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.     

植物病原菌抗药性及其抗性治理策略

随着现代高活性的选择性杀菌剂的研发和广泛使用,病原菌的抗药性问题日趋严重,这已成为植物病害化学保护领域最受关注的问题之一。本文阐释了抗药性相关术语的定义,概述了病原菌的抗药性现状,并从自然选择和诱导突变两种学说的角度分析了抗药性产生的原因。进一步分析了抗药性群体流行与病原菌自身特点、杀菌剂类型和作用机制等影响因子密切相关,综述了抗药性风险评估、抗药性机制、抗药性进化以及抗药性常规和分子检测方法等内容。最后,提出了抗药性治理的目标和策略,即根据抗药病原群体形成的主要影响因素,针对性地设计抗药性治理短期和长期策略,特别是需进一步加强对新药剂和新防治对象开展抗药性风险评估、制定抗药性管理策略、建立再评价机制等。综上,明确植物病原菌抗药性发生发展特点并制定科学合理的抗性治理策略,对进一步开展植物病害的科学防控具有重要的参考价值。     

Chemical control of plant diseases

As the world population increases, we also need to increase food production. Chemical control has been critical in preventing losses due to plant diseases, especially with the development of numerous specific-action fungicides since the 1960s. In Japan, a host-defense inducer has been used to control rice blast since the 1970s without any problems with resistance development in the pathogen. Leaf blast has been controlled using a labor-saving method such as the one-shot application of a granular mixture of fungicide and insecticide to nursery boxes, which became mainstream in the 2000s. However, the need for many choices of fungicides that have several modes of action was demonstrated by the development of resistance to cytalone dehydratase inhibitors. In Europe, many pathogens have threatened cereals since the great increase in cereal production in 1970s, creating a large market for broad-spectrum fungicides. In Brazil, Phakopsora pachyrhizi was distributed to large soybean acreages during 2000s, and the outbreak of soybean rust resulted in a large increase in fungicide use. While the importance of chemical control is recognized, fungicide resistance is an avoidable problem; published guidelines on countermeasure and manuals on testing sensitivity to fungicides are available. Since chemical regulations have become stricter, new fungicides are less likely to be developed. Our task is to maintain the effectiveness and diversity of the present modes of action for fungicides and implement countermeasures against the development of fungicide resistance.     

植物病原菌抗药性遗传研究

 植物病原菌对杀菌剂的抗药性是由遗传基因控制的,抗药基因位于细胞质遗传因子或细胞核染色体基因上,细菌对许多药剂如铜制剂、链霉素等的抗药性和真菌对少数药剂如甲氧丙烯酸酯类药剂的抗性属于前一种情况,而真菌对大多数药剂的抗性则属于后一种情况。核基因控制的抗药性又可分为主效基因(major-gene)抗性和微效多基因(poly-gene)抗性,分别使病菌对药剂的抗性表现质量性状和数量性状。病原菌对苯并咪唑类药剂、春日霉素、羧基酰胺类药剂、苯酰胺类药剂、芳烃类药剂、二甲酰亚胺类药剂等的抗性通常为主效基因控制;使病菌表现微效多基因抗性的杀菌剂主要有多果定、羟基嘧啶类药剂、甾醇合成抑制剂(SBIs)等     

杀菌剂对水稻旱育秧立枯病的控制作用及其对秧苗的生理效应

为了筛选防治水稻旱育秧立枯病的有效药剂,通过秧盘试验和室内致病菌离体毒力测定,结合考察药剂对秧苗生长的影响,对保护性杀菌剂代森锰锌和福美双以及内吸治疗性杀菌剂稻瘟灵、 FDA1 霉灵等进行了系统评价。结果表明, FDA1 霉灵、甲基硫菌灵和稻瘟灵对致病菌离体毒力虽然不高,但由于 FDA1 霉灵和甲基硫菌灵对水稻秧苗生长均有促进作用,稻瘟灵能显著提高秧苗抗低温胁迫能力,这3种药剂对水稻立枯病的防治效果较好,防效达65%~81%;而离体毒力虽然高,但对秧苗生长不利的三唑酮、丙环唑和烯唑醇等三唑类药剂效果稍差,防效为51%~75%。根据试验结果提出稻瘟灵是防治该类病害的一种较理想的单用或混用药剂;针对水稻立枯病的发生受环境影响大、病原种类多等特点,指出在研究防治药剂时应采用活体筛选为主、离体筛选为辅的筛选方法,而且提倡不同杀菌谱品种的混用。     

Fungicide control of mushroom cobweb disease caused by Cladobotryum strains with different benzimidazole resistance profiles

