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WO1996018302A1 - Potentialisation des delta-endotoxines de bacillus thuringiensis par additifs tensioactifs - Google Patents

Potentialisation des delta-endotoxines de bacillus thuringiensis par additifs tensioactifs Download PDF

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Publication number
WO1996018302A1
WO1996018302A1 PCT/US1995/015019 US9515019W WO9618302A1 WO 1996018302 A1 WO1996018302 A1 WO 1996018302A1 US 9515019 W US9515019 W US 9515019W WO 9618302 A1 WO9618302 A1 WO 9618302A1
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Prior art keywords
bacillus thuringiensis
thuringiensis subsp
biopesticide
surfactant
delta
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PCT/US1995/015019
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English (en)
Inventor
David G. Hobbs
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Abbott Laboratories
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Publication date
Application filed by Abbott Laboratories filed Critical Abbott Laboratories
Priority to AU42396/96A priority Critical patent/AU4239696A/en
Publication of WO1996018302A1 publication Critical patent/WO1996018302A1/fr

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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
    • A01N63/50—Isolated enzymes; Isolated proteins

Definitions

  • the present invention relates to a pesticidal composition
  • a pesticidal composition comprising a Bacillus thuringiensis biopesticide and a surfactant. More specifically, the present invention relates to said pesticidal composition wherein the surfactant potentiates the pesticidal activity of a Bacillus thuringiensis biopesticide or reduces the amount of the Bacillus thuringiensis biopesticide typically needed to be a commercially effective biopesticide.
  • Bacillus thuringiensis is a motile, rod-shaped, gram-positive bacterium that is extensively distributed in nature, especially in soil and insect-rich environments. During sporulation, Bacillus thuringiensis produces a parasporal crystal inclusion(s) which is insecticidal upon ingestion to susceptible insect larvae of the order Lepidoptera, Diptera, or Coleoptera. The inclusions may vary in shape, number, and composition. They are comprised of one or more proteins called delta-endotoxin proteins, which may range in size from 27-140 kDa. The insecticidal delta-endotoxin proteins are generally converted by proteases in the larval gut into smaller (truncated) toxic polypeptides, causing midgut destruction, and ultimately, death of the insect.
  • Bacillus thuringiensis crystal delta-endotoxin proteins are the most widely used biopesticide. There are several Bacillus thuringiensis strains that are used as producers of crystal delta-endotoxin proteins for the forestry, agricultural, and public health areas. Bacillus thuringiensis subsp. kurstaki and Bacillus thuringiensis subsp. aizawai produce delta-endotoxin proteins specific for Lepidoptera. A delta-endotoxin protein specific for Coleoptera is produced by Bacillus thuringiensis subsp. tenebrionis. Furthermore, Bacillus thuringiensis subsp. israelensis produces delta-endotoxin proteins specific for Diptera.
  • Bacillus thuringiensis crystal delta-endotoxin proteins are also reportedly pesticidal to nematodes, Acari, Hymenoptera, Phthiraptera, Platyhelminthes, Homoptera, Blattodea, and Protozoa.
  • the delta-endotoxin proteins are encoded by cry (crystal protein) genes which are generally located on plasmids.
  • cry genes have been divided into six classes and several subclasses based on relative amino acid homology and pesticidal specificity.
  • the major classes are Lepidoptera-specific (cry/); Lepidoptera-and Diptera-specific (cry//); Coleoptera-specific (crylll); Diptera-specific (cry V); Coleoptera- and Lepidoptera-specific (cry V genes); and Nematode-specific (cryVI genes).
  • the art has strived to improve the effectiveness and to broaden the host rang of Bacillus thuringiensis delta-endotoxin proteins.
  • Means have included isolating Bacillus thuringiensis strains with improved or new pesticidal activity, engineering presen Bacillus thuringiensis strains, and designing more effective formulations by combining Bacillus thuringiensis crystal delta-endotoxin proteins with new pesticidal carriers.
  • Fluorochemical surfactants such as fluoroalkyl quaternary ammonium iodides, ammonium perfluoroalkyl sulfonates, fluoroaliphatic oxyethylene adducts, fluorinated alkyl alkoxylates, fluorinated alkyl sulfonamides, and fluoroaliphatic polymeric esters are used as wetting and leveling agents for cleaning, rinsing, and polishing products as well as for organic polymer systems.
