Quinone Outside Inhibitor Fungicides Research Articles

Soybean Fungal Endophytes Alternaria and Diaporthe spp. are Differentially Impacted by Fungicide Application.

In field trials in Iowa, we investigated the association of a fungicide applied at early pod set to the diversity and composition of foliar endophytic fungi in presenescent soybeans. The main purpose of our study was to determine whether fungicides affect the microbiome of soybean plants during the pod-fill reproductive stage. In a replicated experiment focused on the impact of a fungicide application including a quinone outside inhibitor (QoI) and a pyrazole-carboxamide spanning two growing seasons, healthy stems and leaves near the tops of soybean were sampled for endophytic fungi. The survey yielded 1,791 isolates belonging to 17 putative species, identified by morphology and sequence analysis of the ribosomal DNA internal transcribed spacer region. Taxa were grouped by genus into operational taxonomic units: Alternaria, Colletotrichum, and Diaporthe were the dominant genera isolated. Plant parts were analyzed separately using a multivariate community analysis of isolate counts per plant. The 14.3% fluxapyroxad and 28.6% pyraclostrobin fungicide spray significantly increased the proportion of Diaporthe isolates over no-spray controls, whereas the inverse occurred for foliar Alternaria isolates. In addition, seed harvested from fields with shorter-season varieties and sprayed with fungicide showed higher percentages of Diaporthe isolates than fields with no fungicide spray. In conclusion, soybean farmers may want to consider that the application of a QoI fungicide in the absence of disease pressure might adversely impact seed quality.

Emergence of Stemphylium Leaf Blight of Onion in New York Associated With Fungicide Resistance.

A complex of foliar diseases affects onion production in New York, including Botrytis leaf blight (Botrytis squamosa), purple blotch (Alternaria porri), Stemphylium leaf blight (SLB; Stemphylium vesicarium), and downy mildew (Peronospora destructor). Surveys were conducted in 2015 and 2016 to evaluate the cause of severe premature foliar dieback in New York onion fields. SLB was the most prevalent disease among fields with the greatest incidence, surpassing downy mildew, purple blotch, and Botrytis leaf blight. Sequencing of the internal transcribed spacer region of ribosomal DNA and the glyceraldedyhe-3-phosphate dehydrogenase and calmodulin genes identified S. vesicarium as the species most commonly associated with SLB. S. vesicarium was typically associated with a broad range of necrotic symptoms but, most commonly, dieback of leaf tips and asymmetric lesions that often extended over the entire leaf. Because of the intensive use of fungicides for foliar disease control in onion crops in New York, the sensitivity of S. vesicarium populations to various fungicides with site-specific modes of action was evaluated. Sensitivity of S. vesicarium isolates collected in 2016 to the quinone outside inhibitor (QoI) fungicide, azoxystrobin, was tested using a conidial germination assay. Isolates representing a broad range of QoI sensitivities were selected for sequencing of the cytochrome b gene to evaluate the presence of point mutations associated with insensitivity to azoxystrobin. The G143A mutation was detected in all 74 S. vesicarium isolates with an azoxystrobin-insensitive phenotype (effective concentrations reducing conidial germination by 50%, EC50 = 0.2 to 46.7 µg of active ingredient [a.i.]/ml) and was not detected in all 31 isolates with an azoxystrobin-sensitive phenotype (EC50 = 0.01 to 0.16 µg a.i./ml). The G143A mutation was also associated with insensitivity to another QoI fungicide, pyraclostrobin. Sensitivity to other selected fungicides commonly used in onion production in New York was evaluated using a mycelial growth assay and identified isolates with insensitivity to boscalid, cyprodinil, and pyrimethanil, but not difenoconazole. The frequency of isolates sensitive to iprodione, fluxapyroxad, and fluopyram was high (93.5 to 93.6%). This article discusses the emergence of SLB as dominant in the foliar disease complex affecting onion in New York and the complexities of management posed by resistance to fungicides with different modes of action.

