Quinone Outside Inhibitor Fungicides Research Articles

Isolates of Corynespora cassiicola, Causal Agent of Target Spot of Soybean, in Kentucky Contain the G143A Mutation Conferring Resistance to Quinone Outside Inhibitor Fungicides

Target spot, caused by Corynespora cassiicola, is a soybean disease of increasing importance in the southern United States. Recently, isolates of C. cassiicola with resistance to quinone outside inhibitor (QoI) fungicides have been confirmed with the G143A mutation in multiple southern states, including Alabama, Arkansas, Mississippi, and Tennessee. From 2017 to 2019, a total of 84 isolates of C. cassiicola were recovered from soybean field in 12 counties in Kentucky. DNA sequencing of the cytochrome b gene of these isolates revealed that 15.5% of the isolates had the G143A mutation that confers resistance to QoI fungicides and were in 50% of the counties in which isolates originated. This represents the first report of QoI fungicide resistance in C. cassiicola isolates from Kentucky soybean fields. Considering these findings, Kentucky soybean growers should adopt target spot management practices which include rotating to non-host crops, planting resistant soybean cultivars, and applying fungicides from different fungicide classes.

Characterization of QoI-Fungicide Resistance in Cercospora Isolates Associated with Cercospora Leaf Blight of Soybean from the Southern United States.

Cercospora leaf blight (CLB) of soybean, caused by Cercospora cf. flagellaris, C. kikuchii, and C. cf. sigesbeckiae, is an economically important disease in the southern United States. Cultivar resistance to CLB is inconsistent; therefore, fungicides in the quinone outside inhibitor (QoI) class have been relied on to manage the disease. Approximately 620 isolates from plants exhibiting CLB were collected between 2018 and 2021 from 19 locations in eight southern states. A novel polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay based on two genes, calmodulin and histone h3, was developed to differentiate between the dominant species of Cercospora, C. cf. flagellaris, and C. cf. sigesbeckiae. A multilocus phylogenetic analysis of actin, calmodulin, histone h3, ITS rDNA, and transcription elongation factor 1-α was used to confirm PCR-RFLP results and identify remaining isolates. Approximately 80% of the isolates collected were identified as C. cf. flagellaris, while 15% classified as C. cf. sigesbeckiae, 2% as C. kikuchii, and 3% as previously unreported Cercospora species associated with CLB in the United States. PCR-RFLP of cytochrome b (cytb) identified QoI-resistance conferred by the G143A substitution. Approximately 64 to 83% of isolates were determined to be QoI-resistant, and all contained the G143A substitution. Results of discriminatory dose assays using azoxystrobin (1 ppm) were 100% consistent with PCR-RFLP results. To our knowledge, this constitutes the first report of QoI resistance in CLB pathogen populations from Alabama, Arkansas, Kentucky, Mississippi, Missouri, Tennessee, and Texas. In areas where high frequencies of resistance have been identified, QoI fungicides should be avoided, and fungicide products with alternative modes-of-action should be utilized in the absence of CLB-resistant soybean cultivars.

Identification of Fungicide Combinations for Overcoming Plasmopara viticola and Botrytis cinerea Fungicide Resistance.

