Movement, persistence and performance of fungitoxicants applied as pruning wound treatments on apple trees

The systemic fungicides triadimenol, fenpropimorph, furmecyclox and carben-dazim, applied to pruning wounds as solutions in a non-aqueous solvent containing N-methyl–2-pyrrolidone moved predominantly towards the trunk. Coriolus versicolor, introduced deliberately, moved both proximally and distally irrespective of whether it was introduced in the dormant or growing season. Natural resistance to damage by chemical or C. versicolor appeared to be greater in the growing than in the dormant season and was inhibited when a cationic surfactant was used to aid penetration of fungitoxicant into the wood. Against C. versicolor the non-systemic fungicide captafol was ineffective, furmecyclox or fenpropimorph gave some protection and triadimenol was very effective. Carbendazim, although inactive against wood-rotting basidiomycetes, moved extensively within the wood and persisted well, and so could be an effective treatment for ascomycete pathogens. It also stimulated callus formation. The mercuric oxide preparation Santar was effective in these experiments and also stimulated callus formation.     

Biological treatments for the control of decay in tree wounds.: Field tests

In an attempt to control fungi causing decay a range of micro-organisms, selected after laboratory tests, was inoculated into wounds made in the trunks of beech trees. The most successful micro-organism was an isolate of Trichoderma viride, which, over a period of 4 years, maintained significantly lower colonisation by decay fungi at both sites - c. 15% of the level found in uninoculated wounds. T. viride could be reisolated from the majority of wounds into which it had been inoculated up to 4 years later. No other antagonist persisted very effectively. Inoculation of T. viride in glycerol improved its establishment but malt extract and sucrose had no such effect. Both the addition of glycerol to wounds and the wrapping of them in black PVC sheet increased natural colonisation by T. viride. The use of PVC but not glycerol reduced numbers of decay fungi but not through the agency of T. viride present by natural infection, as this proved generally ineffective in the control of decay fungi. Another isolate of T. viride persisted considerably better when covered with a wound sealant.     

Gel formulations for the treatment of pruning wounds.

Gels incorporating carbendazim and triadimefon were prepared from sodium alginate, a xanthan gum or an esterified alginic acid and evaluated as wound treatments on apple and plum trees. Fungicide movement and persistence were similar with sodium alginate and xanthan gels but movement was reduced and persistence increased with the esterified material. Callus tissue formation was not inhibited by any formulation. Adding acid to fungicides to form salts had little effect on fungicide movement or persistence and caused unacceptable phytotoxicity. Thiophanate-methyl or thiabendazole were not superior to carbendazim, nor imazalil to triadimefon, for protection against Nectria galligena and Chondroster-eum purpureum respectively. Sealants containing octhilinone, carbendazim plus captan, or mercuric oxide quickly established sufficient fungicide in the wood to eradicate N. galligena but possibly insufficient to protect adequately against C. purpureum. Gel formulations are valid alternatives to sealants as fresh wound pathogen treatments but fungicide persistence is insufficient for them to be recommended for treatment against mature wound pathogens.     

The phylloplane microflora of ripening wheat and effect of late fungicide applications

Flag leaves and ears of spring wheat cv. Timmo (in 1980) and winter wheat cv. Maris Huntsman in 1981 and 1982 were colonised by a variety of micro-organisms whose numbers increased rapidly between anthesis and harvest. The predominant mycoflora were yeasts, yeast-like fungi and filamentous fungi which included Cladosporium spp., Verticillium lecanii, Alternaria alternata, Fusarium spp. and Epicoccum nigrum. Although similar species were isolated, their relative abundance on flag leaves and ears differed. The fungicide captafol was most effective as a protectant and significantly decreased populations of fungi on flag leaves and ears for 6 and 4 wk respectively, compared to untreated controls. Benomyl and Delsene M (carbendazim + maneb) were the most effective of the systemic sprays and formulations. In general, fungicides affected populations of yeasts, yeast-like fungi and Cladosporium spp. most while Alternaria was tolerant of all treatments. Yields of winter wheat were increased in two seasons by an average 0–2 t ha^-1 (2–4%) following a single late fungicide treatment at G.S. 50 or 60 and 0–41 t ha^-1 (5-1%) when this was combined with an early spray against foliar diseases (G.S. 38–40). Individual treatments increased yield by up to 12% with little difference between applications at G.S. 50 or 60. The yield benefit came mainly from increased 1000-grain weights. Germination of the treated grain was increased only slightly.     