The benzimidazole fungicides thiabendazole and carbendazim, and the imidazole fungicide prochloraz-Mn, were tested for their efficacy in controlling cobweb disease of mushrooms caused by two Cladobotryum isolates. Isolate 202A was benzimidazole-sensitive in vitro and cobweb growth on the casing was well controlled by both benzimidazole fungicides in cropping experiments. Carbendazim also controlled the development of spotting symptoms much more effectively than thiabendazole. A second isolate (192B1) was benzimidazole-resistant and was highly resistant to thiabendazole in vitro but it showed some sensitivity to carbendazim in vitro at moderate to high concentrations. Despite this, carbendazim did not control disease symptoms in cropping experiments, confirming that isolate 192B1 is cross-resistant to other benzimidazole fungicides. Both isolates showed some sensitivity to prochloraz-Mn in vitro. This fungicide gave between 45% and 65% control of cobweb growth on the casing caused by either 202A or 192B1 but gave no control of spotting symptoms. Reducing the fungicide application volume did not give enhanced disease control. The emergence of benzimidazole resistance reduces the value of benzimidazoles in the control of mushroom pathogens. However, the lack of effective alternatives means they continue to have utility in cases where pathogens are still sensitive but this requires regular monitoring of pathogen populations for resistance.     

Interaction between phyllosphere yeasts, aphid honeydew and fungicide effectiveness in wheat under field conditions

Aphid honeydew can reduce fungicide effectiveness against necrotrophic pathogens of wheat under controlled conditions. In 1988 and 1989, field experiments were carried out at different locations in The Netherlands, to assess the interference of honeydew with fungicide effectiveness under field conditions, and to establish whether honeydew consumption by naturally occurring saprophytes could prevent this effect. Different combinations of chemicals were used to create differences in saprophytic population densities and aphid populations, which were expected to result in differences in the amount of honeydew and other nutrients on the leaves. In two of the experiments, Septoria nodorum was the only necrotrophic pathogen present; in the other two the only necrotroph was S. tritici . On several sampling dates, the effect of the fungicides was significantly enhanced by addition of insecticide, most often when the saprophytes were reduced by maneb/anilazine. This effect of insecticide is ascribed to prevention of interference of honeydew with fungicide activity, and not to a direct effect of insecticide, as insecticide alone had no effect on necrosis. On the basis of the fact that interference of honeydew with fungicide activity was most apparent when saprophytic population densities were reduced, it is concluded that the naturally occurring saprophytes can generally prevent the effect of honeydew. The same interactions are likely to occur between other nutrients and fungicides, and it is therefore argued that the naturally occurring saprophytes should be protected by elimination of harmful chemicals.     

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。     

Interactions of fungicides and leaf saprophytes

Phylloplane micro-organisms may metabolise substances present on the leaf surface, they may interact with leaf pathogens and they may be involved in the process of senescence. The effects of fungicides on many leaf saprophytes are, however, largely unknown. Studies under both field and laboratory conditions have indicated that wide spectrum protectant fungicides, such as captan, captafol and zineb, are as effective against the saprophytic flora as they are against plant pathogens. If the leaf surface microflora is restricted by fungicides then antagonism towards pathogens will also be reduced, but beneficial results may ensue if the microbial impact on senescence is lessened. Field trials with barley have demonstrated that it is possible to delay senescence by using fungicide sprays.     

A review of the selectivity of fungicides in agriculture and horticulture

Protectant fungicides rely for their selectivity on physical barriers (e.g. the leaf cuticle) between their deposits and the protoplasts of the host plant. Systemic fungicides, by contrast, come into direct contact with host cells and need to be more selective. Nevertheless, important side-effects can occur, involving higher plants, animals and other microorganisms; examples of these are cited. Sometimes mycelium and spores of the target fungus show differing susceptibility; also, activity of a fungicide in vitro does not always reflect its effectiveness in vivo. An extreme form of selectivity is the undesirable emergence of insensitive strains from an otherwise sensitive fungal species.     

Large-scale fungicide spray heterogeneity and the regional spread of resistant pathogen strains

ABSTRACT Most models for the spread of fungicide resistance in plant pathogens are focused on within-field dynamics, yet regional invasion depends upon the interactions between field populations. Here, we use a spatially implicit metapopulation model to describe the dynamics of regional spread, in which subpopulations correspond to single fields. We show that the criterion for the regional invasion of pathogens between fields differs from that for invasion within fields. That is, the ability of a fungicide-resistant strain of a pathogen to invade a field population does not necessarily imply an ability to spread through many fields at the regional scale. This depends upon an interaction between the fraction of fields that is sprayed and the reproductive capacity of the pathogen. This result is of practical significance and indicates that resistance management strategies which currently target within-field processes, such as the use of mixtures and alternations of fungicides, may be more effective if between-field processes also were targeted; for example, through the restricted deployment of fungicides over large areas. We also show that the fraction of disease-free fields is maximized when the proportion of fields that is sprayed is just below the threshold for invasion of the resistant strain.     

