The drug transporter MgMfs1 can modulate sensitivity of field strains of the fungal wheat pathogen Mycosphaerella graminicola to the strobilurin fungicide trifloxystrobin

BACKGROUND: The major facilitator superfamily (MFS) drug transporter MgMfs1 of the wheat pathogen Mycosphaerella graminicola (Fuckel) J Schroeter is a potent multidrug transporter with high capacity to transport strobilurin fungicides in vitro. The data presented in this paper indicate that, in addition to the predominant cause of strobilurin resistance, cytochrome b G143A subsititution, MgMfs1 can play a role in sensitivity of field strains of this pathogen to trifloxystrobin. RESULTS: In a major part of field strains of M. graminicola (collected in the Netherlands in 2004) containing the cytochrome b G143A substitution, the basal level of expression of MgMfs1 was elevated as compared with sensitive strains lacking the G143A substitution. Induction of MgMfs1 expression in wild-type isolates upon treatment with trifloxystrobin at sublethal concentrations proceeded rapidly. Furthermore, in disease control experiments on wheat seedlings, disruption mutants of MgMfs1 displayed an increased sensitivity to trifloxystrobin. CONCLUSION: It is concluded that the drug transporter MgMfs1 is a determinant of strobilurin sensitivity of field strains of M. graminicola.     

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.     

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.     

The effect of dose and mobility on the strength of selection for DMI fungicide resistance in inoculated field experiments

Plots of spring wheat cv. Baldus were inoculated at GS 13 with four Mycosphaerella graminicola isolates, two relatively susceptible and two relatively resistant to DMI fungicides. Changes in the ratio of relatively susceptible to resistant types following fungicide or water sprays were measured. Three fungicides were compared: flutriafol, which is very mobile within leaves, fluquinconazole, which is less so, and prochloraz, which is almost immobile. All are inhibitors of sterol demethylation. In 1996, fungicide-treated plots were sprayed once with half the recommended dose at GS 39–47. In 1997, three doses were used: one-quarter and one-eighth of the recommended dose and a dual application of two one-eighth recommended doses, a week apart. Isolates were classified using a discriminating dose assay and the ratio of relatively susceptible to relatively resistant isolates in each field plot before and after fungicide application calculated. In both years, the numbers of relatively susceptible and relatively resistant isolates were equal just before fungicide application. All fungicides caused significant selection towards resistance, but the strength of selection varied with fungicide, dose and position in the crop canopy. Fluquinconazole selected most strongly and gave the best control of disease. Interactions between fungicide and dose were not significant. Selection was equally strong all along leaves sprayed with prochloraz, but increased smoothly from base to tip of leaves sprayed with fluquinconazole or flutriafol. Averaged over fungicides, reducing the dose of a single fungicide application from one-quarter to one-eighth slightly reduced selection towards resistance on both leaf layers. The dual one-eighth dose caused twice the change of the single one-eighth dose on the flag leaf, but was similar to a single spray on leaf 2.     

Modelling the effects of sub-lethal doses of herbicide and nitrogen fertilizer on crop–weed competition

The effects of sub‐lethal dose of herbicide and nitrogen fertilizer on crop–weed competition were investigated. Biomass increases of winter wheat and a model weed, Brassica napus, at no‐herbicide treatment with increasing nitrogen were successfully described by the inverse quadratic model and the linear model respectively. Increases in weed competitivity (β₀) of the rectangular hyperbola and parameter B in the dose–response curve for weed biomass, with increasing nitrogen were also successfully described by the exponential model. New models were developed by incorporating inverse quadratic and exponential models into the combined rectangular hyperbola with the standard dose–response curve for winter wheat biomass yield and the combined standard dose—response model with the rectangular hyperbola for weed biomass, to describe the complex effects of herbicide and nitrogen on crop–weed competition. The models developed were used to predict crop yield and weed biomass and to estimate the herbicide doses required to restrict crop yield loss caused by weeds and weed biomass production to an acceptable level at a range of nitrogen levels. The model for crop yield was further modified to estimate the herbicide dose and nitrogen level to achieve a target crop biomass yield. For the target crop biomass yield of 1200 g m^?2 with an infestation of 100 B. napus plants m^?2, the model recommended various options for nitrogen and herbicide combinations: 140 and 2.9, 180 and 0.9 and 360 kg ha^?1 and 1.7 g a.i. ha^?1 of nitrogen and metsulfuron‐methyl respectively.     

