WO1996028023A2 - Synergistes de la delta-endotoxine de bacillus thuringiensis - Google Patents

Synergistes de la delta-endotoxine de bacillus thuringiensis Download PDF

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Publication number
WO1996028023A2
WO1996028023A2 PCT/US1996/003386 US9603386W WO9628023A2 WO 1996028023 A2 WO1996028023 A2 WO 1996028023A2 US 9603386 W US9603386 W US 9603386W WO 9628023 A2 WO9628023 A2 WO 9628023A2
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bacillus thuringiensis
synergist
composition according
pesticidal composition
biopesticide
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PCT/US1996/003386
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English (en)
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WO1996028023A3 (fr
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Rodney Kahn
Guy Orgambide
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Abbott Laboratories
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Priority to AU53095/96A priority Critical patent/AU5309596A/en
Publication of WO1996028023A2 publication Critical patent/WO1996028023A2/fr
Publication of WO1996028023A3 publication Critical patent/WO1996028023A3/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
    • A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
    • 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 pesticidal compositions comprising a Bacillus thuringiensis biopesticide and a synergist, wherein the synergist enhances the pesticidal activity of a Bacillus thuringiensis biopesticide and/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-posinve bacte ⁇ um that is widely distributed L ⁇ nature, especially in soil and insect- ⁇ ch environments Dunne sporulanon, Bacillus thuringiensis produces a parasporal crystal inclusions ) which is insec ⁇ cidal upon inges ⁇ on to susceptible msect 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 kD_
  • 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 biopesticids. 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. tenebrioms. Furthermore, Bacillus thuringiensis subsp. israelensis produces delta-endotoxin proteins specific for Diptera.
  • cry crystal protein
  • the major classes are Lepidoptera-specific (cryl); Lepidoptera-and Diptera-specific (cryll); Coleoptera-specif ⁇ c (crylll); Diptera-specific (crylV); Coleoptera- and Lepidoptera-specific (cryV genes); and Nematode-specific (cryVI genes).
  • israeltaki biopesticide against 4 day old Spodoptera frugiperda larvae is mainly additive at levels where individually Neem (25 to 125 ppm) and the biopesticide (0.5%) are lethal, but show some synergism when Neem is present at levels between 25 and 125 ppm with the biopesticide at 0.1% (Hellpap and Zebitz, 1986, Journal of Applied Entomology 101:515-524). Antagonism is observed when Neem is present at a low level of 25 ppm with the biopesticide at 0.5%.
  • Moar and Trumble disclose that there is no synergistic interaction of Neem with a biopesticide from Bacillus thuringiensis subsp.
  • Knauf et al. disclose the synergistic combination of avermectin and a Bacillus thuringiensis biopesticide (Knauf et al, 1987, European Patent Application No. 242 502 A2).
  • Noriyoshi and Hisaki disclose a synergistic composition containing avermectin B l and a Bacillus thuringiensis biopesticide (1984, JP 59199616). Guertin et al.
  • the present invention relates to pesticidal compositions comprising an amount of a Bacillus thuringiensis biopesticide and a synergist in an effective amount for enhancing the pesticidal activity of the Bacillus thuringiensis biopesticide and/or 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 the pesticidal compositions.
  • the present invention is directed to pesticidal compositions comprising an amount of a Bacillus thuringiensis biopesticide and a synergist in an effective amount for enhancing the pesticidal activity of the Bacillus thuringiensis biopesticide to effectively control the 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 the amount of a synergist sufficient to enhance the pesticidal activity of a Bacillus thuringiensis biopesticide by about 20%, preferably by about 40%, more preferably by about 60%, and most preferably by about 100%.
  • the synergist may or may not have pesticidal activity at the rates used to synergize the pesticidal activity of a Bacillus thuringiensis biopesticide.
  • the pesticidal activity of a Bacillus thuringiensis biopesticide and the synergistic effect of a synergist 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 synergist to enhance the pesticidal activity of the Bacillus thuringiensis biopesticide.
