Identifying when it is financially beneficial to increase or decrease fungicide dose as resistance develops

When fungicide efficacy declines due to the development of resistance in the pathogen population, growers have to either change to an alternative mode of action or adjust their treatment programme. Adjustments may include either decreasing (or stopping) use of the mode of action, or increasing the total dose applied (by increasing number of applications and/or dose per application, where permitted) to try to maintain effective disease control. This study explores the circumstances under which increasing/decreasing total applied fungicide is financially optimal. A model based on field data is used to optimize the dose of fungicide applied when fungicide resistance develops in a pathogen population. The model is used to explore contrasting pathosystems and fungicide classes. When qualitative fungicide resistance develops, the shape of the disease–yield loss relationship determines whether the optimal total dose increases or decreases with increasing frequency of resistance in the pathogen population. When quantitative fungicide resistance develops, such that effective control can still be obtained with doses close to the maximum permitted dose, the optimal dose increases with increasing frequency of resistance in the pathogen population.     

Dose and number of applications that maximize fungicide effective life exemplified by Zymoseptoria tritici on wheat – a model analysis

Two key decisions that need to be taken about a fungicide treatment programme are (i) the number of applications that should be used per crop growing season, and (ii) the dosage that should be used in each application. There are two opposing considerations, with control efficacy improved by a higher number of applications and higher dose, and resistance management improved by a lower number of applications and lower dose. Resistance management aims to prolong the effective life of the fungicide, defined as the time between its introduction onto the market for use on the target pathogen, and the moment when effective control is lost due to a build‐up of fungicide resistance. Thus, the question is whether there are optimal combinations of dose rate and number of applications that both provide effective control and lead to a longer effective life. In this paper, it is shown how a range of spray programmes can be compared and optimal programmes selected. This is explored with Zymoseptoria tritici on wheat and a quinone outside inhibitor (QoI) fungicide. For this pathogen–fungicide combination, a single treatment provided effective control under the simulated disease pressure, but only if the application timing was optimal and the dose was close to the maximum permitted. Programmes with three applications were generally not optimal as they exerted too much selection for resistance. Two‐application fungicide programmes balanced effective control with reasonable flexibility of dose and application timing, and low resistance selection, leading to long effective lives of the fungicide.     

Effects of fungicide dose and mixtures on selection for triazole resistance in Mycosphaerella graminicola under field conditions

The effects of varying doses of fungicides, alone or in mixtures, on selection for triazole resistance were examined under field conditions. Two experiments were conducted using the triazole fungicide fluquinconazole with the strobilurin fungicide azoxystrobin as a mixture partner. Inoculated wheat plots with a known ratio of more sensitive to less sensitive isolates of the leaf blotch fungus Mycosphaerella graminicola were sprayed with fungicide and sampled once symptoms had appeared. Selection for fluquinconazole resistance increased in proportion to the dose, up to one-half of the full dose (the maximum tested) in both experiments. At the higher doses of fluquinconazole, the addition of azoxystrobin was associated with a decrease in selection (nonsignificant in the first experiment) for triazole resistance. Control by low doses of fluquinconazole was increased by mixture with azoxystrobin, but at higher doses mixture with azoxystrobin sometimes decreased control, so that reduced selection was obtained at the cost of some reduction in control. The effects on resistance are not necessarily general consequences of mixing fungicides, and suggest that the properties of any specific mixture may need to be demonstrated experimentally. Selection was inversely related to control in the unmixed treatments in both experiments, but the relationship was weaker in the mixtures with azoxystrobin.     

The dose rate debate: does the risk of fungicide resistance increase or decrease with dose?

