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.     

The redistribution of fungicides on plants

In field trials amounts of soluble copper were found in rain water collected form apple trees sprayed with copper fungicides, but the copper was relatively non-toxic. The solution of copper did not appear to be correlated with the number of micro-organisms on the leaves. Rain washings phate. Some evidence for beneficial redistribution by solution is presented. By contrast, in a greenhouse experiment small amounts of copper were washed by very light rain from coffee seedlings sprayed with copper oxychloride and Bordeaux mixture. The soluble fraction of these washings was generally small and did not vary with ageing of the deposit up to 32 days. There was no changes in the toxicity of the washings from deposits aged up to 32 days. Under static laboratory conditions more copper was usually solubilized form standardized deposits of Bordeaux mixture, copper oxychloride, and cuprous oxide on leaves than on glass. The amount of copper solubilized varied with the leaf used, the nature of the deposit, the presence of adjuvants and the length of time water was left in contact with the deposit. Deposits of Bordeaux mixture and Bordeaux mixture was equally toxic on a leaf surface which caused solution and one which did not. Soluble copper extracts leaves were less toxic than equivalent quantities of copper supplied as copper sulphate.     

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.     

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.     

Effect of surfactants and oils on the phytotoxicity of difenzoquat to Avena fatua, barley and wheat

Controlled drop (CDA) and conventional applications of difenzoquat to pot-grown Avena fatua were compared. With the recommended surfactant (0.5% v/v Agral), very low volume CDA was less effective than conventional spray application. However, addition of extra Agral or various blends of paraffinic oil with Agral and the surfactants Burtemul A2 or Burtemul P2 improved the effects of the CDA treatments. When difenzoquat was absent the additives were inactive against A. fatua. They had little direct effect on wheat and barley and did not substantially increase the phytotoxicity of difenzoquat to these crops. Oil/surfactant mixtures were less viscous than high concentrations of Agral, and so easier to spray. In a pot experiment, smaller (150 μm diameter) drops of difenzoquat solution were more active against A. fatua than larger (200 μm-300 μm) drops. Reduction of the spray volume within the range 40 litres/ha to 5 litres/ha also reduced phytotoxicity. An oil/surfactant additive improved the activity of all difenzoquat CDA treatments. There was slightly more effect at the lowest spray volume but interactions between additive and application treatments were not statistically significant. When simulated rain treatments were applied 2 h or 5 h after spraying, difenzoquat controlled drop application was much less phytotoxic than a conventional 150 litres/ha treatment. However, addition of an oil/surfactant mixture markedly improved the performance of CDA. When rain was withheld for 24 h the additive had relatively less effect. In the field an oil/surfactant mixture improved control of A. fatua by difenzoquat with both conventional and controlled drop treatments. The additive did not increase injury to the wheat crop. The oil/surfactant mixtures markedly improved the spreading and wetting properties of sprays. This reduced localised contact injury, which, it is suggested, improved uptake and translocation of difenzoquat.     

Effectiveness of late copper and streptomycin sprays for the control of halo-blight of beans (Pseudomonas phaseolicola)

Two sprays of copper oxychloride or streptomycin sulphate (0–1 % a.i., 675 1/ha) applied to dwarf beans at first flowering and at pod set reduced pod infection by c. 70%. When applied as single sprays at pod set, copper was more effective than streptomycin and reduced pod infection by 50%. Although a copper spray at pod set increased the proportion of pods suitable for processing, more effective control was obtained with a regular spray programme, commencing at the seedling stage.     

