固定和染色方法对小麦胚乳细胞结构显示的影响  

Effect of Fixation and Staining Methods on Structure Observation of Endosperm Cell of Wheat

作　　者：王玲玲[1] 刘智[1] 熊飞[1] 李栋梁[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：《作物学报》2012年第9期1688-1697,共10页Acta Agronomica Sinica

基　　金：国家自然科学基金项目(31071341,31171482);江苏省自然科学基金项目(BK2011445)资助

摘　　要：以扬麦16小麦颖果为材料,采用戊二醛-四氧化锇(GA-OsO4)或高锰酸钾固定,并分别用5种染色方法[甲苯胺蓝-O(TBO)、多色性染液、高碘酸-希夫试剂(PAS)、考马斯亮蓝和PAS-TBO]进行半薄切片染色,对超薄切片进行醋酸双氧铀和柠檬酸铅染色,在光学显微镜和透射电镜下观察比较了小麦胚乳细胞结构显示的差异。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固定,但是在PAS-TBO复染中,高锰酸钾固定制样效果最好。在超薄切片观察中,GA-OsO4固定制样能较好地保存胚乳细胞超微结构,但对膜性结构显示不够清晰,而高锰酸钾固定制样则能较好地固定细胞的膜性结构,并且能够增强膜结构反差,只是对多糖和蛋白质的固定和显色较差。因此,应根据观察对象来选择适当的固定及染色方法。Two fixing methods and five staining agents were used to compare which one is best for optimal observation of en- dosperm cell of wheat under optical and transmission electron microscopes. Caryopsis samples of Yangmal 16 were fixed with glutaraldehyde-osmium tetroxide （GA-OsO4） or potassium permanganate and stained with toluidine blue （TBO）, polychromatic dye, periodic acid schiff （PAS）, coomassie brilliant blue, or PAS-TBO during the preparation of semi-thin sections. The GA-OsO4 fixation showed better overall performance than the potassium permanganate fixation, but the best observation was got by potas- sium permanganate fixation and PAS-TBO counterstaining. The GA-OsO4 fixation had the advantage of preserving endosperm cell intrinsic feature in ultra-thin section but the disadvantage of unclear display of the membranous structure. The potassium permanganate fixation was able to show the structure of the cell endomembrane vividly with satisfactory contrast between background and membranous structure; however, the color stained in polysaccharide and protein was poor. Therefore, appropriate fixation and staining methods should be based on the target and objective of experiment.

关 键 词：小麦 胚乳细胞 固定方法 染色方法 半薄切片 超薄切片 

分 类 号：S512.1[农业科学—作物学]