Fungal diseases, including downy mildew (caused by Plasmopara viticola) and gray mold (caused by Botrytis cinerea), significantly impact the marketable yield of grapes produced worldwide. Cytochrome b of the mitochondrial respiratory chain of these two fungi is a key target for Quinone outside inhibitor (QoI)-based fungicide development. Since the mode of action (MOA) of QoI fungicides is restricted to a single site, the extensive usage of these fungicides has resulted in fungicide resistance. The use of fungicide combinations with multiple targets is an effective way to counter and slow down the development of fungicide resistance. Due to the high cost of in planta trials, in silico techniques can be used for the rapid screening of potential fungicides. In this study, a combination of in silico simulations that include Schrödinger Glide docking, molecular dynamics, and Molecular Mechanism-Generalized Born Surface Area calculation were used to screen the most potent QoI and non-QoI-based fungicide combinations to wild-type, G143A-mutated, F129L-mutated, and double-mutated versions that had both G143A and F129L mutations of fungal cytochrome b. In silico docking studies indicated that mandestrobin, famoxadone, captan, and thiram have a high affinity toward WT cytochrome b of Botrytis cinerea. Although the QoIs mandestrobin and famoxadone were effective for WT based on in vitro results, they were not broadly effective against G143A-mutated isolates. Famoxadone was only effective against one isolate with G143A-mutated cytochrome b. The non-QoI fungicides thiram and captan were effective against both WT and isolates with G143A-mutated cytochrome b. Follow-up in silico docking and molecular dynamics studies suggested that fungicide combinations consisting of famoxadone, mandestrobin, fenamidone, and thiram should be considered in field testing targeting Plasmopara viticola and Botrytis cinerea fungicide resistance.

TaqMan-MGB PCR Method for Rapid Detection of QoI Fungicide Resistance in Chinese Populations of Plasmopara viticola.

Grape downy mildew caused by Plasmopara viticola is one of the most devastating diseases of grapevine worldwide. Quinone outside inhibitor (QoI) fungicides are commonly used for the control of the pathogen in grape fields across China. However, their recurrent use could lead to the emergence of resistance against these compounds. Based on the most common mutation in resistant isolates, a glycine to alanine substitution at amino acid position 143 (G143A) in the cytochrome b protein, a TaqMan-MGB PCR was developed for the rapid detection of resistance to the QoI fungicide azoxystrobin in P. viticola. Specificity and sensitivity of this method showed it could specifically detect the point mutations linked with QoI resistance in P. viticola, and the detection limit was 0.2 pg. It could also quantify the resistance allele even in isolate mixtures containing as little as 5% QoI-resistant P. viticola strains. With this method, a large P. viticola population (n = 2,373) was screened, and QoI-resistant isolates were identified for the first time in China. The average frequencies of the resistant genotype from eight major-grapevine regions were up to 66%. Taken together, the results not only provide a novel tool for the rapid distinction and quantification of the QoI-resistant allele in P. viticola but also provide important references for fungicide selection and application, which will facilitate resistance management of grape downy mildew and improve grape production systems in Chinese vineyards.

Defining Fungicide Resistance Mechanisms in the Corynespora cassiicola Population from Mississippi Soybean.

Target spot, caused by Corynespora cassiicola, is a common lower canopy soybean disease in the southern United States. Recently, target spot has resurged in importance especially following the identification of resistance to the quinone outside inhibitor (QoI) fungicides. As a result, a survey of C. cassiicola from soybean throughout Mississippi began in 2018. A total of 819 C. cassiicola monoconidial isolates were obtained from 228 fields in 75 counties. The molecular mechanism of QoI resistance was determined, which resulted from an amino acid substitution from glycine (G) to alanine (A) at position 143 using a PCR-RFLP method and comparing nucleotide sequences of the cytochrome b gene. Five previously defined geographic regions were used to present the distribution of the G143A substitution and included the Capital, Coast, Delta, Hills, and Pines. The Capital had the greatest proportion of G143A-containing isolates (95.0%), followed by the Coast (92.9%), Delta (89.8%), Pines (78.8%), and Hills (69.4%). In all, 85.8% of the C. cassiicola isolates carried the G143A substitution. In addition, the effective fungicide concentration (EC50) of randomly selected C. cassiicola isolates to azoxystrobin was used to characterize isolates as resistant (n = 14) (based on the presence of the G143A substitution and EC50 values >52 μg/ml) or sensitive (n = 11) (based on the absence of the G143A substitution and EC50 values <46 μg/ml). The EC50 values varied among isolates (P < 0.0001), with QoI-sensitive isolates exhibiting lower EC50 values than QoI-resistant isolates. The current study revealed that a reduction in sensitivity to QoI fungicides has likely resulted based on the percentage of C. cassiicola isolates containing the G143A substitution identified in Mississippi.