  • Silicon surfactants such as silicon polyether, 2-(3-hydroxypropyl)- heptamethyltrisiloxane, and polyalkyleneoxide modified heptamethyltrisiloxane are employed as anti-foam agents in industrial chemical processes, fermentation systems, and aqueous pesticide formulations.
  • Aoki et al. disclose a composition containing a silicon surfactant where the silicon surfactant is an insecticidal or miticidal agent (Aoki et al., 1991, Japanese Patent Application 3/1991-10484).
  • Ionic surfactants such as organic phosphate ester free acids and alkylaryl sulfonates are used as emulsifiers in agricultural formulations, solvents, detergents, and alkaline cleaners.
  • Nonionic surfactants such as nonylphenol polyethylene glycol ether, 2,6,8- trimethyl-4-nonyloxy-polyethyleneoxyethanol, or polyethylene sorbitan monopalmitate ar employed as emulsifying and wetting agents, components of commercial dishwashing detergents, and scouring agents in the textile industry.
  • the present invention relates to a pesticidal composition
  • a pesticidal composition comprising a Bacillus thuringiensis biopesticide in an ineffective amount for controlling a pest and a surfactant in an effective amount for potentiating the pesticidal activity of the Bacillus thuringiensis biopesticide and for reducing the amount of the Bacillus thuringiensis biopesticide typically needed to be a commercially effective biopesticide.
  • the present invention further relates to a method for controlling pest infestation on plants comprising applying to plants said pesticidal composition.
  • the object of this invention is to reduce the amount of a Bacillus thuringiensis biopesticide needed to achieve commercially acceptable control of a pest, thereby reducing the cost of the Bacillus thuringiensis biopesticide. It is a further object of the present invention to improve the efficacy of a Bacillus thuringiensis biopesticide which alone would have been commercially impractical or commercially ineffective as a biopesticide.
  • the present invention is directed to a pesticidal composition
  • a pesticidal composition comprising a Bacillus thuringiensis biopesticide in an ineffective amount for controlling a pest and a surfactant in an effective amount for potentiating the pesticidal activity of said Bacillus thuringiensis biopesticide to effectively control said pest.
  • Pesticidal activity is defined herein as a measure of the amount of activity of a Bacillus thuringiensis biopesticide against a pest through killing or stunting of the growth of the pest or protecting a plant from pest infestation.
  • “Ineffective amount” is defined herein as the amount of a Bacillus thuringiensis biopesticide which provides less than about 35% control of a pest when used alone.
  • Effective amount is defined herein as an amount of a surfactant sufficient to potentiate the pesticidal activity of a Bacillus thuringiensis biopesticide to a level greater than about 10%, preferably greater than about 30%, more preferably greater than about 50% and most preferably about 100% of the pesticidal activity of the biopesticide used alone, to achieve commercially acceptable control of a pest .
  • Patentiate is defined herein as to enhance or increase the pesticidal activity of a Bacillus thuringiensis biopesticide at least about 10%, preferably at least about 30%, more preferably at least about 50%, and most preferably at least about 100%, relative to the pesticidal activity of the biopesticide used alone, wherein the substance causing the enhancing activity has no significant pesticidal activity by itself, e.g., having an LC 50 of more than 3000 ⁇ g/g as assayed by bioassay (LC 50 is the concentration of the substance required to kill 50% of the pests).
  • the pesticidal activity of a Bacillus thuringiensis biopesticide and the potentiating effect of a surfactant on the pesticidal activity of a Bacillus thuringiensis biopesticide may be assayed using procedures known in the art, such as artificial diet incorporation, artificial diet overlay, leaf painting, leaf dip, and foliar spray.
  • a Bacillus thuringiensis biopesticide alone, at certain levels, may provide commercially acceptable control of a pest.
  • the amount of any given Bacillus thuringiensis biopesticide required to achieve commercially acceptable control may be quite high, and, therefore, impractical economically.
  • the amount of a Bacillus thuringiensis biopesticide required in any given composition may be reduced by combining the Bacillus thuringiensis biopesticide with an effective amount of a surfactant to potentiate the pesticidal activity of the Bacillus thuringiensis biopesticide.