Fungicide Sensitivity Monitoring of Alternaria spp. Causing Leaf Spot of Sugarbeet (Beta vulgaris) in the Upper Great Lakes.

Alternaria leaf spot (ALS), caused by Alternaria spp., can occur wherever sugarbeet is grown. Infection by Alternaria spp. and disease management has historically been considered a minor issue in sugarbeet production in the United States. An increase of both incidence and severity in 2016 of ALS high enough to cause yield loss has been observed in Michigan. With a renewed need to consider potential management of this disease, the sensitivity was determined for populations of Alternaria spp. to three classes of fungicides currently labeled for management of leaf spot on sugarbeet, including demethylase inhibitor (DMI), quinone outside inhibitor (QoI), and organo-tin fungicides. Leaves with symptoms of ALS were sampled from sugarbeet fields in east-central Michigan and southwestern Ontario, Canada. Monoconidial isolates were obtained to determine sensitivity to each fungicide class above. A spiral gradient dilution method was used to estimate the fungicide effective concentration (in milligrams per liter) that caused a 50% inhibition of fungal growth in vitro for all isolates. Significant temporal shifts were detected in the frequencies of sensitivity phenotypes to DMI and QoI but not organo-tin fungicides from 2016 through 2017. Individual isolates of Alternaria spp. were recovered with cross-resistance to DMI and multiple resistance to DMI, QoI, and triphenyltin hydroxide fungicides. To our knowledge, this is the first report of a fungus other than Cercospora beticola with resistance to organo-tin fungicides. Fungicide sensitivity monitoring indicates that an effective integrated disease management approach combining fungicide efficacy trials and monitoring pathogen biology is essential for developing effective resistance management recommendations.

Distribution and Stability of Quinone Outside Inhibitor Fungicide Resistance in Populations of Potato Pathogenic Alternaria spp. in Wisconsin.

Quinone outside inhibitor (QoI) fungicides have been an important class in managing potato early blight caused by Alternaria solani and brown spot caused by A. alternata. Because of the single-site mode of action character of QoI fungicides, which are relied on for management of diseases in Wisconsin, and the abundant asexual conidia production of the Alternaria species, pathogen isolates with QoI resistance have been detected after just a few years of QoI fungicide usage in commercial production fields. Resistance to QoIs has been attributed to amino acid substitutions F129L and G143A in cytochrome b of A. solani and A. alternata, respectively, as a result of point mutations. The aim of this study was to assess Alternaria populations in Wisconsin for QoI resistance before and after fungicide applications in order to evaluate resistance stability. A TaqMan single nucleotide polymorphism genotyping assay was designed based on the sequences of the cytochrome b gene from Alternaria isolates collected in Wisconsin to profile QoI resistance in Alternaria populations as well as to explore factors that may influence frequency of QoI resistance in the pathogen populations. This assay successfully identified the mutations conferring QoI resistance in isolates collected from four locations each year from 2015 to 2017. During the course of this study, the frequency of A. solani isolates with the F129L mutation was consistently high and showed primarily the TTA mutation type. The frequency of A. alternata isolates with the G143A mutation started relatively low and increased at the end of the production season in each year (P = 0.0109, P = 0.2083, and P = 0.0159). A potato field managed without use of QoI fungicides showed a significantly lower (P < 0.05) frequency of A. alternata isolates carrying G143A than conventionally managed potato fields. The overall frequency of A. alternata isolates carrying G143A in the four locations was similar over the 3 years (P = 0.2971). The QoI resistance characteristics of the isolates were stable even when QoI selection pressure was removed for at least five subculture transfers, and the mutation types of codons 129 and 143 in the cytochrome b gene in A. solani and A. alternata, respectively, remained the same. This indicated that the application of QoIs in the field is not the sole factor responsible for the variation of the frequency of QoI resistance in the pathogen populations.

