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

针叶树种成熟胚同步培养的不定胚形成

基本信息

批准号：
08660200
负责人：
KUDO Takafumi
金额：
$ 1.47万
依托单位：
Tokyo University of Agriculture and Technology
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1996
资助国家：
日本
起止时间：
1996 至 1997
项目状态：
已结题

项目摘要

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.

1)落叶松（Larix leptolepis）和日本云杉（Picea jezoensis）胚性愈伤组织诱导不定胚的研究（1）落叶松幼胚和成熟胚愈伤组织诱导不定胚的研究将未成熟胚在含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、易碎的胚性愈伤组织。将诱导愈伤组织继代培养1年以上获得的红色愈伤组织，在含有1 μ M阿坝和0.3 μ M BAP的液体培养基中诱导早期不定胚萌发，成熟胚愈伤组织在与固体培养基成分相同的液体培养基中形成胚性组织，但形成率很低。(2)厚松成熟胚愈伤组织诱导不定胚的研究厚松成熟胚在含10 μ M 2，4-D和5 μ M BAP的LP培养基中培养12周，可诱导出白色易碎的胚性愈伤组织（落叶松）将用网眼调节大小为300 μ m至30 μ m的分离的愈伤组织稀释至两种细胞密度，即5.0 × 104个细胞/iota和1.2 × 104个细胞/iota，并在含有7 μ M的2，4-D和3 μ M BAP的培养基中培养14天后，由于菌落增殖和分化能力开始下降，因此将该殖民地在高细胞分裂素低生长素的培养基中进行继代培养。因此，在5.0 × 104个细胞/μ l的细胞密度条件下形成具有高增殖和分化能力的集落。