The microflora of ripening barley grain and the effects of pre harvest fungicide application

Newly-emerged ears of barley soon became colonised by micro-organisms with numbers increasing to 5·8 × 10⁵ viable propagules/g dry weight by harvest. Bacteria were numerically predominant throughout. Yeasts and yeast-like fungi were also numerous during the early stages of grain development but were exceeded by propagules of filamentous fungi during ripening, with Cladosporium spp., Alternaria alternata, Epicoccum purpurascens and Verticillium lecanii most abundant. Numbers of most micro-organisms were greatest in a wet season but Alternaria was equally abundant in hot dry weather. Application of fungicides at or after anthesis modified the grain microflora. Captafol was most effective overall in decreasing numbers of fungi. However, benomyl decreased Cladosporium inoculum more than catpafol but was ineffective against Alternaria which became more numerous on benomyl-treated grain than on untreated. Yield increases of up to 4% were obtained by late fungicide treatment, least in a hot, dry year. Germination was also increased by some treatments by up to 5%.     

The effects of some commonly-used foliar fungicides on Collembola in winter barley: laboratory and field studies

Four commonly-used cereal foliar fungicides were screened for their laboratory toxicity against the symphypleone collembolan, Sminthurinus aureus. A proportional hazards analysis of time-survival curves following the fungicide treatments showed that carbendazim, propiconazole, pyrazophos and triadimenol significantly increased the laboratory mortality of S. aureus. The organophosphorus fungicide pyrazophos caused high levels of mortality of S. aureus in the laboratory so a field evaluation of the effects of this fungicide on a wider range of Collembola was undertaken in winter barley. Comparison of the effects of pyrazophos with those of the broad-spectrum insecticide dimethoate in the field revealed both compounds to have similar activity against some Collembola. Of the 11 species caught only the four symphypleone species exhibited these effects but the numbers of three symphypleone species were reduced to zero 4 wk after treatment with pyrazophos. The effects of pyrazophos and dimethoate were, however, not detectable in individual species after 11 wk.     

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%.     

Integration of metabolomics and in vitro metabolism assays for investigating the stereoselective transformation of triadimefon in rainbow trout

Triadimefon is a systemic agricultural fungicide of the triazole class whose major metabolite, triadimenol, also a commercial fungicide, provides the majority of the actual fungicidal activity, i.e., inhibition of steroid demethylation. Both chemicals are chiral: triadimefon has one chiral center with two enantiomers while its enzymatic reduction to triadimenol produces a second chiral center and two diastereomers with two enantiomers each. All six stereoisomers of the two fungicides were separated from each other using a chiral BGB‐172 column on a GC‐MS system so as to follow stereospecificity in metabolism by rainbow trout hepatic microsomes. In these microsomes the S‐(+) enantiomer of triadimefon was transformed to triadimenol 27% faster than the R‐(?) enantiomer, forming the four triadimenol stereoisomers at rates different from each other. The most fungi‐toxic stereoisomer (1S,2R) was produced at the slowest rate; it was detectable after 8 h, but below the level of method quantitation. The triadimenol stereoisomer ratio pattern produced by the trout microsomes was very different from that of the commercial triadimenol standard, in which the most rat‐toxic pair of enantiomers (known as “Diastereomer A”) is about 85% of the total stereoisomer composition. The trout microsomes produced only about 4% of “Diastereomer A”. Complementary metabolomic studies with NMR showed that exposure of the separate triadimefon enantiomers and the racemate to rainbow trout for 48 h resulted in different metabolic profiles in the trout liver extracts, i.e., different endogenous metabolite patterns that indicated differences in effects of the two enantiomers. Chirality, 2010. © 2009 Wiley‐Liss, Inc.     

