慈姑根尖细胞微管骨架的免疫荧光观察

Immunofluorescent Observation of Micro-tubule Cytoskeleton in Root Tip Cells of Sagittaria sagittifolia L.

分别用考马斯亮蓝染色和间接免疫荧光标记,并运用荧光倒置显微镜和激光共聚焦显微镜,对慈姑根尖固定后酶解获得的去壁细胞和细胞团块以及根尖细胞分裂周期中微管骨架列阵进行详细观察,以探索高等植物微管周期的普遍性。结果表明:慈姑根尖固定后酶解可获得大量结构完整的去壁细胞与细胞团块;考马斯亮蓝染色观察可见,慈姑根尖细胞中丰富的蛋白物质以及处于不同分裂期的细胞核染色体;免疫荧光观察可见,慈姑根尖细胞周期中微管骨架保存较好,主要有周质微管、早前期带微管、纺缍体微管和成膜体微管4种循序变化的排列方式,构成了高等水生植物分裂细胞中典型的微管周期。实验结果证明,高等水生植物与陆生植物微管周期具有相似性,为植物微管周期概念提供了新的实例。

In order to investigate the catholicness of microtubule cycle in higher plants,changes of microtubule orientation in cell division cycle of Sagittaria sagittifolia L. ,root tips were detected by the inverted fluorescence microscope and laser scanning confocal microscope, through the techniques of enzymatic digestion and separation of root tip ceils after fixation, coomassie brilliant blue staining and indirect immunofluorescence microscopy. The results have shown that a large number of cells without cell walls and their aggregates of S. sagittifolia L. root tips can be gained by enzymatic digestion and separation. Abundant protein and nuclear chromosomes at different mitotic stage can be observed by coomassie brilliant blue staining. Under the observation of immunofluorescence microscopy, there are four basic types of mierotubule organization, namely, interphase cortical microtubule, preprophase band microtubule, spindle microtubule and phragmoplast microtubule,which constitute the typical microtubule cycle in dividing cells of higher aquatic plants. The changing similarity between the microtubule cycles of higher aquatic and terrestrial plants has provided a new instance for the notion of plant microtubule cycles.