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应用免疫荧光标记法结合激光扫描共聚焦显微镜术对黑松花粉和花粉管中的微管骨架进行观察。结果表明:黑松萌发花粉内存在一密集的微管网络,在花粉粒中微管呈斜向或横向分布,在伸长的花粉管内微管呈轴向排列,并一直延伸至花粉管顶端。花粉管顶端区域的微管网络荧光特别明亮。在含秋水仙碱的培养基中萌发的黑松花粉,其花粉管形态明显异常,特别是花粉管顶端肿胀的频率显著提高。认为是秋水仙碱破坏了花粉管顶端区域的微管骨架而导致顶端肿胀的产生,这说明分布于裸子植物花粉管顶端的微管网络对于维持正常的花粉管顶端形态具有重要作用。裸子植物花粉管顶端的生长可能与被子植物不同,微管网络参与了引导顶端生长的方向。
Immunofluorescence labeling and laser scanning confocal microscopy were used to observe the microtubule cytoskeletons in pollen and pollen tubes. The results showed that there was a dense network of microtubules in the germinating pollen of Pinus thunbergii. The microtubules in the pollen grains were obliquely or transversely distributed. The microtubules in the elongated pollen tube were arranged axially and extended to the top of the pollen tube. Microtubule network fluorescence at the tip of the pollen tube is particularly bright. In colchicines containing germination of black pine pollen, the pollen tube morphology was abnormal, especially the top of the pollen tube swelling was significantly increased. It is believed that colchicine destroys the microtubule cytoskeleton in the apical region of the pollen tube and leads to apical swelling. This indicates that the microtubule network distributed on the apical side of the pollen tube of gymnosperms plays an important role in maintaining the normal apical morphology of the pollen tube. The growth of the top of the gymnosperm pollen tube may be different from that of the angiosperm. The microtubule network is involved in guiding the growth of the top end.