US 11,807,846 B2
Methods for transforming corn explants
Yurong Chen, Verona, WI (US); Brian J. Martinell, Mt. Horeb, WI (US); Anatoly Rivlin, Brooklyn, WI (US); Yuechun Wan, Madison, WI (US); Edward J. Williams, Madison, WI (US); Xudong Ye, Madison, WI (US); and Ashok Shrawat, Chesterfield, MO (US)
Assigned to Monsanto Technology LLC, St. Louis, MO (US)
Filed by Yurong Chen, Verona, WI (US); Brian J. Martinell, Mt. Horeb, WI (US); Anatoly Rivlin, Brooklyn, WI (US); Yuechun Wan, Madison, WI (US); Edward J. Williams, Madison, WI (US); Xudong Ye, Madison, WI (US); and Ashok Shrawat, Chesterfield, MO (US)
Filed on Apr. 29, 2012, as Appl. No. 13/873,092.
Claims priority of provisional application 61/639,992, filed on Apr. 29, 2012.
Prior Publication US 2022/0340925 A1, Oct. 27, 2022
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 15/00 (2006.01); C12N 15/82 (2006.01)
CPC C12N 15/8275 (2013.01) [C12N 15/8274 (2013.01)] 35 Claims
 
1. A method of producing a transgenic corn plant comprising
a) transforming a dry mature corn seed explant with a heterologous DNA,
b) co-culturing the explant in a co-culture medium for about 3 days to about 6 days, wherein the co-culture medium does not contain a selective agent,
c) transferring the transformed explant to at least a first bud induction medium, wherein the first bud induction medium comprises at least one cytokinin and at least one auxin, wherein the concentration of cytokinin in the first bud induction medium is from 4 mg/L to 10 mg/L, wherein the concentration of auxin in the first bud induction medium is from 1 mg/L to 2.2 mg/L, and wherein the first bud induction medium does not contain a selective agent,
d) culturing the transformed explant in the first bud induction medium for at least about 14 days to produce a transformed explant with multiple buds, and
e) regenerating a transgenic corn plant in a regeneration medium, from the transformed explant with multiple buds, wherein the transgenic corn plant is produced without generating a callus from the transformed explant;
wherein delaying selection by culturing the transformed explant in said first bud induction medium for at least about 14 days results in an increased transformation frequency compared to culturing the transformed explant in a different first bud induction medium that contains a selective agent.