积雪草漆斑病病原鉴定及室内药剂筛选

近年来,在广西各地积雪草种植基地,出现一种为害极重的病害,该病害病原菌主要为害积雪草叶片、叶柄,可致使叶片大面积枯死,该病害发生普遍、侵染期长,严重影响积雪草药材的品质和产量。该研究采用常规组织分离法从积雪草病叶上分离病原菌并根据柯赫氏法则验证其致病性;通过菌落特征、菌株形态及ITS序列分析鉴定病原菌;在室内利用生长速率法测定10种杀菌剂对该菌的抑制活性。结果表明：为害积雪草叶片的病原菌为泛生漆斑菌( Myrothecium inundatum);该菌侵染性强,接种至健康积雪草叶片后24 h即可发病,并迅速扩展引起叶片萎蔫;杀菌剂60％唑醚?代森联水分散粒剂、45％恶霉灵?甲霜可湿性粉剂、1.8％辛菌胺醋酸盐水剂、80％代森锰锌可湿性粉剂室内对积雪草漆斑病病原菌的抑菌率均在98％以上,抑菌效果极强,建议在生产中轮换应用防治积雪草漆斑病。15％咪鲜胺微乳剂、70％甲基硫菌灵可湿性粉剂和10％苯醚菌酯悬浮剂对该病原菌的抑菌率也均在92％以上,抑菌效果良好,在生产中可酌情考虑使用。     

A comparison ofin vitro andin vivo effect of clay minerals,humic acid and micronutrients on the activity of fungicides againstRhizoctonia solani

Activity of nine fungicides against mycelial growth ofRhizoctonia solani in potato dextrose broth and in pot tests as seed treatment against cowpea seedling rot in infested soil was differentially in fluenced by clay minerals, humic acid and micronutrients. Humic acid, extracted from farmyard manure, considerably lowered the activity, bothin vitro andin vivo, of all fungicides except chloroneb. Montmorillonite caused substantial decrease in disease control by fungicides but enhanced the toxicity of 2-methoxyethyl mercury chloride (MEMC), and quintozene in culture. Kaolinite inactivated carbendazim, benomyl and thiophanatemethyl in nutrient broth but had little effect on disease control by these fugicides. The six micronutrients altered the activity of fungicides to varying levels often without definite correlation betweenin vitro andin vivo results. The results ofin vitro growth inhibition tests are largely inapplicable to dieseas control tests in infested soil mainly due to the differences in the ambient conditions of the two systems.     

Evaluation of some fungicides for seed treatment and foliar application in management of damping-off of seedlings and blight of rapeseed caused by Alternaria brassicae

Eighteen fungicides were first evaluated for their effects on growth of Alternaria brassicae and for ascertaining their fungicidal and fungistatic natures in artificial cultures. The chemicals emerging fungicidal in action, were later evaluated for their efficacy as seed treatment and foliar application in the management of damping-off of seedlings and blight of rapeseed separately. Of 18 fungicides tested, six fungicides, viz., Dithane M-45, Dithane Z-78, Ziram, Difolatan-80, Blitox-50 and Benlate completely inhibited the growth of the pathogen and were fungicidal in action. Thiram and Brestan-60, which also caused total growth inhibition, were, however, fungistatic. Benlate (0.1 %) followed by Dithane M-45 was best seed-dressing fungicide for controlling damping-off of seedlings. Dithane M-45 (0.2%) followed by Dithane Z-78 as foliar spray was most effective for controlling the blight and increasing the yield in field trials.     

Impact of fungicides on Metarhizium anisopliae in the rhizosphere, bulk soil and in vitro

