Fungicide resistance: definitions and use of terms

On the basis of an analysis of current use of terms concerning resistance to fungicides, a series of definitions is proposed for discussion. Consistent use of such terms in scientific publications will facilitate understanding and avoid possible misinterpretations.     

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。     

Fungicide resistance in soil-borne Phytophthora species1

Systemic fungicides such as the phenylamides and phosphonates have provided new opportunities for chemical control of soil-borne Phytophthora species. Potential problems in their use include biodegradation in soils, and the emergence of pathogen isolates resistant to these compounds. Resistance to phenylamides is well known in leaf-infecting Oomycetes, but less is known about the development and relative fitness of resistant isolates among populations of soil-borne Phytophthora species. Experiments with P. capsici and P. palmivora have shown that stable resistant and virulent isolates can be selected under laboratory conditions. Some mutants resistant to both metalaxyl and fosetyl-A1 have been recovered, with growth rates and sporulation equivalent to parental wild types. Selected mutants proved able to compete in vivo with fungicide-sensitive wild types in the absence of the compounds. More information is required to fully evaluate the implications of these results for chemical control of soil-borne Phytophthora species in the field. Résistance des Phytophthora du sol aux fongicides     

Fungicide modes of action and resistance in downy mildews

Among oomycetes, Plasmopara viticola on grape and Phytophthora infestans on potato are agronomically the most important pathogens requiring control measures to avoid crop losses. Several chemical classes of fungicides are available with different properties in systemicity, specificity, duration of activity and risk of resistance. The major site-specific fungicides are the Quinone outside inhibitors (QoIs; e.g. azoxystrobin), phenylamides (e.g. mefenoxam), carboxylic acid amides (CAAs; e.g. dimethomorph, mandipropamid) and cyano-acetamide oximes (cymoxanil). In addition, multi-site fungicides such as mancozeb, folpet, chlorothalonil and copper formulations are important for disease control especially in mixtures or in alternation with site-specific fungicides. QoIs inhibit mitochondrial respiration, phenylamides the polymerization of r-RNA, whereas the mode of action of the other two site-specific classes is unknown but not multi-site. The use of site-specific fungicides has in many cases selected for resistant pathogen populations. QoIs are known to follow maternal, largely monogenic inheritance of resistance; they bear a high resistance risk for many but not all oomycetes. For phenylamides, inheritance of resistance is based on nuclear, probably monogenic mechanisms involving one or two semi-dominant genes; resistance risk is high for all oomycetes. The molecular mechanism of resistance to QoIs is mostly based on the G143A mutation in the cytochrome b gene; for phenylamides it is largely unknown. Resistance risk for CAA fungicides is considered as low to moderate depending on the pathogen species. Resistance to CAAs is controlled by two nuclear, recessive genes; the molecular mechanism is unknown. For QoIs and CAAs, resistance in field populations of P. viticola may gradually decline when applications are stopped.     

Fungicide resistance in Czech populations of cucurbit powdery mildews

A total of 108 isolates of cucurbit powdery mildew (CPM) fungi, 78Golovinomyces cichoracearum (Gc) and 30Podosphaera xanthii (Px), originating from nine Czech regions and 22 districts of the Czech Republic were collected in the years 2001–2004. These isolates were screened for tolerance and/or resistance to the three frequently used fungicides (fenarimol, dinocap, benomyl). Fungicide sensitivity was determined by a modified leaf-disc bioassay with five concentrations. Fungicide efficacy differed significantly: fenarimol was the most effective and all isolates of both CPM were controlled by the recommended concentration (36μg a.i. ml⁻¹). Some isolates expressed resistance (profuse sporulation) or tolerance (limited sporulation) to lower concentrations (9.6 and 18μg a.i. ml⁻¹). Specific temporal variation in tolerance/resistance was observed, with some isolates ofGc from 2002 evincing tolerant or resistant reactions to these low fenarimol concentrations, but isolates with similar reactions were not detected during the years 2003–2004. Dinocap showed decreasing efficacy during all 3 years. A shift to more tolerant reactions in the CPM populations was detected forGc in 2001–2002, and forPx in 2001 and 2004. Benomyl was found to be ineffective, because the majority of screened isolates (88%Gc and 97%Px) belonged to the highly resistant strains, with resistant reaction on the recommended concentration (250μg a.i. ml⁻¹) and tolerance or resistance on higher concentrations (500 and 1000μg a.i. ml⁻¹). Sensitivity differed between the CPM species. Whereas practically allPx isolates (except one from 2003) were resistant, 12% ofGc isolates from the years 2001–2003 showed sensitive and/or tolerant reactions. In 2004, only benomyl-resistantGc strains were detected. Variation in tolerance/resistance was detected to all screened fungicides during the course of this study at some repeatedly sampled locations. http://www.phytoparasitica.org posting June 12, 2008.     