Prevalence of QoI resistance and mtDNA diversity in the Irish Zymoseptoria tritici population

Abstract The emergence and spread of Quinone outside Inhibitor (QoI) fungicide resistance in the Irish Zymoseptoria tritici population in the early 2000s had immediate impacts on the efficacy of the entire group of fungicides for the control of septoria tritici blotch. As a result, a dramatic reduction in the quantities applied to winter wheat occurred in the following seasons. Even in the absence of these fungicides, the frequency of the resistance allele, G143A in the pathogens mtDNA has remained exceptionally high (&gt;97%), and as such, it can be anticipated that continued poor efficacy of current QoI fungicides will be observed. Amongst the isolates with G143A, differences in sensitivity to the QoI pyraclostrobin were observed in vitro. The addition of the alternative oxidase (AOX) inhibitor salicylhydroxamic acid increased sensitivity in these isolates, suggesting some continued impairment of respiration by the QoI fungicides, albeit weak. Interestingly, amongst those tested, the strains from a site with a high frequency of inserts in the MFS1 transporter gene known to enhance QoI efflux did not exhibit this increase in sensitivity. A total of 19 mtDNA haplotypes were detected amongst the 2017 strain collection. Phylogenetic analysis confirmed the suggestion of a common ancestry of all the haplotypes, even though three of the haplotypes contained at least one sensitive strain.

Effects of SHAM on the Sensitivity of Sclerotinia sclerotiorum and Botrytis cinerea to QoI Fungicides.

It is a common practice to add salicylhydroxamic acid (SHAM) into artificial medium in the in vitro sensitivity assay of fungal phytopathogens to the quinone outside inhibitor (QoI) fungicides. The rationale for adding SHAM is to inhibit fungal alternative oxidase, which is presumed to be inhibited by secondary metabolites of plants. Therefore, the ideal characteristics of SHAM should be almost nontoxic to phytopathogens and have no significant effect on control efficacy of fungicides. However, this study showed that the average effective concentration for 50% inhibition (EC50) of mycelial growth values of SHAM were 97.5 and 401.4 μg/ml for Sclerotinia sclerotiorum and Botrytis cinerea, respectively. EC50 values of the three QoI fungicides azoxystrobin, kresoxim-methyl, and trifloxystrobin in the presence of SHAM at 20 and 80 μg/ml for S. sclerotiorum and B. cinerea, respectively, declined by 52.7 to 78.1% compared with those without SHAM. For the dicarboximide fungicide dimethachlone, the average EC50 values in the presence of SHAM declined by 18.2% (P = 0.008) for S. sclerotiorum and 35.9% (P = 0.012) for B. cinerea. Pot experiments showed that SHAM increased control efficacy of the three QoI fungicides against the two pathogens by 43 to 83%. For dimethachlone, SHAM increased control efficacy by 134% for S. sclerotiorum and 86% for B. cinerea. Biochemical studies showed that SHAM significantly inhibited peroxidase activity (P = 0.024) of B. cinerea and esterase activity (P = 0.015) of S. sclerotiorum. The strong inhibitions of SHAM per se on mycelial growth of B. cinerea and S. sclerotiorum and significant influences on the sensitivity of the two pathogens to both the QoI fungicides and dimethachlone as well as inhibitions on peroxidase and esterase indicate that SHAM should not be added in the in vitro assay of sensitivity to the QoI fungicides.

Sensitivity of Bipolaris oryzae Isolates Pathogenic on Cultivated Wild Rice to the Quinone Outside Inhibitor Azoxystrobin

