不同固定液对小麦花粉母细胞微丝骨架荧光标记的效果

Results of Fluorescent Labeling of Actin Cytoskeleton in Pollen Mother Cell of Wheat Fixed by Different Fixatives

细胞学特征的研究是解析小麦光温敏雄性不育机理的重要基础,已有研究表明小麦光温敏核雄性不育系BS366的败育可能和细胞骨架异常相关。但是与微管骨架观察相比,通常微丝骨架荧光标记难度大、效果欠佳。通过对比和分析利用FAA固定液和多聚甲醛固定液固定材料的微丝TRITC-phalloidin荧光标记,结果表明利用FAA固定液固定花药组织进行微丝标记的形态效果较好,进一步用植物Actin单克隆抗体做间接免疫荧光标记验证了FAA固定材料的可靠性。试验为研究小麦光温敏雄性不育系减数分裂期微丝行为提供了更简便有效的技术和方法,同时也对其他高等植物花粉母细胞的微丝骨架荧光标记观察具有一定的借鉴作用。

Studying of the cytological characteristics associated with male sterility is the fundamental work for photo-thermo-sensitive male genic sterile line of wheat.There have been reports that aberrant dynamics of cytoskeleton may involved in male sterility of BS366,a photo-thermo-sensitive male genic sterile line of wheat.However,the fluorescent labeling of actin cytoskeleton in pollen mother cell of wheat is difficult.Contrasting and analysising of the results of TRITC-phalloidin fluorescent labeling of the pollen mother cell fixed by FAA fixative and paraformaldehyde,we found that the fluorescent labeling of FAA-fixed material got better effects.The results of immunofluorescence of actin cytoskeleton in FAA-fixed tissue was in accordance with that of TRITC-phalloidin fluorescent staining,which implied the utilization of FAA fixative was reliable.Therefore it was possible to investigate the relationship between organization of actin cytoskeleton during meiosis and male sterility in the photo-thermo-sensitive male genic sterile line.The method could be used in other higher plant in the identification of actin cytoskeleton in pollen mother cell.