Activity and Resistance-Related Point Mutations in Target Protein PcORP1 of Fluoxapiprolin in <i>Phytophthora capsici</i>

Abstract

Fluoxapiprolin is a new piperidinyl thiazole isoxazoline fungicide developed by Bayer Crop Science in 2012, but the sensitivity and resistance mechanism of fluoxapiprolin are unclear. In this study, the sensitivities of 130 <i>Phytophthora capsici</i> isolates to fluoxapiprolin were determined, and a unimodal distribution was observed with a mean EC₅₀ value of 0.00043 μg/mL. Nine stable fluoxapiprolin-resistant mutants were obtained by fungicide adaption. The fitness of mutants was similar to or lower than that of the corresponding parental isolate. Seven heterozygous point mutations in the target protein PcORP1 were found in these mutants. These point mutations were confirmed in PsORP1 of <i>P. sojae</i> homologue positions using the CRISPR/Cas9 system. G770V and N835S+I877F do confer high fluoxapiprolin resistance (resistance factor, RF > 1000), and ΔN835, N767I, and N837T+S910C lead to low resistance (RF < 100). <i>P. sojae</i> transformants containing L733W, S768F, S768Y, Δ<i>G</i>818/F819, N837Y, N837F, P861H, L863W, and I877Y in PsORP1 were also associated with high fluoxapiprolin resistance (RF > 100). In conclusion, the resistance risk of <i>P. capsici</i> to fluoxapiprolin is moderate, and multiple point mutations in PcORP1 could confer different resistance levels to fluoxapiprolin.