Compositions and methods for the suppression of target polynucleotides from lepidoptera
Methods and compositions are provided which employ a silencing element that, when ingested by a pest, such as a Lepidopteran pest, decreases the expression of a target sequence in the pest. In specific embodiments, the decrease in expression of the target sequence controls the pest, thereby limiting damage to a plant. The present invention provides target polynucleotides encoding polypeptides from specific protein families and various target polynucleotides as set forth, or active variants and fragments thereof, wherein a decrease in expression of one or more sequences in the target pest 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 a specific embodiment, the pest is Spodoptera frugiperda . Plants, plant parts, bacteria and other host cells comprising the silencing elements or an active variant or fragment thereof of the invention are also provided.
1. A plant cell having stably incorporated into its genome a heterologous polynucleotide comprising a silencing element, wherein said silencing element comprises a fragment of at least 20 consecutive nucleotides fully complementary to a target sequence of SEQ ID NO: 16, wherein said silencing element, when ingested by a pest from the Lepidoptera order, reduces the level of the target sequence in said pest and thereby controls the pest from the Lepidoptera order.
2. The plant cell of claim 1 , wherein said pest comprises Spodoptera frugiperda.
3. The plant cell of claim 1 , wherein said silencing element comprises
a) a polynucleotide comprising the sense or antisense sequence of the sequence set forth in SEQ ID NO: 183; and
b) a polynucleotide comprising the sequence set forth in SEQ ID NO: 184 and 185.
4. The plant cell of 3 , wherein said silencing element encodes a hairpin RNA.
5. A plant cell having stably incorporated into its genome a heterologous polynucleotide comprising a silencing element, wherein said silencing element comprises, in the following order, a first segment, a second segment, and a third segment, wherein
a) said first segment comprises at least 20 nucleotides fully complementary to a target sequence set forth in SEQ ID NO: 16;
b) said second segment comprises a loop of sufficient length to allow the silencing element to be transcribed as a hairpin RNA; and,
c) said third segment comprises at least 20 nucleotides fully complementary to the first segment, wherein said silencing element, when ingested by a pest from the Lepidoptera order, reduces the level of the target sequence in said pest and thereby controls the pest from the Lepidoptera order.
6. The plant cell of any one of claims 1 - 5 , wherein said silencing element is operably linked to a heterologous promoter.
7. The plant cell of 3 , wherein said plant cell has stably incorporated into its genome a second polynucleotide comprising a suppressor enhancer element comprising the target pest sequence or an active variant or fragment thereof, wherein combined expression of the silencing element and the suppressor enhancer element increases the concentration of an inhibitory RNAi specific for the pest target sequence in said plant cell.
8. The plant cell of claim 3 , wherein said plant cell is from a monocot.
9. The plant cell of claim 8 , wherein said monocot is maize, barley, millet, wheat or rice.
10. The plant cell of 3 , wherein said plant cell is from a dicot.
11. The plant cell of claim 10 , wherein said dicot is soybean, canola, alfalfa, sunflower, safflower, tobacco, Arabidopsis, or cotton.
12. A plant or plant part comprising a plant cell claim 3 .
13. The plant cell of claim 7 , wherein the combined expression of said silencing element and the suppressor enhancer element increases the concentration of an inhibitory RNA specific for the pest target sequence in the phloem of a plant or plant part.
14. A transgenic seed wherein the transgenic seed comprises the silencing element from the plant cell of claim 1 .
15. A method for controlling a Lepidoptera pest comprising feeding to a Lepidoptera pest a composition comprising a silencing element, wherein said silencing element, when ingested by said Lepidoptera pest, reduces the level of a target Lepidoptera pest sequence and thereby controls the Lepidoptera pest and said silencing element comprises a fragment of at least 20 consecutive nucleotides fully complementary to a target sequence of SEQ ID NO: 16.
16. The method of claim 15 , wherein said composition comprises a plant or plant part having stably incorporated into its genome a polynucleotide comprising said silencing element.
17. The method of claim 15 , wherein said Lepidoptera pest comprises Spodoptera frugiperda.
18. The method of claim 15 , wherein said silencing element encodes a hairpin RNA.
19. The method of claim 15 , wherein said silencing element is operably linked to a heterologous promoter.
20. The method of claim 16 , wherein said plant or plant part has stably incorporated into its genome a second polynucleotide comprising a suppressor enhancer element comprising the target pest sequence or an active variant or fragment thereof, wherein combined expression of the silencing element and the suppressor enhancer element increases the concentration of an inhibitory RNAi specific for the pest target sequence in said plant cell.
21. The method of claim 15 , wherein said plant is a monocot.
22. The method of claim 21 , wherein said monocot is maize, barley, millet, wheat or rice.
23. The method of claim 15 , wherein said plant is a dicot.
24. The method of claim 23 , wherein said plant is soybean, canola, alfalfa, sunflower, safflower, tobacco, Arabidopsis, or cotton.