Polynucleotide molecules for gene regulation in plants
This invention provides polynucleotide molecules and methods for regulating genes in plants, e. g., by providing RNA for systemic regulation of genes. Various aspects of the invention provide polynucleotide molecules and methods for regulating endogenous genes and transgenes in a plant cell and polynucleotide molecules.
1. A method of delivering a double-stranded RNA (dsRNA) polynucleotide to the interior of a cell of a plant comprising: applying the dsRNA polynucleotide and a transferring agent selected from an organosilicone surfactant and a cationic lipid to an exterior surface of the plant,
wherein the transferring agent facilitates permeation of the dsRNA polynucleotide from the exterior surface of the plant to the interior of the plant cell without the aid of a physical abrasive, and
wherein the dsRNA polynucleotide is 21 to 700 base pairs in length and comprises a nucleotide sequence that is identical or complementary to at least 21 contiguous nucleotides of an endogenous gene or a transcribed RNA of the plant.
2. The method of claim 1 , wherein the plant is growing in a field.
3. The method of claim 1 , wherein the plant is a pigweed, velvetleaf, waterhemp, prickly lettuce, dandelion, alfalfa, corn, soybean, canola, cotton, sugar beet, sugarcane, wheat, or rice plant.
4. The method of claim 1 , wherein the plant is a weed or volunteer plant.
5. The method of claim 1 , wherein the plant is resistant to one or more of glyphosate, dicamba and sulfonylurea.
6. The method of claim 1 , wherein the cell is a leaf cell.
7. The method of claim 1 , wherein the dsRNA polynucleotide and the transferring agent are applied separately.
8. The method of claim 1 , wherein the dsRNA polynucleotide and the transferring agent are applied concurrently.
9. The method of claim 1 , wherein the organosilicone surfactant is a silicone polyether copolymer.
10. The method of claim 9 , wherein the silicone polyether copolymer is a copolymer of polyalkylene oxide modified heptamethyl trisiloxane and allyloxypolypropylene glycol methylether.
11. The method of claim 1 , further comprising applying an organic or an inorganic salt.
12. The method of claim 11 , wherein the salt is an ammonium salt.
13. The method of claim 12 , wherein the ammonium salt is ammonium sulfate.
14. The method of claim 1 , wherein the dsRNA polynucleotide and the transferring agent are applied onto the exterior surface of the plant by a spray apparatus.
15. The method of claim 1 , wherein the endogenous gene: (i) is an essential gene, (ii) encodes a protein that provides herbicide resistance, or (iii) transcribes to an RNA regulatory agent.
16. The method of claim 1 , wherein the endogenous gene encodes a protein selected from the group consisting of a 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS), an acetohydroxyacid synthase, an acetolactate synthase (ALS), an acetyl-coenzyme A carboxylase (ACCase), a dihydropteroate synthase, a phytoene desaturase (PDS), a protoporphyrin IX oxygenase (PPO), a hydroxyphenylpyruvate dioxygenase (HPPD), a para-aminobenzoate synthase, a glutamine synthase (GS), a glufosinate-tolerant glutamine synthase, a 1-deoxy-D-xylulose 5-phosphate (DOXP) synthase, a dihydropteroate (DHP) synthase, a phenylalanine ammonia lyase (PAL), a glutathione S-transferase (GST), a D1 protein of photosystem II, a mono-oxygenase, a cytochrome P450, a cellulose synthase, a beta-tubulin, and a serine hydroxymethyltransferase.
17. The method of claim 1 , wherein the endogenous gene is a native gene or a recombinant transgene.
18. The method of claim 1 , further comprising applying a non-polynucleotide herbicide.
19. The method of claim 18 , wherein the non-polynucleotide herbicide is selected from the group consisting of glyphosate, dicamba, phosphinothricin, bromoxynil, ioxynil and chlorsulfuron.