Chemical control of Alternaria brown spot on Minneola tangelo in South Africa

The effectiveness of several groups of fungicides was compared with that of copper and mancozeb for their ability to control Alternaria brown spot on Minneola tangelo. Variables used for comparing fungicides in order of importance were: performance index, the number of exportable fruits per tree, the number of fruits per tree free of Alternaria lesions and total number of fruits per tree. Percentage exportable fruits was not a reliable variable for comparing treatment efficiency. During the 1992/1993 evaluations iprodione, difenoconazole, and procymidone showed promising control of the disease. Other than for difenoconazole, members of the triazole group were not effective, especially at dosages below 10 g a.i. 100 litre ^] water. Moderate control of Alternaria brown spot was obtained using copper and mancozeb. During the 1993/1994 season spray programmes with copper oxychloride, mancozeb, maneb plus zinc oxide, procymidone, iprodione, and tebuconazole were compared in field trials, for the control of Alternaria brown spot on Minneola tangelo. Mancozeb applied at 2-weekly intervals was the most effective spray programme, followed in order of efficiency by programmes using procymidone, maneb plus zinc oxide, and mancozeb applied at 4- and at 3-weekly intervals. Iprodione and tebuconazole were not particularly effective in this evaluation, although they performed significantly better than the control treatment. Trees treated with copper oxychloride produced poorly, in spite of a high percentage exportable fruits.     

Field trials of copper fungicides for the control of potato blight; spray retention

Spray-retention estimations were made during four seasons at Dartington in south Devon, on maincrop potatoes, twice-sprayed, as in farm practice, with about 120 gal./acre for the first application and 160 gal. for the second.
Evidence is given concerning the limits of reliability of the disk method of leaf sampling; and an alternative method, batten.' washing, is described.
With from 2,1/2 to 3,1/2 in. of rain over test periods of 3-4 weeks, 1% Bordeaux mixture showed about 40 % spray retention. Cuprous oxide and copper oxychloride sprays at the same copper dosage but compounded with sulphite lye or other water-soluble dispersing agent showed less than 20% retention. Compounding of these materials with an insoluble sticker (bentonite) improved the adhesion.
The percentage retention of 1/2% Bordeaux mixture was found to be less than that of the usual 1 % mixture, but adequate spray deposits could be maintained with this and other low-copper fungicides by more frequent applications.
A rapid method for determining total expanse of foliage in the field is described, and the importance of such determinations in connexion with spray-retention trials, and practical spray timing, is stressed.
Experiments were made to determine whether any copper is absorbed by potato leaves from the spray deposits. Absorption, or acid-resistant adsorption, of the order of 0.02 mg. Cu/120 sq.cm. of leaf area was indicated.     

Effect of relative humidity,temperature and fungicide on germination of conidia of Cercospora canescens caused the Cercospora leaf spot disease in mungbean

The aim of the present investigation was to determine the impact of relative humidity (RH) and temperature on conidial germination, nuclear position and effect of important fungicides on growth and conidial germination of Cercospora canescens. Germination of conidia was observed at RH range 92–100% at 5–35°C. Significant interaction between temperature and RH indicated that higher humidity and high temperature promoted quick germination both in the presence and absence of free moisture. Although in absence of free moisture at 92–95% RH higher temperatures 25–35°C promoted quick evaporation of moisture and no conidial germination. Number of germtube was increased significantly at the optimum temperature 25–30°C and higher humidity (98–100%). But higher temperature 25–35°C with lower RH did not support the conidial germination. This finding is very important for disease forecasting using meteorological data. The spray of Carbendazim as contact fungicide may not be useful since it is not effective against the conidia of C. canescens. Triadimefon did not inhibit the conidia germination but completely inhibited mycelium development at 50 μg/ml. Propriconazole inhibited both conidia germination and mycelial development. Therefore, Propiconazole may be taken as protective as well as curative spray. In non-systemic fungicide, Copper oxychloride gave anticipated result by inhibiting both conidial germination and mycelium development. Therefore, copper oxychloride can be used as protectant fungicides for Cercospora leaf spot caused by C. canescens.     