The entomopathogenic fungus Metarhizium anisopliae (Metchnikoff) Sorokin (Hypocreales: Clavicipitaceae) is registered in the United States and The Netherlands for black vine weevil, Otiorhynchus sulcatus (Coleoptera: Curculionidae) control in container-grown ornamentals. These studies were conducted to determine the compatibility of M. anisopliae (F52) with a wide range of fungicides commonly applied to container-grown ornamentals for the management of soil-borne plant pathogens. The impact of fungicides on spore germination and mycelial growth were determined in vitro. In addition, M. anisopliae persistence in bulk and rhizosphere soil was determined 30 days following dual application of each fungicide at 7–28 days intervals as prescribed. A number of fungicides (thiophanate-methyl, dimethomorph, captan, triflumizole, triflozystrobin, pyraclostrobin, azoxystrobin) inhibited spore germination in vitro. A larger number of fungicides (fosetyl-AI, thiophanate-methyl, dimethomorph, captan, quintozene, triflumizole, fludioxanil, triflozystrobin, pyraclostrobin, fludiox-mefanox, iprodione, azoxystrobin, phosphorus acid/K-salts) inhibited mycelial growth in vitro. Only three fungicides (etridiazole, propamocard and mafanoxam) had no significant impact in vitro on spore germination or mycelial growth. While a number of fungicides had a detrimental impact in vitro, there was no impact on M. anisopliae populations in bulk soil following dual application of any fungicide. However, the fungicides captan and triflumizolet, which have a short reapplication interval, had a detrimental impact on M. anisopliae populations in the rhizosphere. As researchers develop rhizosphere competence as an alternative management strategy for black vine weevil, the fungicides captan and triflumizole should be avoided.     

Iturin A: A potential new fungicide for stored grains

The removal of many synthetic fungicides from the market has created a demand for new, environmentally safe fungicides. Iturin A, a cyclic lipopeptide produced byBacillus subtilis, has strong antifungal properties and low mammalian toxicity. To determine the efficacy of this compound as a potential fungicide on stored feed grains, lots of corn, peanuts and cottonseed were treated with varying concentrations of iturin A. The mycoflora of treated seed was assayed along with that of untreated seed and seed treated with fungicides used commercially for planting seed. Fungal species varied considerably in their sensitivity to iturin A. Significant reductions in total mycoflora occurred in most seed lots tested at iturin A concentrations of 50 to 100 ppm.     

Effect of certain fungicides on the production of enzymes byRhizoctonia solani

The effect of fungicidesviz. Benomyl, Carbendazim, Chloroneb, Kitazin, Edifenphos, Thiabendazole and Wettable ceresan on the production of enzymes byR. solani was studied. The fungicides tested were inhibitory to the production and activity of amylase, invertase and cellulase. The fungicides inhibited the growth of the fungus to a considerable extent.     

Effect of fungicides on survival of Rhizobium on seeds and the nodulation of bean (Phaseolus vulgaris L.)

The survival of Rhizobium leguminosarum biovar phaseoli on seeds of bean was tested, using the cultivar Carioca. The seeds were treated seven days before inoculation with Benlate, Vitavax, Banrot, Difolatan or Ridomil fungicides. The rhizobial strains used were: CIAT 899, CPAC 1135 and CIAT 652. Strain CIAT 899 showed greater survival on the seed with fungicide than the other strains. Two hours after the contact with fungicides strains CIAT 652 and CPAC 1135 had significantly lower numbers of rhizobia than the treatment without fungicide. The Benlate and Banrot fungicides had the greatest effect on survival of rhizobial strains. There was a drastic mortality of the two strains, CIAT 652 and CPAC 1135, on seeds treated with Benlate and Ridomil. Under field conditions, granular inoculation produced fewer nodules, but a similar total nodule weight as seed inoculation. Serological tests (ELISA) showed that seed treatment with Benlate in connection with seed inoculation reduced drastically the occurrence of inoculated strains in nodules, while the same fungicide treatment and inoculation applied in the seed furrow did not affect the survival of the inoculated strain.     

Ethylene formation by germinating,Drechslera graminea — Infected barley (Hordeum sativum) grains: A simple test for fungicides

Grains of barley (Hordeum sativum L.); infected with the parasitic, systemic fungus Drechslera graminea, produce more ethylen than uninfected controls. Treatment of infected grains with mercury-free fungicides yields a differentiated suppression of the ethylene evolution 7 d after the beginning of germination. Suppression of visible symptoms (chlorotic stripes on leaves) appearing six to eight weeks after germination of infected, untreated seeds correlates with the decrease in ethylene formation after treatment with fungicides. The gaschromatographic ethylene determination thus allows for an early and reliable (significance higher than 99.9%) differentiation of fungicidal activities against the barley stripe disease.Abbreviations ACC I-aminocyclopropane-1-carbonic acid - CD critical difference     

Comparative trials on the effects of two fungicides on a predatory mite in the laboratory and in the field

Both laboratory tests with the open glass plate method of Louis & Ufer (1995) and a field study in a vineyard were carried out to test the side effects of the two fungicides, Dithane Ultra WG75 and Polyram Combi WG70, on the predatory mite Typhlodromus pyriScheuten (Acarina: Phytoseiidae). The effects of both fungicides in the residual contact test in the laboratory were more pronounced than the actual effects obtained in the field even after multiple application of the test products. The study results suggest, that for the selected test products the current laboratory guidelines correctly triggered the field test. Methodological problems of the laboratory test due to the repellent effect of the fungicides are discussed.     