Fungicide resistance status and chemical control options for the brassica pathogen Pyrenopeziza brassicae

Pyrenopeziza brassicae causes leaf spot disease of Brassicaceae in Europe/Oceania (lineage 1) and North America (lineage 2). In Europe, fungicides currently used for disease management are sterol 14α-demethylase (CYP51) inhibitors (azoles), quinone outside inhibitors (QoIs), and succinate dehydrogenase inhibitors (SDHIs); methyl benzimidazole carbamates (MBCs) are no longer applied. In this study, in vitro screening revealed European populations (collected 2018–2020) had shifted towards decreased azole sensitivity, but the North American population (2014–2016) was highly sensitive. Genotyping revealed CYP51 substitutions G460S or S508T were prevalent in European populations, often with a CYP51 promoter insert. Compared to wildtype CYP51 isolates, those with G460S plus an insert (44/46/151/210/302 bp) were c.25–32-fold and c.50-fold less sensitive to tebuconazole and prochloraz, respectively; those with S508T plus an insert (44/46/151/233 bp) were c.9–15-fold and c.25–40-fold less sensitive to tebuconazole and prochloraz, respectively. Selection for G460S (quantified via pyrosequencing) under different fungicide regimes was investigated in UK field trials, but G460S levels were high (c.76%) before treatment, so further selection during the trials was unclear. Despite the high G460S frequency and low disease pressure, yield data indicated measurable benefit for both azole- and non-azole-based programmes. In vitro screening against the MBC carbendazim showed European populations were predominantly moderately resistant/resistant; the North American population was sensitive. European and North American populations were sensitive to QoI (pyraclostrobin) and SDHI (penthiopyrad) fungicides. Results support an azole plus QoI/SDHI mixing partner for robust disease control and decreased risk of resistance, with continued sensitivity monitoring to ensure optimal strategies are deployed.     

Fungicide resistance and genetic diversity of Botrytis cinerea isolates from a vineyard in Germany

Journal of Plant Diseases and Protection - To evaluate the genetic diversity and the possibility of changes in the population of the grey mould fungus in a single vineyard, a total of 209 Botrytis...     

Fungicide resistance in diseases of vegetables growing under plastic in Crete1

Serious fungicide resistance problems have been encountered in several major vegetable pathogens in glasshouses in Crete. Botrytis cinerea populations are now widely resistant to either or both benzimidazoles and dicarboximides. Pseudoperonospora cubensis very rapidly became resistant to metalaxyl, with cross resistance to other acylalanines. Didymella bryoniae has shown some benzimidazolc resistance. Sphaerotheca fuliginea, against which a great diversity of fungicides is used, has mostly not given problems. However, strains with resistance to benomyl, triforine and ditalimfos have been found in experimental plots. Ways of managing resistance in Crete are discussed.     

