Expression of Function in Pollen Protoplasts

被子植物的成熟花粉在同一种的柱头上萌发并拉长花粉管，其中生殖核分裂成两个能够受精的精核。当从成熟花粉中分离出具有这两种特定功能的原生质体后，在培养中细胞壁再生后，如何在这些再生的花粉原生质体中表达这些功能是本研究的课题。对被子植物的成熟花粉原生质体进行了分离，43种双子叶植物中只有4种分离成功，而31种单子叶植物中有27种分离成功。成功与否取决于发芽杆的大小。在长花百合（Lilium longiflorum）中，从成熟花粉中分离出的原生质体频率很高（70-90%）。在培养中细胞壁再生后，80%以上的植株在固体萌发培养基上能够伸长花粉管。再生花粉也在百合柱头上萌发，花粉管通过胚珠伸长形成花柱，但受精和种子形成尚未确定。当百合的花粉原生质体被电诱导融合时，无论融合的细胞数量多少，被融合的原生质体基本上都延长了一个花粉管。百合与其他物种之间的花粉原生质体融合也导致了一个花粉管的伸长。在花粉管中，两个物种的生殖核分裂成两个精核。在每一种的柱头上，异核体的融合花粉萌发并拉长花粉管形成花柱。百合花粉与郁金香叶肉细胞融合原生植物中花粉管伸长功能的表达。在试管中，观察百合在固体培养基上萌发1天后的生殖核分裂图和体细胞叶绿体的脱落。

Mature pollen of Angiosperms germinate on the stegma of the same species and elongates pollen tubes, in which the generative nucleus divides into two sperm nuclei capable of fertilization. When protoplasts were isolated from mature pollen which have such two specific functions, after the regeneration of cell wall in culture,how expresses the functions in such regenerated pollen protoplasts was the project of this study.1. The isolation of protoplasts from mature pollen were examined in various species of Angiosperms, and only 4 species within 43 species of dicotyledons, but 27 species within 31 species of monocotyledons were successful. The success depends on the size of the germination pole.2. In Lilium longiflorum, protoplasts from mature pollen were isolated at high frequencies (70-90%). After the regeneration of cell wall in culture, more than 80% of them were able to elongate pollen tube on solid germination medium. The regenerated pollen did also germinate on lily stigmata and the pollen tubes elongated into style through the ovule, but the fertilization and the seed formation were not ascertained.3. When the pollen protoplasts of the lily were electrically induced to fuse, the fused protoplasts elongated essentially one pollen tube, regardless of the cell number of fusion. Fusion of pollen protoplasts between the lily and the other species also resulted in the elongation of one pollen tube. In the pollen tubes, the generative nuclei of both species divided into two sperm nuclei. On stigma of each species, the fused pollen of heterokaryon germinated and elongated pollen tube into style.4. The function of the pollen tube elongation expressed in fused protoplats between lily pollen and tulip mesophyl cells. In the tubes, the dividing figures of the generative nucleus of the lily and the removal of chloroplasts of the somatic cells were observed one day after germination on solid medium.