Splash dispersal of fungus spores and fungicides in the laboratory and greenhouse

A laboratory technique is described for the production of drops of simulated rain in which fungal spores were suspended. When such drops containing conidia of Botrytis fabae impacted on a target leaf the secondary droplets produced infections on receptor broad bean leaves. The capacity of fungicides applied to the target leaf to redistribute in secondary splash droplets was examined in terms of the infectivity of the spores in the droplets. The extent to which a copper fungicide reduced infection on the receptor leaves was related to the level and tenacity of the fungicide deposit on the target leaf. The effect of wetting agents on the redistribution of this fungicide could probably be explained by their influence on the tenacity of the initial deposit. In general the capacity of different fungicides to inhibit infection by the secondary droplets was related to the inherent toxicity of the fungicides to B. fabae. Implications of the dispersal of spores and fungicides by rain splash are briefly considered with reference to field conditions.     

In vitro and In vivo Effects of Some Fungicides against the Chickpea Blight Pathogen, Ascochyta rabiei

The effects of various fungicides on mycelial growth and spore germination of Ascochyta rabiei were determined by incorporating them into potato dextrose agar and measuring colony diameter and observing colony growth and spore germination at 20 ± 2°C. Eight fungicides prevented spore germination of the pathogen at concentrations of 0.125–2 μg/ml, three hindered mycelial growth at 2–4 μg/ml and seven failed to inhibit mycelial growth even at 128 μg/ml. The reference fungicide for the pathogen, chlorothalonil, stopped conidial germination at low rates but did not prevent mycelial growth at 128 μg/ml. Thirteen fungicides were tested against seed infections of the pathogen, and benomyl + thiram, carbendazim and carbendazim + chlorothalonil seed treatments gave more than 85% inhibition on both vacuum‐infiltrated and naturally infected seeds. Coating the seeds with polymers did not increase the effectiveness of fungicides. Three fungicides; (azoxystrobin, chlorothalonil and mancozeb), gave the highest protection in the field but protection decreased with increased inoculum pressure. Addition of humic acid to fungicide suspensions did not affect their performance.     

STUDIES ON SYSTEMIC FUNGICIDES

A number of mono-, di-and trichloro-β-naphthols and the corresponding naphthyloxy- n -carboxylic acids were prepared. They were tested as systemic fungicides against chocolate spot of broad bean ( Vicia faba ) caused by Botrytis fabae and as fungicides in vitro by their effect on the germination of spores of Sclerotinia laxa and the growth of the mycelium of Botrytis fabae.
The systemic fungicide tests showed no difference in the activity of the compounds associated with different side chains, though there were slight but significant effects associated with different chlorine substitutions in the naphthalene nucleus. The 3:4-dichloro-2-naphthol derivatives were more effective than the other compounds tested.
Fungicide tests with the 1:3-dichloro-2-naphthol derivatives suggested that there was some increase in toxicity to the spores of Sclerotinia laxa with increasing length of side chain. The effectiveness of the various chlorine-substituted compounds in suppressing growth of Botrytis fabae was in the following descending order, 1:3:4-trichloro-, 1:3-dichloro-, 1:4-dichloro-and 3:4-dichloro-compounds. The concentrations required in agar culture to induce reductions of B. fabae growth comparable to those found in the leaf could reasonably be expected to occur in the bean plants treated with the 2-naphthyloxyacetic acids at 10 p.p.m.     

Vapour action of fungicides against powdery mildews

Certain fungicides prevented the growth of powdery mildews at distances extending several millimetres beyond the edge of localized deposits on leaves. Most of the materials also acted at a distance when separated from the leaf by glass cover-slips, indicating that the effect was due to the emission of vapour. In addition to sulphur and lime-sulphur, whose vapour action is already known, fungicides based on drazoxolon, oxythioquinox, binap-acryl, dinitro-octyl phenyl crotonates and a phthalimido-phosphonothionate were found to act in this way. Two systemic fungicides, griseofulvin and triamiphos, did not give a detectable vapour action. Tests were conducted mostly on open glasshouse benches. A draught from a fan shifted the zones of inhibition to one side of the deposits, but did not reduce their areas. Vapour effects were similar in extent on plants maintained at 18–25° C and at 27–32° C. Variation in the areas protected was studied in relation to the size and fungicide content of the deposits, to different powdery mildews, to time of incubation and to different types of formulation. Deposits applied to leaves by high- or low-volume sprays at concentrations used in the field gave significant protection at a distance. Vapour effects were demonstrated also on mildew conidia incubated on glass slides bearing a spot of fungicide, and on infected plants placed in beakers coated on the bottom with fungicide. Movement of fungicides in the gaseous state is discussed in relation to the control of foliage diseases in the field.     

