Li Y, Zee S Y, Liu Y M, Huang B Q, Yen L F
College of Biological Sciences, China Agricultural University, Beijing, PR China.
Planta. 2001 Sep;213(5):722-30. doi: 10.1007/s004250100543.
The distribution of and relationship between F-actin and G-actin were investigated in pollen grains and pollen tubes of Lilium davidii Duch. using a confocal laser scanning microscope after fluorescence and immunofluorescence labeling. Circular F-actin bundles were found to be the main form of microfilament cytoskeleton in pollen grains and pollen tubes. Consistent with cytoplasmic streaming in pollen tubes, there were no obvious F-actin bundles in the 10- to 20-microm tip region of long pollen tubes, only a few short F-actin fragments. Labeling with fluorescein isothiocyanate (FITC)-DNase I at first established the presence of a tip-focused gradient of intracellular G-actin concentration at the extreme apex of the tube, the concentration of G-actin being about twice as high in the 10- to 20-microm region of the tip as in other regions of the pollen tube. We also found that the distribution of G-actin was related negatively to that of the F-actin in pollen tubes of L. davidii. Caffeine treatment caused the G-actin tip-focused gradient to disappear, and F-actin to extend into the pollen tube tip. Based on these results, we speculate that the circular F-actin bundles may be the track for bidirectional cytoplasmic streaming in pollen tubes, and that in the pollen tube tip most of the F-actin is depolymerized into G-actin, leading to the absence of F-actin bundles in this region.
利用共聚焦激光扫描显微镜,在荧光和免疫荧光标记后,研究了川百合花粉粒和花粉管中F-肌动蛋白和G-肌动蛋白的分布及其相互关系。发现环状F-肌动蛋白束是花粉粒和花粉管中微丝细胞骨架的主要形式。与花粉管中的胞质环流一致,在长花粉管10至20微米的顶端区域没有明显的F-肌动蛋白束,只有一些短的F-肌动蛋白片段。用异硫氰酸荧光素(FITC)-脱氧核糖核酸酶I标记首先确定了在花粉管顶端存在一个聚焦于顶端的细胞内G-肌动蛋白浓度梯度,在顶端10至20微米区域的G-肌动蛋白浓度约为花粉管其他区域的两倍。我们还发现,在川百合花粉管中,G-肌动蛋白的分布与F-肌动蛋白的分布呈负相关。咖啡因处理导致G-肌动蛋白顶端聚焦梯度消失,F-肌动蛋白延伸至花粉管顶端。基于这些结果,我们推测环状F-肌动蛋白束可能是花粉管中双向胞质环流的轨道,并且在花粉管顶端,大多数F-肌动蛋白解聚为G-肌动蛋白,导致该区域没有F-肌动蛋白束。