Effects of wheat foliar fungicides on the aphid endoparasitoid Aphidius rhopalosiphi DeStefani-Perez (Hym., Aphidiidae) on glass plates and on plants

Abstract: The side-effects of several fungicides used in wheat to control disease at heading growth stage were assessed on the aphid parasitoid Aphidius rhopalosiphi by tests conducted in the laboratory on glass plates and in the greenhouse on young wheat plants. Very few formulations containing only one active ingredient (carbendazim, cyproconazole or epoxyconazole) or combinations of two (carbendazim + cyproconazole, carbendazim + hexaconazole) were harmless to A. rhopalosiphi in the glass-plate tests. There was no apparent synergism between fungicides tested in combinations. The parasitoid mortalities in tests carried out on plants were less and chlorothalonil, epoxyconazole, fenpropidin, fenpropimorph, flusilazole, flutriafol, prochloraz, tebuconazole, tridemorph and a number of combinations (carbendazim + flutriafol, chlorothalonil + cyproconazole, epoxyconazole + tridemorph, chlorothalonil + hexaconazole, chlorothalonil + flutriafol, cyproconazole + prochloraz, epoxyconazole + fenpropimorph, fenpropimorph + propiconazole, propiconazole + tridemorph, triadimenol + tridemorph) were harmless or only slightly harmful to the aphid parasitoid. Several combinations (carbendazim + epoxyconazole, carbendazim + fenpropimorph, carbendazim + flusilazole, carbendazim + tebuconazole, chlorothalonil + fenpropimorph, chlorothalonil + flusilazole, fenpropimorph + fenpropidin, fenpropimorph + prochloraz, fenpropidin + propiconazole, fenpropidin + tebuco nazole, tebuconazole + triadimenol) were toxic for wasps on plants. The parasitoid mortalities were less on plants than on glass plates but the wasps spent less time on treated leaves and in some cases parasitism of aphids was reduced to a large extent. These results suggest that in addition to study of the direct effects of pesticides on beneficial insects (mortalities, reduction of fertility) their effects on the behaviour of the insects should also be studied. Products that induced a repellent effect need further testing in field or semi-field conditions. However, many fungicide combinations that have little or no effect on A. rhopalosiphi can protect wheat against a wide range of diseases and the results obtained in this study indicate that an appropriate and effective protection of wheat at earing growth stage can be achieved with products that have no effects on aphid parasitoids.     

Effects of delaying fungicide treatment on the incidence of gangrene in stored potato tubers

Potato titbers infested with Phoma exigua var. foveata were uniformly wounded and sprayed or dipped in fungicide suspensions either immediately or after periods of up to 21 days' storage at 5, 10, 15 or 20 °C. Tubers were then stored at 5 °C and gangrene assessed after 12 wk. Incidence of gangrene on untreated tubers was progressively decreased by increasing the length of storage at 15 or 20° (curing) but was not affected by 3 days' storage at any temperature. Fungicide treatment immediately after wounding gave best control of the disease; treatment after 3 days' delay was less effective and after 14 or 21 days was usually ineffective. Gangrene was decreased by fungicides more on tubers stored for 3 or 7 days at 5 °C than at higher temperatures. Control of gangrene by curing or fungicides diminished when the amount of inoculum on tubers was increased. Increasing the amount of fungicide applied improved control and fungicides were more effective in decreasing gangrene on cut and crush wounds than on cut wounds. At the arbitrary concentrations used in these experiments imazalil gave better disease control than thiabendazole, prochloraz, carbendazim plus quinolin 8-ol or triadimefon.     

Side effects of propiconazole (tilt 250 ECTM) on non-target soil fungi in a field trial compared with natural stress effects

The present study was performed as a dose-response field experiment using a trade marked formulation of the ergosterol biosynthesis inhibiting fungicide, propiconazole, applied at the recommended and ten times the recommended application rates. The soil dilution plate method was used to isolate fungi from 0–1 and 1–2 cm soil depth. Soil samples were taken 10 times during the period from May 22 to August 11, 1986. The numbers of yeasts and filamentous fungi were enumerated, the latter includingCladosporium andPenicillium, which were also enumerated separately. The fluctuations in fungal occurrence due to the propiconazole treatment were smaller than seasonal fluctuations, these most likely being caused by variations in the soil matric potential and by soil invasion of phyllosphere fungi. Fungicide treatment had significant inhibitory effects on the filamentous fungi, especially on theCladosporium, whereas no significant effects were found on the yeasts and onPenicillium spp. From an ecotoxicological point of view, it is important that the effects were considerably delayed, with respect to the time of fungicide application.     