Fungicide resistance status in French populations of the wheat eyespot fungi Oculimacula acuformis and Oculimacula yallundae

Eyespot, caused by Oculimacula acuformis and Oculimacula yallundae, is the major foot disease of winter wheat in several European countries, including France. It can be controlled by chemical treatment between tillering and the second node stage. The fungicides used include antimicrotubule toxicants (benzimidazoles), inhibitors of sterol 14α‐demethylation (DMIs) or of succinate dehydrogenase (SDHIs), the anilinopyrimidines cyprodinil and the benzophenone metrafenone. Since the early 1980s, a long‐term survey has been set up in France to monitor changes in the sensitivity of eyespot populations to fungicides. Resistance to benzimidazoles has become generalised since the early 1990s, in spite of the withdrawal of this class of fungicides. In the DMI group, resistance to triazoles is generalised, whereas no resistance to the triazolinethione prothioconazole has yet developed. Resistance to the imidazole prochloraz evolved successively in O. acuformis and O. yallundae and is now well established. Specific resistance to cyprodinil has also been detected, but its frequency has generally remained low. Finally, since the early 2000s, a few strains of O. yallundae displaying multidrug resistance (MDR) have been detected. These strains display low levels of resistance to prothioconazole and SDHIs, such as boscalid. Knowledge of the spatiotemporal distribution in France of O. acuformis and O. yallundae field strains resistant to fungicides allows resistance management strategies for eyespot fungi in winter wheat to be proposed.© 2012 Society of Chemical Industry     

Fungicide resistance inBotrytis cinerea isolates from strawberry crops in Huelva (southwestern Spain)

The severity of disease caused byBotrytis cinerea in strawberries is very high and chemical control is common practice; low residue levels of chemical products are required. Thus, it is important to be aware of the development of fungicide resistance in order to choose the best strategies of chemical control. In the present study we evaluated the response of 36B. cinerea isolates against eight different fungicides. The isolates were sampled twice, at the beginning and the end of the season, in 11 commercial strawberry fields located in the area of Huelva (Spain). In addition, two reference isolates, SAS56 and SAS405, were evaluated. The proportion of isolates resistant to benomyl was very high (86%). Resistance to dicarboximides was detected in 44% of the isolates and resistance to pyrimethanil in 25% of the isolates. Different degrees of sensitivity to captan and dichlofluanid were recorded. No resistance was found to diethofencarb plus carbendazim. http://www.phytoparasitica.org posting Sept. 18, 2002.     

Fungicide resistance in Cercospora species causing cercospora leaf blight and purple seed stain of soybean in Argentina

Cercospora species cause cercospora leaf blight (CLB) and purple seed stain (PSS) on soybean. Because there are few resistant soybean varieties available, CLB/PSS management relies heavily upon fungicide applications. Sensitivity of 62 Argentinian Cercospora isolates to demethylation inhibitor (DMI), methyl benzimidazole carbamate (MBC), quinone outside inhibitor (QoI), succinate dehydrogenase inhibitor (SDHI) fungicides, and mancozeb was determined in this study. All isolates were sensitive to difenoconazole, epoxiconazole, prothioconazole, tebuconazole, and cyproconazole (EC₅₀ values ranged from 0.006 to 2.4 µg/ml). In contrast, 51% of the tested isolates were sensitive (EC₅₀ values ranged from 0.003 to 0.2 µg/ml), and 49% were highly resistant (EC₅₀ > 100 µg/ml) to carbendazim. Interestingly, all isolates were completely resistant to azoxystrobin, trifloxystrobin, and pyraclostrobin, and insensitive to boscalid, fluxapyroxad, and pydiflumetofen (EC₅₀ > 100 µg/ml). The G143A mutation was detected in 82% (53) of the QoI-resistant isolates and the E198A mutation in 97% (31) of the carbendazim-resistant isolates. No apparent resistance mutations were detected in the succinate dehydrogenase genes (subunits sdhB, sdhC, and sdhD). Mancozeb completely inhibited mycelial growth of the isolates evaluated at a concentration of 100 µg/ml. All Argentinian Cercospora isolates were sensitive to the DMI fungicides tested, but we report for the first time resistance to QoI and MBC fungicides. Mechanism(s) other than fungicide target-site modification may be responsible for resistance of Cercospora to QoI and MBC fungicides. Moreover, based on our results and on the recent introduction of SDHI fungicides on soybean in Argentina, Cercospora species causing CLB/PSS are insensitive (naturally resistant) to SDHI fungicides. Insensitivity must be confirmed under field conditions.     

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.     

