Technique of Cryopreservation Using Somatic Embryo in Japanese Larch Larix leptolepis and Induction of Embryogenic Culture

摘要

A method for cryopreservation of Japanese larch (Larix leptolepis) and induction of embryogenic tissue is provided to simplify pretreatment by using somatic embryo of Japanese larch (Larix leptolepis) as compared with liquid embryogenic cell method, store it for a long time in liquid nitrogen without use of expensive cell freezer and reduce DNA mutation of restored embryogenic cell by using no DMSO(dimethylsulphoxide). A method for cryopreservation of Japanese larch (Larix leptolepis) and induction of embryogenic tissue comprises the steps of: dehydrating induced somatic embryo, cryopreserving the dehydrated somatic embryo with a cryopreserving tube in liquid nitrogen at -20 to -196 deg. C for 1 week and melting the cryopreserved somatic embryo in water of 30-40 deg. C for 5-15 minutes; and rehydrating the melted somatic cell and transferring and culturing the rehydrated somatic cell into an induction medium in a dark room to induce the embryogenic tissue.