烟草单个花粉原生质体透射电镜样品制备及其超微结构观察

Specimen Preparation of Single Pollen Protoplasts for TEM and Their Ultrastructural Observation in Nicotiana tabacum

本文介绍了烟草花粉原生质体作试验材料，用自制微吸管手工预先挑选单个花粉原生质体，戊二醛微滴固定，然后用低熔点琼脂糖或明胶预包埋，小塑料管顶扣包埋单个花粉原生质体，进行透射电镜样品制备，初步建立了单个原生质体电镜制样的技术流程，通过电镜观察，与群体制样法比较，结果是今人满意的。此项技术的建立，为今后植物性细胞超微结构的研究，提供了一项精细的操作方法。

The single mature pollen protoplasts of Nicotiana tabacum were observed under a transmission electron mi-croscope.Their preparation for electron microscopy was performed by selecting single target pollen protoplastswith a microcapillary and, after short prefixation in glutaraldehyde droplets,embedding in low gelling agrose orgelatin as preliminary steps,As a comparison, the specimen preparation of mass pollen protoplasts was carriedout following the method of Wu and Zhou(1991)wigh little modification.The results from both methods werecomparable with each other pretty well. The pollen protoplasts observed were completely deprived of pollenwall,but kept the internal structure quite intact.This technique for specimen preparation might be used in theultrastructural research on sexual plant cell manipulation not only for pollen protoplasts but also for gametoplas-ts(e.g.isolated egg cells).