Glasshouse experiments on apple scab

Experiments with fungicide mixtures used protectively and curatively against apple scab showed no advantage from adding phenylmercury chloride (PMC) to certain protective fungicides. Sulphur fungicides greatly impaired the curative activity of PMC, especially in mixtures applied 24 hr. after infection, but captan showed little and dodine acetate no such effect. PMC, however, contributed appreciably to the curative effectiveness of mixtures with captan or dodine acetate. Commercially prepared mixtures of dispersible sulphur with phenylmercury dimethyldithiocarbamate and of zineb with mancozeb gave promising results, and an experimental mixture of dodine/glyodin acetates at low rates was effective protectively and especially curatively. Dodine acetate, dichlofluanid, lime-sulphur, and isobutyl-o-coumarate showed little translocated activity against scab when applied after infection, and PMC, which earlier showed strong translocated activity when applied in summer before infection, was much less effective when applied in spring 24 hr. after infection. Uncontrolled powdery mildew early in the scab-incubation period greatly reduced the establishment of scab infection on the test plants (clonal root-stocks); methods of mildew control in such experiments are suggested.     

Effects of fungicide spray regimes on incidence of dicarboximide resistance in grey mould (Botrytis cinerea) on strawberry plants

In tunnel experiments, the efficacy of dicarboximide sprays in controlling grey mould of strawberries was greatly decreased by the presence of dicarboximide-resistant forms of Botrytis cinerea. The use of dichlofluanid, as a tank-mix or in an alternating programme, with a dicarboximide fungicide, procymidone, helped to maintain the efficacy of disease control but failed to prevent an increase in the proportion of dicarboximide-resistant forms of the pathogen. Alternative ‘partner’ fungicides (thiram, chlorothalonil) delayed build-up of resistance to dicarboximides. Build-up of resistance was absent or relatively small in unsprayed plots. Application of dichlofluanid alone was always associated with a substantial increase in dicarboximide resistance, although less than in procymidone-treated plots. Monitoring dicarboximide resistance in the tunnels during the winter, when no further sprays were applied, revealed a gradual decline in the proportion of dicarboximide-resistant forms in all previously treated plots. In laboratory studies on inoculated leaf debris, dichlofluanid treatment induced the build-up of dicarboximide-resistant forms of B. cinerea. Leaf-disc tests revealed cross-resistance of dicarboximide-resistant isolates towards dichlofluanid but not towards thiram or chlorothalonil. Dichlofluanid is widely used for control of B. cinerea and the implications of these results for the practical management of dicarboximide resistance in this pathogen are discussed.     

Induction of rat liver cytochrome P450 isoenzymes CYP 1A and CYP 2B by different fungicides, nitrofurans, and quercetin.

The genotoxic activity of environmental xenobiotics is manifested either in their direct interaction with cellular genetic material or in provoking secondary events, among which reactive oxygen species (ROS) production is a common phenomenon. Both pathways can be mediated by the activity of the cytochrome P450 monooxygenase system. We studied induction of the CYP 1A or CYP 2B monooxygenases in rat liver by the fungicides: thiram, captan, captafol, dodine and the drugs: nitrofurazone, furazolidone and the plant flavonoid: quercetin. A cytochrome P450 induction assay (CYPIA test) was used. S9 prepared from livers of rats treated with the test compounds were used to activate ethidium bromide (EtBr) (CYP 1A isoenzyme) or cyclophosphamide (CPA) (CYP 2B isoenzyme) in the Ames test.It was found that among the tested compounds, the most potent inducer of CYP 1A was furazolidone (3 x 80 mg/kg). Less potent was thiram (1 x 100mg/kg), as well as quercetin (3 x 80 mg/kg), and captafol (1 x 30 mg/kg). On the other hand, thiram (1 x 100 mg/kg), captafol (1 x 30 mg/kg), and quercetin (3 x 80 mg/kg) were most potent in the CYP 2B isoenzyme induction, while furazolidone (3 x 80 mg/kg), and nitrofurazone (3 x 80 mg/kg) appeared to be less potent in this respect. Captan and dodine (3 x 80 mg/kg) did not affect the activity of any of the cytochrome P450 isoenzymes.     