Evaluating in vitro fitness parameters of G143A-containing and wild-type Corynespora cassiicola isolates from Mississippi soybean.

Quinone outside inhibitor (QoI) fungicides have been widely used to manage diseases of soybean including target spot caused by Corynespora cassiicola. However, resistance to QoI fungicides has recently been reported within the C. cassiicola population from Alabama, Arkansas, Mississippi, and Tennessee as a result of isolates in the population containing the G143A amino acid substitution. Therefore, the relative fitness and stability of isolates containing the G143A substitution compared to wild-type C. cassiicola isolates from Mississippi soybean were investigated by analyzing several fitness parameters in vitro. In addition, in vivo virulence assays were conducted in the greenhouse on a target spot-susceptible cultivar. The evaluations of fitness considered the difference between isolates from the wild-type and G143A-containing genotypes by evaluating colony growth parameters following the first and the tenth subcultures on microbiological media. When considered as an average of all G143A-containing isolates, the G143A-containing isolates following the tenth subculture produced 6.2% greater colony diameter growth but produced 2.3% less conidia. Conversely, over the same period, wild-type isolates produced 6.7% less colony growth but produced 10.9% more conidia. Based on our results, the C. cassiicola isolates that contained the G143A substitution appear stable since successive subculturing did not significantly affect the measured fitness parameters. The lack of fitness cost accompanying the genotypic shift to the G143A amino acid substitution which confers fungicide resistance in C. cassiicola indicates that these isolates may have fitness advantages and may remain stable in the population as well as displace wild-type isolates with repeated fungicide applications of QoI-containing products.

Identification of Fungicide Combinations Targeting Plasmopara viticola and Botrytis cinerea Fungicide Resistance Using Machine Learning.

Downy mildew (caused by Plasmopara viticola) and gray mold (caused by Botrytis cinerea) are fungal diseases that significantly impact grape production globally. Cytochrome b plays a significant role in the mitochondrial respiratory chain of the two fungi that cause these diseases and is a key target for quinone outside inhibitor (QoI)-based fungicide development. Since the mode of action (MOA) of QoI fungicides is restricted to a single active site, the risk of developing resistance to these fungicides is deemed high. Consequently, using a combination of fungicides is considered an effective way to reduce the development of QoI resistance. Currently, there is little information available to help in the selection of appropriate fungicides. This study used a combination of in silico simulations and quantitative structure-activity relationship (QSAR) machine learning algorithms to screen the most potent QoI-based fungicide combinations for wild-type (WT) and the G143A mutation of fungal cytochrome b. Based on in silico studies, mandestrobin emerged as the top binder for both WT Plasmopara viticola and WT Botrytis cinerea cytochrome b. Famoxadone appeared to be a versatile binder for G143A-mutated cytochrome b of both Plasmopara viticola and Botrytis cinerea. Thiram emerged as a reasonable, low-risk non-QoI fungicide that works on WT and G143A-mutated versions of both fungi. QSAR analysis revealed fenpropidin, fenoxanil, and ethaboxam non-QoIs to have a high affinity for G143A-mutated cytochrome b of Plasmopara viticola and Botrytis cinerea. Above-QoI and non-QoI fungicides can be considered for field studies in a fungicide management program against Plasmopara viticola- and Botrytis cinerea-based fungal infections.

Effective Management of Citrus Melanose Based on Combination of Ecofriendly Chemicals.

