Microscopic analysis of the effect of azoxystrobin treatments on Mycosphaerella graminicola infection using green fluorescent protein (GFP)‐expressing transformants

Green fluorescent protein (GFP)‐expressing transformants were used to investigate the effects of strobilurin fungicide azoxystrobin on Mycosphaerella graminicola infection. Azoxystrobin treatments (125 or 250 g AI ha^?1) were applied at various stages of the infection process under controlled conditions. GFP transformants showed conserved in vitro sensitivity to azoxystrobin and pathogenicity. Azoxystrobin controlled over 90% of M graminicola infections when applied before or during penetration of the pathogen (15% of the incubation phase). Azoxystrobin also impaired the growth of intercellular hyphae in M graminicola post‐penetration infection stages when applied at up to 50% of the incubation phase. Incubating infections observed in treated leaves were viable, but their growth was impaired and they did not induce necrosis under controlled conditions. Reduction by half of azoxystrobin dosage had little or no effect on azoxystrobin efficiency in controlling M graminicola. The contribution of post‐penetration fungistatic effect to azoxystrobin curative properties toward M graminicola in a field situation is discussed. © 2001 Society of Chemical Industry     

大麦云纹斑病菌对杀菌剂的抗性检测及同工酶谱类型

1993～1994年大麦云纹斑病菌(Rhynchosporium secalis)对杀菌剂多菌灵、三唑醇的抗药性监测表明,该菌对两种杀菌剂的敏感程度发生了变化,并且首次在田间发现了大麦云纹斑病菌的多抗菌株。这类菌株对多菌灵、乙霉威及三唑醇都具有抗药性,且抗性程度很高。与1990～1991年得到的资料相比,随着杀菌剂选择压力的增加,大田群体中三唑醇高抗菌株比例逐年增加。对该菌同工酶分析结果表明,α—酯酶有7种表现型,磷酸葡萄糖变位酶有2种表现型,过氧化氢酶有3种表现型,无论对去甲基抑制剂类(DMI)表现抗性或敏感菌株,无论调节酶还是非调节酶,其酶谱表现型的变异基本相似。这表明大麦云纹斑病菌对DMI类抗药性突变存在于多个菌系的基因型背景群体中,而不是只起源于单一的广泛分布的菌系。     

The effect of timing of fungicide applications on control of frogeye leaf spot and grain yield of soybeans

Soybean cultivar Samsoy 1, and the breeding lines TGx 849-313D and TGx 996-26E, grown in a field with a heavy epidemic of frogeye leaf spot caused byCercospora sojina, were treated with double foliar applications of the fungicide benomyl. The treatments were made using four application schedules at six different growth stages, starting from V₃ (fully developed leaves, beginning with trifoliate nodes) to R₅ (beginning seed_, to determine the effect of the fungucide timing on frogeye leaf spot severity, soybean grain yield and grain quality. Generally, applications at R₁ (beginning bloom) and R₃ (beginning pod) significantly (P<-0.05) reduced disease severity in the 2 susceptible genotypes, Samsoy 1 and TGx 849-313D. Plot yields of these genotypes were also significantly greater than the untreated controls when the fungicide applications were made at R₁ and R₃. There was no significant difference in diseave severity or grain yield, between the untreated control and the different times of application, on the resistant genotype TGx 996-26E. Improved seed germination and lower levels of seed infection byC. sojina occurred for all fungicide timings in the susceptible genotypes. The results suggest that fungicide spraying initiated at R₁ and followed up at R₃ is most effective in frogeye leaf spot control and can also result in higher grain yields, than applications made earlier or later in the season. Control of frogeye leaf spot, however, is best achieved by growing resistant cultivars.     

Field validation of a decision support system for the control of pests and diseases in cereals in denmark

The decision in 1986, on an action plan to reduce pesticide use in Denmark by 50% led to increased research on the potential of reducing dosages. A decision support system (PC-Plant Protection), developed by The Danish Institute for Plant and Soil Science, implements this research. It combines a detailed use of threshold values to support decisions on treatment need, choice of pesticides and the appropriate dosage for actual problems in cereals. The pest and disease module within the system has been available commercially since 1993 and up to the end of April 1995, 2000 licences have been issued for its use at agricultural schools and by advisers and farmers. The recommendation model for pest and disease control has been validated in field trials since 1990. The validation has shown that the model is able to provide recommendations for the control of pests and diseases to a satisfactory level, without affecting farmers' gross margins. The model was able to adjust pesticide use to large yearly variations and the average amount of pesticides in the plots treated according to the model was well below that in the reference plots and in the commonly used strategies in Denmark.     

Polymorphism of 14α-demethylase Gene (CYP51) in the Cereal Eyespot Fungi Tapesia acuformis and Tapesia yallundae

Cereal eyespot fungi Tapesia acuformis and Tapesia yallundae are closely related species which show different behaviours upon treatment with sterol 14-demethylase inhibitors (DMIs). T. acuformis is naturally resistant to DMIs belonging to the triazole family and susceptible to the imidazole ones, whilst T. yallundae is sensitive to both inhibitors. Cloning of the target enzyme gene, CYP51, from the two species revealed an important polymorphism between them. Further sequencing of CYP51 from sixteen T. acuformis and eleven T. yallundae strains with different phenotypes with regards to resistance to DMIs confirmed that at least eleven variations are species related. Among them, a conserved phenylalanine residue at position 180, found both in T. yallundae and in all known CYP51 proteins from filamentous fungi and yeast, was replaced in T. acuformis by a leucine. Therefore, a leucine at 180 could be possibly involved in natural resistance of T. acuformis to triazoles. Other mutations were observed in some resistant strains, sometimes simultaneously, but in contrast to what was reported for other filamentous fungi, where a mutation at the 136 position of the CYP51 gene product seemed to correlate with resistance to DMIs, we did not find a clear relationship between a given mutation and a particular phenotype. This result suggests that resistance to DMIs could have a polygenic nature in Tapesia. We took advantage of species-related variations to develop a PCR-based assay allowing rapid and easy discrimination between field strains of the two species.     

