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CA-3046226-C - INSECTICIDAL PROTEINS AND METHODS FOR THEIR USE

CA3046226CCA 3046226 CCA3046226 CCA 3046226CCA-3046226-C

Abstract

Compositions and methods for controlling pests are provided. The methods involve transforming organisms with a nucleic acid sequence encoding an insecticidal protein. In particular, the nucleic acid sequences are useful for preparing plants and microorganisms that possess insecticidal activity. Thus, transformed bacteria, plants, plant cells, plant tissues and seeds are provided. Compositions are insecticidal nucleic acids and proteins of bacterial species. The sequences find use in the construction of expression vectors for subsequent transformation into organisms of interest including plants, as probes for the isolation of other homologous (or partially homologous) genes. The pesticidal proteins find use in controlling, inhibiting growth or killing Lepidopteran, Coleopteran, Dipteran, fungal, Hemipteran and nematode pest populations and for producing compositions with insecticidal activity.

Inventors

  • JENNIFER KARA BARRY
  • Hua Dong
  • James English
  • Jacob Gilliam
  • KAI M. HILLMAN
  • DANIEL JAMES THORPE
  • Thomas Chad Wolfe
  • Nasser Yalpani

Assignees

  • PIONEER HI-BRED INTERNATIONAL, INC.

Dates

Publication Date
20260505
Application Date
20171218
Priority Date
20161222

Claims (20)

  1. 83 THAT WHICH IS CLAIMED IS: 1. A recombinant polypeptide having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 2, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO: 46, SEQ 5 ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, or SEQ ID NO: 54, wherein the polypeptide has insecticidal activity.
  2. 2. The recombinant polypeptide of claim 1, wherein the polypeptide has insecticidal activity against Western Com Rootworm (Diabrotica virgifera virgifera).
  3. 3. A recombinant polynucleotide encoding the polypeptide of claim 1 or 2. 10
  4. 4. The recombinant polynucleotide of claim 3, wherein the polynucleotide 1s a non-genomic po lynucleoti de.
  5. 5. The recombinant polynucleotide of claim 4, wherein the polynucleotide is synthetic polynucleotide.
  6. 6. The recombinant polynucleotide of claim 5, wherein the polynucleotide has codons optimized for expression in an agriculturally important crop. 15
  7. 7. A transgenic plant cell comprising a polynucleotide encoding a polypeptide having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 2, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, or SEQ ID NO: 54, wherein the polypeptide has insecticidal activity.
  8. 8. A DNA construct comprising a polynucleotide encoding a polypeptide having at least 95% 20 sequence identity to the amino acid sequence of SEQ ID NO: 2, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, or SEQ ID NO: 54, wherein the polypeptide has insecticidal activity.
  9. 9. A transgenic plant cell comprising the DNA construct of claim 8.
  10. 10.
  11. 11. A composition comprising the polypeptide of claim 1 or 2, and an agriculturally acceptable carrier. A fusion protein comprising the polypeptide of claim 1 or 2.
  12. 12. A method for controlling an insect pest population, comprising contacting the insect pest population with a polypeptide having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 2, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, or SEQ ID NO: 54, wherein the polypeptide has 30 insecticidal activity.
  13. 13. A method of inhibiting growth or killing an insect pest, comprising contacting the insect pest with a composition comprising a polypeptide having at least 95% sequence identity to the amino acid sequence Date Re9ue/Date Received 2024-04-09 84 of SEQ ID NO: 2, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, or SEQ ID NO: 54, wherein the polypeptide has insecticidal activity.
  14. 14. The method of claim 13, wherein the insect pest is a Lepidoptera and/or Coleoptera insect pest. 5
  15. 15. A method for controlling an insect pest population, comprising contacting the insect pest population with the transgenic plant cell of claim 7 or 9.
  16. 16. A method of inhibiting growth or killing an insect pest, comprising transforming a plant with the DNA construct of claim 8.
  17. 17. The method of claim 16, further comprising contacting the insect pest with the transgenic plant or 10 plant cell.
  18. 18. The method of claim 16 or 17, wherein the insect pest is Western Com Rootworm (Diabrotica virgifera virgifera).
  19. 19. The method of any one of claims 13-18, wherein the insect pest or insect pest population is resistant to at least one Bt toxin. 15
  20. 20. Use of the polypeptide of claim 1 or 2 to inhibit growth or kill an insect or insect population. Date Re9ue/Date Received 2024-04-09

