多菌灵等5种杀菌剂对烟草立枯病菌的生物活性

采用生物测定方法分析了烟草立枯病菌Rhizoctonia solani在菌丝生长、菌核形成与萌发阶段对5种杀菌剂(多菌灵、代森锰锌、菌核净、异菌脲及苯醚甲环唑)的敏感性,同时通过离体叶片法考察了5种杀菌剂对烟草立枯病的保护和治疗作用。结果表明:供试5种药剂对立枯病菌菌丝生长、菌核形成与萌发均表现出了不同程度的抑制活性,同时对烟草立枯病具有一定的保护和治疗作用。其中,对菌丝生长以及菌核形成与萌发抑制活性最强的均是多菌灵,其抑制菌丝生长的EC50平均值为(0.06 ±0.01) mg/L,0.25和20 mg/L下对菌核形成和萌发的抑制率均为100%;其次为异菌脲[EC50值为(0.35 ±0.15) mg/L,2和100 mg/L下对菌核形成和萌发的抑制率分别为(54.79 ±12.58)%和100%]、苯醚甲环唑[EC50值为(0.55 ±0.53) mg/L,5和200 mg/L下对菌核形成和萌发的抑制率分别为(97.97 ±2.64)%和100%]及菌核净[EC50值为(1.31 ±0.14) mg/L,10和100 mg/L下对菌核形成和萌发的抑制率均为100%];最弱的均是代森锰锌,其抑制菌丝生长的EC50平均值为(6 ±0.20) mg/L,10和200 mg/L下对菌核形成和萌发的抑制率分别为(43.58 ±31.87)%和0。离体试验表明:对烟草立枯病保护作用最强的是多菌灵、异菌脲和菌核净,50 mg/L时防效均 > 99%,其次为代森锰锌(50 mg/L防效为75.83%),最弱的是苯醚甲环唑(50 mg/L防效为39.29%);治疗作用最强的也是多菌灵,50 mg/L防效为93.46%,其次为菌核净、异菌脲和苯醚甲环唑(200 mg/L时防效均 > 84%),最弱的为代森锰锌(800 mg/L时防效为76.40%)。研究结果可为烟草立枯病化学防治药剂筛选提供参考和依据。     

In vitro inhibition of Sclerotinia sclerotiorum by mixtures of azoxystrobin, SHAM, and thiram

The necrotrophic fungal phytopathogen Sclerotinia sclerotiorum (Lib.) de Bary has a broad host range and frequently causes destructive diseases. The extensive use of common fungicides to control these diseases has selected for resistance in populations of S. sclerotiorum. In this study, 105 isolates of S. sclerotiorum from different geographical regions in Jiangsu Province of China were characterized for baseline sensitivity to azoxystrobin, and the average EC₅₀ value was 0.2932 μg/mL for mycelial growth. Of the mixtures of the fungicides thiram and azoxystrobin that were tested using an in vitro mycelial growth assay, the 1:4 ratio provided the greatest inhibition of S. sclerotiorum. When tested against nine isolates, the 1:4 mixture resulted in a mean synergy ratio of 2.31, indicating synergistic inhibition. Mycelial respiration was inhibited for about 2 h by azoxystrobin alone but for 48 h by the mixture of thiram and azoxystrobin. Salicylhydroxamic acid (SHAM, a known inhibitor of alternative respiration) also increased the inhibition of mycelial growth and respiration caused by azoxystrobin. These results suggest the need for further study of effects of combinations of azoxystrobin with thiram or SHAM in planta to evaluate their potential for management of diseases caused by S. sclerotiorum.     

