Isolation of Sperms,Eggs,Synergids and Central Cells of Carrot(Daucus carota L.)
- 生命科学－已发表论文 
[中文文摘]用两个解剖针挤压胡萝卜花粉使其破裂释放出精细胞。用酶解-解剖方法分离胡萝卜胚囊中的卵细胞、助细胞和中央细胞。胡萝卜胚珠先在酶液中酶解40~50min,然后将其转移到不含酶的分离液中用解剖针解剖胚珠。将胚珠的合点端切破,轻轻挤压胚珠的珠孔,卵细胞、助细胞和中央细胞即可逸出。在最佳条件下,20min可从20个胚珠中分离出5个卵细胞。对分离胚囊细胞的渗透压和酶液成分进行了筛选。分离出的卵细胞用显微操作仪收集。胡萝卜精、卵细胞的成功分离为在双子叶植物中进行离体受精探索创造了条件。[英文文摘]Carrot (Daucus carota L.) two sperms could be released by squeezing pollen grain using two dissecting needles. Viable egg cells, synergids and central cells of carrot were isolated using enzymatic digestion and mechanical dissection. The ovules were digested in enzymatic solution for 40–50 min, and then transported into the isolation solution without enzymes to dissect. When dissected ovules were sliced from their chalazal ends,eggs, synergids and central cells could be released by gently pushing ovule micropyle part. In optimal isolation condition, 5 egg cells could be isolated from 20 ovules during 20 min. The osmolality and enzymes in the enzymatic solutions were screened. The isolated egg cells could be collected using micromanipulator for preparation of molecular biology of egg cell of carrot. The isolation of egg cell of carrot will make a great chance for in vitro fertilization in a dicot plant.