Fungitoxic action of dimethirimol and ethirimol

The systemic fungicides dimethirimol and ethirimol were shown to be toxic to spores of Erysiphe graminis and Sphaerotheca fuliginea, in germination tests in vitro. Toxicity of dimethirimol to spores of Botrytis fabae, Phytophthora infestans, Puccinia recondita, Uncinula necator and Venturia inaequalis was relatively feeble or absent. When applied in foliage sprays or in root treatments, both compounds inhibited the emergence of germ tubes from powdery mildew spores on cucumber and barley plants. The specific disease control shown by these fungicides can be explained by their direct fungitoxic action. The effects of dimethirimol and ethirimol on powdery mildew infections and on spore germination were greatly decreased by the presence of riboflavin in the light. A rapid photo-chemical interaction between riboflavin and dimethirimol was demonstrated. Marked reversals both of disease control and of fungitoxicity were also given by folic acid, and it is suggested that folic acid overcomes a metabolic block induced in powdery mildew fungi by the fungicides.     

Utility of six fungicides for selective isolation of Evlachovaea sp. and Tolypocladium cylindrosporum

The effects of the fungicides dodine, benomyl, thiabendazole, mancozeb, cupric sulfate, and copper oxychloride were examined in vitro upon germination and further development of Evlachovaea sp. and Tolypocladium cylindrosporum. Fungicidal activity depended on concentrations and varied among products, fungi and the strains tested. Depending on the fungicidal concentration, germination of conidia was induced but germlings produced neither mycelium nor new conidia. There was a good recovery of both Evlachovaea sp. and T. cylindrosporum from previously sterilized soils with fungicide-supplemented medium. Fungi were resistant to copper oxychloride up to 30 g/l, and this fungicide was found to have no utility for a selective medium. Minimal fungicide concentrations for successful isolations were 1 mg/l for benomyl, 200 mg/l for cupric sulfate, 50 mg/l for dodine, 100 mg/l for mancozeb, and 4 mg/l for thiabendazole. Thiabendazole, which is easy to obtain and can be used in low quantities, showed the greatest utility for a selective medium with these entomopathogenic fungi. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Individual and Combined Effects of Certain Pesticides on Rhizoctonia solani, Sheath Blight Pathogen of Rice

The effect of different pesticides (fungicide, herbicide and insecticide) on the mycelial growth and formation and germination of sclerotia of sheath blight pathogen of rice, Rhizoctonia solani Kühn was studied in nutrient medium and in natural soil. Among the herbicides and insecticides, fenitrothion (insecticide) and thiobencarb (herbicide) were the most effective in inhibiting the mycelial growth and sclerotial formation. These pesticides effected significant inhibition of both mycelialgrowth and sclerotial formation at 7.5 to 10μg/ml concentration while fungicides carbendazim and tolclofosmethyl effected almost complete inhibition at 2.5 μg/ml. Interestingly, a synergistic increase in the toxicity to mycelial growth, sclerotial formation and sclerotial germination was noticed when the fungicide, mancozeb and the herbicide, thiobencarb were applied at subtoxic concentration in combination. Such synergistic interactions between pesticides in a combination leading to increased toxicity suggest that under the current practice of applying fungicides, herbicides and insecticides either simultaneously or in rotation, certain combinations may help in reducing the dose of a fungicide in disease control.     

In vitro compatibility of Beauveria bassiana strain ATCC 74040 with copper fungicides

Copper fungicides and mycoinsecticides based on entomopathogenic fungi Beauveria spp. are the most common pesticides used in organic crop production systems. The in vitro effects of the copper fungicides copper oxide, copper hydroxide, copper oxychloride, copper sulphate, dicopper chloride trihydroxide and tribasic copper sulphate were investigated for mycelial growth, sporulation and conidial germination of the ATCC 74040 commercial strain of Beauveria bassiana. Mycelial growth was evaluated on potato dextrose agar plates with 100%, 75%, 50%, 25%, 12.5%, 6.25% and 0% of the recommended application rates of each fungicide at 15 and 25°C. Sporulation and conidial germination were determined at the recommended field doses of each fungicide at 25°C. All copper fungicides had fungistatic or fungicidal effects on B. bassiana that varied according to the dose. Only in two cases, copper oxide at 15°C and copper hydroxide at 25°C, at the lowest concentration of 6.5%, was mycelial growth not statistically significantly inhibited. Inhibition of mycelial growth depended both on the fungicide and its concentration, and partly on temperature. Both sporulation and conidial germination of B. bassiana were significantly inhibited by all fungicides. All fungicides inhibited the sporulation in a similar way (99.8%–100%). With the exception of copper oxychloride (inhibition, 13.6%), the other fungicides showed high detrimental effects on conidial germination (inhibition, 91.7%–100%). The fungus was strongly affected by some fungicides even at the lowest doses. The biological index used for the B. bassiana with copper fungicides ranged from 0.6 (copper sulphate) to 18.1 (copper oxychloride). Therefore, the tested fungicides were classified into the upper half of the highly toxic (T) category and are considered incompatible with the entomopathogenic fungus B. bassiana strain ATCC 74040 under in vivo experimental conditions. These results need to be further verified in vitro under both greenhouse and open-field conditions.     

