固定和染色方法对水稻胚乳细胞结构观察的影响  

Effect of Fixation and Staining Methods on Observation of Endosperm Cell Structure in Rice

作　　者：刘智[1] 王玲玲[1] 周卫东 陈义芳 王忠[1] 

机构地区：[1]扬州大学江苏省作物遗传生理重点实验室/农业部长江中下游作物生理生态与栽培重点开放实验室,江苏扬州225009 [2]扬州大学分析测试中心,江苏扬州225009

出　　处：《中国水稻科学》2011年第6期667-671,共5页Chinese Journal of Rice Science

基　　金：国家自然科学基金资助项目(31071341)

摘　　要：以扬稻6号花后3～10d的颖果为材料,采用戊二醛-四氧化锇(GA-OsO4)和高锰酸钾两种方法固定样品,Spurr树脂包埋后分别进行半薄切片和超薄切片。对半薄切片分别进行甲苯胺蓝(AMB)染色、高碘酸-希夫(PAS)法染色、多色性染液染色以及PAS-AMB复染,比较了2种固定方法和4种染色方法对水稻胚乳细胞结构观察的影响。结果表明,GA-OsO4固定的半薄切片染色效果整体上优于高锰酸钾,在超薄切片观察中,GA-OsO4固定制样的细胞组织结构清晰,但细胞中内膜性结构不够清晰,而高锰酸钾固定制样则能较好的显示出细胞中内膜结构,但是对多糖和蛋白质的呈色较差。建议在实验中根据观察内容选择固定及染色方法。The caryopses of rice cv.Yangdao 6 were harvested at 3 to 10 days after flowering and prepared for the conventional glutaraldehyde-osmium tetroxide（GA-OsO4） fixation and for the potassium permanganate（KMnO4） fixation,respectively.After being embedded with Spurr resin,the sections were made and stained by ammonium methylbenzene blue staining（AMB）,periodic acid schiff（PAS）,multicolor dye and restained by PAS-AMB,respectively.The different effects of the two fixation and four stained methods were observed and compared.The results revealed that better semi-thin sections could be obtained by GA-OsO4 fixation.The ultra-thin sections by GA-OsO4 fixation presented a clearer cell structure,but the endomembrane structure was not clear enough.For the potassium permanganate fixation method,the endomembrane structure was clear,while polysaccharide and protein were in poor colour.It is recommended that selecting proper fixation and staining methods should be based on the observation of the contents in the experiments.

关 键 词：水稻 胚乳细胞 固定方法 染色方法 半薄切片 超薄切片 

分 类 号：S511[农业科学—作物学]