  • 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, and furthermore, will vary depending on the pest and the crop.
  • the efficacy of a large number of Bacillus thuringiensis biopesticides is well established in the art.
  • the amounts required for commercial utility are either known or are easily determined by the ordinary skilled artisan. For example, in agriculture in the treatment of lettuce, an amount of a Bacillus thuringiensis biopesticide required for commercial utility is that which achieves greater than about 80% control of Spodoptera exigua.
  • the benefit of the present invention may be achieved by determining the amount of a Bacillus thuringiensis biopesticide required alone to achieve commercially acceptable control of a pest.
  • the amount of a synergist 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 synergist 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 synergist 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.
  • synergists have been determined to enhance the pesticidal activity of a Bacillus thuringiensis biopesticide.
  • the synergist is an azadirachtin, e.g., Neem seed extract, azadirachtin A, azadirachtin B, or derivatives and analogs of an azadirachtin.
  • the synergist is zinc oxide or zinc chloride.
  • the synergist is a combination of azadirachtin and a zinc salt. In a more preferred embodiment, the synergist is a combination of azadirachtin and zinc oxide.
  • the synergist is an amphiphatic lipid wherein said amphiphatic lipid is decanoyl-N-methyl-gluc amide, digalactosyl diglyceride. (glycerol moiety esterified with oleic acid and palmitic acid), dodecyl glucopyranoside, dodecyl trimethyl ammonium bromide, triolein, or phosphatidylethanolamine.
  • the synergist is an analgesic wherein said analgesic is acetylsalicylic acid, ibuprofen, or acetaminophen.
  • the synergist is esculin.
  • the synergist is poly-L-lysine.
  • the synergist is alpha-tocopherol.
  • synergists disclosed above may be obtained synthetically or from natural sources.
  • the pesticidal compositions of the present invention may comprise a combination of two or more of the synergists described above.
  • the pesticidal compositions of the present invention may comprise one or more synergists, potentiators, or phagostimulants known in the art.
  • the potentiator disclosed in WO 94/09630 or a pesticidally-acrive salt thereof may be used in the compositions of the present invention:
  • compositions of the present invention comprise the synergist in an amount between about 0.001 and about 100 grams, preferably between about 0.001 and about 50 grams, more preferably between about 0.001 and about 25 grams, more preferably between about 0.001 and about 10 grams, more preferably between about 0.001 and about 2 grams, more preferably between about 0.001 and about 0.2 gram, more preferably between about 0.001 and about 0.1 gram, and most preferably between about 0.001 and about 0.01 gram per gram of Bacillus thuringiensis biopesticide. These amounts are based on a composition in which the biopesticide is a Bacillus thuringiensis subsp.
  • israeltaki 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 ⁇ star Trichoplusia ni using the Bacillus thuringiensis subsp. kurstaki standard HD1-S-1980, USDA, Peona, Illinois). It will be obvious to a person skilled in die art that the amount of a particular synergist preparation needed to enhance a Bacillus thuringiensis biopesticide may vary significantiy depending on the purity of die synergist preparation.
  • the Bacillus thuringiensis biopesticide in the pesticidal compositions of d e 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. colmeri, Bacillus thuringiensis subsp. coreanensis, Bacillus thuringiensis subsp. dakota. Bacillus thuringiensis subsp. darmstadiensis, Bacillus thuringiensis subsp. dendrolimus, Bacillus thuringiensis subsp.
  • Bacillus thuringiensis subsp. finitimus Bacillus thuringiensis subsp. galleriae, Bacillus thuringiensis subsp. Indiana, Bacillus thuringiensis subsp. israelensis, Bacillus thuringiensis subsp. kenyae. Bacillus thuringiensis subsp. kumamotoensis. Bacillus thuringiensis subsp. kurstaki, Bacillus thuringiensis subsp. kyushuensis, Bacillus thuringiensis subsp. japonensis, Bacillus thuringiensis subsp.
