摘要
本研究建立了一种重复性好的陆地棉(Gossypium hirsuium L.cv.Coker 312)“三步法”体细胞胚发生培养程序。在DK培养基上子叶外植体脱分化形成愈伤组织,转移到4NK培养基后产生胚性愈伤组织,后者在无激素的LSCo培养基上发育形成肉眼可见的体细胞胚(胚状体)。细胞组织学观察表明胚性愈伤组织的某些细胞在发生体细胞胚之前细胞壁增厚,30—40细胞之前的原胚发育一直处于厚壁包围之中。在整个体细胞胚发育过程中,有两次淀粉累积高峰。
A reproducible 3-step procedure of somatic embryogenesis of Gossypium hirstutum L. cv. Coker 312 has been developed. Calli were mitiated on LSC medium containing 0.1mg/L 2,4-D plus 0.1mg/L KT from cotyledon tissue of 5-day-old-seedlings, and subcultured on the same medium with 4 mg/L NAA and 1 mg/L KT. Emtryogenic calli and few globular embryos developed at a frequency of 67.5% after 55 days' culture in the latter medium. When the embryogenic calli were transferred to growth regulator-free medium, embryogenesis occured and all stages of normal zygotic embryos, globular-, heart- and arrowhead- or torpedo-shaped embryo, were developed. Cyto-histological study showed that embryogenic calli were very easily distinguished from non-embryogenic calli. Embryoids were mainly initiated from the cells in the peripheral area of embryogenic calli. At the early stage the development of embryoid was limited in a boundary of thicken cell wall. There were 2 peaks of starch accumulation in the process of embryogenesis, one was at the early globular stagt, and the other at the later torpedo-shaped stage.
基金
山东省教育厅资助项目
关键词
陆地棉
体细胞胚发生
细胞组织学
Gossypium hirsutum L.
Somatic embryogenesis
Cytohistochemistry
Starch grain
