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WO2025132389A1 - Formulation - Google Patents

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Publication number
WO2025132389A1
WO2025132389A1 PCT/EP2024/086846 EP2024086846W WO2025132389A1 WO 2025132389 A1 WO2025132389 A1 WO 2025132389A1 EP 2024086846 W EP2024086846 W EP 2024086846W WO 2025132389 A1 WO2025132389 A1 WO 2025132389A1
Authority
WO
WIPO (PCT)
Prior art keywords
insecticidal protein
composition
protein
composition according
plant
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
PCT/EP2024/086846
Other languages
English (en)
Inventor
Marta OMEDES PUJOL
Aya IBRAHIM
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Syngenta Crop Protection AG Switzerland
Original Assignee
Syngenta Crop Protection AG Switzerland
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Syngenta Crop Protection AG Switzerland filed Critical Syngenta Crop Protection AG Switzerland
Publication of WO2025132389A1 publication Critical patent/WO2025132389A1/fr
Pending legal-status Critical Current
Anticipated expiration legal-status Critical

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Classifications

    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
    • A01P7/00—Arthropodicides
    • A01P7/04—Insecticides
    • 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
    • A01N37/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids
    • A01N37/44—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids containing at least one carboxylic group or a thio analogue, or a derivative thereof, and a nitrogen atom attached to the same carbon skeleton by a single or double bond, this nitrogen atom not being a member of a derivative or of a thio analogue of a carboxylic group, e.g. amino-carboxylic acids
    • A01N37/46—N-acyl derivatives