The occurrence of fungal brown spot, caused by Bipolaris oryzae, has increased in cultivated wild rice (Zizania palustris) paddies in spite of the use of azoxystrobin-based fungicides. The active ingredient blocks electron transfer at the quinone outside inhibitor (QoI) site in the mitochondrial cytochrome b within the bc1 complex, thus obstructing respiration. The in vitro averaged EC50 of baseline isolates collected in 2007 before widespread fungicide use was estimated to be 0.394 µg/ml with PROBIT and 0.427 µg/ml with linear regression analyses. Isolates collected during 2008, 2015, and 2016 had a range of sensitivity as measured by relative spore germination (RG) at a discriminatory dose of 0.4 µg/ml azoxystrobin. Isolates with a higher (≥80%) and lower RG (≤40%) had the wild type nucleotides at amino acid positions F129, G137, and G143 of cytochrome b, sites known to be associated with QoI fungicide resistance. Two Group I introns were found in the QoI target area. The splicing site for the second intron was found immediately after the codon for G143. A mutation for fungicide resistance at this location would hinder splicing and severely reduce fitness. B. oryzae expresses an alternative oxidase in vitro, which allows the fungus to survive inhibition of respiration by azoxystrobin. This research indicates that B. oryzae has not developed resistance to QoI fungicides, although monitoring for changes in sensitivity should be continued. Judicious use of QoI fungicides within an integrated disease management system will promote an effective and environmentally sound control of the pathogen in wild rice paddies.

Quantifying the Effects of a G137S Substitution in the Cytochrome bc1 of Venturia effusa on Azoxystrobin Sensitivity Using a Detached Leaf Assay.

The quinone outside inhibitor (QoI) fungicides are known for their inherently high resistance risk owing to substitutions in amino acid residues 129, 137, or 143 of the cytochrome b gene of phytopathogens. In Venturia effusa, cause of pecan scab, an intron adjacent to position 143 likely reduces this risk; however, the effects of a recently discovered substitution at position 137 (G137S) are unknown. Traditional in vitro assays are not useful for determining sensitivity of isolates of V. effusa to the QoI fungicides, owing to the fungitoxic effects of required alternative oxidase inhibitors. A detached leaf assay was developed to quantify the sensitivity of 59 isolates to azoxystrobin: 45 wild-type isolates and 14 carrying G137S. Isolate EC50 values ranged from <0.0001 to 0.3047 µg/ml; EC50 values for wild-type isolates ranged from <0.0001 to 0.2007 µg/ml (median 0.0023 µg/ml), whereas EC50 values for G137S isolates ranged from 0.0033 to 0.3047 µg/ml (median 0.0178 µg/ml). The median EC50 value for G137S isolates was significantly greater than that of the wild-type isolates; however, there was overlap between the two groups. This is the first report of sensitivity of V. effusa isolates to a QoI fungicide and evidence of G137S as a potential mechanism of partial resistance. However, although a complete control failure is unlikely, the impact of this substitution on QoI efficacy in Georgia pecan orchards remains to be determined.

Effects of fungicide applications on root-infecting microorganisms and overwintering survival of perennial stevia

Koehler, A.M. and Shew, H.D. 2018. Effects of fungicide applications on root-infecting microorganisms and overwintering survival of perennial stevia. Crop Protection 117:000–000.Stevia (Stevia rebaudiana) is a perennial species emerging as a new crop in the southeastern United States. In previous studies, application of quinone outside inhibitor (QoI) fungicides enhanced overwintering survival of stevia. Greenhouse and field trials were conducted to assess the effect of multiple fungicides on plant growth, root associated microorganisms, and overwintering survival. In greenhouse trials in the absence of soilborne pathogens, no differences in plant height, shoot weight, or root system ratings were observed among treatments. In two field trials screening seven combination fungicides and one biological control, significant increases in overwintering survival of plants treated with QoI fungicides were observed. Plants were dug from the fields monthly to destructively sample root crowns and species of Fusarium, Ceratobasidium, Pythium, and Macrophomina phaseolina were consistently isolated. Abundant microsclerotia from M. phaseolina were observed on stems of plants that did not survive winter. In seven sample months at each of the two field trials, non-fungicide treated plants had the lowest root weights. This study is one of the first to evaluate interactions between fungicide use and root associated fungi in a perennial crop and provides a framework to further investigate the role of root-associated fungi in overwintering survival.