Citrus melanose, caused by the ascomycete fungus Diaporthe citri, is one of the most important diseases in China that affects not only the production but also the quality of citrus. In China, mancozeb is recommended to control melanose disease at the dose of 1.34 g/liter. However, it is widely applied in practice at the dose of 2.66 g/liter or even 4 g/liter, because reduced efficacy of the recommended dose was observed in regions severely damaged by melanose. In this study, some ecofriendly chemicals for melanose management were evaluated. First, the sensitivity to fungicides was screened in the laboratory based on the inhibition of mycelial growth and conidial germination of D. citri. Results showed that both quinone outside inhibitor (QoI) fungicides kresoxim-methyl and trifloxystrobin inhibited conidial germination of D. citri up to 100% at 0.1 μg/ml. The in vivo control efficacy on detached fruit indicated that treatments with elastic nanocopolymer film at 2 g/liter, mancozeb at 1 g/liter, and kresoxim-methyl at 0.1 g/liter significantly inhibited the infection process compared with the control treatment of mineral oil alone. In field trials, the efficacy of kresoxim-methyl at 0.1 g/liter and elastic nanocopolymer film at 2 g/liter mixed with mancozeb at 1 g/liter was equal to that of mancozeb at 2.66 g/liter. The use of mancozeb could be reduced greatly, and the newly developed fungicide combinations are more environmentally friendly due to the low toxicity of both QoI fungicides and elastic nanocopolymer film. The newly developed method with ecofriendly chemicals should play an important role in the management of citrus melanose in the future.

Determining the Distribution of QoI Fungicide-Resistant Cercospora sojina on Soybean from Indiana.

Frogeye leaf spot (FLS) is a foliar disease of soybean (Glycine max) caused by Cercospora sojina. Application of fungicide products that contain quinone outside inhibitor (QoI) active ingredients has been one of the major tools used in the management of this disease, but, since 2010, QoI-resistant C. sojina isolates have been confirmed in over 20 states in the United States, including Indiana. In summer 2019 and 2020, 406 isolates of C. sojina were collected from 32 counties across Indiana and screened for QoI resistance using a PCR-restriction fragment length polymorphism (RFLP) method. An in vitro fungicide sensitivity test was also performed on a subset of isolates to evaluate their sensitivity to three QoI fungicides: azoxystrobin, pyraclostrobin, and picoxystrobin. A discriminatory dose of picoxystrobin was established as 1 µg/ml by testing five concentrations (0.001, 0.01, 0.1, 1, and 10 µg/ml). QoI-resistant isolates were found in 29 counties, and 251 of the 406 isolates (61.8%) were confirmed to be resistant to QoI fungicides based on PCR-RFLP results. Partial nucleotide sequences of the cytochrome b gene from four resistant and four sensitive isolates corroborated the presence and absence, respectively, of the G143A mutation. Results from the sensitivity assays with discriminatory doses of azoxystrobin (1 µg/ml) and pyraclostrobin (0.1 µg/ml) also supported the findings from the PCR-RFLP assay, because all QoI-resistant isolates were inhibited less than 50% relative to a no-fungicide control when exposed to these doses. Resistant isolates harboring the G143A mutation also exhibited resistance to picoxystrobin. The effective concentrations to inhibit mycelial growth by 50% relative to the nonamended control (EC50) in QoI-sensitive isolates ranged from 0.087 to 0.243 µg/ml, with an overall mean of 0.152 µg/ml, while EC50 values in QoI-resistant isolates were established as >10 µg/ml for picoxystrobin. Results from this study indicated that QoI-resistant C. sojina isolates are spread throughout Indiana and exhibit cross-resistance to QoI fungicides.

Foliar Fungicides Containing FRAC 11 Mitigate Phomopsis Stem Canker in Sunflower (Helianthus annuus).

