ISOLATION OF ZYGOTES AND APICAL AND BASAL CELLS OF BICELLUARE PROEMBRYO FROM TORENIA FOURNIERI

Abstract

合子是被子植物个体发育的第一个细胞。合子分裂的规律性是植物早期胚胎发生的种属甚至是科的特征,也是控制早期个体发育的关键阶段。许多被子植物的合子分裂形成二胞原胚时就呈现出调控个体发育的迹象:顶细胞继续分裂形成胚体,以后发育为胚,而基细胞则分裂数次或不分裂形成胚柄结构[1]。卵细胞和合子发育过程中的特异基因表达是配子体发育转向孢子体发育的一个关键事件之一。Torenia fournieri is a special plant with embryo sac partly protruding through the micropyle of ovule, and the cells of egg apparatus can be clearly observed using light microscope. Zygotes and the cells of bicellular proembryo could be isolated using enzymatic digestion. In the solution containing 0.05% cellulase and 0.05% pectinase, 14—15 zygotes were isolated from 50 ovules in 1h. In the solution containing 0.2% cellulose, 0.4% hemicellulase and 0.2% pectinase, 19 pairs of apical and basal cells of bicellular proembryo could be isolated from 50 ovules in 1 h. The isolated zygotes and epical and basal cells were collected using a micromanipulator up to a number which will prepare for suing molecular methods to probe the mechanism of early embryoge-nesis of higher plants.国家自然科学基金项目(30670126)资

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