Cloning and characterization of the histidine kinase gene<Emphasis Type="Italic"> Dic1</Emphasis> from<Emphasis Type="Italic"> Cochliobolus heterostrophus</Emphasis> that confers dicarboximide resistance and osmotic adaptation

A gene for a putative two-component histidine kinase, which is homologous to os-1 from Neurospora crassa, was cloned and sequenced from the plant-pathogenic fungus Cochliobolus heterostrophus. The predicted protein possessed the conserved histidine kinase domain, the response regulator domain, and six tandem repeats of 92-amino-acids at the N-terminal end that are found in histidine kinases from other filamentous fungi. Introduction of the histidine kinase gene complemented the deficiency of the C. heterostrophus dic1 mutant, suggesting that the Dic1 gene product is a histidine kinase. Dic1 mutants are resistant to dicarboximide and phenylpyrrole fungicides, and they are sensitive to osmotic stress. We previously classified dic1 alleles into three types, based on their phenotypes. To explain the phenotypic differences among the dic1 mutant alleles, we cloned and sequenced the mutant dic1 genes and compared their sequences with that of the wild-type strain. Null mutants for Dic1, and mutants with a deletion or point mutation in the N-terminal repeat region, were highly sensitive to osmotic stress and highly resistant to both fungicides. A single amino acid change within the kinase domain or the regulator domain altered the sensitivity to osmotic stress and conferred moderate resistance to the fungicides. These results suggest that this predicted protein, especially its repeat region, has an important function in osmotic adaptation and fungicide resistance.Communicated by C. A. M. J. J. van den Hondel     

Antifungals: Mold control and safe levels in artificial media forTrichogramma [Hymenoptera: Trichogrammatidae]

Ten chemicals known to be fungicides are tested in artificial media forTrichogramma dendrolimi Matsu. [Hymenoptera: Trichogrammatidae]. Tests are performed in France by using artificial host egg capsules. Most of the fungicides cannot control fungus or are toxic forTrichogramma at effective levels. Geneticin, Amphotericin B and Nystatin, used at proper levels are able to control molds efficiently without modifyingTrichogramma development. These antifungals are also tested in actual conditions of mass production, in Guangzhou (China).      

Potential of genes and gene products fromTrichoderma sp. andGliocladium sp. for the development of biological pesticides

Fungal cell wall degrading enzymes produced by the biocontrol fungiTrichoderma harzianum andGliocladium virens are strong inhibitors of spore germination and hyphal elongation of a number of phytopathogenic fungi. The purified enzymes include chitinolytic enzymes with different modes of action or different substrate specificity and glucanolytic enzymes with exo-activity. A variety of synergistic interactions were found when different enzymes were combined or associated with biotic or abiotic antifungal agents. The levels of inhibition obtained by using enzyme combinations were, in some cases, comparable with commercial fungicides. Moreover, the antifungal interaction between enzymes and common fungicides allowed the reduction of the chemical doses up to 200-fold. Chitinolytic and glucanolytic enzymes fromT. harzianum were able to improve substantially the antifungal ability of a biocontrol strain ofEnterobacter cloacae. DNA fragments containing genes encoding for different chitinolytic enzymes were isolated from a cDNA library ofT. harzianum and cloned for mechanistic studies and biocontrol purposes. Our results provide additional information on the role of lytic enzymes in processes of biocontrol and strongly suggest the use of lytic enzymes and their genes for biological control of plant diseases.     

In vitro sensitivity of Verticillium fungicola to selected fungicides

Twenty isolates of Verticillium fungicola var. fungicola collected from diseased fruit-bodies of Agaricus bisporus from prochloraz-treated crops, were exposed to a range of concentrations of six chemicals (benomyl, chlorothalonil, formaldehyde, iprodione, prochloraz-Mn-complex and prochloraz + carbendazim) in vitro. EC₅₀ values were determined for each fungus-fungicide combination. All isolates were more sensitive to prochloraz-Mn-complex (EC₅₀ values less than 5 mg 1^–1) than to the remainder fungicides, and only seven isolates were moderately sensitive (EC₅₀ values between 5 and 50 mg 1^–1) to prochloraz + carbendazim. All isolates were moderately sensitive to formaldehyde, whereas the majority of isolates were very resistant to the other three fungicides (benomyl, chlorothalonil and iprodione).     