This paper reviews the evidence relating to the question: does the risk of fungicide resistance increase or decrease with dose? The development of fungicide resistance progresses through three key phases. During the ‘emergence phase’ the resistant strain has to arise through mutation and invasion. During the subsequent ‘selection phase’, the resistant strain is present in the pathogen population and the fraction of the pathogen population carrying the resistance increases due to the selection pressure caused by the fungicide. During the final phase of ‘adjustment’, the dose or choice of fungicide may need to be changed to maintain effective control over a pathogen population where resistance has developed to intermediate levels. Emergence phase: no experimental publications and only one model study report on the emergence phase, and we conclude that work in this area is needed. Selection phase: all the published experimental work, and virtually all model studies, relate to the selection phase. Seven peer reviewed and four non‐peer reviewed publications report experimental evidence. All show increased selection for fungicide resistance with increased fungicide dose, except for one peer reviewed publication that does not detect any selection irrespective of dose and one conference proceedings publication which claims evidence for increased selection at a lower dose. In the mathematical models published, no evidence has been found that a lower dose could lead to a higher risk of fungicide resistance selection. We discuss areas of the dose rate debate that need further study. These include further work on pathogen‐fungicide combinations where the pathogen develops partial resistance to the fungicide and work on the emergence phase.     

Predicting effective doses for the joint action of two fungicide applications

A function was derived to predict fungicide efficacy when more than one application of a single active ingredient is made to a crop, given parameters describing the dose–response curves of the component single-spray applications. In the function, a second application is considered to act on that proportion of the total pathogen population which was uncontrollable at the time of the first application (represented by the lower asymptote of the dose–response curve for the first treatment), plus any additional part of the population which survived the first application as a result of a finite dose being applied. Data to estimate the single-spray dose–response curve parameters and validate predictions of two-spray programme efficacy were obtained from separate subsets of treatments in four field experiments. A systemic fungicide spray was applied to wheat at a range of doses, at one or both of two times (t₁ and t₂), in all dose combinations. Observed values of the area under the disease progress curve (AUDPC) for septoria leaf blotch (Mycosphaerella graminicola) were used to construct response surfaces of dose at t₁ by dose at t₂ for each culm leaf layer. Parameters were estimated from single-spray and zero-dose treatment data only. The model predicted a high proportion (R² = 71–95%) of the variation in efficacy of the two-spray programmes. AUDPC isobols showed that the dose required at t₂ was inversely related to the dose at t₁, but the slope of the relationship varied with the relative timings of t₁ and t₂ in relation to culm leaf emergence. Isobols were curved, so the effective dose – the total dose required to achieve a given level of disease suppression – was lower when administered as two applications.     

Real-time PCR法定量检测柑橘绿霉病菌对抑霉唑的抗性频率

 抑霉唑被广泛用来防治由指状青霉菌(Penicillium digitatum)引起的柑橘绿霉病。已有研究表明,柑橘绿霉病菌对抑霉唑的抗性由CYP51基因启动子区5个126-bp转录增强子的简单串联重复和126-bp转录增强子上199-bp的特异性片段插入所引起。基于这2种抗性分子机制,通过设计特异引物和优化条件,建立real-time PCR高通量分子检测技术,用于快速检测柑橘包装贮藏库中绿霉病菌群体对抑霉唑的抗性频率F_R,指导科学用药。     

Integrated control of potato late blight: predicting the combined efficacy of host resistance and fungicides

Integrating cultivars that are partially resistant with reduced fungicide doses offers growers an opportunity to decrease fungicide input but still maintain disease control. To use integrated control strategies in practice requires a method to determine the combined effectiveness of particular cultivar and fungicide dose combinations. Simple models, such as additive dose models (ADM) and multiplicative survival models (MSM), have been used previously to determine the joint action of two or more pesticides. This study tests whether a model based on multiplicative survival principles can predict the joint action of fungicide doses combined with cultivars of differing partial host resistance. Data from eight field experiments on potato late blight (Phytophthora infestans) were used to test the model; the severity of foliar blight was assessed and scores used to calculate the area under the disease progress curve (AUDPC). A subset of data, derived from the most susceptible cultivar, King Edward, was used to produce dose–response curves from which parameter values were estimated, quantifying fungicide efficacy. These values, along with the untreated values for the more resistant cultivars, Cara and Sarpo Mira, were used to predict the combined efficacy of the remaining cultivar by fungicide dose combinations. Predicted efficacy was compared against observations from an independent subset of treatments from the field experiments. The analysis demonstrated that multiplicative survival principles can be applied to describe the joint efficacy of host resistance and fungicide dose combinations.     