  • Bacillus thuringiensis subsp. morrisoni Bacillus thuringiensis subsp. neoleonensis, Bacillus thuringiensis subsp. nigeriae, Bacillus thuringiensis subsp. ostriniae, Bacillus thuringiensis subsp. Pakistani, Bacillus thuringiensis subsp. pondicheriensis, Bacillus thuringiensis subsp. shandongiensis, Bacillus thuringiensis subsp. si/ ⁇ , Bacillus thuringiensis subsp. _ ⁇ tt ⁇ , Bacillus thuringiensis subsp.
  • the biopesticide is derived from a Bacillus thuringiensis subsp. kurstaki strain.
  • the synergist 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 synergist for enhancing the pesticidal activity of a commercially ineffective amount of a Bacillus thuringiensis biopesticide will be the same.
  • the Bacillus thuringiensis biopesticide in the pesticidal compositions 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 group including, but not limited to, Cryl, Cryll . CryHI, CryIN, Cry V, and Cry VI.
  • the Bacillus thuringiensis delta-endotoxin protein or pesticidally-active fragment thereof may include, but is not limited to, Cry ⁇ A(a), Cry ⁇ A(b), Cry ⁇ A(c), CrylB, CrylC, CrylD, CrylE, CrylF, CryllA, CryHB, CryHIA, CrylUB, CrymC, CrylNA, CrylVB, CrylVC, CrylVD, CryV, CryNI, and CytA.
  • 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 synergists 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 californica 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.
  • the synergists 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 pesticidal compositions 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 compositions 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, soy 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, vermiculite, diatomaceous earth), natural clays (attapulgite, bentonite, kaolinite, montmorillonite), and synthetic clays (Laponite).
  • the deposition agent is present in the pesticidal compositions of the present invention in an amount of between about 0.4% w/w and about 50% w/w, preferably between about 1% w/w and about 20% w/w.
  • the pesticidal compositions 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 making the product more efficacious and palatable.
  • the antifreeze/humectant agent is selected from the group consisting of ethylene glycol, pr ⁇ pylene glycol, dipropylene glycol, glycerol, butylene glycols, pentylene glycols and hexylene glycols.
  • the antifreeze/humectant agent is present in the pesticidal compositions of the present invention in an amount of between about 0.5% w/w and about 25% w/w, preferably between about 2% w/w and about 15% w/w.
  • the pesticidal compositions of the present invention may further comprise a surfactant in an amount where it acts as an emulsifying, a wetting, or a dispersing agent.
  • a 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 such
  • 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.
  • surfactants are cationic surfactants such as aliphatic mono-, di-, or polyamine as acetates, naphthenates or oleates; oxygen-containing amines such as an amine 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 surfactant is present in an amount of between about 0.5% w/w and about 25% w/w, preferably between about 1% w/w and about 8% w/w.
  • the pesticidal compositions 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 sugars, starches, or cyclodextrins; or botanical materials such as wood products, cork, powdered corncobs, rice hulls, peanut hulls, and walnut shells.
  • the pesticidal compositions 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 pesticidal compositions of the present invention.
  • the pesticidal compositions 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 may contain 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 synergist.
  • the compositions will be preferably administered at the labeled rate for the commercial product, preferably about 0.01 pound to 5.0 pounds per acre when in dry form and at about 0.01 pint to 25 pints per acre when in liquid form.
  • the pesticidal compositions 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 compositions can be applied by foliar, furrow, broadcast granule, "lay-by", or soil drench application.
  • the compositions 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 compositions can be applied by spraying, dusting, sprinkling, or the like.
  • the spray or dust can conveniently contain another pesticide.
  • the pesticidal compositions of the present invention are preferably applied directly to the plant.
  • the pesticidal compositions 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 (a
  • the synergists of the present invention may be applied separately to a plant previously exposed to a Bacillus thuringiensis biopesticide.
  • the present invention further relates to a method for applying a synergist of the present invention to a transgenic plant, which contains a gene that encodes a Bacillus thuringiensis biopesticide.
