应用水稻花粉离体萌发体系观察花粉管内微丝分布
Visualization of Microfilament Distribution in Pollen Tubes through in vitro Rice Pollen Germination System
-
摘要: 采用非固定、DMSO渗透和异硫氰酸标记的鬼笔环肽(FITC-Ph)染色方法,观察水稻花粉离体萌发过程中花粉管内肌动蛋白微丝的形态和分布。结果表明:(1)水稻花粉水合2 min后即可萌发,花粉管生长速度在6001500μm/h之间。(2)水合而未萌发的花粉粒中,大量较短的梭形微丝束构成微丝网络结构,萌发过程中花粉粒内的梭形微丝束松解,部分微丝转移至萌发的花粉管内沿花粉管纵轴呈束状结构;随着花粉管的伸长,微丝束主要分布在花粉管中前端,但在花粉管顶端区域始终未见明显的微丝束。(3)水合后不能正常萌发的花粉粒内肌动蛋白微丝呈弥散不规则分布,在相同萌发时间生长迟缓的花粉管中,微丝束较少,且主要位于花粉管近萌发孔的部位。表明微丝骨架的形态和分布影响水稻花粉管的萌发和生长。Abstract: Microfilament patterns during pollen germination and pollen tube growth were observed with no pre-fixation using dimethylsulphoxide(DMSO)as a permeabilising agent and FITC-Phalloidin as the stain.The main results were:(1) Pollen germinated after hydration for 2 min and growth speed of the pollen tube was between 600-1500 μm/h.(2) In the hydration of the pollen grains without germination,a number of short fusiform microfilaments constituted a network-like structure.In the germination process,some microfilaments gradually transferred to the pollen tube as the short fusiform microfilaments were released along the pollen tube longitudinally.For pollen tube elongation,microfilaments were mainly in the middle and front region of pollen tube,but there were no obvious microfilaments in the tip of pollen tube.(3) The distribution of microfilaments was diffuse and irregular in certain non-germination pollen.Some pollen germinated at same time but were shorter,with fewer microfilaments gathered in the region near the apertures.These results indicated that the distribution of microfilament cytoskeleton affected the germination and growth of rice pollen tubes.