Welcome to Acta Agronomica Sinica，

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

• TILLAGE & CULTIVATION · PHYSIOLOGY & BIOCHEMISTRY • Previous Articles     Next Articles

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

PDF

2206

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

[1]Buttrose M S. Ultrastructure of the developing wheat endosperm. Aust J Biol Sci, 1963, 16: 305–317

[2]Bechtel D B, Gains R L, Pomeranz Y. Early stages in wheat endosperm formation and protein body initiation. Annu Bot, 1982, 50: 507–518

[3]Parker M L. The relationship between A-type and B-type starch granules in the developing endosperm of wheat. J Cereal Sci, 1985, 3: 271–278

[4]Shewry P R, Napier J A, Tatham A S. Seed storage proteins: structures and biosynthesis. Plant Cell, 1995, 7: 945–956

[5]Langeveld S M J, van Wijk R, Stuurman N, Kijne J W, de Pater S. B-type granule containing protrusions and interconnection between amyloplasts in developing wheat endosperm revealed by transmission electron microscopy and GFP expression. J Exp Bot, 2000, 51: 1357–1361

[6]Shewry P R, Halford N G. Cereal seed storage proteins: structures, properties and role in grain utilization. J Exp Bot, 2002, 53: 947–958

[7]Kumamaru T, Ogawa M, Satoh H, Okita T W. Protein body biogenesis in cereal endosperms. In: Olsen O A ed. Plant Cell Monogry: Endosperm. Heidelberg: Springer-Verlag Berlin, 2007. pp 141–158

[8]Sheng J(盛婧), Guo W-S(郭文善), Zhu X-K(朱新开), Feng C-N(封超年), Peng Y-X(彭永欣). Starch granules development in grain endosperm of wheat for different end uses. Acta Agron Sin (作物学报), 2004, 30(9): 953–954 (in Chinese with English abstract)

[9]Wei C-X(韦存虚), Zhang J(张军), Zhou W-D(周卫东), Chen Y-F(陈义芳), Xu R-G(许如根). Accumulation of storage protein and formation of protein body during wheat endosperm development. J Triticeae Crops (麦类作物学报), 2009, 29(1): 73–78 (in Chinese with English abstract)

[10]Yin Y-A(银永安), Qi J-C(齐军仓), Li W-H(李卫华), Cao L-P(曹连莆), Wang Z-B(王自布). Physico- chemical characteristics of A, B type starch granule in wheat endosperm. Sci Agric Sin (中国农业科学), 2010, 43(11): 2372–2379 (in Chinese with English abstract)

[11]Liu Z(刘智), Wang L-L(王玲玲), Zhou W-D(周卫东), Chen Y-F(陈义芳), Wang Z(王忠). The effect of fixation and staining methods on observed the endosperm cell in rice. Chin J Rice Sci (中国水稻科学), 2011, 25(6): 667–671 (in Chinese with English abstract)

[12]Bechtel D B, Zayas I, Kaleikau L, Pomeranz Y. Size distribution of wheat starch granules during endosperm development. Cereal Chem, 1990, 67: 59–63

[13]Liu D-J(刘冬娟), Zhu S-W(祝素文), Li Y-J(李艳杰). The application of semi-section staining with borax toluidine blue. J Chin Med Univ (中国医科大学学报), 2004, 33(1): 33 (in Chinese)

[14]Wang S(王爽), Wang C-M(王春梅), Chen-D(陈丹), Huang X-F(黄晓峰). Polychromatic dying method on semithin resin sections resin. J Fourth Mil Med Univ (第四军医大学学报), 2002; 23(9): 775 (in Chinese)

[15]Lyon H O, Schulte E K, Prento P, Barer M R, Béné M C. Standardized staining methods: feulgen-rossenbeck reaction for desoxyribonucleic acid and periodic acid -schiff (PAS) procedure. Biotech Histochem, 2002, 77: 121–125

[16]Hu S-Y(胡适宜), Xu L-Y(徐丽云). A cytochemical technique for demonstration of lipids, polysaccharides and protein bodies in thick resin sections. Aata Bot Sin (植物学报), 1990, 32(11): 841–846 (in Chinese with English abstract)

