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以菠萝(Ananas comosus L. Merrill)杂交种MD1无菌苗为材料,通过培养薄细胞层切片(Thin cell layer,TCL)的方法,研究直接再生胚的固体培养体系。结果证明,预培养的设计2(在预培养基PT上黑暗预培养无菌苗1周后,从预培养苗茎顶端切取TCL,在MS+2,4-D2mg·L-1或1mg·L-1胚诱导培养基EI2上黑暗培养1周,然后转到光照条件下继续诱导胚1周,再在成熟胚培养基EM上培养2周,最后转入幼苗培养基PL上培养)是直接诱导获得体胚的最佳途径;诱导的体胚最初均来自于薄细胞层上微管束附近的细胞;picloram有利于芽的诱导,2,4-D有利于直接再生胚,但如果球形胚形成后一直培养在较高浓度的2,4-D培养基中,将阻碍球形胚和心形胚进一步发育为成熟胚,反而促进芽的生成。
The solid culture system of direct regenerated embryo was studied by culturing the thin-layer (TLC) cell of Ananas comosus L. Merrill hybrid MD1. As a result, pre-cultured design 2 (TCL was cut from the top of the stems of the preculture seedlings 1 week after preincubation of the sterile seedlings on the pre-culture medium PT in the medium of MS + 2,4-D2 mg · L-1 or 1 mg · L -1 embryo induction medium EI2 for 1 week in the dark, then continue the induction of embryos 1 week after irradiation, and then cultured on the mature embryo medium EM for 2 weeks and finally transferred to the seedling culture medium PL for culture) is directly induced The best way to get somatic embryos is that the induced somatic embryos are all from the cells near the microtubule bundles on the thin layer. Picloram is helpful for the induction of shoots. 2,4-D is beneficial to regenerate the embryos directly. However, Has been cultured in higher concentrations of 2,4-D medium, will hinder the globular embryos and heart-shaped embryos further developed into mature embryos, but to promote bud formation.