Effects of two sterol biosynthesis inhibitor fungicides (fenpropimorph and fenhexamid) on the development of an arbuscular mycorrhizal fungus

The effects of different concentrations (0.2, 2, 20, 200 mg l⁻¹) of two sterol biosynthesis inhibitor (SBI) fungicides, i.e. fenpropimorph and fenhexamid, were evaluated on the spore germination, germ tube elongation, sporulation, and root colonization of Glomus intraradices grown monoxenically in association with transformed carrot roots. The percentage of germinated spores incubated on the SBI fungicides and the length of the germ tubes decreased with increasing concentrations of both fungicides. However, for spore germination this impact was fungistatic rather than fungicidal. Extraradical mycelium architecture and spore production in contact with the SBI fungicides were also strongly impacted at high concentration (20 mg l⁻¹). Conversely, the colonization of roots developing in the fungicide-free compartment, but interconnected with the extraradical mycelium developing on the SBI fungicides, appeared unaffected. Our results demonstrated that the monoxenic culture system could be used as a standardized, reproducible technique to compare the impacts of different molecules on arbuscular mycorrhizal fungi, and for the initial screening of new candidate molecules before registration.     

Studies in vitro and in vivo with powdery mildew fungicides

Droplets of 4-(i-cycloalkylalkyl)-2,6-dinitrophenols often protect areas of leaf very much greater than that of the initial deposit against powdery mildew. For a given alkyldinitrophenol, this type of protection is greater on apple than on marrow leaves. This zonal protection is not correlated with vapour activity in vitro or in vivo, or with inhibition of conidial germination in vitro, but is correlated with the degree of protection obtained when leaves are sprayed to give good cover. By contrast, crotonic esters of these alkyl-dinitrophenols do not show zonal activity, but nevertheless often exhibit protectant activity comparable with that of the parent phenols. These results suggest that exploitation of favourable hydrogen bonding characteristics in the fungicide molecule may lead to better control of fungal diseases in the field as a result of enhanced movement of fungicide in or on the leaf cuticle (zonal movement) or easier penetration of the fungal conidium.     

INVESTIGATIONS ON FUNGICIDES. I. FUNGICIDAL AND SYSTEMIC FUNGICIDAL ACTIVITY IN CERTAIN ARYLOXYALKANECARBOXYLIC ACIDS

Fifty-four aryloxyalkanecarboxylic acids have been examined as systemic fungicides. Of these, four which produced negligible damage in broad-bean seedlings yet conferred systemic fungicidal protection to broad beans against Botrytis fabae and to tomatoes against Alternaria solanixvere studied in detail. The direct fungicidal activity of twenty compounds was assessed against B. fabae and Pythium ultimum. No correlation between fungicidal and systemic fungicidal action toward B. fabae was established. Other possibilities regarding their mode of action as systemic fungicides are discussed. The systemic fungicidal activity of a-phenoxyziobutyric and 2:4:6-trichlorophenoxyacetic acids in beans and of the latter compound in tomato plants was found to depend upon seasonal factors, but for 3-phenoxybutyric and 5-phenoxycaproic acids such variation was not observed. Results obtained in the bean test by two methods of assessment were found to correlate. The potato petiole test (van Raalte) for assessing systemic fungicidal activity was studied and modified. Evidence was obtained that the results might be unreliable owing to toxic effects produced by the compounds on the petiole tissue. Attempts were made to avoid this complication, but when phytotoxicity was eliminated, little fungicidal activity was shown.     