Impact of fungal drug transporters on fungicide sensitivity, multidrug resistance and virulence

Drug transporters are membrane proteins that provide protection for organisms against natural toxic products and fungicides. In plant pathogens, drug transporters function in baseline sensitivity to fungicides, multidrug resistance (MDR) and virulence on host plants. This paper describes drug transporters of the filamentous fungi Aspergillus nidulans (Eidam) Winter, Botrytis cinerea Pers and Mycosphaerella graminicola (Fückel) Schroter that function in fungicide sensitivity and resistance. The fungi possess ATP-binding cassette (ABC) drug transporters that mediate MDR to fungicides in laboratory mutants. Similar mutants are not pronounced in field resistance to most classes of fungicide but may play a role in resistance to azoles. MDR may also explain historical cases of resistance to aromatic hydrocarbon fungicides and dodine. In clinical situations, MDR development in Candida albicans (Robin) Berkhout mediated by ABC transporters in patients suffering from candidiasis is common after prolonged treatment with azoles. Factors that can explain this striking difference between agricultural and clinical situations are discussed. Attention is also paid to the risk of MDR development in plant pathogens in the future. Finally, the paper describes the impact of fungal drug transporters on drug discovery.     

杀菌剂氟苯醚酰胺的创制

琥珀酸脱氢酶 (succinate dehydrogenase, SDH) 是重要的杀菌剂靶标之一,而很多植物病原菌对靶向SDH的杀菌剂已经产生了较为严重的抗药性,因此新型靶向SDH的杀菌剂设计显得尤为重要。基于药效团连接碎片的虚拟筛选 (PFVS) 是一种独立于生物物理技术的高通量药物发现方法,采用PFVS方法成功获得了靶向SDH的新型杀菌剂候选化合物—氟苯醚酰胺。本文主要从PFVS原理、先导化合物的发现、取代基的修饰以及杀菌活性研究等方面对氟苯醚酰胺的创制进行系统分析。氟苯醚酰胺创制的案例分析可为农药研究工作者提供新思路和新方法。     

New Trends in the Chemical Control of Wheat Diseases1

Following a list of the economically most important fungal wheat diseases, recent trends in chemical seed treatment are discussed. Different fungicide mixtures are now available for the replacement of mercurial dressings, but are generally more expensive. The control of fungal diseases by spraying has become more and more common over the past ten years in certain countries. Advantage has been taken of the curative action of the modern systemic chemicals. The major diseases are powdery mildew, rusts, eyespot, glume-and leaf-blotch. The general application of a fungicide programme is restricted by regional wheat prices, and should also be limited to reduce environmental contamination. Knowledge about economic thresholds and correct timing of control are indispensable, and in special cases the development of warning systems is advisable. The role of Cycocel (chlormequat) is also discussed. A danger of fungicide resistance in populations of plant pathogenic fungi has arisen with the introduction of the systemic fungicides (one-site-inhibitors). As yet, however, no economically important case of fungicide resistance within cereal pathogens is known. Factors favouring the stepwise selection of resistant strains under field conditions are discussed in detail, and experimental data are presented on this subject. Other positive or negative side-effects of the modern fungicides are described.     

Control strategies using systemic fungicides for limiting disease development and resistance buildup: practical implications of a simulation model

A simulation model was used to assess the control efficacy of, and the buildup of resistant populations to, systemic fungicides as affected by preventive vs responsive (curative) treatments with a protectant fungicide, or with a mixture composed of a systemic and a protectant fungicide. The variables introduced in the model were: rate of fungicide weathering, coverage efficacy, and relative fitness of the resistant population of the pathogens. In all simulated epidemics (with different combinations of apparent infection rates, critical disease levels and rates of weathering of the fungicides), preventive treatments, before the critical disease levels were reached, were significantly more effective than responsive (curative) treatments. At low rates of weathering of the protectant fungicide (half-life of the protectant greater than or equal to that of the systemic), the buildup of the resistant population was significantly inhibited when a mixture was used either preventively or responsively. At high rates of weathering of the protectant fungicide (half-life of the protectant less than that of the systemic), both the buildup of the resistant population and the control efficacy were dependent on the control program used and on the different combinations of fungicide weathering, fitness of the resistant population, and coverage efficacy of both the protectant and systemic fungicides.     