Biological control of sclerotinia stem rot of canola using antagonistic bacteria

Stem rot caused by Sclerotinia sclerotiorum is a major fungal disease of canola worldwide. In Australia the management of stem rot relies primarily on strategic application of synthetic fungicides. In an attempt to find alternative strategies for the management of the disease, 514 naturally occurring bacterial isolates were screened for antagonism to S. sclerotiorum. Antifungal activity against mycelial growth of the fungus was exhibited by three isolates of bacteria. The bacteria were identified as Bacillus cereus (SC‐1 and P‐1) and Bacillus subtilis (W‐67) via 16S rRNA sequencing. In vitro antagonism assays using these isolates resulted in significant inhibition of mycelial elongation and complete inhibition of sclerotial germination by both non‐volatile and volatile metabolites. The antagonistic strains caused a significant reduction in the viability of sclerotia when tested in a greenhouse pot trial with soil collected from the field. Spray treatments of bacterial strains reduced disease incidence and yielded higher control efficacy both on inoculated cotyledons and stems. Application of SC‐1 and W‐67 in the field at 10% flowering stage (growth stage 4·00) of canola demonstrated that control efficacy of SC‐1 was significantly higher in all three trials (over 2 years) when sprayed twice at 7‐day intervals. The greatest control of disease was observed with the fungicide Prosaro^® 420SC or with two applications of SC‐1. The results demonstrated that, in the light of environmental concerns and increasing cost of fungicides, B. cereus SC‐1 may have potential as a biological control agent of sclerotinia stem rot of canola in Australia.     

Effect of fungicides on stages of the life cycle of Phytophthora cinnamomi

An assessment was made of the effects of several fungicides on the formation of zoosporangia, the release of zoospores from zoosporangia, zoospore motility, germ tube formation and mycelial growth, by Phytophthora cinnamomi Rands. Compounds active against mycelial growth (such as metalaxyl and furalaxyl), as well as those active against zoospores (such as captafol and zineb), could be detected by a lupin (Lupinus angustifolius L.) assay which has been developed. This assay may be useful as a screening procedure to detect new fungicides.     

铁皮石斛黑斑病菌室内药效试验

通过供试的18种药剂对铁皮石斛黑斑病菌体外抑制作用测试结果表明,除克菌特外,所有药剂对其孢子萌发都有显著的抑制作用,其中代森锰锌、易保、代森锌和炭疽福美可完全抑制孢子萌发,霉克特、扑海英、使百克、新太生、甲基托布津和世高对孢子萌发有较强烈的抑制作用。与对照比较,克菌特促进孢子萌发率高于对照23.34%,克菌特和庄园乐都能促进芽管生长。18种药剂对石斛黑斑病菌菌丝体的抑制作用测定结果显示,所有药剂对菌丝生长都有显著的抑制作用,其中使百克和世高对菌丝体抑制效果最好,抑菌率都达91.28%,其次是施佳乐、福星和扑海英。     

Heat treatment effect on Cadophora luteo-olivacea of kiwifruit

Heat treatment represents an alternative method to fungicides to control latent fruit pathogens, such as Cadophora luteo-olivacea of kiwifruit. This study reports the efficacy of different hot water (HW) and hot air (HA) treatments (45, 50, 55, 65, and 70°C with durations of 10, 15 or 20, 5 or 10, 5, and 3 min, respectively) on the conidial and mycelial growth of fungal isolates by in vitro assays. Both treatments at 70°C were the most effective in inhibiting conidial and mycelial growth of C. luteo-olivacea isolates. Treatment efficacy for HW and HA was on average 88% and 71%, respectively, on mycelial growth, and 100% and 91.3%, respectively, on conidial growth. A significant reduction of xylanase and pectinase enzyme activities of the isolates was detected after HW and HA treatment (70°C × 3 min). In most cases, both treatments showed a total reduction, sometimes varying with the target isolate. HA treatment was the most efficient treatment for reducing the skin pitting incidence in in vivo experiments, showing an average efficacy of 46.6%. Results show that heat treatments are effective against C. luteo-olivacea, but the main challenge will be to develop the optimal time × temperature combinations.     