Phomopsis stem canker reduces yield of sunflower (Helianthus annuus L.) up to or exceeding 40%; however, management recommendations have not been developed for U.S. farmers. Between 2009 and 2020, foliar fungicide trials were conducted in Minnesota, Nebraska, North Dakota, and South Dakota for a total of 49 location-years. Random effects meta-analyses were performed on the disease severity index (DSI) and yield data collected from the foliar fungicide trials to determine the overall and individual effectiveness of the tested fungicides. Effect sizes, Cohen's f or Hedges' g, were calculated as the difference in DSI or yield between the fungicide treatment and nontreated control (NTC) divided by the pooled SD. The pooled Cohen's f for DSI and yield was 0.40 (95% CI = [0.29, 0.42]), indicating a large effect size and that fungicide treatments had a significant effect on DSI and yield (P < 0.0001). Among the fungicide groups, quinone outside inhibitor (QoI) (DSI [k = 45; g = -0.47] and yield [k = 46; g = 0.41]) is moderately effective and premixes of demethylation inhibitors (DMI), succinate dehydrogenase inhibitors (SDHI), and QoI (DMI + SDHI + QoI) (DSI [k = 3; g = -0.79] and yield [k = 3; g = 0.94]) are largely effective in comparison with NTC. Upon performing prediction analyses, the probability of not recovering the fungicide application cost (Ploss) associated with QoI (pyraclostrobin) was <0.35 for a range of sunflower grain prices suggesting a greater probability of return on investment from a single application of fungicide. Overall, our study suggests that the use of QoI fungicides is likely to be profitable in the presence of Phomopsis stem canker (DSI > 5%).

Identification of Quinone Outside Inhibitor Fungicide-Resistant Isolates of Parastagonospora nodorum from Illinois and Kentucky.

Stagonospora leaf and glume blotch, caused by Parastagonospora nodorum, is a major disease of winter wheat (Triticum aestivum) in the United States capable of significantly reducing grain yield and quality. Pathogens such as P. nodorum that overwinter in crop residue are often an increased concern in cropping systems that utilize no-till farming. In addition, the lack of wheat cultivars with complete resistance to P. nodorum has led to the reliance on foliar fungicides for disease management. Quinone outside inhibitor (QoI) fungicides (Fungicide Resistance Action Committee group 11) are one of the major classes used to manage foliar diseases in wheat. Use of the QoI class of fungicides tends to select isolates of fungal pathogens with resistance due to mutations in the fungal cytochrome b gene. Isolates of P. nodorum were collected from Illinois in 2014 and Kentucky in 2018, 2019, and 2020. Amplification and sequencing of a segment of the cytochrome b gene from these isolates revealed a mutation at codon 143 that confers a change from glycine to alanine in the amino acid sequence (known as the G143A mutation). In vitro plate assays and greenhouse trials were used to confirm and characterize the QoI resistance caused by the G143A mutation. The frequency of the tested isolates with the G143A mutation was 46% (57 of 123 isolates) and 5% (3 of 60 isolates) for Kentucky and Illinois, respectively. This research is the first to identify the G143A mutation in P. nodorum isolates with resistance to QoI fungicides in Illinois and Kentucky.

The Toxicity of Salicylhydroxamic Acid and Its Effect on the Sensitivity of Ustilaginoidea virens to Azoxystrobin and Pyraclostrobin.

