Parental RNAI suppression of hunchback gene to control hemipteran pests

Patent metadata

Field	Value
Publication number	US-10047360-B2
Application number	US-201514971417-A
Country	US
Kind code	B2
Filing date	Dec 16, 2015
Priority date	Dec 16, 2014
Publication date	Aug 14, 2018
Grant date	Aug 14, 2018

Abstract

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

First claim

What may be claimed is:

1. An isolated nucleic acid molecule comprising at least one polynucleotide operably linked to a heterologous promoter, wherein the polynucleotide encodes a double-stranded ribonucleic acid (dsRNA) molecule, the polynucleotide comprising:
a first nucleotide sequence selected from the group consisting of: SEQ ID NO:1, the complement of SEQ ID NO:1, at least 23 contiguous nucleotides of SEQ ID NO:1, the complement of at least 23 contiguous nucleotides of SEQ ID NO:1, a native coding sequence of Euschistus heros comprising SEQ ID NO:1, the complement of a native coding sequence of Euschistus heros comprising SEQ ID NO:1, at least 23 contiguous nucleotides of a native coding sequence of Euschistus heros comprising SEQ ID NO:1, and the complement of at least 23 contiguous nucleotides of a native coding sequence of Euschistus heros comprising SEQ ID NO:1;
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 operably linked to the first nucleotide sequence by the second nucleotide sequence.

2. The nucleic acid molecule of claim 1 , wherein the first nucleotide sequence is SEQ ID NO:2 or the complement of SEQ ID NO:2.

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

4. The nucleic acid molecule of claim 1 , wherein the first nucleotide sequence comprises at least 23 contiguous nucleotides of SEQ ID NO:1 or the complement of at least 23 contiguous nucleotides of SEQ ID NO:1.

5. A double-stranded ribonucleic acid (dsRNA) molecule comprising a first, a second, and a third ribonucleotide sequence,
wherein the first ribonucleotide sequence is encoded by a deoxynucleotide sequence selected from the group consisting of SEQ ID NO:1, the complement of SEQ ID NO:1, at least 23 contiguous nucleotides of SEQ ID NO:1, the complement of at least 23 contiguous nucleotides of SEQ ID NO:1, a native coding sequence of Euschistus heros comprising SEQ ID NO:1, the complement of a native coding sequence of Euschistus heros comprising SEQ ID NO:1, at least 23 contiguous nucleotides of a native coding sequence of Euschistus heros comprising SEQ ID NO:1, and the complement of at least 23 contiguous nucleotides of a native coding sequence of Euschistus heros comprising SEQ ID NO:1,
wherein the third ribonucleotide sequence is linked to the first ribonucleotide sequence by the second ribonucleotide sequence, and
wherein the third ribonucleotide sequence is the reverse complement of the first ribonucleotide sequence, such that the first and the third ribonucleotide sequences hybridize to form the stem of a hairpin structure in the molecule.

6. The dsRNA molecule of claim 5 , wherein the first ribonucleotide sequence is encoded by at least 23 contiguous nucleotides of SEQ ID NO:1, or the complement of at least 23 contiguous nucleotides of SEQ ID NO:1.

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

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

9. A plant cell comprising the nucleic acid molecule of claim 1 , wherein the heterologous promoter is functional in the plant cell.

10. A transgenic plant comprising the plant cell of claim 9 .

11. A seed of the plant of claim 10 , wherein the seed comprises the polynucleotide.

12. A commodity product produced from the transgenic plant of claim 10 , wherein the commodity product comprises a detectable amount of the polynucleotide.

13. The plant cell of claim 9 , wherein the cell is a Zea mays cell, a Glycine max cell, or a cell from a Gossypium sp.

14. The transgenic plant of claim 10 , wherein the plant is maize, soybean, or cotton.

15. A method for controlling a Euschistus heros insect pest population, the method comprising feeding an insect of the population with an agent comprising the dsRNA molecule of claim 5 .

16. The method according to claim 15 , wherein the insect pest is a male Euschistus heros insect pest.

17. The method according to claim 16 , wherein the insect pest is a female Euschistus heros insect pest, the method further comprising releasing the female Euschistus heros insect pest into the pest population.

18. The method according to claim 17 , wherein the agent is a sprayable formulation.

19. The method according to claim 18 , wherein the agent is a transgenic host plant of Euschistus heros or a part thereof, comprising a polynucleotide that is expressed in the plant to produce the dsRNA molecule.

20. A method for improving the yield of a crop, the method comprising:
cultivating in the crop the transgenic plant of claim 10 .

21. The method according to claim 20 , wherein the crop is a corn crop, and the transgenic plant is a corn plant.

22. 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 dsRNA molecule.

23. A method for producing a transgenic plant, the method comprising:
regenerating a transgenic plant from the plant cell of claim 9 .

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

25. The nucleic acid molecule of claim 24 , wherein the insecticidal polypeptide from B. thuringiensis is selected from a group comprising Cry1A, Cry2A, Cry3A, Cry11A, and Cry51A.

26. The plant cell of claim 9 , wherein the plant cell comprises a polynucleotide encoding an insecticidal polypeptide from Bacillus thuringiensis, Alcaligenes spp., or Pseudomonas spp.

27. The plant cell of claim 26 , wherein the insecticidal polypeptide from B. thuringiensis is selected from a group comprising Cry1A, Cry2A, Cry3A, Cry 11A, and Cry51A.

28. The transgenic plant of claim 10 , wherein the plant comprises a polynucleotide encoding an insecticidal polypeptide from Bacillus thuringiensis, Alcaligenes spp., or Pseudomonas spp.

29. The transgenic plant of claim 28 , wherein the insecticidal polypeptide from B. thuringiensis is selected from a group comprising Cry1A, Cry2A, Cry3A, Cry11A, and Cry51A.

30. The method according to claim 15 , wherein the method further comprises feeding the insect of the population with an insecticidal polypeptide from Bacillus thuringiensis, Alcaligenes spp., or Pseudomonas spp.

31. The method according to claim 30 , wherein the polypeptide from B. thuringiensis is selected from a group comprising Cry1A, Cry2A, Cry3A, Cry11A, and Cry51A.

Assignees

• Dow Agrosciences Llc
• Univ Nebraska

Inventors

• Siegfried Blair D
• Narva Kenneth E
• Arora Kanika
• Worden Sarah E
• Khajuria Chitvan
• Fishilevich Elane
• Storer Nicholas P
• Frey Meghan
• Hamm Ronda L
• Velez Ana

Classifications

• C12N2310/14

  interfering nucleic acids [NA] · CPC title

• C12N15/113 Primary

  Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; {Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing (when used in plants C12N15/8218)} · CPC title

• C12N15/8218 Primary

  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

  for insect resistance · CPC title