Induced resistance: a tool for fungicide resistance management

Induced resistance to the apple scab fungus Venturia inaequalis was demonstrated in greenhouse tests with 12-day-old seedlings of the apple cultivar Golden Delicious treated with methyl 2,6-dichloro-isonicotinate or 3,5-dichlorosalicylic acid prior to inoculation with the causal fungus. Studies of the dose-response of flusilazole on induced resistant plants revealed synergistic effects between both crop protection principles. Therefore, the use of such resistance-inducing compounds in the field might allow a reduction in the number of fungicide applications, and possibly a reduction in dose, thus resulting in improved efficacy of fungicides. There was also evidence that induced resistance could prove to be a valid strategy for the treatment of pathogen populations with reduced sensitivity to a given fungicide. © 1998 SCI     

梨黑斑病菌抗药性检测及其对啶酰菌胺的敏感性基线

黑斑病是梨的主要病害之一,近年来不少地区反映多菌灵等传统常用杀菌剂对其防治效果已出现下降。作者从浙江、江苏和安徽3省分离了252株梨黑斑病菌Alternaria kikuchiana,采用菌丝生长速率法检测了其抗药性发生情况。结果发现:所检测的黑斑病菌群体(n=252)对苯并咪唑类杀菌剂多菌灵的抗性频率为57.1%,且全部为高水平抗性(HR);对二甲酰亚胺类杀菌剂异菌脲的抗性频率为46.8%,全部为低水平抗性(LR);对甾醇脱甲基抑制剂类杀菌剂苯醚甲环唑的抗性为低水平(LR)及中等水平(MR),抗性频率均为28.6%;表明梨黑斑病菌对常用杀菌剂已产生较为严重的抗性。供试252株梨黑斑病菌对琥珀酸脱氢酶抑制剂啶酰菌胺的EC50值分布在0.12~3.85μg/m L之间,平均EC50值为(1.21±0.12)μg/m L,且其分布呈近似正态的单峰曲线。研究表明,啶酰菌胺可作为潜在的梨黑斑病防治替代药剂,其平均EC50值(1.21±0.12)μg/m L可作为梨黑斑病菌对啶酰菌胺的敏感性基线。     

Target and non-target site mechanisms of fungicide resistance and their implications for the management of crop pathogens

Fungicides are indispensable for high-quality crops, but the rapid emergence and evolution of fungicide resistance have become the most important issues in modern agriculture. Hence, the sustainability and profitability of agricultural production have been challenged due to the limited number of fungicide chemical classes. Resistance to site-specific fungicides has principally been linked to target and non-target site mechanisms. These mechanisms change the structure or expression level, affecting fungicide efficacy and resulting in different and varying resistance levels. This review provides background information about fungicide resistance mechanisms and their implications for developing anti-resistance strategies in plant pathogens. Here, our purpose was to review changes at the target and non-target sites of quinone outside inhibitor (QoI) fungicides, methyl-benzimidazole carbamate (MBC) fungicides, demethylation inhibitor (DMI) fungicides, and succinate dehydrogenase inhibitor (SDHI) fungicides and to evaluate if they may also be associated with a fitness cost on crop pathogen populations. The current knowledge suggests that understanding fungicide resistance mechanisms can facilitate resistance monitoring and assist in developing anti-resistance strategies and new fungicide molecules to help solve this issue. © 2023 Society of Chemical Industry.     