The Control of Fusarium solani f. sp. phaseoli by Fungicide Mixtures

Six fungicides were used either alone or in binary combination to control Fusarium solani f. sp. phaseoli. Of the three methods of application used, seed treatments and soil drenches were the best, although phytotoxicity was evident in some instances. Application as a seed soak for 1 h proved to be the worst method with most treatments showing signs of phytotoxicity. The results are discussed with reference to the use of fungicide mixtures to control foot rot of beans.     

Assessment of the biocontrol potential of a Trichoderma viride isolate: Part II: Protection against soft rot and basidiomycete decay

A previous paper reported on the establishment of a field and fungal cellar trial set up to determine the biocontrol potential of a specific Trichoderma isolate against wood decay fungi. This paper reports on the analyses used to examine the protective effect of the selected isolate, and presents results indicating an initial protective effect against both basidiomycetes and soft rot fungi. The parameters assessed in the field and fungal cellar trials were soft rot decay, basidiomycete decay, Trichoderma colonisation, moisture content and nitrogen content. The results of these analyses show that the introduction of a biological control agent has had a significant effect on moisture content, decay and nitrogen content. A protective effect has been observed against soft rot and basidiomycete decay fungi in field samples.     

Growth and viability of mycorrhizal extraradical mycelia associated with three temperate orchid species

Growth and enzymatic activities of extraradical mycelia (ERM) of native mycorrhizal symbionts associated with three orchid species, Dactylorhiza fuchsii, D. majalis and Platanthera bifolia, were studied. ERM extracted from the mycorrhizosphere of these species showed features typical for fungi that form orchid mycorrhiza. In the first pot experiment, three different treatments were applied on tubers of D. fuchsii transplanted from a natural site: control (no specific treatment), reinoculated (surface-sterilized tubers reinoculated with mycorrhizal fungi-colonised roots), and benomyl (nonsterilized tubers treated with fungicide). However, no significant differences in ERM growth and intensity of root mycorrhizal colonisation at harvest were observed among these treatments. ERM associated with reinoculated D. fuchsii plants showed significantly higher alkaline phosphatase (ALP) enzymatic activity at week 36 than at week 24, but no differences were observed for NADH diaphorase activity. Benomyl application significantly reduced ALP activity in comparison with reinoculated plants at week 36. In the second experiment, plants of all three species were either untreated (control), or repeatedly treated with benomyl. Similarly to the results of the first experiment, benomyl application did not reduce the ERM growth of mycorrhizal symbionts associated with D. majalis and D. fuchsii. The low ERM growth associated with benomyl-treated P. bifolia was probably caused by poor root system development in this treatment. Significantly higher mycorrhizal colonisation was found for D. fuchsii compared to P. bifolia in control treatments at the end of cultivation. The ERM of native symbionts of the three orchid species studied seemed to have a different growth pattern over time and responded differently to fungicide application.     

The disappearance of benomyl from mushroom casing

The failure of Benlate (50% benomyl) to control Mycogone perniciosa on mushroom farms where isolates of the organism were benomyl sensitive was investigated. A comparison was made of carbendazim levels, the major fungitoxic breakdown product of benomyl, in unsterilised casing, autoclaved casing and autoclaved casing mixed with a small proportion (1.7%) of casing previously treated with benomyl but from which all detectable levels of carbendazim had disappeared. Added benomyl was effective in controlling M. perniciosa in autoclaved casing, less effective in unamended casing and ineffective in the amended casing mixture. The recovery of carbendazim from these treatments was directly related to disease occurrence. Studies on farms where benomyl or other carbendazim generating fungicides had been used indicated that a lack of control of M. perniciosa was associated with the disappearance of fungicide from the casing before cropping began. This was not so on a farm where carbendazim generating fungicides had not been used before these investigations. Thiabendazole, a fungicide closely related to the carbendazim generators showed only a slight decline in concentration in casing capable of degrading benomyl. Bacteria isolated from the casing were examined in vitro for their ability to degrade benomyl. Five groups of bacteria were compared and three bacteria, Pseudomonas putida, Pseudomonas aeruginosa and an unidentified member of the Entero-bacteriaceae were most able to degrade benomyl.     