Variation in the response of cashew genotypes to the targeted application of fungicide to flower panicles for control of powdery mildew disease

Application of the fungicide triadimenol (Bayfidan) directly to inflorescences of cashew was investigated as a means of controlling powdery mildew disease caused by Oidium anacardii. Disease development and nut production were studied in 12 cashew genotypes that differed in their susceptibility to mildew. Panicle colonization by O. anacardii reached 100% coverage in all genotypes without fungicide treatment, but rates of infection differed significantly. Triadimenol sprays reduced mildew to less than 9%, even in panicles of highly susceptible genotypes. In the absence of disease, particularly good yield responses with more than nine times more nuts set than untreated controls were achieved by AM6 and AC1, which were categorized as highly susceptible and intermediate, respectively, in reaction to powdery mildew. By contrast, the partially resistant genotypes AZA2 and AC6 both produced yield response ratios of less than 3·0. The targeted treatment of flower panicles to control mildew is recommended, rather than the current practice of wastefully treating whole trees on which all mature leaves are naturally immune to infection.     

防治玉米叶斑病高效药剂筛选及药剂减施增效技术

为有效防治玉米小斑病和弯孢叶斑病并减少化学农药使用量,本研究采用菌丝生长速率法从11种杀菌剂中选取抑菌效果较好的8种药剂进行田间小区药效试验。同时选取对两种叶部病害兼具防效的最佳药剂再与中量元素水溶肥、腐植酸、氨基寡糖素、芸苔素内酯等产品混配开展田间药效试验,确定该药剂田间施用浓度和增效组合。结果显示,各供试药剂对2种病菌菌丝生长具有不同的抑制作用,其中25%丙环唑EC、25%戊唑醇WP、40%氟硅唑EC、30%氟菌唑WP、12.5%烯唑醇WP在供试各浓度下对2种病菌抑制率均为100%,田间小区药效筛选测定中,25%戊唑醇WP防治效果最好,对玉米小斑病和弯孢叶斑病两种叶部病害防效分别为81.07%和72.22%。田间试验中,25%戊唑醇WP防治玉米小斑病和弯孢叶斑病的最佳施用浓度为2 000倍液,防效分别为59.48%和53.96%。在减量50%情况下防效有所下降,配施氨基寡糖素后,防效可提高37.40百分点和22.98百分点,与最佳施用浓度防效相当,达到了减药增效的作用。     

Testing of an integrated regime for effective and sustainable control of invasive Crofton weed (Ageratina adenophora) comprising the use of natural inhibitor species,activated charcoal,and fungicide

Crofton weed is a major invasive species in China. It exhibits superior growth characteristics and can outcompete with native species via allolepathic effects and modulation of the soil fungal microbiome. The simple removal of invading plants will not ensure restoration of the habitat due to the persistence of allelochemicals and viable seeds in the surrounding soil. An orthogonal experimental design was employed to evaluate the effects of three control factors (A, powdered natural inhibitor species to retard growth; B, activated charcoal to absorb allelochemicals; and C, fungicide to reduce fungal modulation effects), applied at three levels, on the growth and competitive ability of Crofton weed against two native species, in a pot‐culture experiment. All treatments reduced all measured growth parameters (P < 0.05) except for a specific leaf area, when compared with control plants. Furthermore, the competitive capacity of Crofton weed was decreased in the treatments while that of the native species was improved. Application to soil of the powdered natural inhibitor species and of activated charcoal significantly inhibited plant growth and competitive ability of Crofton weed (P < 0.05). Application of fungicide was less effective, but significantly reduced the specific leaf area of Crofton weed plants (P < 0.05). The specific combination of factors producing the greatest decrease in plant growth and competitive ability (compared with the control) included the addition of Delavaya toxocarpa powder (37.5 g per kg soil), addition of activated charcoal to soil at a ratio of 1:3 (v/v) (62.5 g per kg soil), and application of fungicide (Thiophanate‐Methyl) (0.28 g per kg soil).