Local redistribution of fungicides on leaves by water

Local redistribution of various fungicides applied as large or small droplets to detached broad-bean leaflets was examined by laboratory bio-assay with Botrytis fabae and by leaf-printing techniques. The results showed that fungicidal particulate and soluble material will spread in water from the initial deposits and that the protected areas on leaves inoculated after washing were often much greater than on unwashed leaves. The extent of redistribution was related to the inherent toxicity, physical form, duration of weathering and tenacity of the fungicides. The effects of formulating copper oxychloride and zineb fungicides with surface-active agents and adhesives seemed due to differences in the tenacity of the deposits. Results indicated that similar types of redistribution would occur in the field, but the possible importance of the electrokinetic theory of redistribution was not proven.     

Signum, a new fungicide for control of leaf diseases in outdoor vegetables

During three years, the new fungicide Signum, containing 6.7% pyraclostrobine + 26.7 % boscalid and developed by BASF. has been evaluated in leek, carrots and cabbages in several outdoor field experiments under practical conditions and during one year in outdoor lettuce. In leek, Phytophthora porri is one of the major leaf diseases causing lesions on differ ent places on the leaves, resulting in at least extra labour costs for trimming or even worse sometimes resulting in complete crop loss. So far, crop protection consists of repeated applications of fungicides especially during autumn and winter. Pyraclostrobin + boscalid has been evaluated in comparison with the fungicides mancozeb, mancozeb + metalaxyl-M and azoxystrobin. The progress of the disease during the growth season is discussed. For all parameters evaluated, pyraclostrobin + boscalid gave comparable or even better results than reference products. Especially during 2003, a small drop of the activity of benalaxyl against P. porri has been observed after repeated applications. In carrots, Erisiphe heraclei and Alternaria dauci are both the most common leaf diseases causing yield and quality loss. During periods of very high pressure of A. dauci, pyraclostrobin + boscalid, applied in a three weeks interval, revealed a superior activity compared with triazole references or compared with azoxystrobin. Against E. heraclei, a good control but also a clear dose response activity have been observed with pyraclostrobin + boscalid. Yield gain was approximately 30 ton /ha compared wih untreated. In Brussels sprouts, good efficacy was obtained against Mycosphaerella spp., Albugo candida and Alternaria spp. In outdoor lettuce Botrytis cinerea and Sclerotinia sclerotiorum are the most important diseases causing crop damage and reducing the quality of the heads. Pyraclostrobin + boscalid was evaluated in comparison with the standard fungicide iprodione. The plant protection was better with the new fungicide pyraclostrobin + boscalid.     

INVESTIGATIONS ON FUNGICIDES

Twenty (aryloxythio)trichloromethanes were examined for in vitro fungicidal activity against six fungi. All compounds showed a direct fungistatic effect and some exhibited a marked fumigant action. Good protectant action against Alternaria solani on tomato and Botrytis fabae on broad bean was obtained with certain compounds, but none was better than the three standard protectants used for comparison. When supplied to plants through their roots, eight conferred significant systemic fungicidal protection against Alternaria solani in tomato, but there was no significant protection against Botrytis fabae in broad bean. In preliminary tests the 2:4:5-trichlorophenoxythio analogue gave promising results when examined as a fumigant fungicide for eradication of downy mildew in lettuce plants, for reducing lenticel rot in stored apples and for preventing blue-mould in oranges.     

Effects of mancozeb and other dithiocarbamate fungicides on <Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis>: the role of mitochondrial petite mutants in dithiocarbamate tolerance

Saccharomyces cerevisiae as model system was used to evaluate the occurrence of resistant mutants and adaptation mechanism to mancozeb (MZ), a widespread fungicide of the dithiocarbamate class with a broad spectrum of action and multiple cell targets. We were unable to isolate mutants resistant to inhibitory concentration of MZ but found an unusually large number of mitochondrial defective petite mutants among cells incubated in the presence of subinhibitory MZ concentration. Similar results were obtained with two other dithiocarbamate fungicides. Comparison of wild type and petite mutants showed that the latter were more resistant to toxic effects of MZ, highlighting the role of mitochondria in MZ-tolerance. The data suggest that petite cells, arising by exposure to sub-inhibitory MZ concentration, are not induced by fungicides but are spontaneous mutants already present in the population before the contact with the fungicide.     

