Cyclobutrifluram is a novel succinate dehydrogenase inhibitor (SDHI) developed by Syngenta and helps to inhibit Fusarium pseudograminearum. Here, the potential for cyclobutrifluram resistance in F. pseudograminearum and the resistance mechanism involved were evaluated. Baseline sensitivity of F. pseudograminearum to cyclobutrifluram was determined with a mean EC 50 value of 0.0248 μg/mL. Fungicide adaption generated five resistant mutants, which possess a comparable or a slightly impaired fitness compared to corresponding parental isolates. This indicates that the resistance risk of F. pseudograminearum to cyclobutrifluram might be moderate. Cyclobutrifluram-resistant isolates also demonstrated resistance to pydiflumetofen but sensitivity to carbendazim, phenamacril, tebuconazole, fludioxonil, or pyraclostrobin. Additionally, point mutations H248Y in FpSdhB and A83V or R86K in FpSdhC 1 were found in cyclobutrifluram-resistant F. pseudograminearum mutants. Molecular docking and overexpression transformation assay revealed that FpSdhB H248Y and FpSdhC 1 A83V or FpSdhC 1 R86K confer the resistance of F. pseudograminearum to cyclobutrifluram.

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 EC 50 value of 0.00035 μg/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.

Background: Gray mold caused by Botrytis cinerea is an airborne plant pathogen with a necrotrophic lifestyle that infects more than 200 crops worldwide. Florylpicoxamid is a second-generation picolinamide fungicide inspired by a natural product. Florylpicoxamid targets the Qi site of the mitochondrial cytochrome bc1 complex and is currently being registered in China for the control of gray mold in a variety of crops. Although a broad spectrum of activity and attributes have been reported for florylpicoxamid, little is known about its effectiveness against gray mold or its protective and curative properties.Results: Florylpicoxamid exhibited substantial inhibitory activity against 12 tested species of plant-pathogenic fungi, with effective concentration for 50% growth inhibition (EC 50 ) values ranging from 0.017 to 2.096 ∼g ml −1 . A total of 129 isolates of B. cinerea from ten regions were tested for their sensitivity to florylpicoxamid, and the mean EC 50 value was 0.04 ± 0.017 ∼g ml −1 . Furthermore, florylpicoxamid was observed to substantially inhibit all developmental stages of B. cinerea, with mycelial development, sclerotium germination, germ tube elongation and conidial germination being restrained with an EC 50 value of 0.051 ± 0.0072, 0.012 ± 0.0069, 0.019 ± 0.0041 and 0.0062 ± 0.0007 ∼g ml −1 , respectively. No crossresistance was observed between florylpicoxamid and quinone outside inhibitor (QoI), methyl benzimidazole carbamates or succinate dehydrogenase inhibitor. Florylpicoxamid also exhibited protective and curative activity against the development of B. cinerea infection in tests on tomato fruits. At application rates of 90, 112.5 and 135 g a.i. ha −1 , florylpicoxamid was also observed to provide more-effective control than boscalid (300 g a.i. ha −1 ). Conclusion:This study demonstrated that the novel fungicide florylpicoxamid exhibits strong inhibitory activity against B. cinerea, regardless of the resistance profiles of those isolates to tested fungicides with different modes of action. This makes florylpicoxamid a powerful new solution to optimize gray mold control and manage fungicide resistance.

Background: Oxathiapiprolin, an oxysterol-binding protein inhibitor (OSBPI), shows unexceptionable inhibitory activity against plant pathogenic oomycetes. FRAC (Fungicide Resistance Action Committee) classifies it into the mode of action group F9 (lipid homeostasis and transfer/storage), but very little is known about the lipid metabolism of oomycete pathogens when subjected to oxathiapiprolin.Results: In this study, seven lipid categories and 1435 lipid molecules were identified in Phytophthora sojae, among which glycerolipids, glycerophospholipids, and sphingolipids account for 30.10%, 50.59%, and 7.28%, respectively. These lipids were categorized into 31 subclasses, which varied to different extents when treated with oxathiapiprolin. A total of 11 lipid subclasses showed significant changes. Among them, 10 lipid subclasses, lysophosphatidylcholine (LPC), lysophosphatidylethanolamine (LPE), phosphatidylcholine (PC), phosphatidylserine (PS), ceramide (Cer), triglyceride (TG), (o-acyl)-1-hydroxy fatty acid, diglycosylceramide, sphingoshine (So), and sitosterol ester, were significantly up-regulated, while digalactosyldiacylglycerol was the only lipid that was significantly down-regulated by a factor of almost three. These lipid molecules were further analyzed at the lipid species level. A total of 542 species were significantly altered when treated with oxathiapiprolin, including 212 glycerolipids [186 TG and 26 diglycerides (DG)], 167 glycerophospholipids (38 PC, 15 LPC, 19 LPE, seven PS, etc.), 156 sphingolipids (146 Cer, four So, etc.), and some other lipid molecules. Finally, from the orthogonal partial least-squares discrimination analysis model, variable importance for the projection score analysis showed that Cer, TG, and some glycerophospholipids contribute to the metabolic disorder when subjected to oxathiapiprolin. Conclusion: Glycerolipids, glycerophospholipids, and sphingolipids in P. sojae undergo significant changes with oxathiapiprolin treatment. These results provided valuable information for further understanding the function of the target protein and the mode of action of OSBPIs in oomycetes.