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为了精确分离油松胚珠雌配子体,获得单一种类的细胞,进一步研究油松胚珠发育分子调控的时空特异性,本文利用激光显微切割技术(LCM)对2年生油松胚珠雌配子体进行了微切割;比较了冷丙酮(4℃)、卡诺固定液、FAA固定液固定及冻藏处理4种不同方法对冰冻切片的影响。结果表明,冻藏处理结合冰冻切片可以保证雌配子体的完整性,采用该方法获得了不同时期油松胚珠雌配子体,为后续油松胚珠雌配子体发育相关分子机制的研究提供了实验材料。
In order to isolate accurately the female gametophyte of Pinus tabulaeformis and obtain a single kind of cell, further study on the space-time specificity of the molecular regulation of ovule developmental regulation in Pinus tabulaeformis, the 2-year-old male pine ovoid female gametophyte was microdissected by laser microdissection (LCM) The effects of cold acetone (4 ℃), Carnot’s fixative and FAA fixative on frozen sections were compared. The results showed that frozen storage combined with frozen sections could ensure the integrity of female gametophyte. The female gametophyte was obtained by this method, which provided the experimental materials for the molecular mechanism of the development of female gametophyte in the next stage.