Synergistic interaction between BABA and mancozeb in controlling<Emphasis Type="Italic">Phytophthora infestans</Emphasis> in potato and tomato and<Emphasis Type="Italic">Pseudoperonospora cubensis</Emphasis> in cucumber

Spray mixtures consisting of the plant activator BABA (DL-3-aminobutyric acid) and the protectant fungicide mancozeb were significantly more effective than BABA or mancozeb alone in controlling late blight (Phytophthora infestans) in potato and tomato and downy mildew (Pseudoperonospora cubensis) in cucumber. A mixture composed of 5 parts BABA and 1 part mancozeb (w/w, a.i.) exhibited a higher synergy factor than the 1+1 or the 1+5 (BABA + mancozeb) mixtures. No synergistic interaction was detected between BABA plus mancozeb in controlling sporangial or cystospore germination, nor mycelial growth ofP. infestans in vitro. The results showed enhanced effect of mancozeb in BABA-induced plants, suggesting, therefore, that lower dosages of this fungicide may be sufficient to control late blight or downy mildew under field conditions. http://www.phytoparasitica.org posting July 15, 2003.     

Diagnosis of dehydratase inhibitors in melanin biosynthesis inhibitor (MBI-D) resistance by primer-introduced restriction enzyme analysis in scytalone dehydratase gene of Magnaporthe grisea

We have established a simple diagnosis method for rice blast fungus resistant to MBI-D. This involves the preparation of PCR templates directly from the lesions in combination with primer-introduced restriction enzyme analysis PCR (PIRA-PCR).     

Genetic Control of Resistance to the Piperidine Fungicide Piperalin in Ustilago maydis

Mutants of Ustilago maydis resistant to the piperidine fungicide piperalin were isolated in a mutation frequency of 2.4 × 10^–5 after UV-irradiation and selection on media containing 50gml^–1 piperalin. Genetic analysis with 15 such mutant isolates resulted in the identifications of two unlinked chromosomal loci, the U/ppl-1 locus with two allelic genes (U/ppl-1A and U/ppl-1B) and the U/ppl-2 locus. The U/ppl-2 and U/ppl-1A mutations are responsible for two levels of moderate and high resistance to piperalin (resistance factor, Rf: 54 and 135, respectively, based on effective concentration causing a 50% reduction in the growth rate, EC₅₀), while the U/ppl-1B mutation gives only a small reduction (approximately 8-fold) in piperalin sensitivity. Cross-resistance studies with other SBIs shows that the major gene (U/ppl-2 and U/ppl-1A) mutants are resistant to fenpropidin (Rf: 43 and 68), fenpropimorph (Rf: 261 and 283) and tridemorph (Rf: 9 and 10), but not to the inhibitors of C-14 demethylase (DMIs) and squalene epoxidase. The minor gene mutation U/ppl-1B codes a low-level of resistance (approximately 5—12-fold) to the above morpholine-type fungicides, but in contrast with the major gene mutations it increases 2–10 times the sensitivity to triazoles: triadimefon, triadimenol, propiconazole and flusilazole. Crosses between mutants carrying the U/ppl-genes with compatible isolates carrying the U/fpd, U/fpm or U/tdm mutations, which have been identified in previous genetic works for resistance to morpholine-type fungicides, yielded, with the exception of U/ppl-2 ×U/fpm-2 cross, a large number of recombinants with wild-type sensitivity, indicating that the mutant genes involved in these crosses, were not allelic. An additive gene effect was observed only between nonallelic minor genes U/ppl-1B and U/fpm-1B or U/tdm-1,2. Studies of the fitness of piperalin-resistant isolates showed that the reduced sensitivity of major gene mutants was not associated with changes on the phytopathogenic fitness determining characteristics, such as growth in liquid culture and pathogenicity on young corn plants. Conversely, the minor gene mutation U/ppl-1B appeared to be pleiotropic, having significantly adverse effects on the phytopathogenic fitness.     

一种用于香蕉叶斑病防治的新型施药技术

试验一种将内吸性农药稀释后灌注于香蕉叶柄基部防治香蕉叶斑病的新型精确定量施药技术。结果表明，这种施药技术与常规施药技术相比，防效相当，且对香蕉的生长无不良影响，而农药成本仅为常规防治的18%，农药利用率提高4倍，值得推广应用。     

Chemical Manipulation of Reproductive Efficiency of Chickpea (Cicer arietinnum L.) by Triadimefon in Relation to Changes in Ethylene

Reproductive efficiency was successfully manipulated by the application of different concentrations of triadimefon at pre‐flowering, flowering and post‐flowering stages in chickpea (Cicer arietinum L.). Triadimefon improved the parameters determining reproductive efficiency. Yield was significantly improved due to reductions in flower and pod abscission by the application of triadimefon, which also lowered the ethylene levels in leaves, flowers and pods and delayed senescence. These findings suggest a relationship between ethylene level and reproductive efficiency in chickpea. Further, they indicate that triadimefon can be used for chemical manipulation of the reproductive efficiency of chickpea under field conditions.