	US 12,256,685 B2
	Cross pollination through liquid-mediated delivery of pollen to enclosed stigmas of flowers from recipient plants
Huachun Wang Larue, Chesterfield, MO (US); and Li Lin, St. Louis, MO (US)
Assigned to Monsanto Technology LLC, St. Louis, MO (US)
Filed by Monsanto Technology LLC, St. Louis, MO (US)
Filed on Mar. 7, 2023, as Appl. No. 18/179,970.
Application 18/179,970 is a division of application No. 16/589,926, filed on Oct. 1, 2019, granted, now 11,653,604.
Prior Publication US 2023/0329166 A1, Oct. 19, 2023
This patent is subject to a terminal disclaimer.
Int. Cl. A01H 1/02 (2006.01); C12Q 1/6876 (2018.01)

CPC A01H 1/02 (2013.01) [C12Q 1/6876 (2013.01); C12Q 2600/158 (2013.01)]

11 Claims

1. A method of producing a pollen suspension injection solution comprising a desired pollen concentration for cross-pollination comprising the steps of:

a) collecting pollen-shedding flowers from a male parent;

b) homogenizing said flowers to release pollen;

c) purifying pollen from said homogenized flowers by removing floral debris;

d) quantifying said purified pollen;

e) suspending said purified pollen in an injection solution to produce the pollen suspension injection solution comprising a desired pollen concentration, wherein the pollen suspension injection solution comprises 10% to 20% sucrose by weight; 0.04% to 0.08% xanthan gum by weight; and 1.5 units/L to about 1500 units/L pectin methylesterase (PME),

f) filling a syringe with the pollen suspension injection solution, and

g) injecting the pollen suspension injection solution into unopened flower buds.