Definitions

  • the present invention relates to a composition comprising an insecticidal protein and maltodextrin, a process for preparing a composition according to the present invention and a method of controlling or preventing infestation of a plant, a part or a locus thereof by an insect pest comprising applying to the plant, the part or the locus thereof a composition according to the present invention.
  • Insect pests are mainly controlled by chemical pesticides.
  • Biological pest control agents for controlling insect pests are receiving increasing attention as alternative chemical control agents.
  • WO2013/122720 discloses an engineered pesticidal polypeptide which has activity against Ostrinia nubilalis (European corn borer).
  • the method according to the present invention comprises controlling an insect pest, wherein the insect pest is a Coleopteran, Hemipteran or Lepidopteran insect pest.
  • the insect pest is a Lepidopteran insect pest, such as Ostrinia nubilalis (European corn borer), Plutella xylostella (diamondback moth), Spodoptera frugiperda (fall armyworm), Spodoptera littoralis (Egyptian cotton leafworm), Spodoptera eridania (Southern armyworm moth), Spodoptera exigua (beet armyworm), Spodoptera litura (tobacco cutworm), Agrotis ipsilon (black cutworm), Agrotis orthogonia (pale western cutworm), Striacosta albicosta (western bean cutworm), Helicoverpa zea (corn earworm), Helicoverpa armigera (cotton bollworm), Heliothis virescens (tobacco budworm),
  • the insect pest is a Ostrinia nubilalis (European corn borer), Plutella xylostella (diamondback moth), Spodoptera frugiperda (fall armyworm), Spodoptera littoralis (Egyptian cotton leafworm), Spodoptera eridania (Southern armyworm moth), Spodoptera exigua (beet armyworm), Spodoptera litura (tobacco cutworm), Agrotis ipsilon (black cutworm), Helicoverpa zea (corn earworm), Helicoverpa armigera (cotton bollworm), Heliothis virescens (tobacco budworm), Trichoplusia ni (cabbage looper), Mamestra brassicae (cabbage moth), Tuta absoluta (tomato leafminer), Lobesia botrana (European grapevine moth), Grapholita molesta (Oriental fruit moth), Eupoecilia ambigu
  • the insect pest is a Spodoptera insect pest, preferably Spodoptera littoralis (Egyptian cotton leafworm).
  • a plant in a method of the invention is a crop plant such as maize, sorghum, wheat, sunflower, tomato, a crucifer, pepper, potato, cotton, rice, soybean, sugar beet, sugarcane, tobacco, barley, oilseed rape plant and the like.
  • the plant comprises soybean and I or cotton.
  • part of a plant comprises seeds, seedlings, saplings, roots, tubers, stems, stalks, foliage, leaves and fruits.
  • locus as used herein means fields in or on which plants are growing, or where seeds of cultivated plants are sown, or where seed will be placed into the soil.
  • the present invention relates to the use of maltodextrin having a DE of 5 to 50 to stabilize an insecticidal protein for at least three days, preferably at least four, five, six or at least seven days.
  • the use comprises stabilizing the insect protein on a plant, a part or a locus thereof.
  • the use further comprises a salt solution at pH 5.0 to 10. All embodiments described above are applicable to the use according to the present invention.
  • Vip3A SEQ ID NO: 1
  • the Vip3D variant protein SEQ ID NO: 3
  • Nucleic acid sequences for the Vip3 proteins were codon optimised for expression in E. coli and were cloned into a standard protein expression vector.
  • the vectors containing the Vip3 encoding sequences were introduced into an E. coli protein expression host via chemical transformation, to provide recombinant strains for Vip3 protein production.
  • the recombinant E.coli strains were cultivated by batch fermentation in a complex liquid medium as disclosed in Estruch et aP and Khan et. at 2 using a stirred tank bioreactor. Expression of Vip3 protein was induced by the addition of IPTG and incubation was continued for a further period followed by the harvesting of biomass by centrifugation.
  • Cell mass was collected from the fermentation broth using a disk stack centrifuge. The cell mass was resuspended in an aqueous sodium phosphate or Tris buffer at variable strength and a 137mM sodium chloride or Tris-buffer unless indicated otherwise. The cells were disrupted using a microfluidics device. Cell debris was removed by centrifugation followed by chemical inactivation of the cells using a biocide (benzisothiazolinone, BIT).
  • a biocide benzisothiazolinone, BIT
  • the clarified material contained soluble protein at a concentration of 2 to 10 mg / ml, measured by SDS- PAGE and densitometry against a purified standard.
  • Spray tank preparation the above solution of solube concentrate (SL) was diluted with water to achieve the correct application rate (g Al/ha), to make up 90% of the spray volume, and lastly the adjuvants (retention aids, wetting agents, rain fastness agents, photoprotectants) were added as a tank mix to achieve the right concentration with the remaining 10% of the spray volume.
  • SL solube concentrate