Baseline sensitivity of Cercospora zeae-maydis to pydiflumetofen, a new succinate dehydrogenase inhibitor fungicide

Gray leaf spot, caused by the fungus Cercospora zeae-maydis, is an important foliar disease of corn (Zea mays) in the United States (U.S.). Application of foliar products that contain quinone outside inhibitor (QoI) and demethylation inhibitor (DMI) fungicides is one way that corn farmers in the U.S. manage this disease. Since DMI and QoI fungicides pose medium to high risks of selecting for resistant fungal pathogens, it is important that fungicides from different classes be registered for use on corn to manage gray leaf spot. The fungicide pydiflumetofen (adepidyn; Syngenta Crop Protection, Greensboro, NC) is a new succinate dehydrogenase inhibitor (SDHI) that currently is being registered for use on corn in the U.S. The objective of this study was to establish the baseline sensitivity of C. zeae-maydis isolates collected from different areas of the U.S. to pydiflumetofen using in vitro assays. Effective concentrations of pydiflumetofen in which C. zeae-maydis isolates were inhibited by 50% (EC50 values) ranged from 0.0008 to 0.0101 μg/ml, with mean and median values of 0.0040 and 0.0038 μg/ml, respectively. These results can be used as part of a fungicide resistance monitoring program for C. zeae-maydis in the U.S.

Resistance to Quinone Outside Inhibitor Fungicides Conferred by the G143A Mutation in Cercospora sojina (Causal Agent of Frogeye Leaf Spot) Isolates from South Dakota Soybean Fields

Resistance to quinone outside inhibitor (QoI) fungicides was detected in Cercospora sojina (causal agent of frogeye leaf spot) isolates collected from soybean (Glycine max) fields in four South Dakota counties during the 2018 growing season. A discriminatory dose assay was used to detect QoI-resistant isolates, and a follow-up polymerase chain reaction assay was used to determine the presence of the G143A mutation in QoI-resistant isolates. This is the first report of resistance to QoI fungicides in C. sojina isolates from South Dakota.

Meta-Analysis of the Effects of QoI and DMI Fungicide Combinations on Fusarium Head Blight and Deoxynivalenol in Wheat.

Field trials were conducted in 17 U.S. states to evaluate the effects of quinone outside inhibitor (QoI) and demethylation inhibitor (DMI) fungicide programs on Fusarium head blight index (IND) and deoxynivalenol (DON) toxin in wheat. Four DMI-only treatments applied at Feekes 10.5.1, five QoI-only treatments applied between Feekes 9 or Feekes 10.5, three QoI+DMI mixtures applied at Feekes 10.5, and three treatments consisting of a QoI at Feekes 9 followed by a DMI at Feekes 10.5.1 were evaluated. Network meta-analytical models were fitted to log-transformed mean IND and DON data and estimated contrasts of log means were used to obtain estimates of mean percent controls relative to the nontreated check as measures of efficacy. Results from the meta-analyses were also used to assess the risk of DON increase in future trials. DMI at Feekes 10.5.1 were the most effective programs against IND and DON and the least likely to increase DON in future trials. QoI-only programs increased mean DON over the nontreated checks and were the most likely to do so in future trials, particularly when applied at Feekes 10.5. The effects of QoI+DMI combinations depended on the active ingredients and whether the two were applied as a mixture at heading or sequentially. Following a Feekes 9 QoI application with a Feekes 10.5.1 application of a DMI reduced the negative effect of the QoI on DON but was not sufficient to achieve the efficacy of the Feekes 10.5.1 DMI-only treatments. Our results suggest that one must be prudent when using QoI treatments under moderate to high risk of FHB, particularly where the QoI is used without an effective DMI applied in combination or in sequence.

Detection and Characterization of QoI-Resistant Phytophthora capsici Causing Pepper Phytophthora Blight in China.