不同杀菌剂对黄瓜靶斑病菌的毒力作用特性比较

为筛选防治黄瓜靶斑病的有效药剂,采用菌丝生长速率法、孢子萌发法和芽管伸长法测定了19种杀菌剂对黄瓜靶斑病菌的毒力,探究了4种毒力较高杀菌剂对病原菌菌丝形态的影响。结果表明:氟啶胺、吡唑萘菌胺、咯菌腈和四霉素对黄瓜靶斑病菌菌丝生长、孢子萌发和芽管伸长各生长发育阶段均有较强的抑制活性,其中,4种药剂抑制菌丝生长的平均EC₅₀值分别为0.516 3、1.538、1.605和0.648 2 mg/L,抑制孢子萌发的平均EC₅₀值分别为0.164 4、1.359、1.127和0.002 5 mg/L,抑制芽管伸长的平均EC₅₀值分别为0.189 6、0.144 7、0.060 4和0.001 5 mg/L;啶酰菌胺、百菌清和肟菌酯3种不同作用机制杀菌剂对黄瓜靶斑病菌孢子萌发和芽管伸长的抑制活性较强,3种药剂抑制孢子萌发的平均EC₅₀值分别为1.564、0.373 0和0.021 3 mg/L,抑制芽管伸长的平均EC₅₀值分别为0.629 5、0.233 4和0.405 0 mg/L,但对菌丝生长的抑制作用较弱,平均EC₅₀大于16 mg/L;氟啶胺、吡唑萘菌胺、咯菌腈和四霉素可引起黄瓜靶斑病菌菌丝不同程度的异常生长,主要表现为菌丝隔膜间距变小、菌丝节间肿大、主菌丝上二次分枝发生频繁、菌丝变形黏连、菌丝新分枝处生长受抑制或分生孢子生长变形。研究表明,氟啶胺、吡唑萘菌胺、咯菌腈和四霉素对黄瓜靶斑病菌具有较高的毒力,在黄瓜靶斑病的田间防治中具有较好的开发应用潜力。     

Effects of some fungicides on <Emphasis Type="Italic">Isaria farinosa</Emphasis>, and <Emphasis Type="Italic">in vitro</Emphasis> growth and infection rate on <Emphasis Type="Italic">Planococcus citri</Emphasis>

The effects of some fungicides used against citrus diseases, on mycelial growth and conidial germination of Isaria farinosa (Holmsk.) Fries [Sordariomycetes: Hypocreales] and also on the pathogenicity of the fungus on citrus mealybug, Planococcus citri (Risso), were determined. Systemic fungicides such as tebuconazole, penconazole and nuarimol were the most effective as regards both conidial germination and mycelial growth. Protective fungicides such as captan, chlorothalonil, mancozeb and propineb inhibited conidial germination at between 1 and 5 μg ml⁻¹ concentration, but captan, chlorothalonil and propineb did not inhibit the mycelial growth at 5,000 μg ml⁻¹. Mancozeb inhibited mycelial growth between 2,500 and 5,000 μg ml⁻¹. Sulphur and copper oxychloride did not inhibit the fungus even at very high concentrations. Sulphur, copper oxychloride, fosetyl-al, chlorothalonil and carbendazim did not decrease the mortality percentage caused by I. farinosa. Tebuconazole, penconazole and mancozeb were the most effective and respectively reduced the mortality from 83% to 33%, 28% and 30% in the ovisacs, from 81% to 29%, 27% and 29% in the 1st instar larvae, and from 84% to 34% in the adult females.     

烟草赤星病菌对苯醚甲环唑和氟环唑敏感性测定及田间防效试验

为明确山东省主要烟草产区烟草赤星病菌Alternaria alternata对苯醚甲环唑和氟环唑的敏感性,采用菌丝生长速率法检测苯醚甲环唑和氟环唑对86株烟草赤星病菌菌株的EC_(50),用Spearman’s秩相关系数法分析供试菌株对2种杀菌剂敏感性的相关性,并评价其对烟草赤星病的田间防效。结果表明,苯醚甲环唑对烟草赤星病菌菌丝生长EC_(50)值介于5.3883～8.0316 mg/L之间,平均值为6.9594 mg/L,敏感性频率分布呈连续性偏正态分布,田间烟草赤星病菌对苯醚甲环唑仍然较为敏感,可建立田间烟草赤星病菌对苯醚甲环唑的敏感基线。氟环唑对烟草赤星病菌菌丝生长的EC_(50)值介于0.1253～0.2517 mg/L,平均值为0.1773 mg/L,敏感性频率分布呈连续性偏正态分布,可建立田间烟草赤星病菌对氟环唑的敏感基线。相关性分析结果表明,烟草赤星病菌对苯醚甲环唑和氟环唑敏感性之间不存在明显的相关性。田间试验结果表明,30%苯醚甲环唑悬浮剂对烟草赤星病具有较好的防效,其有效成分150 g/hm~2处理的防效最高,为73.99%;12.5%氟环唑悬浮剂对烟草赤星病的最高防效为70.57%。表明2种三唑类杀菌剂对烟草赤星病防效较好。     