  • the amount of a Bacillus thuringiensis biopesticide may be reduced by an amount corresponding to the potentiation of the pesticidal activity of the Bacillus thuringiensis biopesticide provided by the surfactant so that the pesticidal activity is equal to the amount typically used by the art in a composition without potentiation to achieve commercially acceptable control of a pest.
  • the amount of a Bacillus thuringiensis biopesticide in a composition of the present invention is reduced between about 10% and about 80%, preferably between about 10% and about 60%, and most preferably between about 10% and about 40%, relative to the amount of the Bacillus thuringiensis biopesticide which would be needed to achieve the same effect in the absence of the surfactant, corresponding to the amount of potentiation provided by the surfactant.
  • the amount of a Bacillus thuringiensis biopesticide needed to achieve commercially acceptable control of a pest will differ from one Bacillus thuringiensis biopesticide to another Bacillus thuringiensis biopesticide, and furthermore, will vary depending on the pest and the crop.
  • Bacillus thuringiensis biopesticides are well established in the art.
  • the amounts required for commercial utility are either known or easily determined by the ordinary skilled artisan.
  • the amount of a Bacillus thuringiensis biopesticide required for commercial utility is that which achieves greater than about 80% control of Spodoptera exigua in the treatment of lettuce.
  • an amount of Bacillus thuringiensis adequate to achieve about 35% control is considered to be effective.
  • an amount which achieves approximately 50% control of Lymantria dispar is needed for a biopesticide is considered to be effective.
  • the benefit of the present invention may be achieved by determining the amount of a Bacillus thuringiensis biopesticide alone required to achieve commercially acceptable control of a pest
  • the amount of a surfactant needed to provide the benefit of reducing the amount of a Bacillus thuringiensis biopesticide used commercially can be determined, for example, by taking the amount of a Bacillus thuringiensis biopesticide required to achieve commercially acceptable control of a pest and progressively reducing that amount of the Bacillus thuringiensis biopesticide, e.g., 10%, and progressively adding a surfactant in increasing amounts until the level of control of the pest equals that which is achieved with the Bacillus thuringiensis biopesticide alone in amounts typically used commercially.
  • a Bacillus thuringiensis biopesticide and a surfactant may be useful in bringing about the commercial use of the Bacillus thuringiensis biopesticide which alone would have been commercially impractical or commercially ineffective as a biopesticide.
  • the surfactant is a fluoroalkyl quaternary ammonium iodide, an ammonium perfluoroalkyl suifonate, a fluoroaliphatic oxyethylene adduct, a fluorinated alkyl alkoxylate, a fluorinated alkyl sulfonamide, or a fluoroaliphatic polymeric ester.
  • the surfactant is a silicon polyether, 2-(3- hydroxypropyl)-heptamethyltrisiloxane, or a polyalkyleneoxide modified heptamethyltrisiloxane.
  • the surfactant is an organic phosphate ester free acid or an alkylaryl suifonate.
  • the surfactant is nonylphenol polyethylene glycol ether, 2,6,8-trimethyl-4-nonyloxy-polyethyleneoxyethanol, or polyethylene sorbitan monopalmitate.
  • the pesticidal composition may comprise a combination of two or more of the surfactants described supra.
  • the effective concentration of the surfactant is in the range of between about 0.05 gram and about 25 gram per gram of Bacillus thuringiensis biopesticide in the pesticidal composition of the present invention. In a more preferred embodiment, the effective concentration of a surfactant is in the range between about about 0.3 gram and about 10 gram per gram of Bacillus thuringiensis biopesticide in the pesticidal composition of the present invention. In a most preferred embodiment, the effective concentration of a surfactant is in the range between about 0.5 gram and about 3 gram per gram of Bacillus thuringiensis biopesticide in the pesticidal composition of the present invention.
  • biopesticide is as a Bacillus thuringiensis subsp. kurstaki whole broth concentrate, where a portion of the water and solubles have been removed from the whole broth by centrifugation and preserved at an acid pH of 4.3 to 5.5, with a potency of 8000 IU (International Units) per mg solids (bioassayed against 3rd instar Trichoplusia ni using the Bacillus thuringiensis subsp. kurstaki standard HD1-S-1980 (USDA, Peoria, Illinois).
