Lepidopteran-active Cry1Da1 amino acid sequence variant proteins

Patent metadata

Field	Value
Publication number	US-10059959-B2
Application number	US-201514884432-A
Country	US
Kind code	B2
Filing date	Oct 15, 2015
Priority date	Oct 16, 2014
Publication date	Aug 28, 2018
Grant date	Aug 28, 2018

Abstract

Engineered Cry1Da amino acid sequences are provided that exhibit improved Lepidopteran insecticidal activity and an enhanced Lepidopteran spectrum compared to the naturally occurring Cry1Da protein toxin. Polynucleotide sequences intended for use in expression of the improved proteins in plants are also provided. Particular embodiments provide compositions containing insect inhibitory amounts of the engineered proteins, as well as recombinant plants, plant parts, and seeds containing polynucleotide constructs encoding one or more of the improved engineered proteins.

First claim

What is claimed is:

1. An engineered insecticidal protein comprising the amino acid sequence as set forth in SEQ ID NO:40.

2. The engineered insecticidal protein of claim 1 , wherein the engineered insecticidal protein exhibits inhibitory activity against an insect species of the order Lepidoptera.

3. The engineered insecticidal protein of claim 2 , wherein the Lepidoptera is selected from the group consisting of a Spodoptera and a Helicoverpa.

4. The engineered insecticidal protein of claim 2 , wherein the Lepidoptera is selected from the group consisting of Helicoverpa zea and Spodoptera frugiperda.

5. The engineered insecticidal protein of claim 1 , encoded by a polynucleotide operably linked to a heterologous promoter.

6. The engineered insecticidal protein of claim 5 , wherein said engineered insecticidal protein is encoded by a polynucleotide comprising a nucleotide sequence that hybridizes to the reverse complement of the polynucleotide sequence as set forth in SEQ ID NO: 39.

7. A host cell comprising the polynucleotide of SEQ ID NO: 39, wherein said polynucleotide is present within a host cell selected from the group consisting of a bacterial host cell and a plant host cell.

8. The host cell of claim 7 , wherein said bacterial host cell is selected from the group consisting of Agrobacterium, Rhizobium, Bacillus, Brevibacillus, Escherichia, Pseudomonas, Klebsiella , and Erwinia.

9. The host cell of claim 7 , wherein said plant host cell is selected from the group of plants consisting of a monocot plant cell and a dicot plant cell.

10. An insect inhibitory composition comprising the engineered insecticidal protein of claim 1 , wherein said protein is present in an insect inhibitory amount.

11. The insect inhibitory composition of claim 10 , wherein said composition further comprises a second insect inhibitory agent different from said engineered insecticidal protein.

12. The insect inhibitory composition of claim 11 , wherein said second insect inhibitory agent is selected from the group consisting of an insect inhibitory protein and an insect inhibitory dsRNA molecule.

13. The insect inhibitory composition of claim 12 , wherein said second pesticidal agent exhibits activity against one or more pest species selected from the group of orders consisting of Lepidoptera, Coleoptera, Hemiptera, Homoptera, and Thysanoptera.

14. A seed comprising the full length polynucleotide set forth in SEQ ID NO: 39, wherein said polynucleotide is operably linked to a heterologous promoter.

15. A method of controlling a Lepidopteran pest comprising contacting said Lepidopteran pest with an insect inhibitory amount of the engineered insecticidal protein of claim 1 .

16. A transgenic plant or plant part comprising the engineered insecticidal protein of claim 1 and a polynucleotide having the nucleotide sequence as set forth in SEQ ID NO:39, wherein said engineered insecticidal protein is present in an insect inhibitory amount.

17. A method of controlling a Lepidopteran pest, said method comprising exposing said pest to the transgenic plant or plant part of claim 16 .

18. A commodity product comprising a detectable amount of the engineered insecticidal protein of claim 1 or a detectable amount of a polynucleotide having the sequence as set forth in SEQ ID NO:39.

19. The commodity product of claim 18 , wherein said commodity product is selected from the group consisting of plant biomass, oil, meal, animal feed, flour, flakes, bran, lint, hulls, and processed seed.

20. A method of producing a seed comprising the engineered insecticidal protein of claim 1 , the method comprising:
a. planting at least one seed comprising a polynucleotide having the sequence as set forth in SEQ ID NO:39;
b. growing a plant from said seed; and
c. harvesting progeny seed from said plant,
wherein said harvested seed comprises said polynucleotide.

Assignees

• Monsanto Technology Llc

Inventors

• Baum James A
• Cerruti Thomas
• Flasinski Stanislaw
• Fu Xiaoran
• Howe Arlene R
• Salvador Sara Ann

Classifications

• A01N47/08 Primary

  the carbon atom having one or more single bonds to nitrogen atoms · CPC title

• C07K14/325 Primary

  Bacillus thuringiensis crystal peptides, i.e. delta-endotoxins · CPC title

• C12N15/8286 Primary

  for insect resistance · CPC title

• A01N63/02

  Human Necessities · mapped topic

• Y02A40/162

  Cross-Sectional Technologies · mapped topic