Detection and characterization of QoI resistance in Pyrenophora tritici-repentis populations causing tan spot of wheat in Argentina

Tan spot, caused by the fungus Pyrenophora tritici-repentis (Ptr), is a disease that has become more prevalent and intense in wheat crops in Argentina in recent years. Failure to control the disease with strobilurin fungicides, which were once effective, has been observed in different zones where wheat is grown. However, whether or not true resistance is present in the pathogen population in the region is not scientifically confirmed. This study evaluated the sensitivity of numerous Ptr isolates to representative QoI fungicides used in Argentina through in vitro and in planta assays, as well as through molecular analysis. Eighty-two monosporic isolates obtained in different locations in the north and south of Buenos Aires province in 2014, 2016, and 2018 were tested to determine sensitivity to selected QoI fungicides in conidial germination and mycelial inhibition assays, as well as in molecular analysis. Conidial germination was not inhibited at 1 µg/ml of azoxystrobin, trifloxystrobin, and pyraclostrobin. On the other hand, mycelial growth was inhibited by 59%, 56%, and 86% at 100 µg/ml of azoxystrobin, trifloxystrobin, and pyraclostrobin, respectively. The molecular analysis detected the G143A mutation in the cytb gene of all the 82 Ptr isolates, but the F129L and G137R substitutions were not present. This study documents the G143A mutation conferring QoI resistance in Ptr in South America. The findings of this study are key for future decisions regarding use of fungicide and rotation in the region.     

Possible methionine biosynthesis inhibition by pyrimidinamine fungicides

The broad-spectrum fungicide CGA 219417 inhibits mycelial growth of Botrytis cinerea Pers ex Fr. Pseudocercosporella herpotrichoides (Fron.) Deighton and Helminthosporium oryzae B. de Haan on defined media lacking amino acids. The growth inhibition of B. cinerea is reversed by the addition of a mixture of 19 amino acids at a concentration of 100 μM each or by the addition of methionine or homocysteine in concentrations of 100 μM or 1 mM, respectively. In the case of B. cinerea, the reversal of growth inhibition by methionine could also be shown for pyrimethanil and mepanipyrim. These findings suggest that the pyrimidinamine fungicides inhibit the biosynthesis of methionine in phytopathogenic fungi.     

Effets de divers herbicides sur la croissance mycélienne de souches de Botrytis cinerea pers et de Penicillium expansum Link, sensibles ou résistantes à certains fongicides

Effects of various herbicides on mycelial growth of strains of Botrytis cinerea. Pers. and Pénicillium expansum Link, sensitive or resistant to certain fungicides Of seventy herbicides tested, thirty-eight were slightly toxic to B. cinerea (CI50, concentration giving 50% inhibition of the speed of mycelial growth, exceeding or equal to 100g of herbicide/ml of nutrient solution) and, of those, ten had no effect even at 1000 g/ml. The most active products (CI50 less than 10 g/ml) were Chlorthiamid, dichlobénil, nitrofen, propyzamide and phenolic derivatives (DNOC, ioxynil, PCP). In most cases P. expansum was less sensitive than B. cinerea except to endothal, propachlor, prynachlor and certain substituted ureas. Strains resistant to the benzimidazole fungicides (carbendazim, thiabendazole etc.) show increased sensitivity to certain carbamade herbicides (barban, chlorbufam etc.), this indicates the existence of a negative cross resistance between these groups of antimitotic pesticides. Strains resistant to the cyclic-imide fungicides (iprodionc, procymidone, vinclozolin) and to various aromatic compounds (biphenyl. chloroneb, dicloran etc.) may also be resistant to bipyridilium, dinitroanilinc and diphenylethcr herbicides, to chlorthamid. dichlobénil and oxadiazon. This cross resistance between pesticides with different biochemical modes of action has yet to be explained.     

