ISOLATION OF SPERM CELLS OF Allium tuberosum Roxb
- 生命科学－已发表论文 
<正>分离精细胞是在高等植物中开展离体受精研究的前提之一,也是开展精细胞分子生物学研究的前提[1,2]。对具三胞花粉的植物,精细胞的分离比较容易,将花粉粒直接撒到含一定渗透压的培养基中,花粉粒爆破释放出精细胞。对于具二胞花粉的Pollen grains of Allium Tuberosum Roxb broke and released their content including generative cells using osmotic shock method. In a medium containing 0.05%CaCl2,0.01%Boric acid,0.01%KH2PO4,15%PEG和10%sucrose (710 mOsmol/kg H2O),86% pollen grains germinated and grew out pollen tubes,which broke after transferred into 6% mannitol solution,and released tube content including generative cell. When pollen grains were cultured in the same medium but adding 0.1% casein,a few generative cells divided into two sperm cells. Stigmas of Allium tuberosum Roxb were pollinated at second day after anthesis and the styles grow 3 h in vivo. Then the styles were cut and cultured in a medium for about 6-8 h,some pollen tubes grew out of the cut end of the style. The cut end of the style was transferred into a solution containing 6% mannitol to burst pollen tubes. Pairs of sperm cells of Allium tuberosum Roxb were released and collected using a micromanipulator.