WUPA nucleic acid molecules that confer resistance to coleopteran and hemipteran pests

Patent metadata

Field	Value
Publication number	US-10344298-B2
Application number	US-201615284922-A
Country	US
Kind code	B2
Filing date	Oct 4, 2016
Priority date	Oct 12, 2015
Publication date	Jul 9, 2019
Grant date	Jul 9, 2019

Abstract

This disclosure concerns nucleic acid molecules and methods of use thereof for control of coleopteran and hemipteran pests through RNA interference-mediated inhibition of target coding and transcribed non-coding sequences in coleopteran and hemipteran pests. The disclosure also concerns methods for making transgenic plants that express nucleic acid molecules useful for the control of coleopteran and hemipteran pests, and the plant cells and plants obtained thereby.

First claim

The invention claimed is:

1. An isolated nucleic acid comprising a polynucleotide operably linked to a heterologous promoter, wherein the polynucleotide comprises at least a first nucleotide sequence that is selected from the group consisting of a native coding sequence of a Diabrotica organism consisting of SEQ ID NO:7 or SEQ ID NO:10; or the complement of a native coding sequence of a Diabrotica organism consisting of SEQ ID NO:7 or SEQ ID NO:10;
a second nucleotide sequence; and,
a third nucleotide sequence that is the reverse complement of the first nucleotide sequence,
wherein the third nucleotide sequence is linked to the first nucleotide sequence by the second nucleotide sequence.

2. The nucleic acid molecule of claim 1 , wherein the Diabrotica organism is selected from the group consisting of D. v. virgifera LeConte; D. barberi Smith and Lawrence; D. u. howardi; D. v. zeae; D. balteata LeConte; D. u. tenella; D. speciosa Germar, and D. u. undecimpunctata Mannerheim.

3. The nucleic acid molecule of claim 1 , wherein the molecule is a plant transformation vector.

4. A polynucleotide encoded by the polynucleotide of the nucleic acid molecule of claim 1 .

5. A double-stranded ribonucleic acid (dsRNA) molecule comprising the polynucleotide of claim 4 .

6. The double-stranded ribonucleic acid molecule of claim 5 , wherein contacting the polynucleotide sequence with a coleopteran insect inhibits the expression of an endogenous nucleotide sequence specifically complementary to the polynucleotide.

7. The double-stranded ribonucleic acid molecule of claim 6 , wherein contacting said double-stranded ribonucleic acid molecule with a coleopteran insect kills or inhibits the growth, viability, and/or feeding of the insect.

8. The nucleic acid molecule of claim 1 , wherein the polynucleotide encodes a double stranded RNA (dsRNA molecule) the polynucleotide further comprising a second nucleotide sequence and a third nucleotide sequence, nucleotide sequence is linked to the first nucleotide sequence by the second polynucleotide sequence, and wherein the third nucleotide sequence is substantially the reverse complement of the first nucleotide sequence, such that the first and the third nucleotide sequences hybridize under moderate stringency conditions when transcribed to form a stem structure in the dsRNA molecule.

9. The dsRNA molecule encoded by the polynucleotide of the nucleic acid molecule of claim 8 , comprising a first, a second, and a third ribonucleotide sequence, wherein the first ribonucleotide sequence is encoded by the first nucleotide sequence, wherein the second ribonucleotide sequence is encoded by the second nucleotide sequence, wherein the third ribonucleotide sequence is encoded by the third nucleotide sequence, wherein the third ribonucleotide sequence is linked to the first ribonucleotide sequence by the second ribonucleotide sequence, and wherein the first and the third ribonucleotide sequences hybridize under moderate stringency conditions into the dsRNA molecule.

10. The nucleic acid molecule of claim 8 , wherein the heterologous promoter is functional in a plant cell.

11. A cell comprising the nucleic acid molecule of claim 1 .

12. The cell of claim 11 , wherein the cell is a prokaryotic cell.

13. The cell of claim 11 , wherein the cell is a eukaryotic cell.

14. A transgenic plant cell comprising the nucleic acid molecule of claim 8 , wherein the heterologous promoter is functional in the plant cell.