Effects of 20 fungicides on the infectivity of conidia of the aphid entomopathogenic fungus Erynia neoaphidis

The effect of 20 fungicides on theinfectivity of conidia of the entomopathogenicfungus, Erynia neoaphidis, were assessedin the laboratory. After projection on broadbean leaves, conidia were treated withfungicides applied at their recommended fieldrate. Afterwards, the infectivity of theseinocula was assessed using an aphid bioassay.Four fungicides, carbendazim, kresoxym-methyl, nuarimol and thiophanate-methyl reduced the infectivity of the conidia by less than 25% and can be considered harmless for this aphid pathogen. Propiconazole was a little more toxic, with 37% reduction. Other products reducedinfectivity by between 50% and 100%. These are, from the least to the most toxic:flutriafol, prochloraz, epoxyconazole,iprodione, hexaconazole, triadimenol,azoxystrobine, cyproconazole, cyprodynil,flusilazole and tridemorph. Chlorothalonil,fenpropimorph, spiroxamine and tebuconazoletotally inhibited infectivity of the fungi. Analysis of the results according to chemicalclass showed that the benzimidazoles were theleast toxic for E. neoaphidis and themorpholines the most toxic. Effects oftriazoles and strobilurines were variable, withreduction ranging from 37% to 100% fortriazoles and from 17% to 68% forstrobilurines.     

Effects of selected pesticides used against glasshouse tomato pests on colony growth and conidial germination of Paecilomyces fumosoroseus

Two isolates of a potential biological control agent, Paecilomyces fumosoroseus, for greenhouse whitefly, Trialeurodes vaporariorum, on tomato in greenhouses were tested for their compatibility with five fungicides (captan, dichlofluanid, iprodione, benomyl, and carbendazim) and six insecticides (malathion, carbaryl, primicarb, pymetrozine, cypermethrin, and deltamethrine) which are used on glasshouse tomato. Conidial germinations after 24 h and radial growths after 14 days of inoculation were determined in the presence of each pesticide at three concentrations (recommended rate, 0.1 and 10 times of the recommended rate by its manufacturer) and resultant variations were evaluated. Our results suggest that the tested pesticides, especially the fungicides, have potential to interfere with the success of P. fumosoroseus if they are to be employed together. The recommended concentration rate of primicarb and pymetrozine, and the lower concentrations of cypermethrin and deltamethrin were less hazardous than the others.     

Laboratory method for testing side-effects of pesticides on the rove beetleAleochara bilineata — Adults

Tests were conducted in glass cells containing moistened sand sprayed with the concentration recommended for the pesticide or with demineralised water (controls). Adult females (1–2 weeks old) ofA. bilineata were placed on the fresh residue in the cells and provided daily with fresh fly eggs as food. The duration of the test was 5 days, mortality being recorded every day. The side effects of the pesticide were expressed as reduction in egg production ofA. bilineata during the whole test period. Results permitted classification of the pesticides according to the 4 categories of harmfulness used by the IOBC working group. 23 pesticides were tested, and the reproducibility of the test appeared to be satisfactory. Generally, insecticides/acaricides had a pronounced effect on the beetles whereas fungicides and herbicides were less detrimental. Exceptions were the insecticides Tedion V 18 and Kilval, classified as harmless, and the fungicides Morestan, Pomarsol forte and Afugan rated as moderately harmful. Among growth regulators Prosevor 85 was highly detrimental while Cycocel Extra and Rhodofix had no effects. Excepting Morestan, Pomarsol forte and Ustinex PA the pesticides did not affect the hatching of eggs. The work was financed by the Danish Environmental Protection Agency.     