Evaluation of Biological and Chemical Treatments for Control of Crown Gall on Young Apple Trees in the Kootenay Valley of British Columbia

Field trials at Creston in 1988, 1989, and 1990 on silty loam soil naturally infested with Agrobacterium tumefaciens showed that Agrobacterium radiobacter strain K84 did not control crown gall on young Antonovka apple trees when apphed as a root-dip treatment. Copper oxychloride applied as a root-dip treatment at 2.5 and 5.0 g ai/1 and sewage sludge applied at 260 g per tree as broadcast were effective in reducing crown gall infection but these treatments were toxic to young apple trees in 1989. Lower rates of copper oxychloride, 0.3, 0.6 and 0.9 g ai/1, and sewage sludge at 130 g per tree, did not control crown gall in the 1990 field trial. The biological treatment with the strain AB8 of Bacillus subtilis applied as a root-dip effectively controlled crown gall and was not phytotoxic to young Antonovka apple trees. These results suggest that strain AB8 of B. subtilis has the potential to control crown gall on young apple trees under field conditions.     

Field evaluation of a tunnel sprayer and effects of spray volume at constant drop size on spray deposits and efficacy of disease control on apple

An air assisted tunnel sprayer (the Noric Joco EX2) incorporating CDA rotary atomisers (Volume Median Diameter (VMD) =c. 140 μm, volume rate = 50 litre ha^-1 forward speed = 3.9 km h^-1) gave at best equal but in some cases significantly poorer control of apple powdery mildew and scab than the standard commercial practice of spraying with an axial fan sprayer with Micron XI rotary atomisers (VMD =c. 90 μm, volume rate = 50 litre ha^-1, forward speed = 7.2 km h^-1). Approximately 30% of the spray volume applied was collected for recycling with the tunnel sprayer. Increasing spray volumes at approximately constant drop size (c. 140 μm) from 50 to 100 to 200 litre ha^-1 with the tunnel sprayer consistently improved efficiency of mildew and scab control. Bulk spray deposits on leaves, and their gross distribution in the tree, was similar with the different spraying methods and volumes. The mean percentage leaf area covered with spray deposit increased with spray volume as did the number of spray deposits per unit area. The tunnel sprayer at 50 litre ha^-1 gave a similar though less variable mean level of cover than the axial fan sprayer at the same volume rate. It gave 2–3 times more cover on upper than on lower leaf surfaces. The axial fan sprayer gave approximately equal cover on upper and lower leaf surfaces. The main limitations of the tunnel sprayer were its slow maximum forward speed and the restricted tree size and shape on which it can be used.     

Studies on the Mode of Action of Tomatine as a Fungitoxic Agent

Removal of 1 or more sugar residues from the alpha-tomatine molecule markedly decreased its fungitoxicity. While partial hydrolysis of alpha-tomatine did not greatly affect its surfactant properties, it did destroy the ability of this alkaloid to form a complex with cholesterol. Only unprotonated alpha-tomatine was capable of binding cholesterol; the protonated form did not. Since alpha-tomatine was far more toxic at a high pH than at a low pH, this suggests that the unprotonated alkaloid is the active form and that it acts by complexing with fungal sterols.     

Synergism and phytotoxicity: The effects of tank-mix additives on the biological efficacy of Cu2+ against Venturia inaequalis and Podosphaera leucotricha