Occurrence of azoxystrobin‐resistant isolates in Passalora fulva,the pathogen of tomato leaf mould disease

Since 2007, serious damage to tomato from leaf mould caused by Passalora fulva has frequently been observed in commercial greenhouses in Gifu Prefecture, Japan. One of the factors relating to this damage was suspected to be a decrease in azoxystrobin sensitivity of the pathogen. Biological and molecular studies were conducted to characterize fungicide resistance. In in vitro sensitivity tests using mycelial homogenate placed on fungicide‐amended medium, the minimum inhibitory concentrations (MIC) of azoxystrobin for mycelial growth of the isolates divided into two ranges, 0.031–0.5 mg L^?1 and 8–32 mg L^?1. Isolates with MICs within the two ranges were considered as sensitive and resistant, respectively, to azoxystrobin because, in in vivo tests, the percentage protection conferred by this fungicide (100 mg a.i. L^?1) against these isolates was 89.7–100% and 4.5–31.1%, respectively. Resistant isolates had a replacement of phenylalanine with leucine at codon 129 (F129L) in cytochrome b. Forty‐five percent of the 271 isolates collected from 63 tomato greenhouses from 2007 to 2008 were resistant to azoxystrobin. In many greenhouses where the isolation frequency of resistant isolates was 80% or more, azoxystrobin had been used twice per crop for approximately 6 years. In 2012, 27% of the 405 isolates collected were resistant to azoxystrobin, and there was a marked difference in the frequency of occurrence of resistant isolates in the field populations between the three locations sampled. The occurrence of azoxystrobin‐resistant P. fulva isolates (F129L mutants) inflicted considerable damage on greenhouse tomatoes.     

植物病原真菌对杀菌剂抗性的研究进展

科学施用杀菌剂是植物病害综合治理的重要措施之一, 然而由于杀菌剂的长期使用, 病菌抗药性问题逐渐加重, 严重影响药剂的防治效果和使用寿命。近年来, 随着分子生物学技术的快速发展, 人们对杀菌剂抗性机制有了更深入的理解, 并开发出了病菌抗药基因型快速检测的方法。本文总结了植物病原真菌对苯并咪唑类杀菌剂(BZD)、肌球蛋白合成抑制剂、甾醇脱甲基抑制剂(DMI)、QoI类抑制剂、琥珀酸脱氢酶抑制剂（SDHI）和二甲酰亚胺类杀菌剂(DC)的抗药性现状与抗性机制。在此基础上, 介绍了聚合酶链反应(PCR)、限制性片段长度多态性(RFLP)、等位基因特异性PCR和环介导等温扩增(LAMP)技术在杀菌剂抗性快速检测方面的研究进展。此外, 对抗药性治理对策进行了讨论和展望。     

赣南脐橙绿霉病菌对常用杀菌剂抗性监测

 本文研究了来自赣南7个县的柑橘绿霉病菌(Penicillium digitatum)种群对该地区常用杀菌剂抑霉唑、咪鲜胺、甲基硫菌灵和百可得的抗性频率、抗性水平和对抑霉唑的抗性分子机制。结果表明:病菌对抑霉唑和咪鲜胺存在基本一致的抗性;2011和2012年病菌种群对抑霉唑和咪鲜胺的抗性频率分别为82%和90%,平均抗性倍数为51.5倍,抗性分子机制均属于IMZ-R3,即CYP51B基因启动子区发生199 bp插入的突变;病菌种群对甲基硫菌灵的抗性频率分别为82%和91%;病菌种群对百可得均表现敏感。本研究为采后柑橘病害防治药剂选择提供了科学的依据。     

The effect of fungicide dose on the composition of laboratory populations of barley powdery mildew