Phytophthora capsici is a highly destructive plant pathogen that has spread worldwide. To date, the quinone outside inhibitor (QoI) azoxystrobin has been the choice of farmers for managing this oomycete. In this study, the sensitivity of 90 P. capsici isolates collected from Yunnan, Fujian, Jiangxi, Zhejiang, and Guangdong in southern China to azoxystrobin was assessed based on mycelial growth, sporangia formation, and zoospore discharge. Furthermore, the mitochondrial cytochrome b (cytb) gene from azoxystrobin-sensitive and -resistant P. capsici isolates was compared to investigate the mechanism of QoI resistance. The high values for effective concentration to inhibit 50% of mycelial growth and large variation factor obtained provide strong support for the existence of azoxystrobin-resistant subpopulations in wild populations. The resistance frequency of P. capsici to azoxystrobin was greater than 40%. Sensitive P. capsici isolates were strongly suppressed on V8 medium plates containing azoxystrobin supplemented with salicylhydroxamic acid at 50 µg ml-1, whereas resistant isolates grew well under these conditions. Multiple alignment analysis revealed a missense mutation in the cytb gene that alters codon 137 (GGA to AGA), causing an amino acid substitution of glycine to arginine (G137R). The fitness of the azoxystrobin-sensitive isolate is similar to that of the G137R mutant. Additionally, the P. capsici isolates used in this study exhibited decreased sensitivity to two other QoI fungicides (pyraclostrobin and famoxadone). Necessary measures should be taken to control this trend of resistance to QoI that has developed in P. capsici in southern China.

Heteroplasmy for the Cytochrome b Gene in Podosphaera xanthii and its Role in Resistance to QoI Fungicides in Spain.

In Spain, management of the cucurbit powdery mildew pathogen Podosphaera xanthii is strongly dependent on chemicals such as quinone outside inhibitor (QoI) fungicides. In a previous report, widespread resistance to QoI fungicides in populations of P. xanthii in south-central Spain was documented, but the molecular mechanisms of resistance remained unclear. In this work, the role of the Rieske-FeS (risp) and the cytochrome b (cytb) gene mutations in QoI resistance of P. xanthii were examined. No point mutations in the risp gene were found in the three QoI-resistant isolates analyzed. For cytb, sequence analysis revealed the presence of a G143A substitution that occurs in many QoI-resistant fungi. This mutation was always detected in QoI-resistant isolates of P. xanthii; however, it was also detected in sensitive isolates. To better understand the role of heteroplasmy for cytb in QoI resistance of P. xanthii, an allele-specific quantitative PCR was developed to quantify the relative abundance of the G143 (sensitive) and A143 (resistant) alleles. High relative abundance of A143 allele (70%) was associated with isolates resistant to QoI fungicides; however, QoI-sensitive isolates also carried the mutated allele in frequencies ranged from 10 to 60%. Our data suggest that G143A mutation in cytb is the primary factor involved in QoI resistance of P. xanthii but the proportion of G143 and A143 alleles in an isolate may determine its QoI resistance level.

Fitness, Competitive Ability, and Mutation Stability of Isolates of Colletotrichum acutatum from Strawberry Resistant to QoI Fungicides.

Quinone-outside inhibitor (QoI) fungicides are used to manage anthracnose of strawberry, caused by Colletotrichum acutatum. However, selection for resistance to QoI fungicides was first reported in 2013 in Florida and, subsequently, in strawberry nurseries and production areas across the United States and Canada. C. acutatum resistance to QoIs is associated with the G143A point mutation in the cytochrome b gene. This mutation is known to be associated with field resistance even at high rates of QoI. In this study, we investigated the relative fitness and competitive ability of QoI-resistant and -sensitive C. acutatum isolates. A fitness comparison did not indicate any difference between resistant and sensitive isolates in aggressiveness, spore production, and mycelial growth at different temperatures. Additionally, in the absence of selection pressure, resistant and sensitive isolates were equally competitive. Cultivation of QoI-resistant and QoI-sensitive isolates for four culture cycles in vitro in the absence of azoxystrobin showed that QoI resistance was stable. The observed lack of fitness penalties and stability of the G143A mutation in QoI-resistant C. acutatum populations suggest that the interruption and further reintroduction of QoI fungicides might not be an option for strawberry nurseries and fruit production areas. Further investigation of alternative chemical and nonchemical C. acutatum control practices, in addition to the integration of multisite fungicides, is needed to reduce the occurrence and distribution of QoI-resistant populations in strawberry fields.

