蝴蝶兰精细胞的分离和收集

 以蝴蝶兰（Phalaenopsis hybrid）为材料，观察授粉后花粉的萌发，并进行精细胞的分离和收集。观察发现，蝴蝶兰花粉管萌发后营养核退化，生殖细胞进入花粉管，授粉后5 d，生殖细胞尚处于分裂期。通过授粉使花粉管在子房内发育，再利用5% ~ 12%的甘露醇将花粉管直接爆破，成功分离出成对的蝴蝶兰精细胞，用显微操作仪30 min 可收集精细胞4 ~ 5 对。成对的两个精细胞在直径、荧光强度方面均有较大差异，预示两个精细胞具有不同的前途。蝴蝶兰精细胞分离的成功为进一步开展兰花离体受精研究打下了良好的基础。

The Isolation and Collection of Sperm Cells in Phalaenopsis

The germination of pollens and the isolation and collection of sperm cells were studied by using a Phalaenopsis hybrid in this paper. The results show that the nucleus of generative cell degenerated after the pollen germinated and the generative cell moved into the pollen tube. The generative cell was still in cell division after pollinated 5 days. Pollen tubes were induced in the ovary after manual pollination and the sperm cells were isolated from pollen tubes by immediately blowing up in a broken solution containing 5%–12% mannitol. Four to five pairs of sperms could be collected within 30 mins by using a micromanipulator. The isolated pairs of two sperm cells shown significant differences in sizes and fluorescent intensities,that suggested both sperms have different futures. The successful isolation of sperm cells from Phalaenopsis would provide a solid basis for the studies of in vitro fertilization in orchid in the future.