Institute of Biological Sciences, The University of Tsukuba, Sakura-mura, Ibaraki 305, Japan.
Plant Physiol. 1985 Sep;79(1):90-4. doi: 10.1104/pp.79.1.90.
A method for the induction of a high rate of cell division and embryogenesis of Nicotiana rustica pollen was developed. Binucleate pollen grains were fractionated by Percoll density gradient (35/45%) centrifugation and cultured in 0.4 molar mannitol at 30 degrees C (the first culture). After 3 days in culture pollen was recollected by a second Percoll fractionation (0/30%) and transferred to and cultured in a medium containing the Murashige-Skoog macro-elements, 0.4 molar mannitol, 40 millimolar galactose, 3 millimolar glutamine, and 5 micromolar ABA for 10 days (the second culture). The cell population consisting of about 80% dividing pollen was transferred to a Murashige-Skoog medium containing 0.4 molar mannitol, 3 millimolar glutamine, and no phytohormone (the third culture), where about 40% of dividing pollen developed into embryos or embryogenic calli.
建立了一种诱导普通烟草花粉高细胞分裂率和胚胎发生的方法。使用 Percoll 密度梯度（35/45%）离心分离双核花粉粒，并在 30°C 的 0.4 摩尔甘露醇中培养（第一培养）。培养 3 天后，通过第二次 Percoll 分级（0/30%）收集花粉，并转移到含有 Murashige-Skoog 大量元素、0.4 摩尔甘露醇、40 毫摩尔半乳糖、3 毫摩尔谷氨酰胺和 5 微摩尔 ABA 的培养基中培养 10 天（第二培养）。约 80%分裂花粉的细胞群体被转移到含有 0.4 摩尔甘露醇、3 毫摩尔谷氨酰胺和无植物激素的 Murashige-Skoog 培养基中（第三培养），其中约 40%的分裂花粉发育成胚胎或胚性愈伤组织。