Rice false smut (RFS) caused by Ustilaginoidea virens has been one of the most severe rice diseases. Fungicide-based chemical control is a significant measure to control RFS. In the sensitivity determination of quinone outside inhibitor (QoI) fungicide in vitro, salicylhydroxamic acid (SHAM) has been commonly added to artificial culture media in order to inhibit alternative oxidase of phytopathogenic fungi. However, some studies showed that artificial media should not include SHAM due to its toxicity. Whether SHAM should be added in the assay of U. virens sensitivity to QoI fungicide remains unknown. In this study, two appropriate media, potato sucrose agar (PSA) and minimal medium (MM), were selected to test SHAM toxicity and sensitivity of U. virens to azoxystrobin and pyraclostrobin. The mycelial growth and sensitivity to azoxystrobin and pyraclostrobin had no significant difference between on PSA and MM. SHAM could significantly inhibit mycelial growth, conidial germination, peroxidase (POD) and esterase activity of U. virens. Average effective concentration for inhibiting 50% (EC50) values of SHAM against mycelial growth of ten U. virens were 27.41 and 12.75 μg/mL on PSA and MM, respectively. The EC50 values of SHAM against conidial germination of isolates HWD and JS60 were 70.36 and 44.69 μg/mL, respectively. SHAM at 30 μg/mL significantly inhibited POD and esterase activity of isolates HWD and JS60, and even SHAM at 10 μg/mL significantly inhibited POD activity of isolate HWD. In addition, SHAM significantly reduced EC50 values and EC90 values of azoxystrobin and pyraclostrobin on both PSA and MM. Even in the presence of SHAM at 10 μg/mL, average EC50 values of ten U. virens isolates for azoxystrobin decreased 1.7-fold on PSA and 4.8-fold on MM, and for pyraclostrobin that decreased 2.8-fold on PSA and 4.8-fold on MM. Therefore, these results suggest that SHAM should not be included in artificial media in the assay of U. virens sensitivity to QoI fungicides.

Baseline Sensitivity and Control Efficacy of Two Quinone Outside Inhibitor Fungicides, Azoxystrobin and Pyraclostrobin, Against Ustilaginoidea virens.

Rice false smut caused by the filamentous fungus Ustilaginoidea virens is a devastating grain disease in rice. Fungicides have been an important measure for the control of this disease. In this study, baseline sensitivities of 179 isolates of U. virens to the quinone outside inhibitor (QoI) fungicides azoxystrobin and pyraclostrobin were established. The distribution of the 50% effective concentration (EC50) values of each fungicide was unimodal. The frequency distribution of logarithmically transformed EC50 values fit or fit closer to a normal distribution. The ranges of EC50 values for azoxystrobin and pyraclostrobin were 0.001 to 0.864 and 0.001 to 0.569 μg/ml, with means and standard errors of the mean values of 0.203 ± 0.012 and 0.079 ± 0.006 μg/ml, respectively. There was a statistically significant and moderately positive correlation (n = 100, r = 0.469, P = 0.001) in sensitivity between these two fungicides. No cross-resistance was found between azoxystrobin, pyraclostrobin, and carbendazim or sterol demethylation inhibitor fungicides. Each fungicide had a significantly higher mean preventive efficacy compared with its curative efficacy. Field assays showed that the control efficacy of pyraclostrobin against rice false smut was greater than that of azoxystrobin. Pyraclostrobin had the best control of rice false smut in three rice varieties, with the control efficacy ranging from 81.5 to 95.5%, whereas azoxystrobin decreased the disease index by 64.1 to 69.2% under the same conditions. These results provide us a reference point in the management of U. virens and future QoI fungicide resistance monitoring programs.

Identification of Septoria glycines Isolates from Soybean with Resistance to Quinone Outside Inhibitor Fungicides.

Brown spot, caused by Septoria glycines, is a common foliar disease of soybean (Glycine max). Applications of fungicide products that contain quinone outside inhibitor (QoI) active ingredients to soybean fields have contributed to the selection and development of QoI-resistant populations of S. glycines. We investigated the molecular mechanisms of QoI-resistance in these populations through targeted analysis of the cytochrome b gene. Isolates of S. glycines collected from several soybean fields over different seasons varied in sensitivity to QoI fungicides. Characterization of the cytochrome b gene revealed a mutation that changed an amino acid from glycine to alanine at codon 143 - one that is generally associated with QoI fungicide resistances. A PCR assay was developed that allowed successful discrimination of QoI-sensitive and -resistant isolates based on the G143A mutation. Results of this study demonstrated that 47.5% of S. glycines isolates tested were resistant to QoI fungicides. Accurate monitoring of this mutation will help slow the spread of QoI resistance and will be important for fungicide resistant management in this pathosystem.