  • biopesticide when other form of the biopesticide are used, for example, purified crystals and/or spores, whole microorganisms, or a less crude broth concentrate, these values can readily be adjusted to the activity, if different, of the chosen form of biopesticide.
  • the Bacillus thuringiensis biopesticide in the pesticidal composition of the present invention may be derived from, but not limited to, Bacillus thuringiensis subsp. aizawai, Bacillus thuringiensis subsp. alesti, Bacillus thuringiensis subsp. americansis, Bacillus thuringiensis subsp. darmstadiensis, Bacillus thuringiensis subsp. dendrolimus, Bacillus thuringiensis subsp. entomocidus, Bacillus thuringiensis subsp. finitimus, Bacillus thuringiensis subsp. galleriae, Bacillus thuringiensis subsp.
  • the biopesticide is derived from a Bacillus thuringiensis subsp. kurstaki strain.
  • the Bacillus thuringiensis biopesticide in the pesticidal composition of the present invention may be also derived from a cell wherein a gene, which encodes a Bacillus thuringiensis delta-endotoxin protein or pesticidally-active fragment thereof, has been inserted. Furthermore, within the scope of the present invention, the Bacillus thuringiensis biopesticide or pesticidally-active fragment thereof may be derived from a transconjugate strain wherein a plasmid containing a gene, which encodes the Bacillus thuringiensis delta- endotoxin protein or pesticidally-active fragment thereof, has been transferred by cell-cell conjugation.
  • the Bacillus thuringiensis biopesticide i.e., a delta-endotoxin protein or a pesticidally-active fragment thereof, may be selected from the groups including, but not limited to, Cryl, Cryll, Crym, CrylV, Cry V, and Cry VI.
  • the Bacillus thuringiensis delta-endotoxin protein or pesticidally-active fragment thereof may include, but is not limited to, CryIA(a), CryIA(b), CryIA(c), CrylB, CrylC, CrylD, CryHA, CryHB, CryfflA, CrylUB, CryHIC CrylVA, CrylVB, CrylVC, CrylVD, CryV, and CryVI. It is also within the scope of the present invention that the Bacillus thuringiensis biopesticide may also comprise a spore derived from the Bacillus thuringiensis strain.
  • the Bacillus thuringiensis delta-endotoxin protein is CrylA.
  • the delta-endotoxin protein is Cry ⁇ A(a), Cry ⁇ A(b), or Cry ⁇ A(c).
  • the pesticidal composition of the present invention may also be used with an entomopathogenic virus alone or in combination with a Bacillus thuringiensis biopesticide.
  • entomopathogenic viruses include, but are not limited to, Autographa califomica nuclear polyhedrosis virus (NPV), Syngrapha falcifera NPV, Cydia pomonella granulosis virus (GV), Heliothis zea NPV, Lymantria dispar NPV, Orgyia pseudotsugata NPV, Spodoptera exigua NPV, Neodiprion lecontei NPV, Neodiprion sertifer NPV, Harrisina brillians NPV, and Endopiza viteana Clemens NPV.
  • NPV Autographa califomica nuclear polyhedrosis virus
  • GV Cydia pomonella granulosis virus
  • GV Heliothis zea NP
  • the pesticidal composition of the present invention may further be used with a chemical pesticide alone or in combination with a Bacillus thuringiensis biopesticide.
  • chemical pesticides include, but are not limited to, insect growth regulators, carbamates, organophosphates, pyrethroids, inorganic fluorines, pyrazoles, pyrroles, and avermectins.
  • the surfactant may be combined either with the Bacillus thuringiensis product in a tank mix or in the formulated product. In either case, the effective concentration of the applied surfactant for potentiating the pesticidal activity of an ineffective amount of a Bacillus thuringiensis biopesticide will be the same.
  • the composition of the present invention may comprise a surfactant and a pesticidal carrier.
  • This composition may be supplied separately to a plant.
  • the composition may be applied to a transgenic plant which contains and encodes a Bacillus thuringiensis biopesticide gene.
  • the composition may be applied to a plant previously exposed to a Bacillus thuringiensis biopesticide.
