Methods and compositions for the modification of plants
Disclosed herein are plant-modifying compositions including a plurality of plant messenger packs (PMPs) (e.g., including a plant extracellular vesicle (EV), or segment, portion, or extract thereof), wherein the PMPs include a plant-modifying agent, and wherein the plant-modifying agent comprising the heterologous RNA. The compositions herein are useful in methods for modifying plants such as to increase plant fitness, wherein the increase in plant fitness comprising an increase in development, growth, yield, resistance to abiotic stressors, or resistance to biotic stressors.
1. A method of administering at least one heterologous RNA to an intact plant, the method comprising non-viral delivery to the intact plant of a composition comprising a plurality of lipid-reconstituted plant messenger packs (LPMPs), each of the plurality comprising the at least one heterologous RNA, an exogenous cationic lipid, and at least one plant protein extracellular vesicle marker, wherein the exogenous cationic lipid increases the RNA loading efficiency as compared to the loading efficiency of an LPMP not comprising the at least one heterologous cationic lipid, and wherein the composition is delivered at an effective concentration to increase plant fitness.
2. The method of claim 1 , wherein the at least one heterologous RNA is a guide RNA (gRNA).
3. The method of claim 2 , wherein the gRNA is a component of a ribonucleoprotein complex (RNP), and wherein each of the plurality of LPMPs comprise the RNP.
4. The method of claim 1 , wherein the exogenous cationic lipid is DC-Cholesterol or DOTAP.
5. The method of claim 1 , wherein the at least one heterologous RNA is encapsulated by the LPMPs.
6. The method of claim 1 , wherein the LPMPs in the composition are at a concentration effective to modify a plant trait, wherein the modification increases the fitness of the plant.
7. The method of claim 1 , wherein the increase in plant fitness is an increase in development, growth, yield, resistance to abiotic stressors, or resistance to biotic stressors.
8. The method of claim 1 , wherein the increase in plant fitness is a modification to flowering time.
9. The method of claim 4 , wherein the DC-cholesterol comprises between 20-40% of total lipids in the LPMP by weight.
10. The method of claim 4 , wherein the DOTAP comprises between 25-40% of total lipids in the LPMP by weight.
11. The method of claim 1 , wherein the at least one heterologous RNA is siRNA or tracrRNA.