Inhibitory Effects of Fungicides on Hydrolysis of Urea and Nitrification of Urea Nitrogen in Soil

The influence of 1 and 50 mg active ingredient (AI) kg^-1 soil of 17 fungicides on transformations of urea nitrogen in soil was studied by determining the amounts of urea hydrolysed and the amounts of nitrate and nitrite produced when samples of two coarse-textured and two fine-textured soils were incubated aerobically for various times after treatment with urea. When applied at the rate of 1 mg AI kg^-1 soil, anilazine, benomyl, captan, chloranil, mancozeb and thiram retarded urea hydrolysis in the two coarse-textured soils and maneb retarded urea hydrolysis in all four of the soils used. Most of the fungicides tested retarded nitrification of urea nitrogen in the two coarse-textured soils when applied at the rate of 1 mg AI kg^-1 soil, but only etridiazole markedly retarded nitrification of urea nitrogen in all of the soils used when applied at this rate. When the fungicides were applied at the rate of 50 mg AI kg^-1 soil, anilazine, captan, chloranil, fenaminosulf, folpet, maneb, mancozeb and thiram retarded urea hydrolysis in the four soils studied, and all fungicides tested except chloroneb, fenarimol and iprodione retarded nitrification of urea nitrogen in these soils. One-way analysis of variance and correlation analyses indicated that the inhibitory effects of the 17 fungicides tested on nitrification of urea nitrogen in soil increased with decrease in the organic-matter content and increase in the sand content of the soil. © of SCI.     

Efficacy of phosphonate in controlling white powdery rot of fig caused by <Emphasis Type="Italic">Phytophthora palmivora</Emphasis>

Pre- and post-infection activities of phosphonate and fungicides (azoxystrobin, chlorothalonil, cyazofamid, copper hydroxide, and copper sulfate) against white powdery rot on fig leaves caused by Phytophthora palmivora were determined. Phosphonate and fungicides were applied at 3, 7, or 14 days before inoculation and 16, 24, or 40 h after inoculation. Phosphonate and four fungicides (except for chlorothalonil) had protectant activity for up to 14 days before inoculation. Protectant activity between phosphonate and the four fungicides did not differ significantly. These fungicides did not have a curative activity, whereas only phosphonate had a curative activity when applied 16 h after inoculation. The pre-infection activity of phosphonate was determined in fig fields. Phosphonate provided excellent protectant activity, and its activity did not differ significantly from that of the copper fungicides. The activity of phosphonate in the life cycle of P. palmivora was compared with that of the five fungicides. Half-maximal effective concentration of the inhibition of mycelial growth was low for phosphonate, chlorothalonil, and the two copper fungicides. Mycelial growth was least affected by azoxystrobin and cyazofamid, but n-propyl gallate, an alternative oxidase inhibitor, in combination with these fungicides, significantly inhibited mycelial growth. Sporangium formation was sensitive to all compounds. Germination of encysted zoospores was sensitive to azoxystrobin, cyazofamid, and chlorothalonil and least sensitive to phosphonate and the two copper fungicides. The activity of phosphonate on mycelial growth and sporulation of P. palmivora suggests it has potential for controlling the disease.     

六种杀菌剂对烟草赤星病菌菌丝生长和分生孢子萌发的抑制作用

采用菌丝生长速率法和孢子萌发法测定了烟草赤星病菌Alternaria alternata对6种杀菌剂(氟硅唑、异菌脲、菌核净、嘧菌酯、醚菌酯及多菌灵)的敏感性。结果表明:6种杀菌剂对烟草赤星病菌菌丝生长和分生孢子萌发均表现出了不同程度的抑制活性。其中,对菌丝生长抑制活性最强的是氟硅唑,EC50平均值为(0.34±0.11)mg/L,其次为菌核净[(1.13±0.68)mg/L]和异菌脲[(2.08±0.24)mg/L];对分生孢子萌发抑制活性最强的是嘧菌酯协同100 mg/L水杨肟酸,EC50平均值为(0.28±0.09)mg/L,其次分别为单独处理的嘧菌酯[(5.79±2.76)mg/L]、醚菌酯[(11.93±3.93)mg/L]、菌核净[(19.95±15.94)mg/L]和异菌脲[(23.33±11.33)mg/L]。多菌灵对烟草赤星病菌菌丝生长和分生孢子萌发的抑制活性均最差,不宜用于烟草赤星病的防治,而异菌脲、氟硅唑、嘧菌酯和醚菌酯均有必要进一步针对烟草赤星病进行田间防效试验。     

Frequency distribution of sensitivity of Ustilaginoidea virens to four EBI fungicides, prochloraz, difenoconazole, propiconazole and tebuconazole, and their efficacy in controlling rice false smut in Anhui Province of China