  • the pesticidal composition of the present invention may further comprise a deposition agent which assists in preventing the composition from drifting from the target area during application (e.g., as it is sprayed from a plane), or from being blown away from the plant once it has been deposited.
  • the deposition agent in the composition of the present invention is a proteinaceous material, which has the added benefit of being palatable to the insect. Any animal or vegetable protein is suitable for this purpose, in dry or in liquid form. Examples of useful sources of protein which can be conveniently and economically added to the composition include, but are not limited to, so protein, potato protein, soy flour, potato flour, fish meal, bone meal, yeast extract, and blood meal.
  • Alternative deposition agents include modified cellulose (carboxymethylcellulose), botanicals (grain flours, ground plant parts), non-phyllosilites (talc, vermiticulite, diatomaceous earth), natural clays (attapulgite, bentonite, kaolinite, montmorillonite), and synthetic clays (Laponite).
  • the deposition agent is present in the pesticidal composition of the present invention in an amount of between about 0.4% w/w and about 50% w/w, more preferably in an amount between about 1% w/w and about 20% w/w.
  • the pesticidal composition of the present invention may further comprise an antifreeze/humectant agent which suppresses the freeze point of the product and helps minimize evaporation when sprayed and which maintains deposit texture (soft and chewey making the product more efficacious and palatable.
  • the antifreeze/humectant agent is selected from the group consisting of ethylene glycol, propylene glycol, dipropylene glycol, glycerol, butylene glycols, pentylene glycols and hexylene glycols.
  • the antifreeze/humectant agent is present in the pesticidal composition of the present invention in an amount of between about 0.5% w/w and about 25% w/w, more preferably in an amount between about 2% w/w and about 15% w/w.
  • the pesticidal composition of the present invention may further comprise a second surfactant in an amount where it acts as an emulsifying, a wetting, or a dispersing agent.
  • a second surfactant in an amount where it acts as an emulsifying, a wetting, or a dispersing agent.
  • anionic surfactants such as carboxylates, for example, a metal carboxylate of a long chain fatty acid; N-acylsarcosinates; mono or di- esters of phosphoric acid with fatty alcohol ethoxylates or salts of such esters; fatty alcohol sulphates such as sodium dodecyl sulphate, sodium octadecyl sulphate or sodium cetyl sulphate; ethoxylated fatty alcohol sulphates; ethoxylated alkylphenol sulphates; lignin sulphonates; petroleum sulphonates; alkyl aryl sulphonates
  • non-ionic surfactants such as condensation products of fatty acid esters, fatty alcohols, fatty acid amides or fatty-alkyl- or alkenyl-substituted phenols with ethylene oxide, block copolymers of ethylene oxide and propylene oxide, acetylenic glycols such as 2,4,7,9-tetraethyl-5-decyn-4,7-diol, or ethoxylated acetylenic glycols.
  • non-ionic surfactants such as condensation products of fatty acid esters, fatty alcohols, fatty acid amides or fatty-alkyl- or alkenyl-substituted phenols with ethylene oxide, block copolymers of ethylene oxide and propylene oxide, acetylenic glycols such as 2,4,7,9-tetraethyl-5-decyn-4,7-diol, or ethoxylated acetylenic glycol
  • surfactants are cationic surfactants such as aliphatic mono-, di-, or polyamine as acetates, naphthenates or oleates; oxygen-containing amines such as an arnine oxide of polyoxyethylene alkylamine; amide-linked amines prepared by the condensation of a carboxylic acid with a di- or polyamine; or quaternary ammonium salts.
  • the second surfactant is preferably present in an amount of between about 0.5% w/w and about 25% w/w, and more preferably in an amount of between about 1 % w/w and about 8% w/w.
  • the pesticidal composition of the present invention may further comprise an inert material.
  • inert materials include inorganic minerals such as diatomaceous earth, kaolin, mica, gypsum, fertilizer, phyllosilicates, carbonates, sulfates, or phosphates; organic materials such as sugar, starches, or cyclodextrms; or botanical materials such as wood products, cork, powdered corncobs, rice hulls, peanut hulls, and walnut shells.