The effect of the use of different doses of the fungicide fenpropimorph on populations of barley powdery mildew Blumeria ( Erysiphe ) graminis f. sp. hordei was investigated in a laboratory selection experiment. A sample from the Danish aerial population of powdery mildew was split into populations, and these were kept separately for 31 generations on susceptible barley seedlings treated with fungicide at two concentrations, as well as on a control. Samples from these populations were tested for their resistance to fenpropimorph and their virulence spectra. There was a large amount of environmental variation in the ED₅₀ values used to measure fungicide resistance. In both treated populations, the average level of fungicide resistance increased, this increase being faster and greater in the population treated with the high dose. The diversity of pathotypes of the treated populations decreased, with the decline being more rapid in the population treated with the high dose, where one pathotype dominated the population after 31 generations. This pathotype was apparently not the fittest in the population treated with the low dose. This implies that knowledge of ED₅₀ is not sufficient to predict pathotype evolution under different fungicide treatments. The dominant pathotype in the high-dose treatment may not have been clonal, as there was evidence of two levels of fungicide resistance. The large environmental variation observed in estimated ED₅₀ values for resistance towards fenpropimorph may help to explain why this resistance has evolved at a slower rate than resistance towards other fungicides.     

Relationship between leaf emergence and optimum spray timing for leaf blotch (Rhynchosporium secalis) control on winter barley

For wheat, the optimum time to apply fungicide to control disease on a given leaf layer is usually at, or shortly after, full leaf emergence. Data from field experiments on barley were used to investigate whether the same relationship was applicable to control of leaf blotch on barley. Replicated plots of winter barley were sown in the autumns of 1991, 1992 and 1993 at sites in southwest England with high risk of Rhynchosporium secalis infection. Single fungicide treatments at four doses (0·25, 0·5, 0·75 or 1·0 times the label rate) were applied at one of eight different spray times, starting in mid-March in each year, with intervals of 10–11 days between spray timings. Disease was assessed every 10–11 days and area under the disease progress curve (AUDPC) values were used to construct fungicide dose by spray time response surfaces for each of the upper four leaves, for each year. Spray timings shortly before leaf emergence were found to minimize the AUDPC for each year and leaf layer, and also the effective dose (the dose required to achieve a specified level of control), similar to wheat. Fungicide treatments on barley were effective for a longer period before leaf emergence than afterwards, probably because treatments before emergence of the target leaf reduced inoculum production on leaves below. This partly explains why fungicides tend to be applied earlier in the growth of barley compared with wheat.     

Steps in predicting the relationship of yield on fungicide dose

ABSTRACT A set of hypothetical steps has been defined, which links fungicide dose to marketable yield, whereby (i) increasing dose decreases symptom area, according to a dose-response curve, (ii) decreased symptom area increases crop green area index (GAI), (iii) increasing GAI increases fractional interception of photosynthetically active radiation, (iv) increased fractional interception increases crop dry matter accumulation, and (v) yield increases, depending on the partitioning of dry matter to the marketable fraction. One equation represented all five steps. By integrating this equation for light interception during the yield forming period and differentiating with respect to the ratio of fungicide cost over yield value, an analytical solution was obtained for the economic optimum dose. Taking published ranges of parameter values for the Septoria tritici wheat pathosystem as an example, yield-response curves and optimum doses were biologically plausible when compared with data from four field experiments. The analytical and empirical results imply that the dose required to optimize economic return will vary substantially between sites, seasons, and cultivars. Sensitivity analyses identified parameters describing specific facets of disease severity, fungicide efficacy, and assimilate partitioning as most influential in determining the dose optimum.     

草莓褐色轮斑病病原鉴定及室内药效研究*

草莓褐色轮斑病近年越来越严重,尤其在草莓育苗阶段。从草莓发病叶片、匍匐茎上分离得到病原菌,对病原菌进行形态特征观察、生物学特性研究、ITS序列分析以及室内药效试验。结果表明：该病原菌为Sphaeronaemella fragariae。该菌菌丝生长最适温度范围是25～28 ℃;适宜pH为6;在供试的几种碳、氮源中,最适的碳源是蔗糖,最适的氮源是酵母浸出液。在供试的9种药剂中,以咪鲜胺1 000倍液对病菌的抑制作用最好。     