Induction of rat liver cytochrome P450 isoenzymes CYP 1A and CYP 2B by different fungicides, nitrofurans, and quercetin

The genotoxic activity of environmental xenobiotics is manifested either in their direct interaction with cellular genetic material or in provoking secondary events, among which reactive oxygen species (ROS) production is a common phenomenon. Both pathways can be mediated by the activity of the cytochrome P450 monooxygenase system. We studied induction of the CYP 1A or CYP 2B monooxygenases in rat liver by the fungicides: thiram, captan, captafol, dodine and the drugs: nitrofurazone, furazolidone and the plant flavonoid: quercetin. A cytochrome P450 induction assay (CYPIA test) was used. S9 prepared from livers of rats treated with the test compounds were used to activate ethidium bromide (EtBr) (CYP 1A isoenzyme) or cyclophosphamide (CPA) (CYP 2B isoenzyme) in the Ames test.It was found that among the tested compounds, the most potent inducer of CYP 1A was furazolidone (3× 80 mg/kg). Less potent was thiram (1× 100 mg/kg), as well as quercetin (3× 80 mg/kg), and captafol (1× 30 mg/kg). On the other hand, thiram (1× 100 mg/kg), captafol (1× 30 mg/kg), and quercetin (3× 80 mg/kg) were most potent in the CYP 2B isoenzyme induction, while furazolidone (3× 80 mg/kg), and nitrofurazone (3× 80 mg/kg) appeared to be less potent in this respect. Captan and dodine (3× 80 mg/kg) did not affect the activity of any of the cytochrome P450 isoenzymes.     

The persistence in soil of the fungicidal seed dressings captan and thiram

The persistence in soil of captan and thiram was investigated by means of a technique in which the fungicidal content of soil was assayed by incubating plugs of soil containing fungicide on agar plates seeded with spores of Myrothecium verrucaria and measuring the diameter of the zone of fungal inhibition that was produced. When the fungicides were well distributed in soil they showed extremely low persistence, both fungicides having a half-life of between 1 and 2 days. In contrast, when the fungicides were added to soil in the form of dressings on the surface of glass beads they persisted well in soil, little change from their initial concentration occurring even after 21 days. These results suggest that captan and thiram persist far longer in soil when localized in high concentrations than when uniformly distributed through soil. If a glass bead is regarded as a reasonable simulation of a seed these results help to explain the effectiveness of these fungicides as seed dressings despite their apparently low persistence in soil.     

Laboratory tests of fungicides against Peronospora parasitica (Pers. ex Fr.) Fr

Methods were investigated of culturing Peronospora parasitica on detached cotyledons in plastic box moist chambers in constant-temperature cabinets. Light was essential and a temperature of 15 °C was most suitable for infection and survival of cotyledons. Using such cotyledons the protectant and eradicant actions of various fungicides against P. parasitica were compared with those of zineb. Captafol, Daconil 2787, dichlofluanid and propineb were markedly superior to zineb as protectants, being highly effective at 0·025 % a.i. None of these compounds (at 0·2 % a.i.) checked fungal development when applied 5 h after inoculation, during which time the fungus had penetrated the cotyledons, and it was concluded that the fungicides had thus no internal effect. Internal fungistasis was shown, however, by a maneb-nickel sulphate fungicide and also by nickel sulphate. The latter (at 0·4% NiSO₄.6H₂O) gave some disease control when applied up to 72 h after inoculation and appeared to be translocated within the tissue of the cotyledon.     

The control of apple canker (Nectria galligena) in a young orchard with established infections

In a factorial experiment on canker control the efficiency of phenylmercurie nitrate (PMN) applied at leaf fall, before bud burst or both was compared with the application of dodine, dithianon, triforine, thiabendazole, benomyl or carbendazim in May and June. Of the total number of cankers which developed in unsprayed trees 76–78% resulted from infections in April to August of each year of the experiment. Infection was reduced significantly by all of the fungicides applied in summer. Of these carbendazim was outstanding, controlling both summer and autumn infections. Sporulation throughout the summer was suppressed by carbendazim and to a lesser extent benomyl, but whereas the suppressant effect of carbendazim persisted until long after leaf fall that of benomyl was evident only until August. Dodine, dithianon, triforine and thiabendazole had no significant effect on spore production. Dithianon and dodine showed highest toxicity to the germination of Nectria galligena spores of all fungicides used in summer. All of the fungicides controlled apple scab (Venturia inaequalis) although thiabendazole was the least effective. Autumn applications of PMN reduced canker incidence but their value alone was not as great as carbendazim, dithianon, benomyl or dodine applied in summer. PMN applied in spring reduced the number of cankers in trees receiving no other fungicides but tended to increase the incidence of infections, particularly in autumn, when used in conjunction with fungicides applied in May and June. PMN applied before bud burst reduced sporulation of N. galligena for a few weeks, after which production resumed and in late summer and autumn actually exceeded the controls. None of the fungicides had any direct effect on leaf fall. Infection of the crotch and basal leaf scars was more common in summer than in autumn whereas infection of leaf scars above the basal region was more common in autumn than in summer. The rootstocks of many of the trees became infected via callus tissue associated with adventitious root development and this was controlled by the carbendazim treatment.     

