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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5d,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30min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。
Phalaenopsis hybrid was used as the material to observe the pollen germination after pollination and to isolate and collect the sperm cells. The observation showed that after the germination of Phalaenopsis pollen tube, the vegetative nucleus degenerated and the germ cells entered the pollen tube. The germ cells were still in the dividing stage 5 days after pollination. The pollen tube was developed in the ovary by pollination, the pollen tube was directly exploded with 5% ~ 12% mannitol, and the pair of Phalaenopsis spp. Cells were successfully isolated. Using a micromanipulator for 30 min, sperm cells of 4 ~ 5 pairs. Pairs of two sperm cells in the diameter, fluorescence intensity are quite different, indicating two sperm cells have different prospects. The successful isolation of Phalaenopsis spermatophore laid a good foundation for the further study of orchid in vitro fertilization.