Genetic correlations in resistance to morpholine and piperidine fungicides inPyrenophora teres populations

Highly significant genetic variation (P<0.001) in resistance to the morpholine fungicides fenpropimorph, tridemorph and dodemorph and the piperidine fungicide, fenpropidin was found in different populations ofPyrenophore teres in North America and Europe which had not been previously exposed to these fungicides. Resistance phenotypes were continuously distributed for each fungicide in each population. Cross resistance relationships were determined by estimating genetic correlation coefficients in resistance to all pairwise combinations of fungicides. The majority of the correlation coefficients were highly positive for all fungicide combinations in all populations; eight of 36 (22%) coefficients were not significantly different from 1 (P>0.05). This result is consistent with the hypothesis that many of the same genes, or genes in gametic disequilibrium, control resistance to more than one fungicide in most populations ofP. teres and that these fungicides comprise a single cross resistance group. Three of 36 (8%) correlation coefficients were not significantly different from 0 (P>0.05) indicating that, in these populations, independent genes controlled resistance to these fungicides. The results of this study indicate that although most of the same genes control resistance to morpholine and piperidine fungicides inP. teres, differences in frequencies of these genes among populations can result in different cross resistance relationships from one population to another.     

Effect of dose rate and mixtures of fungicides on selection for QoI resistance in populations of Plasmopara viticola

Resistance to QoI fungicides (strobilurins, famoxadone and fenamidone) in populations of Plasmopara viticola (Berk & Curt) Berlese & de Toni developed soon after their introduction in France and Italy. Current resistance management strategies include limitation of the number of applications, use of mixtures and alternation of fungicides with different modes of action. The selection pressure resulting from QoI fungicides applied alone or in mixtures with non-QoI fungicides was investigated in whole plant experiments under controlled conditions. QoI-resistant populations of P. viticola gradually reverted to full sensitivity following consecutive transfers to untreated plants, suggesting that resistant phenotypes were less competitive than sensitive ones. When cycled on QoI-treated plants, reduction in sensitivity was greater for the QoI fungicide which had greater intrinsic activity on P. viticola. Sensitivity decreased at each subsequent cycle, resulting in almost full resistance after four generations. Mixture experiments indicated that selection pressure was affected most by the dose of the QoI fungicide and the nature of the partner fungicide. Folpet delayed selection pressure most effectively when it was associated with famoxadone or azoxystrobin. Mancozeb was least effective at reducing the rate of selection compared with the QoI alone, and fosetyl-aluminium was intermediate. Higher rates of selection were recorded when the dose of the QoI fungicide, solo or in a mixture, was increased from 1 to 4 microg ml(-1). Increasing the dose of the non-QoI partner fungicide in the mixture from 10 to 30 microg ml(-1) resulted in reduced selection pressure. These results suggest that the choice of the fungicide partner and its dosage in the mixture can significantly affect the success of QoI resistance management strategies under practical conditions.     

Resistance to Alternaria solani in hybrids between a Solanum tuberosum haploid and S. raphanifolium

Early blight of potato (Solanum tuberosum), caused by the foliar fungal pathogen Alternaria solani, is a major cause of economic loss in many potato-growing regions. Genetic resistance offers an opportunity to decrease fungicide usage while maintaining yield and quality. In this study, an early blight resistant clone of the diploid wild species S. raphanifolium was crossed as a male to a haploid (2n=2x) of cultivated potato. Hybrids were backcrossed to both parents. Eight families were created and evaluated for early blight resistance in the field. Families created by backcrossing to the wild species parent exhibited significantly lower relative area under the disease progress curve means than those from backcrossing to the cultivated parent, leading to the conclusion that S. raphanifolium contributes genes for early blight resistance. The mechanism of resistance in S. raphanifolium is unique because A. solani could not be recovered from lesions. Clones were identified with high levels of resistance and adaptation to the photoperiod of a temperate production region.