Nucleic acid molecules to control insect pests

Patent metadata

Field	Value
Publication number	US-11046972-B2
Application number	US-201615166623-A
Country	US
Kind code	B2
Filing date	May 27, 2016
Priority date	May 29, 2015
Publication date	Jun 29, 2021
Grant date	Jun 29, 2021

Abstract

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

First claim

What is claimed is:

1. An isolated nucleic acid molecule comprising a polynucleotide operably linked to a heterologous promoter, wherein the polynucleotide encodes a hairpin ribonucleic acid (hpRNA) molecule with a stem and a loop structure that kills WCR with an LC 50 of less than 15 ng/cm 2 when ingested by the insect, the polynucleotide comprising:
a sense nucleotide sequence that comprises SEQ ID NO:3; and
the antisense nucleotide sequence of SEQ ID NO:3, wherein the sense nucleotide sequence and the antisense nucleotide sequence encode ribonucleotide sequences in the stem structure of the hpRNA molecule, the polynucleotide further comprising:
a spacer nucleotide sequence comprising SEQ ID NO:78 between the sense and antisense nucleotide sequences,
wherein the loop structure of the hpRNA molecule comprises the ribonucleotide sequence encoded by the spacer nucleotide sequence.

2. An hpRNA molecule encoded by the polynucleotide of the nucleic acid molecule of claim 1 , wherein the hpRNA molecule kills WCR when ingested by the insect.

3. The nucleic acid molecule of claim 1 , wherein the promoter is functional in a plant cell.

4. A prokaryotic cell comprising the nucleic acid molecule of claim 1 .

5. A eukaryotic cell comprising the nucleic acid molecule of claim 1 .

6. A transgenic plant cell comprising the nucleic acid molecule of claim 3 .

7. A plant comprising the hpRNA molecule of claim 2 .

8. A transgenic plant commodity product comprising a detectable amount of the polynucleotide of the nucleic acid molecule of claim 1 .

9. A transgenic plant or seed thereof comprising the nucleic acid molecule of claim 3 , wherein the heterologous promoter is functional in the plant.

10. The transgenic plant or seed thereof of claim 9 , wherein the plant is Zea mays.

11. A method for controlling a Diabrotica virgifera insect population, the method comprising feeding insects of the population with the hpRNA molecule of claim 2 .

12. The method according to claim 11 , wherein the agent is a sprayable formulation.

13. A method for controlling a Diabrotica virgifera insect population, the method comprising feeding insects of the population with plant material from a transgenic Diabrotica virgifera host plant comprising the nucleic acid molecule of claim 3 , wherein the hpRNA molecule is expressed in the plant material.

14. A method for producing a transgenic plant cell, the method comprising:
transforming a plant cell with the nucleic acid molecule of claim 3 ;
culturing the transformed plant cell under conditions sufficient to allow for development of a plant cell culture comprising a plurality of plant cells comprising the polynucleotide; and
selecting a transgenic plant cell that has integrated the polynucleotide into its genome, and that expresses the hpRNA molecule.

15. The nucleic acid molecule of claim 1 , further comprising a polynucleotide encoding an insecticidal polypeptide from Bacillus thuringiensis, Alcaligenes spp., or Pseudomonas spp.

16. The nucleic acid molecule of claim 15 , wherein the insecticidal polypeptide is from B. thuringiensis , and is selected from the group consisting of Cry1B, Cry3, Cry34, Cry35, Cry1I, Cry2A, Cry3, Cry7A, Cry8, Cry9D, Cry14, Cry18, Cry22, Cry23, Cry34, Cry35, Cry36, Cry37, Cry43, Cry55, Cyt1A, and Cyt2C.

17. The transgenic plant or seed thereof of claim 9 , wherein the plant or seed thereof comprises a polynucleotide encoding an insecticidal polypeptide from Bacillus thuringiensis, Alcaligenes spp., or Pseudomonas spp.

18. The transgenic plant or seed thereof of claim 17 , wherein the insecticidal polypeptide is from B. thuringiensis , and is selected from the group consisting of Cry1B, Cry3, Cry34, Cry35, Cry1I, Cry2A, Cry3, Cry7A, Cry8, Cry9D, Cry14, Cry18, Cry22, Cry23, Cry34, Cry35, Cry36, Cry37, Cry43, Cry55, Cyt1A, and Cyt2C.

19. The plant of claim 7 , wherein the plant comprises an insecticidal polypeptide from Bacillus thuringiensis, Alcaligenes spp., or Pseudomonas spp.

20. The plant of claim 19 , wherein the insecticidal polypeptide is from B. thuringiensis , and is selected from the group consisting of Cry1B, Cry3, Cry34, Cry35, Cry 1I, Cry2A, Cry3, Cry7A, Cry8, Cry9D, Cry14, Cry18, Cry22, Cry23, Cry34, Cry35, Cry36, Cry37, Cry43, Cry55, Cyt1A, and Cyt2C.

21. The method according to claim 11 , wherein the method further comprises feeding insects of the population with an insecticidal polypeptide from Bacillus thuringiensis, Alcaligenes spp., or Pseudomonas spp.

22. The method according to claim 21 , wherein the insecticidal polypeptide is from B. thuringiensis , and is selected from the group consisting of Cry1B, Cry3, Cry34, Cry35, Cry 1I, Cry2A, Cry3, Cry7A, Cry8, Cry9D, Cry14, Cry18, Cry22, Cry23, Cry34, Cry35, Cry36, Cry37, Cry43, Cry55, Cyt1A, and Cyt2C.

23. A method for producing a transgenic plant, the method comprising:
regenerating a transgenic plant from the transgenic plant cell of claim 6 .

24. The plant of claim 7 , wherein the plant is a transgenic plant expressing the hpRNA molecule.

25. The transgenic plant of claim 24 , wherein the plant is Zea mays.

26. A method for improving the yield of a corn crop, the method comprising:
cultivating in the corn crop the transgenic plant or seed thereof of claim 9 .

27. A plant material comprising the hpRNA molecule of claim 2 .

28. The plant material of claim 27 , wherein the plant material is a transgenic plant material expressing the hpRNA molecule.

29. The transgenic plant material of claim 28 , wherein the plant material is a Zea mays plant material.

30. A transgenic plant material comprising the nucleic acid molecule of claim 3 .

31. The transgenic plant material of claim 30 , wherein the plant material is a seed.

Assignees

• Dow Agrosciences Llc
• Fraunhofer Ges Forschung

Inventors

• Narva Kenneth E
• Worden Sarah E
• Frey Meghan
• Rangasamy Murugesan
• Gandra Premchand
• Lo Wendy
• Fishilevich Elane
• Vilcinskas Andreas
• Knorr Eileen

Classifications

• Y02A40/146

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

• C12Q2600/178

  miRNA, siRNA or ncRNA · CPC title

• C07K14/325

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

• A01N57/16

  containing heterocyclic radicals · CPC title

• C12Q2600/158

  Expression markers · CPC title