Polynucleotide Molecules for Gene Regulation in Plants

• US 20140057789A1
• Filed: 08/30/2013
• Published: 02/27/2014
• Est. Priority Date: 03/08/2010
• Status: Active Grant

• Current Assignee: Monsanto Technology LLC (Bayer AG)
• Original Assignee: Monsanto Technology LLC (Bayer AG)
• Inventors: Sammons, Robert D, Ivashuta, Sergey, Liu, Hong, Wang, Dafu, Feng, Paul C. C., Kouranov, Andrei Y., Andersen, Scott E.
• Granted Patent: US 9,988,634 B2
• Time in Patent Office: Days
• US Class Current: 504/206
• CPC Class Codes: A01N 2300/00 Combinations or mixtures of active ingredients covered by classes A01N27/00 - A01N65/48 with other active or formulation relevant ingredients, e.g. specific carrier materials or surfactants, covered by classes A01N25/00 - A01N65/48
• CPC Class Codes: A01N 33/12 Quaternary ammonium compounds
• CPC Class Codes: A01N 57/16 containing heterocyclic radicals
• CPC Class Codes: A01N 57/20 containing acyclic or cycloaliphatic radicals
• CPC Class Codes: A01N 57/34 having phosphorus-to-halogen bonds; Phosphonium salts
• CPC Class Codes: A01N 63/60 Isolated nucleic acids
• CPC Class Codes: C12N 15/113 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 Class Codes: C12N 15/1137 against enzymes viral enzymes C12N15/1131; receptors C12N15/1138
• CPC Class Codes: C12N 15/8206 by physical or chemical, i.e. non-biological, means, e.g. electroporation, PEG mediated
• CPC Class Codes: C12N 15/8207 by mechanical means, e.g. microinjection, particle bombardment, silicon whiskers
• CPC Class Codes: C12N 15/8218 Antisense, co-suppression, viral induced gene silencing [VIGS], post-transcriptional induced gene silencing [PTGS]

Abstract

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.

Claims

1. A method of delivering a non-transcribable polynucleotide to the interior of a cell of a plant comprising:

applying the non-transcribable polynucleotide and an organosilicone surfactant to an exterior surface of the plant cell, wherein the non-transcribable polynucleotide is conveyed from the exterior surface to the interior of the plant cell.

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 non-transcribable polynucleotide is selected from the group consisting of ssDNA, dsDNA, ssRNA, dsRNA and RNA/DNA hybrids.

8. The method of claim 1, wherein the non-transcribable polynucleotide and the organosilicone surfactant are applied separately.

9. The method of claim 1, wherein the non-transcribable polynucleotide and the organosilicone surfactant are applied concurrently.

10. The method of claim 1, wherein the organosilicone surfactant is a silicone polyether copolymer.

11. The method of claim 10, wherein the silicone polyether copolymer is a copolymer of polyalkylene oxide modified heptamethyl trisiloxane and allyloxypolypropylene glycol methylether.

12. The method of claim 1, wherein the organosilicone surfactant is BREAK-THRU®

S 321, BREAK-THRU®

S 200, BREAK-THRU®

OE 441, BREAK-THRU®

S 278, BREAK-THRU®

S 243, SILWET L-77®

, SILWET®

HS 429, SILWET®

HS 312, or BREAK-THRU®

S 233.

13. The method of claim 1, further comprising applying an organic or an inorganic salt.

14. The method of claim 13, wherein the salt is an ammonium salt.

15. The method of claim 14, wherein the ammonium salt is ammonium sulfate.

16. The method of claim 1, wherein the non-transcribable polynucleotide and the organosilicone surfactant are applied onto the exterior surface of the plant cell by a spray apparatus.

17. The method of claim 1, wherein the non-transcribable polynucleotide comprises a sequence that is essentially identical or essentially complementary to a coding or non-coding sequence of an endogenous gene of the plant cell.

18. The method of claim 17, 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.

19. The method of claim 17, wherein the endogenous gene encodes a protein that provides herbicide and said protein is 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.

20. The method of claim 17, wherein the endogenous gene is a native gene or a recombinant transgene.

21. The method of claim 1, further comprising applying a non-polynucleotide herbicide.

22. The method of claim 21, wherein the non-polynucleotide herbicide is selected from the group consisting of glyphosate, dicamba, phosphinothricin, bromoxynil, ioxynil and chlorsulfuron.