Resistance Risk and Novel Resistance-Related Point Mutations in Target Protein PiORP1 of Fluoxapiprolin in Phytophthora infestans

Fluoxapiprolin is a new oxysterol binding protein inhibitor (OSBPI), which showed excellent inhibitory activity to plant pathogenic oomycetes. Its resistance risk and mechanism in Phytophthora infestans are unclear. In the current study, the sensitivities of 103 P. infestans isolates to fluoxapiprolin were investigated, and a unimodal distribution with a mean EC50 value of 0.00035 mu g/mL was observed. Four types of resistant mutants, with a resistance factor from 14 to more than 1000, and point mutations S768I+N837I, S768I+L860I, S768I, and I877F in PiORP1, were acquired using fungicide adaption. The fitness of the mutants was similar to or lower than that of the corresponding parental isolate. Positive cross-resistance was detected between fluoxapiprolin and oxathiapiprolin. The point mutations were verified in P. sojae homologue positions using the CRISPR/Cas9 genome editing system. Transformants containing S768I+N837I or S768I+L860I, showed high fluoxapiprolin resistance (RF > 1000). In conclusion, the risk of P. infestans resistance to fluoxapiprolin is moderate, and novel point mutation types S768I+N837I or S768I+L860I could cause high fluoxapiprolin resistance in P. infestans.

Automated summary

In this study, the sensitivity of Phytophthora infestans to fluoxapiprolin was investigated and a moderate resistance risk was observed. Four different types of resistant mutants were obtained using fungicide adaption. Point mutations in the PiORP1 gene were identified as the mechanism of resistance. Cross-resistance between fluoxapiprolin and oxathiapiprolin was detected. The point mutations were verified using the CRISPR/Cas9 genome editing system and two mutants showed high resistance to fluoxapiprolin.