  • the pesticidal composition according to the present invention may further comprise a preservative, a feeding stimulant, an attractant, an encapsulating pesticide, a binder, a dye, a UN. protectant, a buffer, a flow agent, or other component to facilitate product handling and application for particular target pests.
  • the present invention is also directed to a method for controlling pest infestation on plants comprising applying to a plant a pesticidal composition of the present invention.
  • the pesticidal composition of the present invention can be applied in a dry or liquid form, e.g., a suspension, a solution, an emulsion, a dusting powder, a dispersible granule, a wettable powder, an emulsifiable concentrate, an aerosol or impregnated granule, or a concentrate or primary composition which requires dilution with a suitable quantity of water or other diluent before application.
  • concentrations of each component in the composition will vary depending on the Bacillus thuringiensis biopesticide and mode of application. The Bacillus thuringiensis biopesticide concentration will vary depending upon the nature of the particular composition, specifically, whether it is a concentrate or to be used directly.
  • the composition contains about 1% to about 98% of a solid or liquid inert carrier, about 1% to about 30% of a Bacillus thuringiensis biopesticide, and about 0.5% to about 25%, preferably about 1% to about 8% of the second surfactant.
  • the composition will be preferably administered at the labeled rate for the commercial product, preferably about 0.01 pound to about 5.0 pounds per acre when in dry form and at about 0.01 pint to about 25 pints per acre when in liquid form.
  • the pesticidal composition of the present invention can be applied directly to a plant by, for example, spraying or dusting at the time when the pest has begun to appear on the plant or before the appearance of pests as a protective measure.
  • the pesticidal composition can be applied by foliar, furrow, broadcast granule, "lay-by", or soil drench application.
  • the composition of the present invention can also be applied directly to ponds, lakes, streams, rivers, still water, and other areas subject to infestation by pests of concern to public health.
  • the composition can be applied by spraying, dusting, sprinkling, or the like.
  • the spray or dust can conveniently contain another pesticide if this is thought necessary.
  • the pesticidal composition of the present invention is applied directly to the plant.
  • the pesticidal composition of the present invention can be applied to protect a number of different plant types, including, but not limited to, cereals (wheat, barley, rye, oats, rice, sorghum and related crops), beets (sugar beet and fodder beet), drupes, pomes and soft fruit (apples, pears, plums, peaches, almonds, cherries, strawberries, raspberries, and blackberries), leguminous plants (alfalfa, beans, lentils, peas, soybeans), oil plants (rape, mustard, poppy, olives, sunflowers, coconuts, castor oil plants, cocoa beans, groundnuts), cucumber plants (cucumber, marrows, melons), fibre plants (cotton, flax, hemp, jute), citrus fruit (oranges, lemons, grapefruit, mandarins), vegetables (spinach, lettuce, asparagus, cabbages and other brassicae, carrots, onions, tomatoes, potatoes), lauraceae (avoc
  • transgenic plants which contain and encode a Bacillus thuringiensis biopesticide gene.
  • the pesticidal composition of the present invention can be used in the treatment or prevention of infestations of a number of different insect types. It is particularly preferred to use the composition of the present invention to eliminate pests of the order Lepidoptera, e.g., Achroia grisella, Acleris gloverana, Acleris variana, Adoxophyes orana, Agrotis ipsilon, Alabama argillacea, Alsophila pometaria, Amyelois transitella, Anagasta kuehniella, Anarsia lineatella, Anisota senatoria, Antheraea pernyi, Anticarsia gemmatalis, Archips sp., Argyrotaenia sp., Athetis mindara, Bombyx mori, Bucculatrix thurberiella, Cadra cautella, Choristoneura sp.,
  • the pesticidal composition of the present invention may also be effective against insect pests of the order Diptera, e.g., Aedes sp., Andes vittatus, Anastrepha ludens, Anastrepha suspensa, Anopheles barberi, Anopheles quadrimaculatus, Armigeres subalbatus, Calliphora stygian, Calliphora vicina, Ceratitis capitata, Chironomus tentans, Chrysomya rufifacies, Cochliomyia macellaria, Culex sp., Culiseta inornata, Dacus oleae, Delia antiqua, Delia platura, Delia radicum, Drosophila melanogaster, Eupeodes corollae, Glossina austeni, Glossina brevipalpis, Glossina fuscipes, Glossina morsitans centralis
  • Example 1 Cultivation of Bacillus thuringiensis subsp. kurstaki HD1
  • Bacillus thuringiensis subsp. kurstaki HD1 is cultivated for 72 hours at 30°C in a medium with the following composition:
  • Each surfactant is dissolved or suspended in 75 ml of deionized water and then diluted to 100 ml by the addition of 25 ml of IX egg agar to give a final surfactant concentration in the range of 0.1% to 0.7% w/v.
