In search of a new method to illustrate the microsporogenesis,indirect immunofluorescence microscopy with DAPI (4′,6-diamidino-2-phenylindole) staining was conducted to observe the microtubule cytoskeleton pattern and chromosomes behavior during microsporogenesis of Populus euphratica.The results showed that:(1)Meiotic chromosome behavior is normal during microsporogenesis,with univalent at diplotene and lagging chromosome at metaphase Ⅰ and the varied number of nucleoli.(2)Microtubule cytoskeleton was involved in many key processes of microsporogenesis.Spindles of metaphase Ⅱ were mutually parallel or tripolar.No typically phragmoplasts were observed at telophase Ⅱ,while simultaneous-type cytokinesis show an inward-oriented infurrowing of the callosic parental wall to partition the meiotic cytoplasm,mediated by the radial microtubules systems.The tetrad was shaped in a tetrahedral or isobilateral pattern.(3)Abnormal meiosis were also observed during microsporogenesis,such as fused spindles at metaphase Ⅱ,asynchronous division of the second meiosis,furrow occurred between the mutually parallels at the second division,as well as triad and dyad were found together with tetrad at tetrad stage,which led to formation of natural 2n pollen and stuck pollen.