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.     

Evaluation of the effects of chemical versus biological control on Botrytis cinerea agent of gray mould disease of strawberry

This study investigates on effects of four fungicide and six isolate from Trichoderma and Gliocladium on Botrytis cinerea agent grey mold of strawberry under library and greenhouse condition. The effect of four fungicides i.e. benomyl, dichlofluanid, captan and triadimenol on B. cinerea was studied in the laboratory condition by method mixed poison to culture medium. It was shown that the fungicide including benomyl, triadimenol, dichlofluanid and captan were able to inhibit mycelial growth of B. cinerea on PDA plate with EC50 of 0.16, 1.42, 3.40 and 7.73 ppm respectively. These fungicides delayed myceliogenic germination of sclerotia at 1000 ppm, while exhibiting no fungicidal effect. Moreover, the antagonistic effects of six fungi including Trichoderma koningii (T21), T. viride (T4), T. harzionum (T5), T. viride (T2), G. virens (G2), G. virens (G8) on B. cinerea were assessed. This assessment was done under library condition and its results as follows: The antagonistic mechanism occurred through branching at the end of B. cinerea hyphae, hyphal contact, coiling, vacuolization and lyses. Volatile metabolites of T. koningii (T21) and non-volatile metabolites of G. virens (G2 and G8) and T. koningii (T21) caused maximum inhibition of the fungal growth. Trichoderma spp and G. virens were able to colonize and sporulate on sclerotia and caused their lysis within 7-21 days. In greenhouse, a completely randomized design with 11 treatments (4 chemical and 6 biological and one untreated control) each replicated five times were used for the comparison. Greenhouse studies revealed that application of fungicides i.e. captan, dichlofluanid, triadimenol and benomyl reduces disease severity by 42, 45, 48 and 52% respectively. The fungal antagonists reduce the grey mold disease severity between 5-42%. All treatments caused a decline in post harvest disease, as the most effective treatment of chemical control was benomyl with 68.33% and for the biological treatment this was T. koningii (T21) with 56%.     

A Cucumber Disease Caused by Alternaria alternata and its Control

A severe leaf spot disease of cucumber caused by a pathotype of Alternaria alternata (Fr.) Keissler was recorded recently in plastic houses in Crete. Lesions ranged in size of a pin point to over 5 cm in diameter, with necrotic tissue on most of their area and a surrounding yellow zone. The pathogen grew satisfactorily on PDA at temperatures between 5 °C–40 °C and spore germination occurred in the range less than 10 °C to over 37 °C. Optimum temperature in both cases was near 26 °C. Of,13 fungicides tested in vitro, sodium omadine, etem, dichlofluanid, captan and folpet were the most inhibitory on spore germination, and iprodione, sodium omadine and dichlofluanid on mycelial growth. Of 25 fungicides applied on two leaf cucumber plants 24 h before inoculation, maneb, etem, dichlofluanid and chlorothalonil were the most effective. When the last fungicides, plus mancozeb, were applied 24 h after inoculation only maneb was effective. In greenhouse experiments, iprodione, prochloraz-manganese-complex, chlorothalonil, dichlofluanid, guazatine, maneb and etem were the most effective for disease control, while mancozeb was less effective. The local cucumber cv. Knossos and the Dutch F₁ hybrids Evadan, Frella, Herta, Malfa, Mazourka, Pepinex 69 and Renova were all susceptible to infection.     

Control of Botrytis cinerea on glasshouse tomato by high volume sprays

The new fungicides iprodione, vinclozolin and prochloraz, and also a mixture containing carbendazim and maneb, were compared with the established protectant fungicides dichlofluanid and chlorothalonil for effectiveness against grey mould ( Botrytis cinerea ) in an unheated tomato crop. Iprodione and vinclozolin gave the best control of lesions on leaves and stems but dichlofluanid was the most effective in controlling ghost-spotting of fruit. The carbendazim/maneb mixture was effective against a carbendazim-sensitive isolate but not against an insensitive isolate of B. cinerea . When applied 1 or 2 days after inoculation, all five protectant fungicides controlled the insensitive isolate better than the sensitive isolate.     

