ADVENTITIOUS EMBRYO FORMATION BY SYNCHRONOUS CULTURE FROM MATUR EMBRYO IN CONIFEROUS SPECIES

1)卡拉松落叶松（Larix leptolepis）和云杉（Picea jezoensis）胚愈伤组织诱导不定胚(1)卡拉松未成熟胚和成熟胚愈伤组织诱导不定胚在含7 μ M 2,4- d和3 μ M BAP的CD培养基中培养12周，获得白色和脆性胚性愈伤组织。将诱导愈伤组织继代培养1年得到的红色愈伤组织，在含有1 μ M ABA和0.3 μ M BAP的液体培养基中诱导出早期不定胚并发芽。成熟胚愈伤组织在与固体培养基成分相同的液体培养基中可形成胚性组织，但形成率很低。(2)骨松成熟胚愈伤组织诱导不定胚在骨松成熟胚中，用10 μ M的2,4- d和5 μ M的BAP在LP培养基中培养12周，可多次诱导出白色脆性胚性愈伤组织。2)以松松落叶松（Larix leptolepis）游离愈伤组织的未成熟胚为材料，以300 μ m ~ 30 μ m的网眼调节大小，将其稀释为5.0*104个细胞/iota和1.2*104个细胞/iota两种细胞密度，在含7 μ m 2,4- d和3 μ m BAP的液体CD培养基中诱导集落形成不定胚。但在培养14天后，由于菌落的繁殖能力和分化能力开始下降，于是将菌落传代到含有高细胞分裂素和低生长素的培养基中。因此，在细胞密度为5.0*104个/iota的条件下，形成了具有较高繁殖和分化能力的菌落。

1) Induction of adventitious embryo from callus of embryo in karamatsu (Larix leptolepis) and ezomatsu (Picea jezoensis)(1) Induction of adventitious embryo from callus of immature and mature embryo in karamatsuWhite and friable embryogenic callus were derived from immature embryo by culture in CD medium with 7 mu M of 2,4-D and 3 mu M of BAP for 12 weeks. Red callus, obtained by subculturing the induction callus over 1 year, induced the early adventitious embryo, resulting in the germination, in liquid medium containing 1 mu M of ABA and 0.3 mu M of BAP.From callus of mature embryo, embryogenic tissues were formed in liquid medium, which was and identical component to the solid, but the rate of formation was very low.(2) Induction of adventitious embyo from callus of mature embryo in ezomatsuIn mature embryo of ezomatsu, white friable embryogenic callus were frequently induced by culture in LP medium with 10 mu M of 2,4-D and 5 mu M of BAP for 12 weeks.2) An attempt for adventitious embryo formation by suspension culture from callus of immature embryo in karamatsu (Larix leptolepis)Dissociated callus, regulated in size of 300 mu m to 30 mu m by meshes, was diluted to two cell densites, as 5.0*104 cells/iota and 1.2*104 cells/iota, and colony was induced in liquid CD medium with 7 mu M of 2,4-D and 3 mu M of BAP.However, as abilities of the propagation and the differentiation of colony begun to decrease after culturing for 14 days, the colony, then, was subcultured in medium containing high cytokinin and low auxin. Consequently, colony with high ability of the propagation and the differentiation was formed in cell density condition of 5.0*104 cells/iota.