Compositions and methods to control insect pests
Methods and compositions are provided which employ a silencing element that, when ingested by a pest, such as a Coleopteran plant pest or a Diabarotica 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.
1. An isolated polynucleotide comprising a heterologous promoter operably linked to a polynucleotide sequence encoding a double stranded RNA, wherein the double stranded RNA targets a Coleopteran plant pest target sequence, and wherein the Coleopteran plant pest target sequence comprises:
(a) the nucleotide sequence comprising SEQ ID NO: 73 or the full length complement thereof; or
(b) the nucleotide sequence comprising at least 90% sequence identity to SEQ ID NO: 73 or the full length complement thereof; and 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 , wherein said double stranded RNA comprises a hairpin RNA.
5. 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 polynucleotide.
6. A host cell comprising a heterologous expression cassette of claim 3 .
7. A plant cell having stably incorporated into its genome a heterologous polynucleotide 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 as set forth in SEQ ID NO: 73 or the full length complement thereof; or,
b) a nucleotide sequence comprising at least 90% sequence identity to SEQ ID NO: 73 or the full length complement thereof;
wherein the double stranded RNA, when ingested by a Coleopteran plant pest, reduces the level of a target sequence in said Coleopteran plant pest and thereby controls the Coleopteran plant pest.
8. The plant cell of claim 7 , wherein the Coleopteran plant pest is a Diabrotica plant pest.
9. The plant cell of claim 7 , wherein said plant cell comprises an expression cassette capable of expressing the heterologous polynucleotide encoding the dsRNA.
10. The plant cell of claim 7 , wherein said double stranded RNA is a hairpin RNA.
11. The plant cell of claim 7 , wherein said heterologous polynucleotide is operably linked to a heterologous promoter.
12. The plant cell of claim 7 , wherein said plant cell is from a monocot.
13. The plant cell of claim 12 , wherein said monocot is maize, barley, millet, wheat or rice.
14. The plant cell of claim 7 , wherein said plant cell is from a dicot.
15. The plant cell of claim 14 , wherein said plant cell is soybean, canola, alfalfa, sunflower, safflower, tobacco, Arabidopsis , or cotton.
16. A plant or plant part comprising a plant cell of claim 7 .
17. A transgenic seed comprising the heterologous polynucleotide comprising the double stranded RNA of claim 7 .
18. A method for controlling a Coleopteran plant pest comprising feeding to a Coleopteran plant pest a composition comprising a double stranded RNA, wherein said double stranded RNA targets a Coleopteran plant pest having a nucleotide sequence comprising at least 90% sequence identity to SEQ ID NO: 73, or the full length complement thereof, and wherein said double stranded RNA has insecticidal activity against said Coleopteran plant pest.
19. The method of claim 18 , wherein said Coleopteran plant pest is a Diabrotica plant pest.
20. The method of claim 19 , wherein said Diabrotica plant pest is D. virgifera virgifera, D. speciosa, D. barberi , or D. undecimpunctata howardi.
21. The method of claim 18 , wherein said composition comprises a plant or plant part having stably incorporated into its genome a polynucleotide encoding the double stranded RNA.
22. The method of claim 18 , wherein said double stranded RNA comprises a hairpin RNA.
23. The method of claim 21 , wherein said polynucleotide is operably linked to a heterologous promoter.
24. The method of claim 21 , 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.
25. The method of claim 21 , wherein said plant is a monocot.
26. The method of claim 25 , wherein said monocot is maize, barley, millet, wheat or rice.
27. The method of claim 21 , wherein said plant is a dicot.
28. The method of claim 27 , wherein said plant is soybean, canola, alfalfa, sunflower, safflower, tobacco, Arabidopsis , or cotton.