Description

1 INSECTICIDAL PROTEINS AND METHODS FOR THEIR USE CROSS-REFERENCE TO RELATED APPLICATIONS This Application claims the benefit of U.S. Provisional Application No. 62/438,179 filed on 5 December 22, 2016 . REFERENCE TO SEQUENCE LISTING SUBMITTED ELECTRONICALLY The official copy of the sequence listing is submitted electronically via EFS~W eb as an ASCII formatted sequence listing with a file named "6729WOPCT Sequence_Listing" created on November 30, 10 2017, and having a size of 107 kilobytes and is filed concurrently with the specification. The sequence listing contained in this ASCII formatted document is part of the specification . FIELD This disclosure relates to the field of molecular biology. Provided are novel genes that encode pesticidal proteins. These pesticidal proteins and the nucleic acid sequences that encode them are useful in preparing pesticidal formulations and in the production of transgenic pest-resistant plants. BACKGROUND Biological control of insect pests of agricultural significance using a microbial agent, such as fungi, bacteria or another species of insect affords an environmentally friendly and commercially attractive alternative to synthetic chemical pesticides. Generally speaking, the use of biopesticides presents a lower risk of pollution and environmental hazards and biopesticides provide greater target specificity than is characteristic of traditional broad-spectrum chemical insecticides. In addition, biopesticides often cost less 25 to produce and thus improve economic yield for a wide variety of crops. Certain species of microorganisms of the genus Bacillus are known to possess pesticidal activity against a range of insect pests including Lepidoptera, Diptera, Coleoptera, Hemiptera and others. Bacillus thuringiensis (Bt) and Bacillus popilliae are among the most successful biocontrol agents discovered to date. Insect pathogenicity has also been attributed to strains of B. larvae, B. lentimorbus, B. sphaericus and 30 B. cereus. Microbial insecticides, particularly those obtained from Bacillus strains, have played an important role in agriculture as alternatives to chemical pest control. Date Rer;ue/Date Received 2024-04-09 WO 2018/118811 PCT/0S2017/067107 2 Crop plants have been developed with enhanced insect resistance by genetically engineering crop plants to produce pesticidal proteins from Bacillus. For example, corn and cotton plants have been genetically engineered to produce pesticidal proteins isolated from strains of Bacillus thuringiensis. These genetically engineered crops are now widely used in agriculture and have provided the farmer with an 5 environmentally friendly alternative to traditional insect-control methods. While they have proven to be very successful commercially, these genetically engineered, insect-resistant crop plants may provide resistance to only a narrow range of the economically important insect pests. In some cases, insects can develop resistance to different insecticidal compounds, which raises the need to identify alternative biological control agents for pest control. Accordingly, there remains a need for new pesticidal proteins with different ranges of insecticidal activity against insect pests, e.g., insecticidal proteins which are active against a variety of insects in the order Lepidoptera and the order Coleoptera, including but not limited to insect pests that have developed resistance to existing insecticides. SUMMARY In one aspect compositions and methods for conferring pesticidal activity to bacteria, plants, plant cells, tissues and seeds are provided. Compositions include nucleic acid molecules encoding sequences for pesticidal and insecticidal polypeptides, vectors comprising those nucleic acid molecules, and host cells comprising the vectors. Compositions also include the pesticidal polypeptide sequences and antibodies to 20 those polypeptides. Compositions also comprise transformed bacteria, plants, plant cells, tissues and seeds. In another aspect isolated or recombinant nucleic acid molecules are provided encoding IPD101 polypeptides including amino acid substitutions, deletions, insertions, and fragments thereof. Provided are isolated or recombinant nucleic acid molecules capable of encoding IPD101 polypeptides of SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 25, 26, 28, 29, 30, 32, 46, 48, 50, 52, 54, 56, 58, and 60, as well as 25 amino acid substitutions, deletions, insertions, fragments thereof, and combinations thereof. Nucleic acid sequences that are complementary to a nucleic acid sequence of the embodiments or that hybridize to a sequence of the embodiments are also encompassed. The nucleic acid sequences can be used in DNA constructs or expression cassettes for transformation and expression in organisms, including microorganisms and plants. The nucleotide or amino acid sequences may be synthetic sequences that have 30 been designed for expression in an organism including, but not limited to, a microorganis