Possible explanations of synergism in fungicide mixtures against Phytophthom infestans

Single foliar sprays of oxadixyl, mancozeb or cymoxanil alone or the mixtures oxadixyl/mancozeb and oxadixyl/cymoxanil were applied to tomato plants (cv. Baby) one day before inoculating with a sensitive or a phenylamide-resistant strain of Phytophthora infestans. Fungicide mixtures were far more effective in controlling both sensitive and resistant strains compared to the individual components alone, representing significant levels of synergistic interactions. When analysed by the Abbott method, synergy ratios continuously decreased with increasing amounts of fungicides in the mixture. All fungicide mixtures showed higher synergy ratios against resistant strains when using the Abbott method, whereas only mixtures containing cymoxanil showed this phenomenon when using the Wadley method for interpretation of interactions. Synergism significantly decreased when the components were used as foliar split applications with intervals of 48 and 72 h. A combination of a drench application of oxadixyl and a single spray application of mancozeb or cymoxanil resulted in a high level of synergism. The explanation of synergism may be that the exposure of sporangia to sublethal concentrations of one fungicide affects them to an extent that sublethal doses of the other fungicide becomes detrimental.     

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     

Reduced sensitivity of tomato early blight pathogen (Alternaria solani) isolates to protectant fungicides,and implication on disease control

The sensitivity of Alternaria solani isolates to the fungicides mancozeb and chlorothalonil was evaluated, to determine if inadequate disease management by these fungicides could be attributed to reduced sensitivity of A. solani isolates to these fungicides. The sensitivity of 60 isolates of A. solani was assessed using the inhibition of radial mycelial growth (RG) method, using fungicide concentrations of 0, 1.0, 10, 100, 500 and 1000 μg a.i ml^?1 medium. EC₅₀ was calculated for each isolate and fungicide combination. The EC₅₀ values of different A. solani isolates to mancozeb ranged from 9.05 to 712.65 μg ml^?1. EC₅₀ values of different isolates to chlorothalonil ranged from 4.25 to 849.4 μg ml^?1. The percentage of isolates with reduced sensitivity was 46.7 and 53.3% for mancozeb and chlorothalonil, respectively. Results of the in vivo tests demonstrated decline in disease control by the two fungicides with the reduced-sensitivity isolates compared to the sensitive ones.     

Management of the late blight (Phytophthora infestans) disease of potato in the southern hills of India

Late blight of potato is considered to be the most devastating problem causing severe yield losses in potato worldwide. Among the different management strategies, the use of resistant cultivars is the most viable option, but the non‐availability of enough quantity of quality seed materials of resistant cultivars forces the farmers to grow susceptible cultivars with proper fungicide scheduling. Therefore, in the present study, chemical control using fungicide has been attempted with newer molecules in the susceptible cultivar along with a resistant cultivar as a positive control. All the tested fungicides were found safe, and no phytotoxicity was observed with any chemical at the applied rate. In resistant cultivar, no late blight was appeared in both the years, whereas maximum AUDPC was observed in the untreated control (276.3) and minimum (41.7) in mancozeb‐cymoxanil + mancozeb based scheduling which was found on par with chlorothalonil‐famoxadone + cymoxanil (51.3) and chlorothalonil‐ametoctradin + dimethomorph (53.5) based scheduling. Among the treatments, resistant cultivar, Kufri Girdhari followed by chlorothalonil‐ametoctradin + dimethomorph and mancozeb‐cymoxanil + mancozeb based fungicidal scheduling were proven as the best treatments for both the crop seasons resulting in the highest yield parameters. The disease severity showed a strong negative correlation with the tuber yield of potatoes in both the years. Based on overall observations including BC ratio, it can be concluded that, wherever seed material of resistant cultivar is available farmers should grow the same or else with susceptible cultivars the fungicidal scheduling based on mancozeb‐cymoxanil + mancozeb or chlorothalonil‐ametoctradin + dimethomorph can be followed to obtain the maximum returns with effective management of late blight at the southern hills of India.     

Effect of wax and lanolin emulsions on the efficiency of potato-blight fungicides in laboratory tests

Four potato-blight fungicides (copper oxychloride, tetrachloroisophthalo-nitrile, zineb and fentin acetate) were formulated either by adding emulsions of paraffin wax or lanolin to solid fungicide dispersed in water, or by dissolving the fungicide in wax or lanolin and then emulsifying the solution; all emulsions were non-creaming and extremely stable. These formulations were compared in the laboratory with commercial wettable powders of the same fungicides for their ability to protect detached potato leaflets against infection by Phytophthora infestans (Mont.) de Bary; in these and in other laboratory tests, the quality of rain-fastness, or tenacity, was involved directly or indirectly. Most of the wax or lanolin formulations gave better protection than the wettable powders. It is suggested that the wax and lanolin acted as viscous stickers, suitably increasing the rain-fastness of deposits on leaflets; and that when the fungicide was dissolved in the wax or lanolin before emulsification, the particle size of the dispersed fungicide was extremely small, a quality that increases biological action.