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Acta Agron Sin ›› 2012, Vol. 38 ›› Issue (09): 1688-1697.doi: 10.3724/SP.J.1006.2012.01688


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

WANG Ling-Ling1,**,LIU Zhi1,**,XIONG Fei1,LI Dong-Liang1,ZHOU Wei-Dong2,CHEN Yi-Fang2,WANG Zhong1,*   

  1. 1Key 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 of Ministry of Agriculture, Yangzhou 225009, China; 2Testing Center, Yangzhou University, Yangzhou 225009, China?
  • Received:2012-02-10 Revised:2012-04-20 Online:2012-09-12 Published:2012-07-03
  • Contact: 王忠, E-mail: wangzhong@yzu.edu.cn

Abstract: Two fixing methods and five staining agents were used to compare which one is best for optimal observation of endosperm cell of wheat under optical and transmission electron microscopes. Caryopsis samples of Yangmai 16 were fixed with glutaraldehyde-osmium tetroxide (GA-OsO4) or potassium permanganate and stained withtoluidine 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 potassium permanganate fixationand 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 endomembranevividly 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.

Key words: Wheat, Endosperm cell, Fixation method, Stain method, Semi-thin section, Ultra-thin section

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