  • the pesticidal compositions 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 compositions 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., Athens mindara, Bombyx mori, Bucculatrix thurberiella, Cadra cautella, Choristoneura sp., Cochylis hospes, Colias eurytheme, Corcyra cephalonica, Cy
  • the pesticidal compositions 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, Cera ⁇ tis 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
  • 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 adjusted to pH 7.0:
  • the whole broth is centrifuged to recover the crystals/spores (and other insoluble substances).
  • the pelleted material or whole broth concentrate is used to characterize the effect of various synergists on the insecticidal activity of the crystal/spore preparation horn Bacillus thuringiensis subsp. kurstaki HD1.
  • the ability of a synergist to enhance the pesticidal activity of the Bacillus thuringiensis subsp. kurstaki whole broth concentrate from EXAMPLE 1 is determined using a Spodoptera exigua bioassay.
  • the Spodoptera exigua bioassay is performed in a 240 well tray where each well is filled with 0.5 ml of insect diet (10 wells for each dose).
  • the synergist is tested in combination with three concentrations of Bacillus thuringiensis subsp. kurstaki whole broth concentrate: 0.5, 0.25, and 0.12 mg per ml.
  • the three concentrations of whole broth concentrate are also run without the synergist
  • the synergist is also bioassayed alone in the absence of the whole broth concentrate.
  • a score of 15/30 results when there is no synergistic effect When fewer than 15 wells have surviviors, this indicates there is synergism provided that there is no mortality in the zero dose of the whole concentrate. Some synergists can cause stunting at the zero dose of the whole concentrate, but not cause immediate mortality.
  • a volume of 50 ⁇ l of each concentration of diluted whole broth concentrate is first loaded into 10 wells (per dose of the whole broth concentrate), then 50 ul of the synergist solution are added to 40 wells and the mixture is allowed to dry. Once the samples are dried, 3-5 Spodoptera exigua eggs are placed into each well.
  • Azadirachtin (MARGOSAN-OTM, 0.25% azadirachtin, W.R. Grace & Co., Boca Raton, FL) is evaluated for its synergistic ability using the bioassay described in EXAMPLE 2 to enhance the pesticidal activity of the Bacillus thuringiensis subsp. kurstaki whole broth concentrate of Example 1.
  • Zinc oxide and zinc chloride are evaluated, using the bioassay described in EXAMPLE 2, for their synergistic ability to enhance the pesticidal activity of the Bacillus thuringiensis subsp. kurstaki whole broth concentrate of EXAMPLE 1.
  • Zinc Oxide (5 mg/ml) 0 2 2 10 1.5 4 no synergist 4 5 8 10 4.0 17
  • azadirachtin MARGOSAN-OTM
  • zinc oxide Sigma Chemical Company, St. Louis, MO
  • the concentration of each in the combination can be significantly reduced while achieving a similar synergistic effect compared to the the compounds alone at higher concentrations.
  • the combination of azadirachtin and zinc oxide is assayed at the same concentrations in the absence of the whole broth concentrate, the combinations produce significant stunting, but have no lethal effect
  • MARGOSAN-OTM (0.01 mg/ml) 0 2 0 10 1.0 2
  • Zinc Oxide (5 mg/ml) 0 2 2 10 1.5 4
  • Zinc Oxide (2.5 mg/ml) 1 2 2 10 2.0 5
  • MARGOSAN-OTM/Zinc Oxide 0 0 0 10 1.0 0 (0.005/2.5 mg/ml)
  • Decanoyl-N-methyl-glucamide, digalactosyl diglyceride (in which the glycerol moiety is esterified with oleic acid and palmitic acid), dodecyl glucopyranoside, dodecyl trimethyl ammonium bromide, triolein, and phosphatidylethanolamine (Sigma Chemical Company, St. Louis, MO) are evaluated, using the bioassay described in EXAMPLE 2, for their synergistic ability to enhance the pesticidal activity of the Bacillus thuringiensis subsp. kurstaki whole broth concentrate of EXAMPLE 1.