Effect of Fusarium Head Blight Management Practices on Mycotoxin Contamination of Wheat Straw.

Management of Fusarium graminearum-associated mycotoxins in wheat grain has been extensively evaluated, but little is known about management of mycotoxins in straw. Two research trials were conducted at four locations from 2011 to 2014. The objective of the first trial was to determine the efficacy of fungicides, and the objective of the second trial was to evaluate the use of integrated disease management strategies, for the control of Fusarium head blight (FHB) and reducing the concentration of the Fusarium mycotoxins deoxynivalenol, 3-acetyl-deoxynivalenol, and 15-acetyl-deoxynivalenol in straw. In the first trial, it was determined that demethylation inhibitor (DMI) fungicides did not offer significant (P ≤ 0.05) reductions of mycotoxin concentrations in the straw compared with a no-fungicide control treatment, but significant (P ≤ 0.05) reductions in mycotoxin concentration were observed in the control when compared with treatments with the application of quinone outside inhibitor (QoI)-containing fungicides. In the second trial, mycotoxin concentrations in the straw were significantly (P ≤ 0.05) reduced in the moderately resistant cultivar compared with the susceptible cultivar, but were not affected by the use of a fungicide. The practices typically used to manage Fusarium mycotoxins in wheat grain, especially the selection of resistant cultivars and not using a QoI fungicide, may be an effective means to reduce mycotoxin concentrations in the straw.

Phylogenetic relationship and fungicide sensitivity of members of the Colletotrichum gloeosporioides species complex from apple

The members of the Colletotrichum gloeosporioides species complex (CGSC), the dominant pathogens of apple bitter rot in Nagano prefecture, Japan, were reidentified and the relationship between the species and fungicide sensitivity was revealed. Based on phylogenetic analysis of the ApMat locus with the neighbor-joining (NJ) method, isolates from apple contained three species of the CGSC; C. fructicola, C. aenigma, C. siamense, and three clades of the CGSC: Clade V, S and K. Colletotrichum fructicola and Clade S dominated in Nagano Prefecture. Isolates of C. siamense, C. aenigma and Clade V, S and K remained sensitive to benomyl and quinone outside inhibitor (QoI) fungicides, while C. fructicola often developed resistance to benomyl and QoI fungicides. These results suggest that the development of fungicide resistance differs among members of the CGSC.

The Detection and Characterization of QoI-Resistant Didymella rabiei Causing Ascochyta Blight of Chickpea in Montana.

Ascochyta blight (AB) of pulse crops (chickpea, field pea, and lentils) causes yield loss in Montana, where 1.2 million acres was planted to pulses in 2016. Pyraclostrobin and azoxystrobin, quinone outside inhibitor (QoI) fungicides, have been the choice of farmers for the management of AB in pulses. However, a G143A mutation in the cytochrome b gene has been reported to confer resistance to QoI fungicides. A total of 990 isolates of AB-causing fungi were isolated and screened for QoI resistance. Out of these, 10% were isolated from chickpea, 81% were isolated from field peas, and 9% isolated from lentil. These were from a survey of grower’s fields and seed lots (chickpea = 17, field pea = 131, and lentil = 21) from 23 counties in Montana sent to the Regional Pulse Crop Diagnostic Laboratory, Bozeman, MT, United States for testing. Fungicide-resistant Didymella rabiei isolates were found in one chickpea seed lot each sent from Daniels, McCone and Valley Counties, MT, from seed produced in 2015 and 2016. Multiple alignment analysis of amino acid sequences showed a missense mutation that replaced the codon for amino acid 143 from GGT to GCT, introducing an amino acid change from glycine to alanine (G143A), which is reported to be associated with QoI resistance. Under greenhouse conditions, disease severity was significantly higher on pyraclostrobin-treated chickpea plants inoculated with QoI-resistant isolates of D. rabiei than sensitive isolates (p-value = 0.001). This indicates that where resistant isolates are located, fungicide failures may be observed in the field. D. rabiei-specific polymerase chain reaction primer sets and hydrolysis probes were developed to efficiently discriminate QoI- sensitive and - resistant isolates.