[17]Dang J M, Copeland L. Studies of the fracture surface of rice grains using environmental scanning electron microscopy. J Sci Food Agric, 2004, 84: 707–713

[18]Buttrose M S. Submicroscopic development and structure of starch granules in cereal endosperms. J Ultrastructure Res, 1960, 4: 231–257

[19]Fu H-L(付洪兰). Practical Electronic Microscope Technology (实用电子显微镜技术). Beijing: Higher Education Press, 2004. pp 12–13 (in Chinese)

[20]Song J-D(宋今丹). The membrane structure of cultured cell by potassium permanganate fixed. J Chin Electron Microscopy Soc (电子显微学报), 1990, 9(3): 1 (in Chinese)

[21]Loussert C, Popineau Y, Mangavel C. Protein bodies ontogeny and localization of prolamin components in the developing endosperm of wheat caryopses. J Cereal Sci, 2008, 47: 445–456

[22]Hughes C E. The Developing Endosperm of Triticum aestivum (L.): an Ultrastructural and Morphometric Study. PhD Dissertation of University of Nottingham, 1976

[23]Wei C-X(韦存虚), Zhang J(张军), Zhou W-D(周卫东), Chen Y-F(陈义芳), Xu R-G(许如根). Accumulation of storage protein and formation of protein body during barley endosperm development. Acta Agron Sin (作物学报), 2009, 35(2): 334–340 (in Chinese with English abstract)

