|Budget Amount *help
¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1997: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1996: ¥1,800,000 (Direct Cost: ¥1,800,000)
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 karamatsu
White 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 ezomatsu
In 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.