云南红豆杉小孢子发生和雄配子发育的研究方法比较

 以云南红豆杉为实验材料,比较了石蜡切片法、洋红简易染色法和整体染色透明法研究小孢子发生过程的效果,分析了用石蜡切片法及离体培养法研究雄配子体发育优缺点.在离体培养中,对比了不同的蔗糖质量浓度、培养温度及染色方法对实验结果的影响,从而为针叶树种雄性生殖系统发育过程研究提供参考.结果显示:整体透明染色法操作简单,能够反应小孢子发育过程中各个阶段的细胞结构,是用于小孢子发育过程研究的比较理想的方法;对于雄配子体发育过程的研究,需要用石蜡切片法与离体培养法相结合,在离体培养中,采用雄球花表面灭菌,自然散粉,低糖低温培养有利于防止污染,洋红染色法方便快捷,不需要特别的设备,是离体培养时比较理想的染色方法.

A comparison and evaluation of methods for sporogenesis and male gametogenesis studying in Taxus yunnanensis

By using Taxus yunnanensis as experiment material,different methods including paraffin embed,squashed with aceto-camine and stain-clearing,methods of sporogenesis,paraffin embedded samples and cultured in vitro methods for microgametogenesis were used and evaluated.In cultured in vitro methods,the influence of culture temperature,the sucrose concentration of medium and staining methods on the results was studied.The results can be summarized as follows:stain-clearing technique is a better method for sporogenesis due to its simple operation and better results.Raffin embedded samples and cultured in vitro are necessary methods in gametogenesis,and in vitro,sterilizing on surface and releasing microspores by itself,low temperature and low sucrose concentration are helpful to eliminate contamination.It is convenient that male gametogenesis in vitro be stained by aceto-carmine.