  • the Bacillus thuringiensis subsp. kurstaki crystal delta-endotoxin protein/spore preparation from Example 1 is suspended in 0.1% TweenTM 20 to a concentraton of 10% w/v.
  • the 10% solution is further diluted in 25% egg agar, and then 0.2 ml of each diluted crystal delta-endotoxin protein/spore solution is blended with 9.8 ml of the 25% egg agar solution containing each surfactant. Controls without crystal delta- endotoxin protein/spore or surfactant are also prepared.
  • a 50 ⁇ l aliquot of each surfactant/crystal delta-endotoxin protein/spore solution and each control solution is applied to the surface of a 0.5 ml insect diet plug in a 1.2 ml plastic well of an assay tray.
  • the final concentration of Bacillus thuringiensis subsp. kurstaki crystal delta-endotoxin protein/spore preparation is 1000, 500, 250, 125, 62, and 31 ⁇ g per gram of solution.
  • the final concentration of surfactant is in the range of 700 ⁇ g to 100 ⁇ g per gram of solution applied.
  • the solutions are allowed to dry overnight at room temperature.
  • fluorochemical surfactants fluoroalkyl quaternary ammonium iodide FLUORADTM FC-135, 3M Chemical Company, St. Paul, Minnesota
  • ammonium perfluoroalkyl suifonate FLUORADTM FC-129, 3M Chemical Company, St. Paul, Minnesota
  • silicon surfactants silicon polyether (2-[3-hydroxypropyl]- heptamethyltrisiloxane; (SYLGARDTM 309, Dow Chemical Company, Midland, Michigan), polyalkyleneoxide modified heptamethyltrisiloxane (SBLWETTM L-77, Union Carbide, Danbury, Connecticut), and a blend of silica, silicon, and polyethylene glycol dioleate (DREW PLUSTM L-768, Drew Chemical Company, Boonton, New Jersey) are tested for their ability to potentiate the insecticidal activity of the Bacillus thuringiensis subsp. kurstaki crystal delta-endotoxin protein/spore preparation as described in Example 2.
  • T-MULTZTM 565, and T-MULTZTM 734-2, Harcross Chemical Co., Kansas City, Kansas) and calcium alkylaryl suifonate (NINATETM 401, Stepan Chemical Company, Chicago, Illinois) are tested for their ability to potentiate the insecticidal activity of the Bacillus thuringiensis subsp. kurstaki crystal delta-endotoxin protein/spore preparation as described in Example 2.
  • T-MULTZTM 565 , T- MULTZTM 734-2 and NINATETM 401 relative to the control, i.e., the Bacillus thuringiensis subsp. kurstaki crystal delta-endotoxin protein/spore preparation alone, potentiated the insecticidal activity of the Bacillus thuringiensis subsp. kurstaki crystal delta-endotoxin protein/spore preparation.
  • the non-ionic surfactant 2,6,8-trimethyl-4-nonyloxypolyethylene-oxyethanol (TERGITOLTM TMN-6, Union Carbide, Danbury, Connecticut) is tested for its ability to potentiate the insecticidal activity of the Bacillus thuringiensis subsp. kurstaki crystal delta-endotoxin protein/spore preparation.
  • a Spodoptera exigua bioassay is performed in a 240 well tray where each well is filled with 0.5 ml of insect diet.
  • the surfactant is tested at 0.01% and 0.1% w/w in combination with three concentrations of Bacillus thuringiensis subsp.
  • israeltaki crystal delta-endotoxin protein/spore preparation 0.5, 0.25, and 0.12 mg per ml.
  • a volume of 50 ul of each protein concentration is first loaded into 10 wells, then 50 ul of the surfactant solution are added and the mixture is allowed to dry. Once the samples are dried, Spodoptera exigua eggs are placed into each well, and the tray is incubated at 30 * C for 7 days prior to scoring of the results.

