Compositions and methods for liquid-mediated delivery of pollen
The invention provides novel compositions and methods for liquid-mediated delivery of pollen to a female reproductive part of a recipient plant. The pollen suspension solutions provided include a surfactant, an oil, a solute, or an aqueous solution, and about 2% to about 20% pollen by weight. The methods provided include producing a liquid pollen suspension solution comprising pollen from a donor plant and spraying the solution onto at least a first female reproductive part of a recipient plant, thereby pollinating the female reproductive part with the pollen from the donor plant.
1. A pollen suspension solution comprising:
a) an isoparaffin oil, about 2% to about 20% monocot pollen by weight, and a surfactant present in an amount equal to or less than about 3.0% by weight; or
b) an aqueous solution, about 2% to about 20% monocot pollen by weight, and a surfactant present in an amount of less than about 1.0% by weight, wherein the surfactant is selected from the group consisting of a modified cellulose polymer and a block copolymer of ethylene oxide and propylene oxide;
wherein said pollen suspension solution is capable of producing a substantially equivalent number of seeds when sprayed onto at least a first female reproductive part of a recipient monocot plant compared to the number of seeds produced using a conventional pollination technique.
2. The pollen suspension solution of claim 1 , wherein the pollen suspension solution comprises from about 2% to about 15% monocot pollen by weight.
3. The pollen suspension solution of claim 1 , wherein the pollen suspension solution comprises an isoparaffin oil, about 2% to about 20% monocot pollen by weight, a surfactant present in an amount equal to or less than about 3.0% by weight, and an aqueous solution.
4. The pollen suspension solution of claim 1 , wherein said pollen suspension solution comprises an aqueous solution, about 2% to about 20% monocot pollen by weight, and a surfactant present in an amount of less than about 1.0% by weight, wherein the surfactant is selected from the group consisting of a modified cellulose polymer and a block copolymer of ethylene oxide and propylene oxide.
5. The pollen suspension solution of claim 1 , wherein the pollen suspension solution comprises less than about 1.0% surfactant by weight.
6. The pollen suspension solution of claim 4 , wherein said surfactant is a block copolymer of ethylene oxide and propylene oxide.
7. The pollen suspension solution of claim 6 , wherein said block copolymer of ethylene oxide and propylene oxide further comprises a terminal alkyl group.
8. The pollen suspension solution of claim 4 , wherein said surfactant is a modified cellulose polymer.
9. The pollen suspension solution of claim 4 , wherein said pollen suspension solution comprises an isoparaffin oil, about 2% to about 20% monocot pollen by weight, and a surfactant present in an amount equal to or less than about 3.0% by weight.
10. A method for liquid-mediated delivery of pollen to a female reproductive part of a recipient monocot plant comprising the steps of:
a) obtaining the pollen suspension solution of claim 1 ; and
b) spraying said pollen suspension solution onto at least a first female reproductive part of the recipient monocot plant, thereby pollinating the female reproductive part with the pollen.
11. The method of claim 10 , wherein the recipient monocot plant is a) selected from the group consisting of a corn plant, a wheat plant, and a rice plant; or b) male sterile at the time of said pollinating.
12. The method of claim 10 , further comprising repeating said steps a), and b) on two or more consecutive days.
13. The method of claim 10 , wherein a) the recipient monocot plant is genetically male sterile; b) the recipient monocot plant is treated with a gametocide; or c) the male reproductive parts or parts thereof of the recipient monocot plant have been removed.
14. The method of claim 10 , wherein said pollen suspension solution comprises an aqueous solution, about 2% to about 20% monocot pollen by weight, and less than about 1.0% surfactant by weight, wherein the surfactant is selected from the group consisting of a modified cellulose polymer and a block copolymer of ethylene oxide and propylene oxide.
15. The method of claim 14 , wherein said surfactant is a block copolymer of ethylene oxide and propylene oxide.
16. The method of claim 15 , wherein said block copolymer of ethylene oxide and propylene oxide further comprises a terminal alkyl group.
17. The method of claim 14 , wherein said surfactant is a modified cellulose polymer.
18. The method of claim 10 , wherein said pollen suspension solution comprises an isoparaffin oil, about 2% to about 20% monocot pollen by weight, and a surfactant present in an amount equal to or less than about 3.0% by weight.
19. The method of claim 10 , wherein said pollen suspension solution comprises from about 2% to about 15% pollen by weight.
20. The method of claim 10 , wherein said liquid pollen suspension is produced less than about 20 minutes prior, less than about 5 minutes prior, or less than about 30 seconds prior to said spraying.
21. The method of claim 10 , wherein said method produces a substantially equivalent number of seeds compared to the number of seeds produced using a conventional pollination technique.
22. The method of claim 10 , wherein a) said spraying comprises air-assisted spraying; or b) said liquid pollen suspension is produced using agitation of said pollen suspension solution.
23. The method of claim 22 , wherein a) said spraying produces droplets with a volume weighted mean droplet diameter of less than about 300 μM; b) liquid cannot be observed visually at a distance of about 36 inches from the female reproductive part of the recipient monocot plant following said air-assisted spraying; or c) said agitation comprises vortexing or sparging said pollen suspension solution.
24. The method of claim 10 , further comprising collecting seed resulting from said pollinating.
25. The method of claim 24 , further comprising crossing a progeny plant grown from said seed with itself or a second plant.
26. The method of claim 10 , the method further comprising the steps of:
c) harvesting seed produced from said pollination; and
d) identifying hybrid progeny.
27. The pollen suspension solution of claim 1 , wherein said pollen suspension solution further comprises at least one solute at a concentration of about 0.5M to about 3.0M to increase the tonicity of the solution, wherein said solute is impermeable to said pollen in said solution.
28. The pollen suspension solution of claim 27 , wherein the solute is selected from the group consisting of a monosaccharide, a disaccharide, a polysaccharide, a polyhydric alcohol, a polyethylene glycol solute, glucose, fructose, galactose, sucrose, lactose, maltose rehalose, cellobiose, chitobiose, kojibiose, nigerose, isomaltose, trehalose, sophorose, laminaribiose, gentiobiose, trehalulose, turanose, maltulose, leucrose, isomaltulose, gentiobiulose, mannobiose, melibiose, melibiulose, rutinose, rutinulose, xylobiose, maltotriose, melezitose, nigerotriose, maltotriulose, raffinose, kestose, maltodextrin, starch, glycogen, galactogen, cellulose, chitin, pectin, peptidoglycan, mannitol, sorbitol, xylitol, lactitol, isomalt, maltitol, sodium chloride, polyethylene glycol, and glycerol ethoxylate.
29. The pollen suspension solution of claim 27 , wherein the solute is present in the solution at a) a concentration of between about 0.5M and about 3.0M; or b) about 5% to about 50% solute (weight/volume).
30. A method for liquid-mediated delivery of pollen to a female reproductive part of a recipient monocot plant comprising the steps of:
a) obtaining the pollen suspension solution according to claim 27 ; and
b) spraying said pollen suspension solution onto at least a first female reproductive part of the recipient monocot plant, thereby pollinating the female reproductive part with the pollen.
31. The method of claim 30 , wherein said liquid pollen suspension solution is produced less than about 6 hours or less than about 30 minutes prior to said spraying.
32. The pollen suspension solution of claim 1 , wherein the monocot pollen is selected from the group consisting of corn pollen, wheat pollen, and rice pollen.