15. A transgenic plant material comprising the nucleic acid molecule of claim 10 .

16. The transgenic plant material of claim 15 , wherein the plant material is a seed or plant.

17. A commodity product produced from the plant of claim 15 , wherein the commodity product comprises a detectable amount of the polynucleotide.

18. The transgenic plant cell of claim 14 , wherein the cell is a Zea mays cell.

19. The transgenic plant material of claim 15 , wherein the plant material is a Zea mays plant material.

20. The transgenic plant or seed of claim 16 , wherein the plant or seed is a Zea mays plant or seed.

21. A method for controlling a coleopteran insect population, the method comprising feeding insects of the population with an agent comprising the dsRNA molecule of claim 5 .

22. A method for controlling a coleopteran insect population, the method comprising feeding insects of the population with an agent comprising the dsRNA molecule of claim 9 .

23. The method according to claim 22 , wherein the agent is a transgenic plant material expressing the dsRNA molecule.

24. The method according to claim 22 wherein the agent is a sprayable formulation.

25. The method according to claim 22 , wherein the agent is a transgenic plant material expressing the dsRNA molecule.

26. The method according to claim 21 , wherein the agent is a sprayable formulation.

27. A method for improving the yield of a crop, the method comprising:
cultivating in the crop the transgenic plant or seed of claim 16 , such that the dsRNA molecule is expressed.

28. The method according to claim 27 , wherein the plant is Zea mays.

29. A method for producing a transgenic plant cell, the method comprising:
transforming a plant cell with the nucleic acid molecule of claim 10 ;
culturing the transformed plant cell under conditions sufficient to allow for development of a plant cell culture; and
selecting a transgenic plant cell that has integrated the polynucleotide in its genome and expresses the dsRNA molecule.

30. A method for producing a coleopteran insect resistant transgenic plant, the method comprising:
regenerating a transgenic plant from the transgenic plant cell of claim 14 .

31. The nucleic acid molecule of claim 1 , further comprising a polynucleotide encoding a polypeptide from Bacillus thuringiensis or a PIP-1 polypeptide.

32. The nucleic acid molecule of claim 31 , wherein the polynucleotide encodes a polypeptide from B. thuringiensis that is selected from a group comprising Cry3, Cry34, and Cry35.

33. The transgenic plant cell of claim 14 , wherein the cell comprises a polynucleotide encoding a polypeptide from Bacillus thuringiensis or a PIP-1 polypeptide.

34. The transgenic plant cell of claim 33 , wherein the polynucleotide encodes a polypeptide from B. thuringiensis that is selected from a group comprising Cry3, Cry34, and Cry35.

35. The transgenic plant of claim 15 , wherein the plant comprises a polynucleotide encoding a polypeptide from Bacillus thuringiensis or a PIP-1 polypeptide.

36. The transgenic plant of claim 35 , wherein the polynucleotide encodes a polypeptide from B. thuringiensis that is selected from a group comprising Cry3, Cry34, and Cry35.

37. The method according to claim 29 , wherein the transgenic plant cell comprises a polynucleotide encoding a polypeptide from Bacillus thuringiensis or a PIP-1 polypeptide.

38. The method according to claim 37 , wherein the polynucleotide encodes a polypeptide from B. thuringiensis that is selected from a group comprising Cry3, Cry34, and Cry35.

Assignees

• Dow Agrosciences Llc

Inventors

• Narva Kenneth E
• Fishilevich Elane
• Rangasamy Murugesan
• Frey Meghan L
• Lo Wendy
• Worden Sarah E
• Gandra Premchand

Classifications

• C12N15/8218

  Antisense, co-suppression, viral induced gene silencing [VIGS], post-transcriptional induced gene silencing [PTGS] · CPC title

• A01N57/16

  containing heterocyclic radicals · CPC title

• C12N15/8286 Primary

  for insect resistance · CPC title

• A01N37/46

  N-acyl derivatives · CPC title

• Y02A40/146

  Genetically Modified [GMO] plants, e.g. transgenic plants · CPC title