NaCl, 2 mmol/L,4 mmol/L oxidative pressure agent hydrogen peroxide(H2O2) and 0.01%,0.02% genomic damage agent methyl mesylate(MMS) were added to the Yeast extract Glucose Minimum(YGM)+UU medium to analyze the stress response of the mutant strains. Results A. flavus mutant strains about ΔAgo1, ΔAgo2, ΔRDRP1, ΔRDRP3 were successfully constructed and its growth and development were normal. The ΔAgo1 and ΔAgo2 strains reduced the stress effects on cell wall and osmotic pressure compared to the control. Ago1 gene deletion reduced the effect of H2O2, and conversely RDRP3 gene deletion increased the inhibition of H2O2. The Ago2 and RDRP1 strains reduced the effect on genetic damage agent. In addition, ΔRDRP1 increased the effect of osmotic stress. Conclusion The Ago1, Ago2, RDRP1 and RDRP3 genes of A. flavus are not involved in the regulation of growth rate and asexual reproduction and can participate in the regulating of the host stress response to the environment.