不同的固定方法对激光显微切割分离油松胚珠雌配子体的影响  

Effects of Different Fixing Methods on Isolation of Ovules in Female Gameto-phyte of Pinus Tabulaeformis Carr. by Laser Capture Microdissection

作　　者：王英旗[1] 姚阳[1] 陈彬丽[1] 张淑静[1] 李芳[1] 郑彩霞[1] 

机构地区：[1]北京林业大学生物科学与技术学院,北京100083

出　　处：《植物生理学报》2013年第2期195-199,共5页Plant Physiology Journal

基　　金：国家自然科学基金(31170563);教育部高等学校博士学科点专项科研基金(20110014110014)

摘　　要：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 separate the ovules in female gametophyte accurately, acquire single cell type and further study the molecular mechanisms of the ovules in female gametophyte during development, laser capture micro- dissection （LCM） was used to isolate the ovules in female gametophyte of 2-year old Pinus tabulaeformis. And the effects of four methods, including cold acetone fixation （4 ~C）, Carnoy＇s fixation, FAA fixation and frozen storage, on frozen sections were compared. The results showed that frozen storage combined with frozen sec- tions could ensure the intact morphology of female gametophyte, and obtain the female gametophytes from dif- ferent developing stages. The method could provide experimental materials for investigating the molecular mechanisms of the development of ovules in female gametophyte ofPinus tabulaeformis.

关 键 词：油松胚珠 雌配子体分离 冰冻切片 激光显微切割 

分 类 号：S791.254[农业科学—林木遗传育种]