微丝骨架动态参与花粉萌发的研究

Study on pollen germination regulated by dynamics of actin cytoskeleton

利用改进的Alexphalloidin活细胞染色方法和激光共聚焦显微镜技术,观察川百合(LiliumdavidiiDuch)花粉原生质体极性形成及萌发过程中微丝骨架的列阵变化。结果表明,花粉原生质体从贮存状态,经过水合、极性形成至萌发花粉管,其微丝结构从短小的梭形体,经过形成均匀的网状结构、向细胞边缘汇集的平行排列的束状结构,逐渐变成多层连续环绕细胞的微丝束结构。用酪氨酸磷酸酶抑制剂苯胂化氧(PAO)处理花粉原生质体,在微丝的汇合处,肌动蛋白聚集成小的团块,花粉的萌发受到抑制;而利用酪氨酸磷酸激酶抑制剂genistein处理细胞,微丝结构的列阵变化与对照相似。结果说明,在川百合花粉萌发过程中,有某种酪氨酸磷酸酶参与了反应。

The formation of the polarity of Lilium davidii Duch pollen protoplast and the dynamics of actin cytoskeleton were observed by non-fixation, Alex-phalloidin probing and confocal laser scanning microscopy. The results showed that the protoplast obtained from stored pollen contained the numerous crystalline fusiform bodies to constitute a storage form of actin. When stored pollen was hydrated, the actin cytoskeleton in the protoplast formed a fine network spreading uniformly. In the process of polarity formation and germination of pollen protoplast, actin filaments marshaled slowly to the brim, and then formed multilayer continuous actin bundles surrounding the cell. Furthermore, during the process of pollen germination, pollen was not sensitive to genistein, the inhibitor of tyrosine kinases, but another inhibitor of tyrosine phosphatase, phenylarsine oxido （PAO）, can effectively disturbed pollen germination. When the pollen protoplast was treated with PAO, the actin gathered into a little mass at actin filaments junction, the pollen germination was inhibited. The results show that some tyrosine phosphatase is involved in pollen germination.