Bacillus thuringiensis gene with lepidopteran activity

Patent metadata

Field	Value
Publication number	US-10669555-B2
Application number	US-201716072698-A
Country	US
Kind code	B2
Filing date	Jan 25, 2017
Priority date	Jan 26, 2016
Publication date	Jun 2, 2020
Grant date	Jun 2, 2020

Abstract

The disclosure provides nucleic acids, and variants and fragments thereof, obtained from strains of Bacillus thuringiensis encoding polypeptides having pesticidal activity against insect pests, including Lepidoptera . Particular embodiments of the disclosure provide isolated nucleic acids encoding pesticidal proteins, pesticidal compositions, DNA constructs, and transformed microorganisms and plants comprising a nucleic acid of the embodiments. These compositions find use in methods for controlling pests, especially plant pests.

First claim

That which is claimed:

1. An isolated nucleic acid molecule selected from:
(a) a polynucleotide encoding an insecticidal polypeptide having at least 97% amino acid sequence identity to SEQ ID NO: 2; and
(b) a polynucleotide encoding an insecticidal polypeptide having at least 97% amino acid sequence identity to SEQ ID NO: 4, wherein the polynucleotide encoding the insecticidal polypeptide is operably linked to a heterologous regulatory element.

2. The isolated nucleic acid molecule of claim 1 , wherein said nucleotide sequence is a synthetic sequence that has been designed for expression in a plant.

3. A DNA construct comprising the nucleic acid molecule of claim 1 operably linked to a heterologous regulatory element.

4. The DNA construct of claim 3 , further comprising a nucleic acid molecule encoding a Cry crystal forming domain, a heterologous signal sequence or a heterologous transit peptide operably linked to the insecticidal polypeptide.

5. A host cell comprising the DNA construct of claim 3 .

6. The host cell of claim 5 that is a bacterial cell.

7. The host cell of claim 5 that is a plant cell.

8. A transgenic plant comprising the DNA construct of claim 3 .

9. The transgenic plant of claim 8 , wherein said plant is selected from the group consisting of maize, sorghum, wheat, cabbage, sunflower, tomato, crucifer, pepper, potato, cotton, rice, soybean, sugar beet, sugarcane, tobacco, barley, and oilseed rape.

10. The transgenic plant of claim 8 , wherein the polynucleotide encoding the insecticidal polypeptide is stacked with a polynucleotide encoding a Cry protein.

11. The transgenic plant of claim 10 , wherein the Cry protein is selected from a Cry1B, a Cry9, and a Cry1Ia14 protein.

12. Transformed seed of the plant of claim 8 , wherein the seed comprise the DNA construct.

13. A recombinant insecticidal polypeptide operably linked to a heterologous signal sequence or transit polypeptide, wherein the insecticidal polypeptide comprises an amino acid sequence selected from:
(a) an amino acid sequence having at least 97% sequence identity to SEQ ID NO: 2; or
(b) an amino acid sequence having at least 97% sequence identity to SEQ ID NO: 4.

14. The insecticidal polypeptide of claim 13 , further comprising a heterologous Cry crystal forming domain operably linked to the insecticidal polypeptide.

15. A composition comprising the insecticidal polypeptide of claim 14 .

16. The composition of claim 15 , wherein said composition is a powder, dust, pellet, granule, spray, emulsion, colloid, or solution.

17. The composition of claim 16 , wherein said composition is prepared by desiccation, lyophilization, homogenization, extraction, filtration, centrifugation, sedimentation, or concentration of a culture of Bacillus thuringiensis cells.

18. The composition of claim 15 , comprising from 1% to 99% by weight of the insecticidal polypeptide.

19. The composition of claim 15 , wherein the composition further comprises an additional insecticidal polypeptide.

20. The composition of claim 15 , wherein the additional insecticidal polypeptide is selected from a Cry1B, a Cry9, and a Cry1Ia14 protein.

21. A method for controlling a Lepidopteran pest population comprising contacting said population with a pesticidally-effective amount of the polypeptide of claim 14 .

22. A method for killing a Lepidopteran pest comprising contacting said pest with, or feeding to said pest, a pesticidally-effective amount of the polypeptide of claim 14 .

23. A method for producing a polypeptide with pesticidal activity, comprising culturing the host cell of claim 4 under conditions in which a nucleic acid molecule encoding the polypeptide is expressed, wherein said polypeptide being selected from the group consisting of:
(a) a polypeptide having at least 97% sequence identity to the amino acid sequence of SEQ ID NO: 2; and
(b) a polypeptide having at least 97% sequence identity to the amino acid sequence of SEQ ID NO: 4.

24. A method for protecting a plant from an insect pest, comprising introducing into said plant or cell thereof at least one expression vector comprising a nucleotide sequence that encodes a pesticidal polypeptide, wherein said nucleotide sequence is selected from the group consisting of:
(a) a polynucleotide encoding a polypeptide having at least 97% sequence identity to the amino acid sequence of SEQ ID NO: 2; and
(b) a polynucleotide encoding a polypeptide having at least 97% sequence identity to the amino acid sequence of SEQ ID NO: 4.

25. The method of claim 24 , wherein the plant is sugarcane.

26. The method of claim 25 , wherein the insect pest is a Lepidopteran.

27. The method of claim 26 , wherein the insect pest is sugarcane borer ( Diatraea sacchralis ).

Assignees

• Pioneer Hi Bred Int

Inventors

• Abad Andre Roger
• Cerf David Charles
• Diehn Scott Henry
• Dong Hua
• Shi Xiaomei
• Wolfe Thomas Chad
• Zhou Lan

Classifications

• A01N37/46

  N-acyl derivatives · CPC title

• C12N15/8286 Primary

  for insect resistance · CPC title

• C07K2319/02

  containing a signal sequence · CPC title

• C07K2319/036

  targeting to the medium outside of the cell, e.g. type III secretion · CPC title

• Y02A40/162

  Cross-Sectional Technologies · mapped topic