  • the total number of wells with survivors for the three doses of the whole broth concentrate in the presence of digalactosyl diglyceride is 5, while in the absence of digalactosyl diglyceride the total number is 17.
  • the total number of wells with survivors for the three doses of the whole broth concentrate in the presence of dodecyl glucopyranoside is 7, while in the absence of dodecyl glucopyranoside the total number is 17.
  • the total number of wells with survivors for the three doses of the whole broth concentrate in the presence of dodecyl trimethyl ammonium bromide is 6, while in the absence of dodecyl trimethyl ammonium bromide the total number is 16.
  • the total number of wells with survivors for the three doses of the whole broth concentrate in the presence of triolein is 5, while in d e absence of triolein the total number is 16.
  • the total number of wells with survivors for the three doses of the whole broth concentrate in the presence of phosphatidylethanolamine is 7, while in the absence of phosphatidylethanolamine the total number is 17.
  • Triolein (9.74 mg/ml) 0 1 4 9 3.0 5 No Synergist 2 7 7 10 4.0 16
  • Acetyl salicylic acid, ibuprofen, and acetaminophen are evaluated, using the bioassay described in EXAMPLE 2, for their synergistic ability to enhance the pesticidal activity of Bacillus thuringiensis subsp. kurstaki whole broth concentrate of EXAMPLE 1.
  • the total number of wells with survivors for the three doses of the whole broth concentrate in the presence of acetaminophen is 7, while in the absence of acetaminophen the total number is 16.
  • the total number of wells with survivors for the three doses of the whole broth concentrate in the presence of ibuprofen is 6, while in the absence of ibuprofen the total number is 16.
  • Acetyl Salicylic Acid (4.51 mg/ml) 0 2 0 10 4.0 2
  • Esculin Sigma Chemical Company, St. Louis, MO is evaluated, using the bioassay described in EXAMPLE 2, for its synergistic ability to enhance the pesticidal activity of Bacillus thuringiensis subsp. kurstaki whole broth concentrate of EXAMPLE 1.
  • Poly-L-lysine (Sigma Chemical Company, St Louis, MO) is evaluated, using the bioassay described in EXAMPLE 2, for its synergistic ability to enhance the pesticidal activity of Bacillus thuringiensis subsp. kurstaki whole broth concentrate of EXAMPLE 1.
  • Alpha-tocopherol (Sigma Chemical Company, St Louis, MO) is evaluated, using the bioassay described in EXAMPLE 2, for its synergistic ability to enhance the pesticidal activity of Bacillus thuringiensis subsp. kurstaki whole broth concentrate of EXAMPLE 1.
  • alpha-tocopherol at a concentration of approximately 17 mg to 72 mg per mg of whole broth concentrate enhances the pesticidal activity of the Bacillus thuringiensis subsp. kurstaki whole broth concentrate.
  • alpha-tocopherol When alpha-tocopherol is assayed at the same concentration in the absence of the whole broth concentrate, it minimally stunts insect growth (stunt score of 3.5), but has no lethal effect
  • the total number of wells with survivors for the three doses of the whole broth concentrate in the presence of alpha-tocopherol is 2, while in the absence of alpha : tocopherol the total number is 16.
  • a lethal effect by alpha-tocopherol is observed at a concentration equal to or greater than 50 mM.
  • Alpha-Tocopherol (8.61 mg/ml) 0 1 1 10 3.5 2 No Synergist 4 6 6 10 4.0 16

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  • Life Sciences & Earth Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Health & Medical Sciences (AREA)
  • Zoology (AREA)
  • Plant Pathology (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Dentistry (AREA)
  • Wood Science & Technology (AREA)
  • Agronomy & Crop Science (AREA)
  • Environmental Sciences (AREA)
  • Mycology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Pest Control & Pesticides (AREA)
  • Virology (AREA)
  • Agricultural Chemicals And Associated Chemicals (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

Compositions antiparasitaires renfermant une certaine quantité de bio-pesticides Bacillus thuringiensis ainsi qu'un synergiste, sous une quantité efficace pour accroître l'activité antiparasitaire de ce bio-pesticide Bacillus thuringiensis destiné à la destruction des nuisibles. Ce synergiste permet de réduire la quantité de bio-pesticides Bacillus thuringiensis normalement nécessaire pour constituer un bio-pesticide commercialement rentable. L'invention porte en outre sur un procédé de destruction des nuisibles consistant à soumettre lesdits nuisibles à l'action des compositions antiparasitaires faisant l'objet de l'invention.