  • Spray tank preparation the above soluble powder (SP) was added and dissolved in water to achieve the correct application rate (g Al/ha) to make up 90% of the spray volume, and lastly the adjuvants (eg. retention aids, wetting agents, rain fastness agents, or photoprotectants) were added as a tank mix where needed to achieve the right concentration with the remaining 10% of the spray volume.
  • the adjuvants eg. retention aids, wetting agents, rain fastness agents, or photoprotectants
  • Soybean (Glycine max) or cotton plants (Gossypium hirsuturri) were sprayed with diluted test solutions in an application chamber. The plants were incubated in the greenhouse under controlled conditions (temperature at 22°C during day and 20°C during night and relative humidity of 65%).
  • the leaves were then cut and placed in petri dishes with wetted filter paper at different instances.
  • the petri dishes were infested (few hours after the application or 1 , 3, 4 or 5 days after the application day (DAA), with the Egyptian cotton leafworm, Spodoptera littoralis (8-10 L2 larvae per dish) and covered with a fabric filter and plastic lids. The larvae were assessed 5 days after infestation for mortality in each of the infestation instances.
  • DAA application day
  • Soluble concentrate (SL) type formulations were prepared by adding a solution of Vip3 clarified lysate into a 10% maltodextrin or disaccharide solutions.
  • Soybean plants were treated as described above with a solution of Vip3A and maltodextrin at 75 ppm (Table 2) and a solution of Vip3A and different disaccharides at 37.5 ppm (Table 3).
  • Cotton plants were treated with 200 ppm of Vip3A and polysaccharides (maltodextrin) and disaccharides (maltose and lactose) (Table 4).
  • Karate (lambda cyhalothrin) was taken as a positive control.
  • results in Table 3 show that the presence of disaccharides protected the Vip3A protein and provided control of Spodopetra litoralis L2 larvae on soybean leaves for only 1 day. Control against S. litoralis was very weak when larvae were infested on leaves 3 days after the application of a composition of disaccharides and Vip3A.
  • results in Table 4 show that the addition of maltodextrin to Vip3A resulted in a longer persistence of Vip3A on cotton leaves (up to 4 DAA) as compared to maltose and lactose.
  • Soluble concentrate (SL) type formulations prepared by adding a solution of clarified Vip3 lysate into a 10% maltodextrin solution, were used in this example.
  • Soybean plants were treated as described above with solutions of Vip3A or Vip3D containing maltodextrin Glucidex 29 at 75 ppm (Table 5).
  • Karate (lambda cyhalothrin) was taken as a positive control.
  • Soluble concentrate (SL) type formulations prepared by adding a solution of Vip3 clarified lysate into a 10% sugar solution (potato starch, methyl cellulose, glucose and Glucidex 29), were used in this example.
  • Soybean plants were treated as described above with a solution of Vip3A and in the presence of various types of sugars (see Table 1 and Table 6) at 37.5 and 9.375 ppm (Table 6).
  • Karate (lambda cyhalothrin) was used as a positive control.
  • the results in Table 6 show that amongst all the sugars tested, maltodextrin Glucidex 29 D stabilized Vip3 both applied at 37.5 and 9.375 ppm, providing persistent control against S. littoralis up to 5 days after the application on soybean plants.
  • Potato starch and methyl cellulose did not provide good persistent control against S. littoralis larvae at a concentration of 37.5 and 9.375 ppm and glucose was less effective against S. littoralis larvae at 9.375 ppm as compared to maltodextrin Glucidex 29.
  • Table 6 Mortality of Spodopetra littoralis L2 larvae, exposed at 0, 3 and 5 DAA, assessed 5 days after infestation on Vip3A-treated soybean leaves, with or without the addition of Glucidex 29 and other sugars.
  • the biomass collected from the fermenter was re-suspended in 50mM sodium phosphate or 50mM Tris-buffer solutions at pH 8 and 137mM sodium chloride.
  • the cells were lysed, and the cell debris removed via centrifugation.
  • Soluble powder type formulations were prepared by mixing the Vip3 clarified lysate in a 20% Glucidex 29 solution followed by lyophilization to remove the water.
  • Soybean plants were treated by dissolving the above powder, at a 75 ppm Vip3A dose rate (Table 7 and 8).
  • Karate (lambda cyhalothrin) was used as a positive control.
  • Tables 7 and 8 show that a solution of sodium phosphate resulted in increased stability of Vip3A in comparison with Tris-buffer. Incorporating Glucidex 29 with Vip3A in a sodium phosphate buffer provided control against S. littoralis larvae up to 5 DAA (table 7).
  • Table 7 Mortality of Spodopetra littoralis L2 larvae, infested at 0, 3 and 5 DAA, assessed 5 days after infestation on Vip3A-treated soybean leaves, with incorporated Glucidex 29 at different ratios in a sodium phosphate buffer.
  • Table 8 Mortality of Spodopetra littoralis L2 larvae, infested at 0, 3 and 5 DAA, assessed 5 days after infestation on Vip3A-treated soybean leaves, with incorporated Glucidex 29 D at different ratios in a Tris buffer.

