地钱在离体条件下的无性繁殖及脱分化与再分化的研究

IN VITRO PROPAGATION,DEDIFFERENTIATION AND REDIFFERENTIATION OF MARCHANTIA POLYMORPHA L.

地钱(Marchantia polymorpha L.)的胞芽和配子体先在 MS 培养基上补加1mg/l 2,4-D和3%蔗糖,经过启动部分脱分化后,再移入1/2KNOP 培养基补加4—8mg/l 2,4-D,0.25～0.5mg/l BA 与 MS 的铁盐,20g 蔗糖,此后愈伤组织肉眼可见,但仍伴有假根。最后移入 White 培养基添加丙酮酸、延胡索酸与柠檬酸三者混合物(5mmol/l)及1mg/l 2,4-D与4%葡萄糖后,始呈现彻底的脱分化状态,愈伤组织才能正常生长。整个脱分化时间长达10个月。而再分化成配子体却比高等植物容易,甚至移入不含激素的 MS 基本培养基即可形成正常的配子体。

The gemma and gametophyte of Marchantia polymorpha were propagated in vitro.De- differentiation and redifferentiation as well as the media used and cultural conditions reguir- ed were described. Since the differentiation of bryophytes was very difficult,it was necessary to culture the tissue through initiation of partial dedifferentiation on MS agar medium supplemented with 1 mg/l 2,4-D and 3% sucrose,and then subsequently the tissue was transplanted onto 1/2 Knop agar medium with addition of 4—8 mg/l 2,4-D,0.25—0.5 mg/l BA and Fe salt of MS medium. The formed calli were visual but still contained rhizoid,in this stage.The small calli finally were subcultured in white agar medium supplemented with mixture of pyruvic acid,citric acid and fumaric acid(5 mmol/l);1mg/l 2,4-D and 4% glucose.They could be differentiated thoroughly into normal tissue.The time of the total process for differentiation requied as long as 10 months. The redifferentiation and regeneration of thalli were far easier than those of higher plants even if they were transplanted onto MS phytohormone-free medium.