False smut, caused by Ustilaginoidea virens, is an important emerging disease of rice (Oryza sativa L.) in China. Up to now, as most varieties with high yielding and good quality are susceptible or even highly susceptible to false smut in most rice-growing ecological regions, especially in Anhui Province, chemical control with fungicides would be an important measure for the control of this disease. The ergosterol biosynthesis inhibitor (EBI) fungicides, such as prochloraz, difenoconazole, propiconazole and tebuconazole, are extensively used in China for the control of rice diseases, such as rice sheath blight and rice blast. In this study, a total of 102 U. virens isolates (from Anhui Province of China) were tested for their sensitivity to these four EBI fungicides during the stage of mycelial growth. The EC₅₀ ranges of values for prochloraz, difenoconazole, propiconazole and tebuconazole inhibiting mycelial growth of the 102 U. virens isolates were 0.04–0.75, 0.04–1.08, 0.04–0.38 and 0.03–0.57 μg?ml^?1, with the average EC₅₀ values of 0.32?±?0.08, 0.45?±?0.08, 0.19?±?0.03 and 0.21?±?0.06 μg?ml^?1, respectively. These values suggested that the tested U. virens isolates were very sensitive to these four EBI fungicides. Results of field trials showed that two sprays of three of the fungicides exhibited greater control efficacy than a single spray for the control of rice false smut. Two sprays of each was better than a single spray for the control of rice sheath blight. Two sprays of 50% propiconazole EC at 300 g a.i. ha^?1 gave the best control of rice false smut at both two sites during the two consecutive years, 2010 and 2011, with the control efficacy ranging from 71.5 to 74.3%. Sensitivity of the field U. virens isolates to EBI fungicides should be monitored. Mixtures, as well as alternation with other fungicides with different modes of action, should be tested.     

Baseline and differential sensitivity of Peronophythora litchii (lychee downy blight) to three carboxylic acid amide fungicides

Downy blight, caused by Peronophythora litchii, is an important disease of lychee (litchi) plants in China. The in vitro sensitivities of various asexual stages of P. litchii to the three carboxylic acid amide (CAA) fungicides dimethomorph, flumorph and pyrimorph were studied with four single‐sporangium isolates. None of the three fungicides affected zoospore discharge from sporangia, but they strongly inhibited mycelial growth (mean EC₅₀ values of 0·075, 0·258 and 0·115 mg L^?1, respectively); sporangial production (mean EC₅₀ values of 0·085, 0·315 and 0·150 mg L^?1, respectively); germination of cystospores (mean EC₅₀ values of 0·140, 0·150 and 0·645 mg L^?1, respectively); and germination of sporangia (mean EC₅₀ values of 0·203, 0·5 and 0·743 mg L^?1, respectively). As mycelial growth was the most sensitive stage to dimethomorph and pyrimorph, it was chosen to test baseline sensitivities to the three fungicides. In 2007, from 131 isolates collected in Fujian, Guangdong and Guangxi provinces, 127, 116 and 113 isolates were used to establish baseline sensitivity for dimethomorph, flumorph and pyrimorph respectively. Isolates from different provinces exhibited similar baseline sensitivity to the same fungicide. Baseline sensitivities to dimethomorph, flumorph and pyrimorph were distributed as unimodal curves, with mean EC₅₀ values of 0·082 (± 0·01), 0·282 (± 0·047), and 0·115 (± 0·032) mg L^?1, respectively. This information will serve as a baseline for tracking future changes in sensitivities of P. litchii populations to these three CAA fungicides.     

Detection and characterization of benzimidazole resistance of <Emphasis Type="Italic">Botrytis cinerea</Emphasis> in greenhouse vegetables

From 2003 to 2006, a total of 426 single-conidial isolates of B. cinerea collected from greenhouse vegetables in China were characterized for resistance to benzimidazole fungicides and diethofencarb according to inhibition of mycelial growth. Rapid development of double-resistance to benzimidazoles and diethofencarb was observed. Three types of benzimidazole-resistant isolates, Ben R1, Ben R2 and Ben R3 were detected. A new phenotype, Ben R3, which showed low level of resistance to benzimidazole fungicides and resistance to diethofencarb, was detected with frequencies of 6.8%, 10.0%, 13.2% and 12.4% from 2003 to 2006, respectively. Further studies indicated that Ben R3 was caused by a point mutation from GAG in sensitive(S) isolates to GTG at codon 198 in the β-tubulin gene, predicted to cause a change from glutamic acid to valine. Ben R3 isolates had comparable growth, sporulation and pathogenicity ability as isolates of other phenotypes but were more sensitive at lower temperatures.     