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Environmental Sciences (AREA)
  • Pest Control & Pesticides (AREA)
  • Plant Pathology (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Health & Medical Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Dentistry (AREA)
  • Agronomy & Crop Science (AREA)
  • Insects & Arthropods (AREA)
  • Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Agricultural Chemicals And Associated Chemicals (AREA)

Abstract

La présente invention concerne une composition comprenant une protéine insecticide et de la maltodextrine ayant un DE de 5 à 50, la composition ayant un pH de 5,0 à 10. La présente invention concerne en outre un procédé de préparation d'une composition selon la présente invention et un procédé de lutte ou de prévention contre l'infestation d'une plante, d'une partie ou d'un locus de celle-ci par un insecte nuisible, consistant à appliquer à la plante, une partie ou un locus de celle-ci, une composition selon la présente invention.
PCT/EP2024/086846 2023-12-18 2024-12-17 Formulation Pending WO2025132389A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
EP23217598 2023-12-18
EP23217598.4 2023-12-18

Publications (1)

Publication Number Publication Date
WO2025132389A1 true WO2025132389A1 (fr) 2025-06-26

Family

ID=89223950

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP2024/086846 Pending WO2025132389A1 (fr) 2023-12-18 2024-12-17 Formulation

Country Status (1)

Country Link
WO (1) WO2025132389A1 (fr)

Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1994025611A1 (fr) * 1993-04-23 1994-11-10 Sandoz Ltd. Segment d'adn d'integration comprenant un gene codant une proteine insecticide
WO2010081815A1 (fr) 2009-01-13 2010-07-22 Universiteit Gent Compositions aqueuses coacervées comprenant des agents biologiquement actifs faiblement solubles dans l'eau
WO2013012272A2 (fr) 2011-07-20 2013-01-24 엘지전자 주식회사 Procédé de commutation de canal dans un système de communication sans fil et appareil pour ledit procédé
WO2013122720A2 (fr) 2012-02-16 2013-08-22 Syngenta Participations Ag Protéines pesticides modifiés
US20130296165A1 (en) 2010-01-28 2013-11-07 Advanced Bionutrition Corporation Stabilizing Composition for Biological Materials
WO2020182994A1 (fr) 2019-03-13 2020-09-17 Certis Europe B.V. Composition
WO2020247465A2 (fr) 2019-06-05 2020-12-10 Syngenta Participations Ag Lutte contre spodoptera
CA3218556A1 (fr) * 2021-07-02 2023-01-05 Pivot Bio, Inc. Souches bacteriennes genetiquement modifiees pour une fixation d'azote amelioree

Patent Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1994025611A1 (fr) * 1993-04-23 1994-11-10 Sandoz Ltd. Segment d'adn d'integration comprenant un gene codant une proteine insecticide
WO2010081815A1 (fr) 2009-01-13 2010-07-22 Universiteit Gent Compositions aqueuses coacervées comprenant des agents biologiquement actifs faiblement solubles dans l'eau
US20130296165A1 (en) 2010-01-28 2013-11-07 Advanced Bionutrition Corporation Stabilizing Composition for Biological Materials
WO2013012272A2 (fr) 2011-07-20 2013-01-24 엘지전자 주식회사 Procédé de commutation de canal dans un système de communication sans fil et appareil pour ledit procédé
WO2013122720A2 (fr) 2012-02-16 2013-08-22 Syngenta Participations Ag Protéines pesticides modifiés
WO2020182994A1 (fr) 2019-03-13 2020-09-17 Certis Europe B.V. Composition
WO2020247465A2 (fr) 2019-06-05 2020-12-10 Syngenta Participations Ag Lutte contre spodoptera
CA3218556A1 (fr) * 2021-07-02 2023-01-05 Pivot Bio, Inc. Souches bacteriennes genetiquement modifiees pour une fixation d'azote amelioree

Non-Patent Citations (6)

* Cited by examiner, † Cited by third party
Title
BEHLE ET AL., J. OF ECONOMIC ENTOMOLOGY, vol. 90, no. 6, 1997, pages 1561 - 1566
ESTRUCH JJWARREN GWMULLINS MANYE GJCRAIG JAKOZIEL MG: "Vip3A, a novel Bacillus thuringiensis vegetative insecticidal protein with a wide spectrum of activities against lepidopteran insects", PROC NATL ACAD SCI U S A., vol. 93, no. 11, 28 May 1996 (1996-05-28), pages 5389 - 94, XP002071759, DOI: 10.1073/pnas.93.11.5389
M. H. KHANG. JANDERZ. MUKHTARM. ARSHADM. SARWARS. ASAD: "Comparison of in Vitro and in Planta Toxicity of Vip3A for Lepidopteran Herbivores", JOURNAL OF ECONOMIC ENTOMOLOGY, vol. 113, December 2020 (2020-12-01), pages 2959 - 2971
R W BEHLE: "Effects of Sunlight and Simulated Rain on Residual Activity of Bacillus thuringiensis Formulations", JOURNAL OF ECONOMIC ENTOMOLOGY, vol. 90, no. 6, 1 December 1997 (1997-12-01), pages 1560 - 1566, XP093158448, DOI: 10.1093/jee/90.6.1560 *
SYED ET AL., TOXINS, vol. 12, 2020, pages 522
W. S. ABBOTT: "A Method of Computing the Effectiveness of an Insecticide", JOURNAL OF ECONOMIC ENTOMOLOGY, vol. 18, 1 April 1925 (1925-04-01), pages 265 - 267

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