Fungicidal management of wilt complex of scented geranium

Systemic fungicides, carbendazim + mancozeb and carbendazim alone completely inhibited the mycelia growth even at 100 and 200 ppm, whereas among non-systemic fungicides, thiram was found to be the most effective and gave 52.77, 62.77 and 85.00% inhibition at 500, 1000 and 1500 ppm concentrations, respectively. Thiram was found to be most effective fungicides in inhibiting the both fungi (68.84 and 58.69%) followed by the indofil M 45 (47.59 and 51.38%). In pot house condition, carbendazim + mancozeb (0.2%) and carbendazim (0.1%) were most effective fungicides in reducing cent percent disease incidence and severity, but it was at par with thiram (82.35; 87.50). In field condition, mulching with transparent polyethylene mulch (25 μm) resulted in highest survival rate (95.83%) and least mortality rate (4.16%) of rooted geranium cuttings after 30 days of germination. The application of carbendazim (Bavistin) at 0.1% when applied individually was also found almost complete control as is evident from low disease incidence (1.04%) and severity (0.52%), considered to be second best treatment. Apparent infection rate (0.0) was observed in carbendazim + mancozeb and carbendazim followed by copper oxychloride (0.006) and carboxin (0.009). Area under disease progress curve was less than 100 in cabendazim + mancozeb and carbendazim.     

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.     

Synthesis and Anti-Oomycete Activity of 1-Sulfonyloxy/Acyloxydihydroeugenol Derivatives

Endeavor to discover biorational natural products-based fungicides, two series (26) of novel 1-sulfonyloxy/acyloxydihydroeugenol derivatives ( 3a – p and 5a – j ) were prepared and assessed for their fungicidal activity against P. capsici Leonian, in vitro. Results of fungicidal activity revealed that, among all compounds, especially compounds 3a , 5c , and 5e displayed the most potent anti-oomycete activity against P. capsici with EC₅₀ values of 69.33, 68.81, and 67.77 mg/L, respectively. Overall, the anti-oomycete activities of 1-acyloxydihydroeugenol derivatives ( 5a – j ) were higher than that of 1-sulfonyloxydihydroeugenol derivatives ( 3a – p ). It is proved that the introduction of the acyl group at hydroxy position of dihydroeugenol is more beneficial to improve its anti-oomycete activity than that of the sulfonyl group. These preliminary results will pave the way for further modification of dihydroeugenol in the development of potential new fungicides.     

The control of fungal seed–borne diseases by means of a thiram seed soak

A method of soaking seeds in an 0·2% aqueous suspension of thiram for 24 h at 30 °C, which had previously been shown to eradicate infection by several fungi, was tested against a further thirteen seed-borne pathogens. Eleven of these were completely controlled, one was almost completely controlled and one was not adequately controlled by the treatment. The thiram soak treatment was much more effective than dust treatments with fungicides for the control of many internal fungal pathogens. It was also generally more effective and less damaging than hot-water treatment. The ways of applying the method commercially have been investigated and it is being adopted for the treatment of celery, brassicas and red beet.     

Tomato leaf mould: its assessment and effects on yield

When the natural incidence of Cladosporium fulvum Cooke, occurring at a relatively late stage of cropping, was controlled with fungicides, comparisons with unsprayed control plants showed that there was an interval of c. 6 weeks between the incidence of severe infection, when the fungus was colonizing at least 50 % of the host's leaf area, and fruit yield decreases. Eye estimates made on separate leaves usually over-estimated the percentage of infection. Correction factors were derived from the regression of eye estimates on infected areas calculated from leaf tracings. Whole-plant infection indices were obtained by averaging corrected values of percentage infection separately assessed on leaves immediately above each fruit truss. Although dichlofluanid and tank-mixed zineb decreased the incidence of C. fulvum more than captan sprays, tomato yields (including green fruit) were increased similarly by all three fungicides from 2·99 to an average of 3·51 kg/plant during c. 3 months' picking. Decreasing the intervals between successive sprays from 21 to 7 days greatly decreased leaf mould infection but did not significantly affect yields. Restricting applications to plant tops (namely foliage from the fourth-youngest truss to the mainstem apex) gave yield benefits equal to those gained from sprays applied to whole plants.     