Spiral gradient dilution, a rapid method for determining growth responses and 50% effective concentration values in fungus-fungicide interactions

ABSTRACT A new technique, the spiral gradient dilution method, was evaluated for determining 50% effective concentrations (EC(50) values) of fungicides for the inhibition of mycelial growth and conidial germination of various fungi. In this method, an agar medium is plated with a fungicide solution by means of a spiral plater, which applies the fungicide in a 2.5-log dilution in a continuous radial concentration gradient. Fungal inoculum is then placed along radial lines across the gradient. After incubation of the plates, distinct growth shapes were observed in different fungus-fungicide interactions. Mycelial growth responses to increasing fungicide concentrations ranged from abrupt to gradual transitions. Conidial germination responses were similar; in addition, distinct zones of confluent growth, nonconfluent growth, and outlier colonies were also identified, depending on the fungus-fungicide interaction. The fungicide concentration at the radial distance at which 50% reduction of growth or spore germination occurred, compared with growth or germination on unamended media, was calculated by a computer program. EC(50) values were obtained for mycelial growth in 22 fungus-fungicide interactions and for conidial germination in five interactions. The fungi evaluated were members of the Zygomycota, Ascomycota, Basidiomycota, and Deuteromycota. Nine fungicides, belonging to six different chemical classes, were tested. EC(50) values were compared with those obtained by the traditional agar dilution method. In linear regression analyses of the two methods, the models were highly significant (P < 0.01), and coefficients of determination (r(2)) were 0.92 for the mycelial growth assays and 0.94 for the conidial germination assays. Regression slopes were not significantly different from 1 (P > 0.05) with optimal program settings in the software. Estimated bias, coefficients of variation, and actual confidence intervals for the regression slope were 13.5%, 6.5%, and 1.14 +/- 0.14 for the mycelial growth assays and 7.5%, 14.5%, and 1.08 +/- 0.37 for the conidial germination assays. These analyses indicate that the spiral gradient dilution method is accurate and precise compared with the agar dilution method for calculating EC(50) values of fungicides in continuous growth responses to fungicide concentration gradients.     

四种杀菌剂对烟草灰霉病菌的毒力及对烟草灰霉病的抑制作用

为筛选防治烟草灰霉病的有效药剂,采用菌丝生长速率法和孢子萌发法测定氟啶胺、咪鲜胺、苯醚甲环唑及代森锰锌4种杀菌剂对烟草灰霉病菌Botrytis cinerea的毒力,同时通过离体叶片评价这4种杀菌剂对烟草灰霉病的保护和治疗作用。结果表明,氟啶胺和咪鲜胺对菌丝生长活性抑制最强,有效抑制中浓度ECjs50平均值分别为0.02、0.03 mg/L,苯醚甲环唑次之,代森锰锌最弱,EC_(js50)平均值分别为0.39、7.86 mg/L;氟啶胺对孢子萌发活性抑制最强,代森锰锌次之,有效抑制中浓度EC_(mf50)平均值分别为0.06、0.16 mg/L,咪鲜胺和苯醚甲环唑最弱,ECmf50平均值均大于25.00 mg/L。离体试验表明,氟啶胺对烟草灰霉病保护作用最强,浓度为50 mg/L时,防治效果为100.00%,咪鲜胺和苯醚甲环唑次之,防治效果分别为88.62%和76.46%,代森锰锌最弱,浓度为1 000 mg/L时防治效果仅为75.81%;氟啶胺对烟草灰霉病治疗作用最强,浓度为100 mg/L时的防治效果为85.75%,咪鲜胺和苯醚甲环唑次之,浓度为200 mg/L时的防治效果分别为85.47%和76.48%,代森锰锌最弱,浓度为1 000 mg/L时防治效果为70.24%。表明氟啶胺和咪鲜胺更适合烟草灰霉病的防治。