The wetting behaviour of the spray and the biological efficacy of Cu²⁺ active ingredients in agrochemical formulations may be enhanced by tank-mix additives. We investigated how three BREAK-THRU® additives (BT301: biodegradable, BT133 and BT420: bio-based and biodegradable) tank-mixed with commercial copper preparations influenced the spray distribution, leaf uptake and biological efficacy of copper additive mixtures against apple scab and apple powdery mildew under controlled conditions. We quantified the synergetic effects of these additives in foliar applications. In addition, we determined the phytotoxic potential and evaluated their impacts on photosynthetic activity, non-photochemical quenching and ROS activity. The additives BT301 and BT420 strongly reduced surface tension and contact angle of copper treatments. The fluorescence observations revealed that BT301 achieved better spreading of copper formulation with more complete coverage of the leaf surface than BT420 and BT133, whereas ‘coffee-ring’ spreading was observed with BT133. The additive BT301 showed an increase in relative fluorescence area, indicating higher ROS production as a signal of intra-cellular tissue activity. The photochemical efficiency of photosystem II (Fv/Fm) was not negatively influenced by copper or additive treatment. Thus, we observed no phytotoxic effects of copper-additive mixtures on apple leaves at treatment doses of 4 g Cu²⁺ L⁻¹. All copper treatments reduced apple scab infections significantly, by 53%–76%. Interestingly, addition of BT301 to copper preparations showed the strongest biological efficacy (83% reduction) against Venturia inaequalis, whereas addition of BT420 showed the strongest effect against Podosphaera leucotricha (89% infection reduction). The synergetic effects of additives on the biological efficacy without phytotoxic effects on plants may have potential for reducing copper loads in horticultural production systems.     

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.     

DETERMINATION OF METHYLENE BLUE BIOSORPTION BY Rhizopus arrhizus IN THE PRESENCE OF SURFACTANTS WITH DIFFERENT CHEMICAL STRUCTURES

Methylene blue (MB) biosorption properties of Rhizopus arrhizus were investigated in the presence of surfactants. The effects of cationic and anionic surfactants on MB removal by dead biomass (1 g L^?1) were determined. MB removal was tested as a function of initial pH (2–12), contact time (5–1440 min), and dye (37.4–944.7 mg L^?1) and surfactant (0–10 mM) concentrations. The opposite charged anionic surfactant dodecylbenzenesulfonic acid sodium salt (DBS) enhanced sorption of cationic MB by biomass dramatically. Maximum biosorption capacity was 471.5 mg g^?1 at pH 8 with 0.5 mM DBS at 944.7 mg L^?1 MB concentration. The surfactant-stimulated fungal decolorization method may provide a highly efficient, inexpensive, and time-saving procedure in biological wastewater treatment technologies.     

Dilution of protein‐surfactant complexes: A fluorescence study

Dilution of protein–surfactant complexes is an integrated step in microfluidic protein sizing, where the contribution of free micelles to the overall fluorescence is reduced by dilution. This process can be further improved by establishing an optimum surfactant concentration and quantifying the amount of protein based on the fluorescence intensity. To this end, we study the interaction of proteins with anionic sodium dodecyl sulfate (SDS) and cationic hexadecyl trimethyl ammonium bromide (CTAB) using a hydrophobic fluorescent dye (sypro orange). We analyze these interactions fluourometrically with bovine serum albumin, carbonic anhydrase, and beta‐galactosidase as model proteins. The fluorescent signature of protein–surfactant complexes at various dilution points shows three distinct regions, surfactant dominant, breakdown, and protein dominant region. Based on the dilution behavior of protein–surfactant complexes, we propose a fluorescence model to explain the contribution of free and bound micelles to the overall fluorescence. Our results show that protein peak is observed at 3 mM SDS as the optimum dilution concentration. Furthermore, we study the effect of protein concentration on fluorescence intensity. In a single protein model with a constant dye quantum yield, the peak height increases with protein concentration. Finally, addition of CTAB to the protein–SDS complex at mole fractions above 0.1 shifts the protein peak from 3 mM to 4 mM SDS. The knowledge of protein–surfactant interactions obtained from these studies provides significant insights for novel detection and quantification techniques in microfluidics.     

A comparison of fungicide spray volumes for the control of coffee berry disease

A continuation of previous trials in which three sprayers were used to apply 8.4 g. of Perenox in a range of spray volumes from 0.2 to 1.7 l. per tree confirmed that the volume of spray did not affect greatly the deposit retained on the branch surface. Branches were able to retain up to about 250 mg. Cu/m.² and it is considered that a volume equivalent to 100 gal./acre would provide the greatest margin of safety to compensate for inaccuracies of application, while avoiding serious run-off. Where coffee was well pruned, intensive copper spraying raised the annual yield by about 2 cwt. of clean coffee per acre. Copper spraying did not provide an adequate control of coffee berry disease and, under certain circumstances, increased incidence.     

