Activity and Resistance-Related Point Mutations in Target Protein PcORP1 of Fluoxapiprolin in Phytophthora capsici
Activity and Resistance-Related Point Mutations in Target Protein PcORP1 of Fluoxapiprolin in Phytophthora capsici
复制标题
辣椒疫霉氟氧哌丙林靶蛋白 PcORP1 活性及抗性相关点突变
DOI：
10.1021/acs.jafc.0c05119
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发表时间：
2021-03-25
影响因子：
6.1
通讯作者：
Liu, Xili
中科院分区：
文献类型：
--
作者：
Miao, Jianqiang;Li, Chengcheng;Liu, Xili
氟唑吡脯林是拜耳作物科学公司于2012年开发的新型哌啶基噻唑类异恶唑啉杀菌剂，但其敏感性和抗性机制尚不清楚。在这项研究中，130个辣椒疫霉菌分离物对氟恶吡脯林的敏感性进行了测定，并观察到单峰分布，平均EC 50值为0.00043 μ g/mL。通过药剂驯化获得了9株稳定的氟恶吡啉抗性突变体。突变体的适合度与相应的亲本菌株相似或低于亲本菌株。在这些突变体中发现了靶蛋白PcORP 1的7个杂合点突变。使用CRISPR/Cas9系统在大豆疫霉同源物位置的PsORP 1中证实了这些点突变。G770 V和N835 S + I877 F确实赋予高的氟恶匹普林抗性（抗性因子，RF > 1000），并且Delta N835、N767 I和N837 T + S910 C导致低抗性（RF < 100）。在PsORP 1中含有L733 W、S768 F、S768 Y、Delta G818/F819、N837 Y、N837 F、P861 H、L 863 W和I877 Y的大豆疫霉转化体也与高氟吡脯氨酸抗性（RF > 100）相关。总之，辣椒疫霉对氟恶匹普林的耐药风险为中等，PcORP 1的多个点突变可赋予对氟恶匹普林不同的耐药水平。
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 Phytophthora capsici isolates to fluoxapiprolin were determined, and a unimodal distribution was observed with a mean EC50 value of 0.00043 mu g/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 P. sojae homologue positions using the CRISPR/Cas9 system. G770V and N835S+I877F do confer high fluoxapiprolin resistance (resistance factor, RF > 1000), and Delta N835, N767I, and N837T+S910C lead to low resistance (RF < 100). P. sojae transformants containing L733W, S768F, S768Y, Delta G818/F819, N837Y, N837F, P861H, L863W, and I877Y in PsORP1 were also associated with high fluoxapiprolin resistance (RF > 100). In conclusion, the resistance risk of P. capsici to fluoxapiprolin is moderate, and multiple point mutations in PcORP1 could confer different resistance levels to fluoxapiprolin.