Metyltetraprole activity against plant pathogens with relatively rare cytochrome b haplotypes for azoxystrobin resistance

Metyltetraprole is a novel quinone outside inhibitor (QoI) fungicide designed to avoid cross-resistance in cytochrome b G143A-harboring QoI-resistant phytopathogenic fungi. The resistance factors of G143A-harboring fungal isolates for metyltetraprole are around 1, but > 200 for the reference QoI fungicide azoxystrobin. In this study of metyltetraprole activities against azoxystrobin-resistant isolates carrying G137R, G137S, L299F, N256S + L299F, or L275F + L299F in cytochrome b, metyltetraprole had potent activity against all isolates with these cytochrome b haplotypes. The resistance factors ranged from 0.7 to 2.9 for metyltetraprole and from 3.0 to 175.1 for azoxystrobin. We revealed unique metyltetraprole inhibitory activities against QoI-resistant plant pathogens.

Sensitivity of Colletotrichum acutatum Species Complex from Strawberry to Fungicide Alternatives to Quinone-Outside Inhibitors.

Colletotrichum acutatum is a species complex that causes anthracnose fruit rot and root necrosis on strawberry. The major and minor species within the complex that affect strawberry production are C. nymphaeae and C. fioriniae, respectively. The disease can significantly reduce yield under conducive weather, and its management has greatly relied on quinone-outside inhibitor fungicides (QoI). However, due to the emergence of resistant isolates, such products are no longer effective. Therefore, alternative fungicides were investigated. C. nymphaeae and C. fioriniae isolates were collected from multiple strawberry fields in the United States from 1995 to 2017. The sensitivity of benzovindiflupyr, penthiopyrad, pydiflumetofen, fluazinam, fludioxonil, and cyprodinil was assessed by invitro and invivo assays. Both Colletotrichum species were sensitive to benzovindiflupyr, penthiopyrad, fluazinam, and fludioxonil based on mycelial growth assays. Interestingly, of these products, only penthiopyrad did not inhibit conidial germination at 100 µg/ml. For cyprodinil, C. nymphaeae was sensitive based on the mycelial growth, whereas C. fioriniae was not inhibited. Neither species was inhibited by pydiflumetofen in mycelial growth, conidial germination, nor detached fruit assays. The prepackaged mixtures fludioxonil + cyprodinil and fludioxonil + pydiflumetofen were effective in a field trial; however, their use should be carefully considered because of the lack of efficacy of one of the compounds in the mixture. This study sheds light on the potential registration of products alternative to QoIs, such as benzovindiflupyr and fluazinam, which could improve the management of strawberry anthracnose.

Evaluation of the risk of development of resistance to QoI fungicide ZJ0712 in Podosphaera xanthii under greenhouse conditions

Greenhouse-grown cucumbers were monitored over two successive growing seasons to investigate the effects of successive application of ZJ0712, a new quinone outside inhibitor (QoI) fungicide, on the development of resistance in cucumber powdery mildew (Podosphaera xanthii). Resistant P. xanthii isolates were detected after nine successive applications of ZJ0712, although the control efficacy of this fungicide against cucumber powdery mildew at that time was still higher than 80%. Seven ZJ0712-resistant P. xanthii isolates with resistance factor values greater than 180 were obtained, which exhibited a stable resistance to ZJ0712. These resistant P. xanthii isolates had similar pathogenicity to the wild-type isolate on cucumber plants. The ZJ0712 showed significant cross-resistance with azoxystrobin, enostrobilurin, or chlorothalonil, but not with the azole fungicide triadimefon. Furthermore, the most commonly reported G143A mutation in Cyt b associated with QoI resistance was found in five of the seven resistant isolates. These findings suggest that there is a high risk of resistance development associated with using ZJ0712 for controlling cucumber powdery mildew under greenhouse conditions, and the underlying resistance mechanisms in different P. xanthii isolates are not consistent and need to be further unraveled.