Genetic diversity, QoI fungicide resistance, and mating type distribution of Cercospora sojina-Implications for the disease dynamics of frogeye leaf spot on soybean.

Frogeye leaf spot (FLS), caused by Cercospora sojina, causes significant damage to soybean in the U.S. One control strategy is the use of quinone outside inhibitor (QoI) fungicides. QoI resistant isolates were first reported in Tennessee (TN) in 2010. To investigate the disease dynamics of C. sojina, we collected 437 C. sojina isolates in 2015 from Jackson and Milan, TN and used 40 historical isolates collected from 2006–2009 from TN and ten additional states for comparison. A subset of 186 isolates, including historical isolates, were genotyped for 49 single nucleotide polymorphism (SNP) markers and the QoI resistance locus, revealing 35 unique genotypes. The genotypes clustered into three groups with two groups containing only sensitive isolates and the remaining group containing all resistant isolates and a dominant clonal lineage of 130 isolates. All 477 C. sojina isolates were genotyped for the QoI locus revealing 344 resistant and 133 sensitive isolates. All isolates collected prior to 2015 were QoI sensitive. Both mating type alleles (MAT1-1-1 and MAT1-2) were found in Jackson and Milan, TN and recovered from single lesions suggesting sexual recombination may play a role in the epidemiology of field populations. Analysis of C. sojina isolates using SNP markers proved useful to investigate population diversity and to elaborate on diversity as it relates to QoI resistance and mating type.

Erratum to: Resistance of wheat pathogen Zymoseptoria tritici to DMI and QoI fungicides in the Nordic-Baltic region - a status

Septoria tritici blotch (STB) caused by the ascomycete Zymoseptoria tritici (Z. tritici) is currently the most prevalent foliar disease in wheat in the Nordic-Baltic region. Fungicide availability in this region differs greatly and is generally more limited than in other European regions. Monitoring of fungicide sensitivity is an essential tool to survey changes in fungal populations in order to react and be able to adapt recommendations for fungicide use. In this study the authors give an overview of the current situation of 14α-demethylation inhibitor (DMI) and quinone outside inhibitor (QoI) sensitivity of Z. tritici from Scandinavia and the Baltic countries. A total of 985 isolates from the Nordic-Baltic region were investigated for EC50 of DMI epoxiconazole and prothioconazole. Fungicide sensitivity remains at a high level with values ranging from 0.07 to 0.48 mg L−1 for epoxiconazole and 1.17 to 9.47 mg L−1 for prothioconazole. Point mutation I381V in the DMI target gene CYP51 was dominant throughout the region, but mutations D134G, V136A/C and S524T were also detected in the population in 2014. Screening for inserts in the CYP51 promoter region revealed that a ~ 1000 bp insert is predominant in the entire region. Only a single isolate was found in Denmark, harbouring the 120 bp insert, known to reduce fungicide sensitivity. Two Danish isolates which had elevated resistance levels were associated with an enhanced efflux. Significant differences were found across the area for the presence of G143A, conferring QoI resistance. As there is only limited access to results from this area, these findings can serve as reference for future fungicide sensitivity investigations and for evaluation of changes in the Northern European Z. tritici population.