Compositions and methods for delivery of a polynucleotide into a plant
The present disclosure provides compositions and methods for delivering a polynucleotide from the exterior surface of a plant or plant part into the interior of a plant cell. More specifically, the present disclosure relates to compositions comprising at least one polynucleotide and at least one agent that is able to disrupt at least one barrier of the plant or plant part.
1. A method for delivering a polynucleotide from the exterior surface of a plant or plant part into the interior of a plant cell, comprising
a) applying onto the surface of the plant or plant part at least one agent that is able to disrupt at least one barrier of the plant or plant part, and
b) applying onto the surface of the plant or plant part one or more polynucleotides,
wherein the at least one agent comprises a lipase enzyme,
wherein steps a) and b) are carried out concurrently or sequentially in any order, and
wherein the method further comprises applying onto the surface of the plant or plant part one or more osmolytes selected from the group consisting of sorbitol, mannitol, D-proline, and L-proline.
2. The method of claim 1 , wherein the method further comprises applying onto the surface of the plant or plant part one or more surfactants, and
wherein the polynucleotides, the enzyme, the osmolytes, and the surfactants are applied concurrently, or sequentially in any order and grouped in any combination thereof.
3. The method of claim 1 , wherein the polynucleotide is a non-transcribable polynucleotide.
4. The method of claim 3 , wherein the polynucleotide is selected from the group consisting of single-stranded DNA, single-stranded RNA, double-stranded DNA, double-stranded RNA, and an RNA/DNA hybrid.
5. The method of claim 1 , wherein the polynucleotide comprises a sequence that is identical to, or complementary to, 21 or more contiguous nucleotides of a target sequence or an RNA transcribed from the target sequence.
6. The method of claim 1 , wherein the polynucleotide encodes a site-specific enzyme or one or more RNA components of an RNA-guided nuclease.
7. The method of claim 1 , wherein the at least one agent further comprises one or more enzymes selected from the group consisting of cellulase, hemicellulase, pectinase, cutinase, and any combination thereof.
8. The method of claim 2 , wherein the one or more surfactants are nonionic surfactants.