Chinese Journal OF Rice Science

Chinese Journal of Rice Science ›› 2011, Vol. 25 ›› Issue (6): 667-671.DOI: 10.3969/j.issn.10017216.2011.06.015

• Experimental Techniques • Previous Articles     Next Articles

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

LIU  Zhi1, WANG  Lingling1, ZHOU  Weidong2, CHEN  Yifang2, WANG  Zhong 1,*   

  1. 1 Key Laboratory of Crop Genetics and Physiology of Jiangsu Province/Key Laboratory of Crop Physiology, Ecology and Cultivation in Middle and Lower Reaches of Yangtze River, Ministry of Agriculture, Yangzhou 225009, China; 2 Testing Center， Yangzhou University,  Yangzhou 225009, China;
  • Received:2010-12-30 Revised:2011-03-11 Online:2011-11-10 Published:2011-11-10
  • Contact: WANG Zhong1,*

固定和染色方法对水稻胚乳细胞结构观察的影响

刘智1 王玲玲1周卫东2陈义芳2王忠1,*   

  1. 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；
  • 通讯作者: 王忠1,*
  • 基金资助:

    国家自然科学基金资助项目（31071341）

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Abstract: 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.

Key words: rice, endosperm cell, fixation method, staining method, semithin section, ultrathin section

摘要： 以扬稻6号花后3~10 d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。

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

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