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Abstract

La présente invention se rapporte à une composition pesticide comprenant un biopesticide Bacillus thuringiensis en une quantité inefficace pour éliminer des insectes, et un tensioactif en une quantité efficace pour potentialiser l'activité pesticide du biopesticide Bacillus thuringiensis afin d'éliminer les insectes ou de réduire la quantité du biopesticide Bacillus thuringiensis habituellement nécessaire pour qu'il soit commercialement rentable. La présente invention se rapporte également à un procédé d'élimination d'insectes consistant à exposer ceux-ci à ladite composition pesticide.
PCT/US1995/015019 1994-12-13 1995-11-28 Potentialisation des delta-endotoxines de bacillus thuringiensis par additifs tensioactifs WO1996018302A1 (fr)

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AU42396/96A AU4239696A (en) 1994-12-13 1995-11-28 Potentiation of bacillus thuringiensis delta-endotoxins with surfactant additives

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US35430094A 1994-12-13 1994-12-13
US08/354,300 1994-12-13

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WO1998013497A1 (fr) * 1996-09-26 1998-04-02 Ecogen, Inc. COMPOSITIONS A BASE DE CryET29 TIRE DE BACILLUS THURIGIENSIS TOXIQUES POUR LES COLEOPTERES ET LES CTENOCEPHALIDES SPP
US6130186A (en) * 1996-10-25 2000-10-10 Monsanto Company Composition and method for treating plants with exogenous chemicals
WO2007002988A1 (fr) * 2005-06-30 2007-01-11 Microbial Products Pty Ltd Formulation de pesticides microbiens
WO2010039652A3 (fr) * 2008-10-01 2011-04-28 Vestaron Corporation Formulation de toxine peptidique

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Cited By (14)

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US7572587B2 (en) 1996-09-26 2009-08-11 Monsanto Technology Llc Bacillus thuringiensis CryET29 compositions toxic to coleopteran insects and Ctenocephalides spp
US6093695A (en) * 1996-09-26 2000-07-25 Monsanto Company Bacillus thuringiensis CryET29 compositions toxic to coleopteran insects and ctenocephalides SPP
US6537756B1 (en) 1996-09-26 2003-03-25 Monsanto Technology, Llc Bacillus thuringiensis CryET29 compositions toxic to coleopteran insects and Ctenocephalides SPP
US6686452B2 (en) 1996-09-26 2004-02-03 Monsanto Technology Llc Bacillus thuringiensis CryET29 compositions toxic to coleopteran insects and ctenocephalides SPP
EP1681351A1 (fr) * 1996-09-26 2006-07-19 Monsanto Technology LLC Compositions de Bacillus thuringiensis CryET29 toxiques pour les insectes Coleopteran et Ctenocephalides Spp.
WO1998013497A1 (fr) * 1996-09-26 1998-04-02 Ecogen, Inc. COMPOSITIONS A BASE DE CryET29 TIRE DE BACILLUS THURIGIENSIS TOXIQUES POUR LES COLEOPTERES ET LES CTENOCEPHALIDES SPP
US7186893B2 (en) 1996-09-26 2007-03-06 Monsanto Technology Llc Plants transformed with CryET29-encoding nucleic acids
US6130186A (en) * 1996-10-25 2000-10-10 Monsanto Company Composition and method for treating plants with exogenous chemicals
WO2007002988A1 (fr) * 2005-06-30 2007-01-11 Microbial Products Pty Ltd Formulation de pesticides microbiens
WO2010039652A3 (fr) * 2008-10-01 2011-04-28 Vestaron Corporation Formulation de toxine peptidique
US8217003B2 (en) 2008-10-01 2012-07-10 Vestaron Corporation Peptide toxin formulation
US8703910B2 (en) 2008-10-01 2014-04-22 Vestaron Corproation Peptide toxin formulation
US9352022B2 (en) 2008-10-01 2016-05-31 Vestaron Corporation Peptide toxin formulation
EP3066924A1 (fr) * 2008-10-01 2016-09-14 Vestaron Corporation Formulation de toxine peptidique

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