Compatibility of Chemical Disease and Insect Management Practices Used in New York Vineyards with Biological Control by Anagrus spp. (Hymenoptera: Mymaridae), Parasitoids of Erythroneura Leafhoppers

Toxicity to Anagrus spp. of fungicides and insecticides used in grape production was assessed with laboratory and field bioassays. Field-equivalent rates of fungicides were relatively nontoxic to Anagrus spp. adults in laboratory bioassays. In bioassays with field-weathered residues, sulfur (9600 ppm) caused elevated mortality of adults for 14 to 21 days posttreatment. Residues of microencapsulated methyl parathion (600 and 1200 ppm) increased mortality relative to the control up to 43 days posttreatment. Duration of elevated mortality of adults exposed to carbaryl was dose dependent and ranged from 14 (at 1200 ppm) to >43 days (at 4800 ppm). Residues of carbaryl and methyl parathion applied over parasitized eggs had little effect on emergence, but may have delayed development. In a field trial, adults trapped in carbaryl-treated plots were significantly reduced starting 3 weeks after treatment. Subsequent lower trap catches may have been related to effects of residues on Anagrus spp. adults or to lower leafhopper egg densities in treated plots.     

Specific patterns of changes in wheat gene expression after treatment with three antifungal compounds

The two fungicides azoxystrobin and fenpropimorph are used against powdery mildew and rust diseases in wheat (Triticum aestivumL). Azoxystrobin, a strobilurin, inhibits fungal mitochondrial respiration and fenpropimorph, a morpholin, represses biosynthesis of ergosterol, the major sterol of fungal membranes. Although the fungitoxic activity of these compounds is well understood, their effects on plant metabolism remain unclear. In contrast to the fungicides which directly affect pathogen metabolism, benzo(1,2,3) thiadiazole-7-carbothioic acid S-methylester (BTH) induces resistance against wheat pathogens by the activation of systemic acquired resistance in the host plant. In this study, we monitored gene expression in spring wheat after treatment with each of these agrochemicals in a greenhouse trial using a microarray containing 600 barley cDNA clones. Defence-related genes were strongly induced after treatment with BTH, confirming the activation of a similar set of genes as in dicot plants following salicylic acid treatment. A similar gene expression pattern was observed after treatment with fenpropimorph and some defence-related genes were induced by azoxystrobin, demonstrating that these fungicides also activate a defence reaction. However, less intense responses were triggered than with BTH. The same experiments performed under field conditions gave dramatically different results. No gene showed differential expression after treatment and defence genes were already expressed at a high level before application of the agrochemicals. These differences in the expression patterns between the two environments demonstrate the importance of plant growth conditions for testing the impact of agrochemicals on plant metabolism.     

In vitro studies on the effects of fungicides on beneficial fungi of peach twig mycoflora

In vitro studies were carried out to investigate a possible integrated use of chemical and biological means to control the peach twig blight pathogen,Monilinia laxa. Three fungal antagonists ofM. laxa (Penicillium purpurogenum, Penicillium frequentans andEpicoccum nigrum) and six fungicides (vinclozolin, iprodione, thiram, captan, benomyl and thiophanate-methyl) were used in the study. Sensitivity of the fungal isolates to the fungicides was determined in vitro by calculating ED50 values. Benomyl and thiophanate-methyl were the most fungitoxic compounds and captan was the least fungitoxic.M. laxa andP. purpurogenum were the most sensitive to all chemicals tested, whileE. nigrum andP. frequentans presented bigger differences in their sensitivity to chemicals compared toM. laxa. E. nigrum was consistently less sensitive to benomyl (ED50=2.26 ppm), thiophanate-methyl (ED50=9.61 ppm) and vinclozolin (ED50=3.89 ppm) than the other fungi.P. frequentans was less sensitive to captan, vinclozolin, iprodione, thiophanate-methyl and thiram thanM. laxa (8, 7, 5, 4 and 2 times respectively). These results suggest thatE. nigrum andP. frequentans could be successfully used in an integrated control programme that combines biological and chemical methods.     

Effect of triadimenol,imazalil and nuarimol seed treatment on subcrown internode length,coleoptile-node-tillering and common root rot in spring wheat

Summary The effect of seed treatment with triadimenol at 0.175 and 0.3, imazalil at 0.15 and 0.2, and nuarimol at 0.1 and 0.15 g a.i./kg seed on subcrown internode (SCI) length, occurrence of coleoptile-node-tillers (CNTs), and common root rot severity was studied in two spring wheat cultivars at three locations in Saskatchewan. All three fungicides showed similar effects on both Cypress and Neepawa cultivars. All fungicides significantly reduced severity of common root rot and SCI length, promoted the early development of CNTs and increased (P<0.01) the proportion of CNTs that produced fertile heads of grain.Contribution from Agr. Canada Res. Stn. No. 869.