A strategy for controlling Cercospora leaf spot, caused by Cercospora beticola, by combining induced host resistance and chemical pathogen control and its implications for sugar beet yield

Cercospora beticola populations resistant to methyl benzimidazole carbamate (MBC), quinone outside inhibitor (QoI), and demethylation inhibitor (DMI) fungicides have been reported in Egypt. This pathogen causes a difficult to control variant of Cercospora leaf spot (CLS) on sugar beet. We conducted field experiments over two years to investigate treatment efficacies on MBC- and QoI-resistant, but partially DMI-sensitive, C. beticola from a research farm in Sakha, Egypt. DMI fungicides showed moderate efficacy with half the maximal effective concentration (EC50). Efficacy was 67.2–69.1% and 63.4–63.6% for epoxiconazole (EPO) and propiconazole (PRO), respectively. When each fungicide was mixed with salicylic acid (SA) at EC50 (EPO + SA) and EC50 (PRO +SA), efficacy was 77.5–79.1% and 77.0–78.2%, respectively. The QoI fungicide, azoxystrobin, showed lower efficacy (46.4–49.5%) when applied alone, but enhanced efficacy when combined with SA at EC50/EC50 (77.0–78.2%). Carbendazim alone was only 47.5–45.1% effective, but when mixed with SA, had a 67.1% co-toxicity factor in vitro, indicating antagonism between components. An Artemisia cina aqueous extract (WEA) combined with SA (WEA/SA) showed 76.0–78.2% efficacy, compared with 69.0–71.0% and 49.0–52.7% for WEA and SA, respectively. Single treatments were classified into five sensitivity classes from sensitive (<1) to resistant (>100) based on their EC50 values. Significant increases in insensitivity were observed from 2015/16 to 2017/18. All treatments increased total yield, sucrose percentage, and recoverable white sugar compared with the untreated control. A combination of control strategies could allow for effective disease and fungicide resistance management.

First Detection of Frogeye Leaf Spot in Soybean Fields in North Dakota and the G143A Mutation in the Cytochrome b Gene of Cercospora sojina

Frogeye leaf spot, caused by Cercospora sojina, is an important foliar disease of soybean (Glycine max) in the United States. Application of quinone outside inhibitor (QoI) fungicides has been an important tool available to farmers to help manage this disease, but in 2010, C. sojina isolates with resistance to QoI fungicides were first discovered in Tennessee and then additional states in the years to follow. During the 2020 growing season, C. sojina isolates collected from North Dakota soybean fields were tested for QoI resistance using laboratory and molecular assays. The results of these assays showed that QoI fungicide-resistant C. sojina isolates are present in North Dakota. Similar to previous findings in other states, these QoI-resistant C. sojina isolates contain the G143A mutation. Soybean farmers in North Dakota will need to use an integrated approach of cultural practices, genetic resistance, and fungicides with multiple modes of action to manage this disease in light of QoI-resistant C. sojina isolates being present in the state.

The G143A Mutation in the Cytochrome b Gene Is Associated with Quinone Outside Inhibitor Fungicide Resistance in Cercospora sojina from Soybean Fields in Wisconsin

Frogeye leaf spot, caused by Cercospora sojina, is an important foliar disease of soybean (Glycine max) in the United States. Application of quinone outside inhibitor (QoI) fungicides has been an important management tool available to farmers to help manage this disease, but in 2010, C. sojina isolates with resistance to QoI fungicides were first discovered in Tennessee and then additional states in the years to follow. During the 2020 growing season, C. sojina isolates collected from Wisconsin soybean fields were tested for QoI resistance using laboratory and molecular assays. The results of these assays showed that QoI fungicide-resistant C. sojina isolates are present in Wisconsin. Similar to previous findings in other states, these QoI-resistant C. sojina isolates contain the G143A mutation. Soybean farmers in Wisconsin will need to use an integrated approach of cultural practices, genetic resistance, and using fungicides with multiple modes of action to manage this disease in light of QoI-resistant C. sojina isolates being present in the state.