杀菌剂毒力及其生物测定

杀菌剂毒力是化合物具有开发和应用价值的固有生物学性状,不仅是农药登记部门予以注册登记的前置条件,而且也是反映杀菌剂生物活性及其安全性和有效性的关键技术参数。本文明确了杀菌剂毒力术语的定义,强调了杀菌剂可能的未知作用方式及通过与病原菌、寄主和环境因子相互作用的病害及流行防控效力在杀菌剂毒力评价中的重要性。深入分析了化合物与受体蛋白相互作用的结构特异性和精确性及干扰靶标蛋白功能的毒理学机制,揭示了杀菌剂毒力的选择性原理。综述了杀菌剂毒力生物测定及其结果分析方法,分析了可能影响毒力测定结果评价的常见因素。本文所述内容可为从事农药创制、加工、应用、毒理与抗性及分子互作科研工作者提供参考。     

Integration of host resistance and fungicide use1

The current strategies for using host resistances and fungicides encourage the increase of pathogen strains with matching pathogenicity or insensitivity. Diversification of cultivars and fungicides is recommended, but involves only diversification between crops and does not include integration of the two methods of control. A strategy of complete or total diversification requires diversification within as well as between crops. For example, the system of using cultivar mixtures can provide assured high yields, even though control of pathogen development is incomplete. Within-crop diversification of fungicides applied to seed is becoming a practical possibility with the development of film-coating of carriers that can adhere different materials firmly to different seeds whilst maintaining uniform seed-flow characteristics. An ideal system would involve integrated total diversification of host cultivars and fungicides. Field data indicate that this can provide excellent profit margins and disease control, while maximising evolutionary problems for the pathogen. The system is technically feasible using developed methods of cultivar mixing together with the new technique of film-coating.     

Fungicide strategies and resistance of Ramularia collo-cygni to demethylation and succinate dehydrogenase inhibitors in Austrian winter barley (Hordeum vulgare)

Journal of Plant Diseases and Protection - Ramularia collo-cygni B. Sutton and J.M. Waller is a major disease in Austrian barley-growing regions. To date, fungicide application is the most...     

枇杷炭疽病病原菌鉴定及防治药剂筛选

[目的]枇杷是我国重要的经济作物,种植广泛,具有重要的食用和药用价值.炭疽病是枇杷上的一种重要病害,严重影响枇杷果实的产量和品质.引起枇杷炭疽病的病原菌种类多样,防治困难.[方法]为明确江西省宜春市枇杷炭疽病的病原种类,获得有效的防治药剂,研究采用形态学和分子生物学方法对分离的病原菌进行了鉴定,利用菌丝生长速率法测定了6种杀菌剂对枇杷炭疽病菌的室内毒力.[结果]引起江西宜春上高县枇杷炭疽病的病原菌为果生炭疽菌(Colletotrichum fructicola).所选的6种药剂对枇杷炭疽病菌的生长均有一定的抑制作用,其中申嗦霉素抑菌效果最好,EC50为0.014 mg/L,其次为30％苯甲·嘧菌酯、50％咪鲜胺锰盐、400 g/L氟硅唑,其EC50分别为0.020,0.021,0.026 mg/L.[结论]研究结果可为枇杷炭疽病菌的致病机理及病害的综合防治提供理论基础.     

三种三唑类杀菌剂对映体生物活性的研究

采用高效液相色谱法(HPLC)拆分了戊唑醇(tebuconazole)、粉唑醇(flutriafol)和己唑醇(hexaconazole)3种杀菌剂的外消旋体,并且制备了对映体的纯品。以花生褐斑病菌Cercospora arachidicola、番茄早疫病菌Alternaria solani、油菜菌核病菌Sclerotinia sclerotiorum、苹果轮斑病菌Alternaria mali、甜菜褐斑病菌Cercospora beticola等5种植物病原菌为供试菌,采用琼脂稀释法测定了3种杀菌剂的外消旋体及对映体的生物活性,比较了对映体之间抗菌活性的差异。结果表明:(-)-戊唑醇的抗菌活性优于(+)-戊唑醇;(+)-粉唑醇的抗菌活性优于(-)-粉唑醇;(-)-己唑醇的抗菌活性优于(+)-己唑醇。