Compositions and methods for silencing gene expression
A method of introducing naked dsRNA into a seed is provided. The method comprising contacting the seed with the naked dsRNA under conditions which allow penetration of the dsRNA into the seed, thereby introducing the dsRNA into the seed.
1. A method of introducing into a monocotyledon seed a naked double-stranded RNA (dsRNA) that is capable of down-regulating a target gene, the method comprising shaking the monocotyledon seed in a solution comprising the naked dsRNA from 1 hour up to 60 hours, thereby introducing the dsRNA into the monocotyledon seed, wherein said naked dsRNA is not associated with a cationic oligopeptide.
2. A method of down-regulating expression of a target gene, the method comprising:
(a) shaking a monocotyledon seed in a solution comprising a naked dsRNA from 1 hour up to 60 hours, wherein the naked dsRNA down-regulates expression of the target gene, wherein said naked dsRNA is not associated with a cationic oligopeptide; and optionally
(b) generating a monocotyledon plant of the monocotyledon seed.
3. The method of claim 2 , wherein said down-regulating expression of the target gene improves tolerance of the monocotyledon plant to abiotic or biotic stress.
4. The method of claim 2 , further comprising drying said monocotyledon seed following said shaking.
5. The method of claim 2 , wherein said target gene is an endogenous gene.
6. The method of claim 2 , wherein said target gene is a gene of a viral pathogen.
7. The method of claim 1 , wherein said introducing comprises penetration to an endosperm, and alternatively or additionally, an embryo of said monocotyledon seed.
8. The method of claim 1 , wherein said naked dsRNA does not integrate into the genome of said monocotyledon seed.
9. The method of claim 2 , wherein said introducing results in presence of said naked dsRNA in the monocotyledon plant for at least 10 days following germination.
10. The method of claim 1 , further comprising priming said monocotyledon seed prior to said shaking, wherein said priming is effected by:
washing the seed prior to said contacting; and
(ii) drying the seed following step (i).
11. The method of claim 10 , wherein said washing is effected for 2-6 hours.
12. The method of claim 10 , wherein said drying is effected at 25-30° C. for 10-16 hours.
13. The method of claim 1 , wherein said naked dsRNA is present in the solution at a final concentration of 0.001-100 μg/μL.
14. The method of claim 13 , wherein said naked dsRNA is present in the solution at a final concentration of 0.001-0.5 μg/μL.
15. The method of claim 1 , further comprising treating the seed with an agent selected from the group consisting of a pesticide, a fungicide, an insecticide, a fertilizer, a coating agent and a coloring agent following said contacting.