PCT/US1996/003386 1995-03-13 1996-03-12 Synergistes de la delta-endotoxine de bacillus thuringiensis WO1996028023A2 (fr)

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FR2772557A1 (fr) * 1997-12-23 1999-06-25 Novartis Ag Utilisation de macrolides dans la lutte contre les nuisibles
WO1999035913A1 (fr) * 1998-01-16 1999-07-22 Novartis Ag Utilisation de neonicotinoides sur des plantes transgeniques
WO1999035910A2 (fr) * 1998-01-16 1999-07-22 Novartis Ag Utilisation d'insecticides dans la lutte contre les insectes nuisibles
WO2002030205A2 (fr) * 2000-10-06 2002-04-18 Monsanto Technology, Llc Traitement de graines de mais transgenique au moyen d'un pesticide pour reduire les degats infliges a la plante par des ravageurs
US6733802B1 (en) 1997-12-17 2004-05-11 Fortune Bio-Tech Limited Natural azadirachtin composition
US6844339B2 (en) 1998-01-16 2005-01-18 Syngenta Crop Protection, Inc. Use of neonicotinoids in pest control
US6875727B2 (en) 1997-12-23 2005-04-05 Syngenta Crop Protection, Inc. Use of macrolides in pest control
AT501406A1 (de) * 1997-12-23 2006-08-15 Syngenta Participations Ag Verfahren zur bekämpfung von mollusken
WO2007053760A2 (fr) * 2005-11-01 2007-05-10 Dow Agrosciences Llc Compositions pesticides presentant une activite amelioree
WO2010116264A2 (fr) * 2009-04-07 2010-10-14 Taminco, Naamloze Vennootschap Agent phytoprotecteur
WO2014067663A1 (fr) * 2012-11-03 2014-05-08 Clariant International Ltd Compositions adjuvantes aqueuses
US8765697B2 (en) 2009-04-30 2014-07-01 Dow Agrosciences, Llc. Pesticide compositions exhibiting enhanced activity
US8785379B2 (en) 2009-04-30 2014-07-22 Dow Agrosciences, Llc. Pesticide compositions exhibiting enhanced activity
US8796476B2 (en) 2009-04-30 2014-08-05 Dow Agrosciences, Llc Pesticide compositions exhibiting enhanced activity and methods for preparing same
CN107467088A (zh) * 2017-06-26 2017-12-15 浦江县昂宝生物技术有限公司 海棠用杀虫剂及其制备方法
CN108935505A (zh) * 2017-05-18 2018-12-07 河北昊阳化工有限公司 一种杀灭地下害虫的农药混配组合物
JP2019519567A (ja) * 2016-06-29 2019-07-11 クラリアント・インターナシヨナル・リミテツド 微生物を阻害するための組成物
US10813862B2 (en) 2012-05-30 2020-10-27 Clariant International Ltd. Use of N-methyl-N-acylglucamines as solubilizers
US10864275B2 (en) 2012-05-30 2020-12-15 Clariant International Ltd. N-methyl-N-acylglucamine-containing composition
US10920080B2 (en) 2015-10-09 2021-02-16 Clariant International Ltd. N-Alkyl glucamine-based universal pigment dispersions
US10961484B2 (en) 2015-10-09 2021-03-30 Clariant International Ltd. Compositions comprising sugar amine and fatty acid
US11220603B2 (en) 2016-05-09 2022-01-11 Clariant International Ltd. Stabilizers for silicate paints
US11425904B2 (en) 2014-04-23 2022-08-30 Clariant International Ltd. Use of aqueous drift-reducing compositions

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