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

1)日本落叶松(Larix Letolepis)和日本云杉(Picea Jezoensis)胚愈伤组织的诱导(1)红松幼胚和成熟胚愈伤组织的诱导。未成熟胚经CD+2，4-D 7 mM+BAP 3 mM培养12周，获得白色、易碎的胚性愈伤组织。继代培养1年以上的红色愈伤组织，在含有1MU的ABA和0.3MM的BAP的液体培养基中诱导早期不定胚萌发。成熟胚愈伤组织在液体培养基中形成胚性组织，其成分与固体成分相同，但形成率很低。(2)以成熟胚愈伤组织为外植体诱导出不定胚。在含有10 mU M的2，4-D和5 mU M的BAP的LP培养基中培养12周，常能诱导出白色的脆性胚性愈伤组织。2)以日本落叶松幼胚愈伤组织为试材，悬浮培养形成不定胚，经筛网调节大小为300~30µm的愈伤组织，稀释成5.0×104个/iota和1.2×104个/iota两种细胞密度，在含有7 mU M的2，4-D和3 mM的BAP的CD液体培养基中诱导出集落。培养14d后，随着菌落增殖能力和分化能力的下降，在高细胞分裂素和低生长素的培养基上进行继代培养。在细胞密度为5.0×104个/IOTA的条件下，可形成增殖和分化能力较强的菌落。

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.