Effects of fungicide timing on control of Rhynchosporium secalis in barley plants

Sprays of captafol, carbendazim, carbendazim + tridemorph + maneb, diclobutrazol, triadimefon or triadimefon + carbendazim all completely protected barley plants in a glasshouse against R. secalis for at least 30 days. However, their effectiveness in preventing disease development when applied after inoculation differed: triadimefon, traidimefon + carbendazim, or diclobutrazol were the most effective, completely preventing symptom development when applied up to 5 days after inoculation to plants grown above 16 °C, and up to 8 days below 8 °C. All the fungicides decreased the number of viable conidia produced by leaf blotch lesions, and when applied to infected plants at G. S. 30, greatly decreased the upward spread of the disease under simulated rain conditions; the most effective fungicides in these respects were triadimefon and triadimefon + carbendazim. The above fungicides and fungicide mixtures, together with the recently introduced materials fenpropimorph and propiconazole were applied to diseased winter barley crops in winter or in spring. All treatments decreased leaf blotch development and increased yields. In most cases, a winter application was more effective than spring applications, particularly if applied in mid-November. The most effective fungicides were triadimefon and propiconazole. The field trials data fitted well with the predictions of performance indicated by the glasshouse investigations.     

Interactions between indigenous arbuscular mycorrhizal fungi and <Emphasis Type="Italic">Aphanomyces euteiches</Emphasis> in field-grown pea

This is the first reported study of the interactions between indigenous arbuscular mycorrhizal fungi (AMF) and Aphanomyces euteiches in pea under field conditions. A. euteiches was applied to the soil by adding oospores produced in vitro. Attempts were made to create a non-mycorrhizal control by incorporating carbendazim (Derosal Fl) in the topsoil before sowing. However, all carbendazim-treated plants showed approximately 20% root colonisation with AMF. Pea plants not treated with carbendazim showed a wide variation in AMF colonisation of 35-70% at the full flowering stage. In these control plots, root length infected with oospores of A. euteiches and colonisation by AMF were negatively correlated. Application of carbendazim increased the percent root length infected with oospores by 50-70%, depending on inoculum density of A. euteiches. Despite the lower levels of AMF colonisation in these treated plots, a negative correlation with oospore-containing root length was still observed. No correlation was found between AMF colonisation and disease severity, disease incidence or pathogen enzymatic activity (glucose-6-phosphate dehydrogenase). Thus, AMF do not seem to influence the vegetative stage of pathogen development during which cortical root rotting takes place, but rather the reproductive stage when oospores are produced. The results of this study underline the importance of field experiments for validating the significance of mycorrhizal fungi for plant health.     

The integration of host resistance with fungicides in the control of oat powdery mildew

Treatment of seed with the systemic fungicides triadimenol plus fuberidazole as Baytan significantly decreased powdery mildew in field experiments involving eight oat cultivars differing widely in their resistance to the disease. This was so up to growth stage G.S. 61 (Zadoks) even in a year of high mildew incidence, although the effect diminished after flowering. Significant (P≥ 0.001) interactions between fungicide treatments and cultivars occurred in each of three years at nearly all assessment times. In years of high mildew incidence (1983 and 1984), susceptible cultivars developed similar levels of mildew in seed-treated and untreated plots by about flowering time, although cultivars with adult plant resistance (APR) had significantly less mildew when seed-treated than when left untreated. Later, as the APR was fully expressed and the fungicide effectiveness was declining, similar mildew levels were recorded on treated and untreated plots. Untreated APR cultivars generally had less mildew than treated susceptible cultivars and in a year of late and light mildew, APR alone provided good protection. Seed treatments combined with foliar sprays of tridemorph as Calixin almost completely controlled mildew except on very susceptible cultivars late in the season in high mildew years. Over all cultivars, seed treatment gave yield advantages of 5.3% (1983) and 6.6% (1984) but in 1982, a year of low mildew, the response was small. The possible influence of integrated host resistance and fungicides in stabilising the pathogen population at economically unimportant levels and the environmental benefit of using host resistance to minimise fungicide usage is discussed.