[24]Luo Y-Y(罗玉英). Ultrastructural study on the division of amyloplast in pollen grain of Polygouatum samizui. J Chive Electr Microse Soc (电子显微学报), 1995, 3: 166–169 (in Chinese with English abstract)
[1] Zhai Sheng-Nan, Cao Xin-You, Li Hao-Sheng, Li Ji-Hu, Li Fa-Ji, Liu Jin-Dong, Xia Xian-Chun, Lyu Ying-Ying, Ma Rui-Feng, Wang Ying, Geng Hong-Wei, Liu Jian-Jun. Analysis of the genetic effects of allelic variation at the Pod-A1, Pod-D1, and Pod-2D loci on peroxidase activity in wheat grains [J]. Acta Agronomica Sinica, 2026, 52(6): 1593-1603.
[2] Xi Qian-Hui, Xu Zi-Yuan, Liu Meng-Meng, Wang Hong-Yi, Lang Kai-Lin, Jing Zhen-Hai, Chen Feng, Zhao Lei. Genome-wide association study and candidate gene prediction of grain copper content in wheat [J]. Acta Agronomica Sinica, 2026, 52(6): 1604-1617.
[3] Mao Jia-Qi, Huang Peng-Yu, Zhao Jia-Jia, Zheng Xing-Wei, Wu Bang-Bang, Hao Yu-Qiong, Qu Fei, Liu Cheng, Ma Peng-Tao, Zheng Jun. Evaluation of powdery mildew resistance in wheat cultivars and molecular detection of resistance genes in Shanxi province, China [J]. Acta Agronomica Sinica, 2026, 52(6): 1669-1681.
[4] Hu Chuan, Zhao Kai-Nan, Huang Xiu-Li, Wu Jin-Zhi, Ren Kai-Ming, Wang He-Zheng, Fu Guo-Zhan, Huang Ming, Li You-Jun. Effects of tillage methods and nitrogen rates on yield and quality of dryland wheat under one-off irrigation [J]. Acta Agronomica Sinica, 2026, 52(6): 1830-1846.
[5] Chen Xue-Yan, He Hua-Chuan, Li Zheng-Jia, Dong Xin-Pan, Li Ou-Qi, Liu Xiao-Yun, Li Dan-Ping, Chen Zhi-Wei, Liu Guo-Xia, Lyu Sheng-Yuan, Wu Yin-Ying, Zhao Zhen-Dong, Cao Xin-You, Wan He-Ping. Dynamic changes in root organic acid secretion and its transcriptional regulatory mechanisms in ‘Jimai 60’ seedlings under combined salinity-alkalinity stress in hydroponics [J]. Acta Agronomica Sinica, 2026, 52(6): 1859-1875.
[6] Gao Pei-Yang, Li Jin-Xuan, Dong Yu-Kui, Shi Yu, Zhang Zhen, Zhang Yong-Li. Response of wheat tillering and spike formation to nitrogen rate under supplementary irrigation based on soil moisture content [J]. Acta Agronomica Sinica, 2026, 52(6): 1847-1858.
[7] Zhang Xian-Feng, Guo Li-Jian, Li Kang-Chun, Kong Bin-Xue, Liu Yu-Fang, Che Zhuo, Yang De-Long. Identification of the ABHD6 gene family and development of functional markers for grain weight in wheat [J]. Acta Agronomica Sinica, 2026, 52(6): 1711-1727.
[8] Wang Zhuang-Zhuang, Wu Zi-Jun, Zhang Yong-Xin, Zhang Xin-Yuan, Yuan Li-Xue, Chen Ru-Xue, Liu Shi-Ju, Duan Jian-Zhao, Feng Wei, Wang Tong-Chao, Wang Yong-Hua. Optimized water-nitrogen synergy enhances winter wheat yield and nitrogen use efficiency in clay-loam fluvo-aquic soils of southeastern Henan, China [J]. Acta Agronomica Sinica, 2026, 52(5): 1501-1521.
[9] He Wan-Long, Geng Hong-Wei, Zhang Fei-Fei, Mikereayi·Ababaikere , Luo Zi-Yang, Li Peng-Cheng, Zhou Zhao-Yu, Cheng Yu-Kun. Development of a deep learning-based image recognition system for major wheat diseases [J]. Acta Agronomica Sinica, 2026, 52(5): 1401-1417.
[10] Zhang Zhen, Feng Lian-Jie, Shi Yu, Yu Zhen-Wen, Zhang Yong-Li. Yield formation of wheat with different ear types under water-saving supplementary irrigation conditions [J]. Acta Agronomica Sinica, 2026, 52(5): 1522-1535.
[11] Hou Si-Yu, Wang Guo-Cui, Wei Jin-Gui, Xie Wei-Xin, Yin Wen, Fan Zhi-Long, Chai Qiang, Hu Fa-Long. Effects of green manure combined with chemical nitrogen fertilizer on dry matter accumulation and yield formation of wheat in arid irrigation areas of northwestern China [J]. Acta Agronomica Sinica, 2026, 52(4): 1208-1219.
[12] Shang Yun-Qiu, Zhao Zhu, Chen Huan, Ding Yong-Gang, Qiao Yu-Qiang, Li Wei, Zhang Xiang-Qian, Cao Cheng-Fu, Du Shi-Zhou. Effects of long-term tillage practices on grain-filling and yield formation in rain-fed wheat [J]. Acta Agronomica Sinica, 2026, 52(4): 1236-1250.
[13] Qiao Yu-Xin, Li Cheng-Yue, Kang Xiao-Yu, Zhang Xin-Qi, Jia Shao-Hui, Liu Qian, Cao Ya-Li, Shi Xin-Rui, Hao Xing-Yu, Li Ping. Study on the effects of long-term no-tillage straw mulching on wheat yield improvement in dryland areas based on the APSIM model [J]. Acta Agronomica Sinica, 2026, 52(4): 1181-1192.
[14] Li Can, Zhang Xi-Wei, Zhu Bo-Tao, Zhang Pei-Pei. Functional characterization of wheat GSK kinase TaSK41 and screening for interacting proteins [J]. Acta Agronomica Sinica, 2026, 52(3): 677-687.
[15] Hou Jie, Fu Duo-Duo, Wu Hai-Feng, Hao Yu-Qiong, Zheng Xing-Wei, Wu Bang-Bang, Zhou Kai, Li Xiao-Hua, Zheng Jun, Zhao Jia-Jia. Chromosome diversity and its effects in wheat landraces from Shanxi province, China [J]. Acta Agronomica Sinica, 2026, 52(3): 746-763.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
Add: No. 80 Xueyuan South Rd, Beijing 100081, P. R. China E-mail: zwxb301@caas.cn
Supported by Beijing Magtech Co.Ltd