The effect of varying rates of glyphosate and an organosilicone surfactant on the control of gorse

The herbicidal effect of glyphosate applied to gorse (Ulex europaeus L.) was improved by the addition of increasing amounts (0.5–20 g/litre) of Silwet L-77, an organosilicone surfactant. Increasing the rate of herbicide also enhanced control. There was a highly significant interaction between surfactant rate and herbicide dosage; as the amount of Silwet L-77 was increased the rate of glyphosate could be reduced without loss of herbicide efficacy. However, without any added organosilicone surfactant, glyphosate did not provide more than 73% control of gorse at any rate up to 6.5 kg a.i./ha. With the addition of Silwet L-77, complete mortality of all plants could be achieved with 2.2 kg glyphosate/ ha.     

Efficacy of different copper compounds in the control of Xanthomonas citri subsp. citri pathotypes A and A*

Citrus canker disease is one of the most devastating diseases that attacks citrus, especially limes in the Southern parts of Iran, and is caused by Xanthomonas citri subsp. citri (Xcc). The efficacy of several formulations of copper compounds including Bordeaux mixture, copper oxychloride and copper sulphate in controlling Xcc in Key lime was estimated in vitro and in planta using artificial inoculation. Specific primers were used to detect copper-resistant genes copA, copB and copL in 30 isolates of Xcc. The copA and copL genes were present in all isolates, and copB was detected only in 6 strains. In this study, we observed a very good in vitro growth inhibition activity of copper compounds against Xcc pathotype A. S14 strain (pathotype A^*) was the sole isolate that grew on media amended with 2/4 mM of Bordeaux mixture, copper oxychloride and copper sulphate. All other strains (pathotype A) failed to grow on media amended with this concentration. Bordeaux mixture exhibited high efficacy in controlling Xcc in both conditions. However, there were no significant differences in the efficacy of copper oxychloride and copper sulphate at 1.2 mM concentration in planta. A significantly minimum canker necrotic spot and highest disease control was achieved with Bordeaux mixture and copper oxychloride. There was a significant difference in disease severity of the type strain LMG9322 (pathotype A) and Xcc strain S14 (pathotype A^*). Our experiments showed that Bordeaux mixture exhibited satisfactory efficacy in controlling the causal agent of citrus canker.     

Microstructures formed in aqueous solutions of a hydrophobically modified nonionic cellulose derivative and sodium dodecyl sulfate: a fluorescence probe investigation

The association behavior of hydrophobically modified ethyl hydroxyethyl cellulose (HM-EHEC) and its interaction with the anionic surfactant sodium dodecyl sulfate (SDS) has been studied in the dilute concentration regime. Steady-state fluorescence probe techniques have been utilized to obtain microstructural information of the system properties and combined with macroscopic bulk information from equilibrium dialysis experiments in order to determine binding isotherms of SDS to HM-EHEC. HM-EHEC was found to self-associate and form polymeric micelles in semi-dilute aqueous solutions. c^* for the self-association process was determined to be approximately 0.4%. The microviscosity of the polymeric micelles is much higher, and the micropolarity slightly higher, than that of ordinary SDS micelles. The onset of interaction between HM-EHEC and SDS was evidenced by a simultaneous strong increase in microviscosity and decrease in micropolarity upon successive addition of SDS. There is a minor, noncooperative SDS binding to the HM-EHEC starting from low concentrations of SDS (<5 mM) followed by a highly cooperative binding region at SDS concentrations ≥5 mM. The polymer–surfactant aggregates are rigid and hydrophobic with a maximum in microviscosity in the noncooperative binding region at a very low degree of SDS-adsorption.