不同杀菌剂对苹果炭疽叶枯病的防治效果

为了筛选出防治苹果炭疽叶枯病的有效杀菌剂, 采用室外先接种后施药和先施药后接种的方法, 测试了6种药剂的内吸治疗效果和8种药剂的保护效果。在病菌侵染后的72 h内使用吡唑醚菌酯, 或在病菌侵染后的24 h内使用咪鲜胺对病斑的显症有一定的治疗效果。波尔多液在喷施后18 d, 对炭疽叶枯病菌侵染的抑制效果仍达50%, 肟菌?戊唑醇、烯酰?吡唑酯和唑醚?代森联3种药剂在施药后的第11天, 其保护效果与对照仍有显著差异, 持效期达11 d, 代森锰锌、甲基硫菌灵、氢氧化铜和咪鲜胺4种保护剂的持效期只能维持6 d。炭疽叶枯病的防治应以波尔多液为主, 并与吡唑醚菌酯等有机杀菌剂交替使用, 有机铜制剂不能替代波尔多液。     

Seed treatments to control seedborne fungal pathogens of vegetable crops

Vegetable crops are frequently infected by fungal pathogens, which can include seedborne fungi. In such cases, the pathogen is already present within or on the seed surface, and can thus cause seed rot and seedling damping‐off. Treatment of vegetable seeds has been shown to prevent plant disease epidemics caused by seedborne fungal pathogens. Furthermore, seed treatments can be useful in reducing the amounts of pesticides required to manage a disease, because effective seed treatments can eliminate the need for foliar application of fungicides later in the season. Although the application of fungicides is almost always effective, their non‐target environmental impact and the development of pathogen resistance have led to the search for alternative methods, especially in the past few years. Physical treatments that have already been used in the past and treatments with biopesticides, such as plant extracts, natural compounds and biocontrol agents, have proved to be effective in controlling seedborne pathogens. These have been applied alone or in combination, and they are widely used owing to their broad spectrum in terms of disease control and production yield. In this review, the effectiveness of different seed treatments against the main seedborne pathogens of some important vegetable crops is critically discussed. © 2013 Society of Chemical Industry     

杀菌剂抗性监测研究进展

使用现代选择性杀菌剂防治植物病害是保证农作物高产、稳产的重要措施之一,但是现代杀菌剂很容易出现抗性而导致化学防治失败．使农业生产蒙受巨大损失。本文从杀菌剂的抗性检测和治理技术等方面概述杀菌剂的抗性研究进展。     

Microscopy reveals disease control through novel effects on fungal development: a case study with an early-generation benzophenone fungicide

The benzophenones are a new class of agricultural fungicides that demonstrate protectant, curative and eradicative/antisporulant activity against powdery mildews. The chemistry is represented in the marketplace by the fungicide metrafenone, recently introduced by BASF and discussed in the following paper. The benzophenones show no evidence of acting by previously identified biochemical mechanisms, nor do they show cross-resistance with existing fungicides. The value of microscopy in elucidating fungicide mode of action is demonstrated through identification of the effects of an early benzophenone, eBZO, on mildew development. eBZO caused profound alterations in the morphology of powdery mildews of both monocotyledons and dicotyledons, affecting multiple stages of fungal development, including spore germination, appressorial formation, penetration, surface hyphal morphology and sporogenesis. Identification of analogous effects of eBZO on sporulation in the model organism Aspergillus nidulans (Eidam) Winter provides a unique opportunity to elucidate important morphogenetic regulatory sites in the economically important obligate pathogens, the powdery mildews. Benzophenones provide a further example of the benefits of whole-organism testing in the search for novel fungicide modes of action.     

Quantifying the effect of interactions between disease control,nitrogen supply and land use change on the greenhouse gas emissions associated with wheat production

A method for calculating the effect of disease control on greenhouse gas (GHG) emissions associated with wheat production, reported previously, was developed further to account for effects of disease control on the amount of fertilizer nitrogen (N) which should be applied and on changes in land use. Data from nine randomized and replicated field experiments from the UK and Denmark showed that the economic optimum N input to winter wheat was greater if diseases were controlled by fungicides, than for untreated wheat. The GHGs associated with this additional N largely negated the benefit to emissions per tonne of grain resulting from disease control. However, the mean grain yield obtained without fungicide treatment was 6·71 t ha^?1, compared to 8·88 t ha^?1 with fungicide treatment, if N input was optimal for each situation. In the absence of disease control by fungicides, and assuming that the optimum N rate was used, an additional 481 kha of wheat would be required to maintain UK wheat production at the current level. If the additional land area came from converting temperate grassland to arable production, the GHG emissions caused by ploughing grassland would cause emissions to rise from 503 to 713 kg CO₂e per tonne of grain produced. This would result in an additional 3·15 Mt CO₂e per year to produce the typical UK annual production of 15 Mt. This analysis reinforces the importance of winning the ‘arms race’ against pathogen evolution towards fungicide insensitivity and virulence.