Compositions and methods to control insect pests

Patent metadata

Field	Value
Publication number	US-9686995-B2
Application number	US-201313791596-A
Country	US
Kind code	B2
Filing date	Mar 8, 2013
Priority date	Aug 28, 2009
Publication date	Jun 27, 2017
Grant date	Jun 27, 2017

Abstract

Methods and compositions are provided which employ a silencing element that, when ingested by a pest, such as a Coleopteran plant pest or a Diabrotica plant pest, decrease the expression of a target sequence in the pest. In specific embodiments, the decrease in expression of the target sequence controls the pest and thereby the methods and compositions are capable of limiting damage to a plant. The present invention provides various target polynucleotides set forth in any one of SEQ ID NOS: 1-236 or active variants and fragments thereof, wherein a decrease in expression of one or more the sequences in the target pest controls the pest (i.e., has insecticidal activity). Further provided are silencing elements which when ingested by the pest decrease the level of the target polypeptide and thereby control the pest. In specific embodiment, the pest is D. virgifera virgifera, D. barberi, D. speciosa , or D. undecimpunctata howardi . Plants, plant part, bacteria and other host cells comprising the silencing elements or an active variant or fragment thereof of the invention are also provided.

First claim

That which is claimed:

1. An isolated polynucleotide comprising a heterologous promoter operably linked to a nucleotide sequence encoding a double stranded RNA, wherein the double stranded RNA targets a Coleopteran plant pest target nucleotide sequence, wherein the Coleopteran plant pest target nucleotide sequence comprises:
a) the nucleotide sequence set forth in SEQ ID NO: 8, or the full length complement thereof; or
b) the nucleotide sequence comprising at least 95% sequence identity to SEQ ID NO: 8, or the full length complement thereof;
wherein said double stranded RNA has insecticidal activity against a Coleopteran plant pest.

2. The isolated polynucleotide of claim 1 , wherein said Coleopteran plant pest is a Diabrotica plant pest.

3. An expression cassette comprising the polynucleotide of claim 1 .

4. The expression cassette of claim 3 comprising the construct of SEQ ID NO: 237.

5. The expression cassette of claim 3 , wherein said double stranded RNA comprises a hairpin RNA.

6. The expression cassette of claim 3 , wherein said polynucleotide is flanked by a first operably linked convergent promoter at one terminus of the polynucleotide and a second operably linked convergent promoter at the opposing terminus of the polynucleotide, wherein the first and the second convergent promoters are capable of driving expression of the double stranded RNA.

7. A host cell comprising a heterologous expression cassette of claim 3 .

8. A plant cell having stably incorporated into its genome a heterologous polynucleotide comprising a nucleotide sequence encoding a double stranded RNA, wherein the double stranded RNA targets a Coleopteran plant pest target nucleotide sequence, wherein the Coleopteran plant pest target nucleotide sequence comprises:
a) the nucleotide sequence set forth in SEQ ID NO: 8, or the full length complement thereof; or,
b) the nucleotide sequence comprising at least 95% sequence identity to SEQ ID NO: 8, or the full length complement thereof;
wherein said double stranded RNA has insecticidal activity against a Coleoptera plant pest.

9. The plant cell of claim 8 , wherein the Coleopteran plant pest is a Diabrotica plant pest.

10. The plant cell of claim 8 , wherein said double stranded RNA comprises a hairpin RNA.

11. The plant cell of claim 8 , wherein said polynucleotide is operably linked to a heterologous promoter.

12. The plant cell of claim 8 , wherein said plant cell is from a monocot.

13. The plant cell of claim 8 , wherein said plant cell is from a dicot.

14. A plant or plant part comprising a plant cell of claim 8 .

15. A transgenic seed comprising the heterologous polynucleotide encoding the double stranded RNA of claim 8 .

16. A double stranded RNA comprising a nucleotide sequence complementary to:
a) the nucleotide sequence set forth in SEQ ID NO: 8 or
b) a nucleotide sequence comprising at least 95% sequence identity to the nucleotide sequence set forth in SEQ ID NO: 8.

17. The double stranded RNA of claim 16 , wherein said Coleoptera plant pest is a Diabrotica plant pest.

18. The double stranded RNA of claim 16 , wherein the double stranded RNA comprises a hairpin RNA.

19. The double stranded RNA of claim 16 , further comprising an agriculturally acceptable carrier.

20. The double stranded RNA of claim 16 , wherein the double stranded RNA is expressed in a plant, plant part, or plant cell.

21. The double stranded RNA of claim 16 , wherein the double stranded RNA is expressed in a microorganism.

Assignees

• Du Pont
• Pioneer Hi-Bred Int
• Du Pont

Inventors

• Broglie Karen E
• Muthalagi Mani
• Kriss Kevin
• Lu Albert L
• Presnail James K

Classifications

• A01N65/20

  Fabaceae or Leguminosae [Pea or Legume family], e.g. pea, lentil, soybean, clover, acacia, honey locust, derris or millettia · CPC title

• C12N15/113

  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

• C12N2310/11

  Antisense · CPC title

• C12N15/8218

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

• C12N2310/531

  Stem-loop; Hairpin · CPC title