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US 6,143,563 AGranted
Cryopreservation of embryogenic callus
AssigneePioneer Hi-Bred International, Inc.
InventorDavid Jay Peterson
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Abstract
A method for cryopreserving plant callus without the use of cryoprotectants or programmable freezers. The callus is spread on a physical support, grown on a media supplemented with an osmoticum, desiccated under controlled conditions, placed directly into cold storage, and can be revived by thawing. Actively growing callus from a number of species can be cryopreserved using the instant invention.
Claims3
A method of cryopreserving plant callus comprising the steps of:
- A. spreading the plant callus evenly on a glass fiber disc;
- B. culturing the plant callus on a maintenance medium supplemented with mannitol or sucrose, and proline;
- C. desiccating the plant callus by exposure to a saturated solution of Ca(NO.sub.3).sub.2 4H.sub.2 O or (NH.sub.4).sub.2 SO.sub.4 ;
- D. placing the desiccated plant callus of (C) free of a cryoprotectant solution in cold storage by immersion in liquid nitrogen;
- and
- E. reviving the stored plant callus by thawing the plant callus at room temperature, wherein said stored plant callus shows normal growth and morphology.
3 more elements
Dependencies are inferred from the claim text.