Development and stratification of pine somatic embryos using a liquid system
AssigneeWeyerhaeuser NR Company
InventorsPramod K. Gupta, Diane G. Holmstrom, Bonnie Larson, Judith Zucati
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Abstract
The invention provides methods for producing pine somatic embryos using a liquid development medium and/or a liquid stratification medium. In a first aspect, the methods comprise the step of culturing embryogenic cells in, or on, a liquid development medium to produce cotyledonary pine somatic embryos. In another aspect, the methods comprise the step of culturing pine cotyledonary somatic embryos in, or on, a liquid stratification medium to produce stratified cotyledonary somatic embryos. The invention also provides methods for producing pine somatic embryos in bioreactors.
Claims20
A method for producing stratified cotyledonary pine somatic embryos, comprising:
- (a) culturing pine embryogenic cells in, or on, a maintenance medium to multiply the pine embryogenic cells;
- (b) culturing the pine embryogenic cells multiplied in step (a) in, or on, a liquid development medium comprising abscisic acid and polyethylene glycol to produce cotyledonary pine somatic embryos;
- and
- (c) culturing the cotyledonary pine somatic embryos produced according to step (b) in, or on, a liquid stratification medium to produce stratified cotyledonary pine somatic embryos, wherein the liquid stratification medium has an osmolality between about 100 mM/kg and about 250 mM/kg.
Dependencies are inferred from the claim text.