Tests of fungicides against Alternaria brassicicola and studies on some factors enhancing attack by the fungus

When various fungicides were sprayed on detached cabbage cotyledons in the laboratory and on young plants in the glasshouse, propineb and mancozeb at 0·2 % a.i. consistently gave good protection against subsequent infection by spores of Alternaria brassicicola. Copper oxychloride, zineb, thiram and maneb showed promise in the laboratory, but were less successful in the glasshouse. Two fentin compounds were active at low concentrations against the fungus but were phytotoxic. Increased pathogenicity of A. brassicicola in the presence of cabbage pollen and of downy mildew infection in the laboratory is reported.     

Sensitivity of Clonostachys rosea and Trichoderma spp. as Potential Biocontrol Agents to Pesticides

Clonostachys rosea 47 (CR47), Trichoderma atroviride 59 (TA59), T. atroviride 312 (TA312), Trichoderma harzianum 24 (TH24), Trichoderma longibrachiatum 9 (TL9), T. longibrachiatum 144 (TL144) and Trichoderma viride 15 (TV15) were tested to evaluate their in vitro sensitivity towards five fungicides (carboxin, guazatine, prochloraz, thiram and triticonazole) and four herbicides (chlorsulfuron, chlorotoluron, flufenacet and pendimethalin). All antagonists showed low sensitivity to carboxin and thiram and high sensitivity to prochloraz. For mycelial radial growth, TV15 was highly sensitive to guazatine, prochloraz and triticonazole and TH24 moderately insensitive to carboxin, guazatine and thiram. For conidial germination TL144 was the most sensitive to the fungicides, for mycelial radial growth and conidial germination CR47 was the least sensitive. None of the antagonists showed any mycelial radial growth inhibition in presence of the herbicides at field dose, except for TL144. Most antagonists did not show any conidial germination inhibition by the herbicides. The in vitro toxicity of prochloraz, guazatine and triticonazole towards the antagonists was confirmed by light and scanning electron microscope showing hyphal disruptions and extrusion of cytoplasmic content. A mixture of CR47 and/or TA312 with carboxin, thiram and triticonazole, applied to wheat seeds, was able to control Fusarium culmorum artificially inoculated to wheat seedlings in growth chambers. In the field, the antagonists applied along with triticonazole or thiram, at 1/10 of the field dose to seeds naturally infected by F. culmorum, gave a disease control comparable to that induced by triticonazole at full field dose. Our results demonstrate how an integration of microorganisms with pesticides makes the control of wheat foot rot possible.     

黑杨水培中营养液抑菌处理研究

水培条件下进行快速繁殖是大量生产黑杨种苗的途径之一 ,营养液的抑菌处理是水培的一个重要方面。该研究进行了二硫氰甲烷、中生菌素、敌磺钠、链霉素、青霉素等五种杀细菌剂和多菌灵、福美双、三唑酮等三种杀真菌剂的抑菌试验 ,结果表明 ,杀细菌剂对微生物生长的抑制率要高于杀真菌剂 ,二硫氰甲烷和中生菌素能较好地抑制营养液中微生物的生长 ,浓度分别为 2 0mg/L和 1 5mg/L时 ,抑制率分别为 75 .60 %和78 94%。杀真菌剂和杀细菌剂复配对营养液中微生物生长的抑制表现出较好的加和作用 ,其中以中生菌素(1 5mg/L)与福美双 (5mg/L)复配的效果最好 ,抑制率为 87.48%。     

Studies on the survival and infective ability of dicarboximide-resistant strains of Botrytis cinerea

Few dicarboximide-resistant strains of Botrytis cinerea produce macroconidia either on culture media or on natural substrates, although, the viability of such conidia was similar to those of sensitive strains. The ability of dicarboximide-resistant strains to infect strawberries was similar to that of sensitive strains and most of the resistant strains competed successfully with sensitive strains when inoculated in equal numbers onto detached strawberries and onto flowers of field-grown plants, especially after treatment with dicarboximide fungicides. All resistant strains tested survived for at least 9 months on inoculated strawberry leaf litter although their incidence was ‘diluted’ by wild type sensitive strains. Sporulation and dispersal of resistant strains from the litter was very limited resulting in a low incidence of fruit infection even after treatment with dicarboximides. Consequently, there was no significant increase of resistant strains in the plantation and control of infection was maintained with iprodione and dichlofluanid. Poor sporulation of dicarboximide-resistant strains of B. cinerea is considered to be the single most important factor in limiting their development in the field.