植物细胞壁组织化学定位染色方法和技术的比较研究

doi:10.7525/j.issn.1673-5102.2017.01.019

摘要/Abstract

摘要： 利用光学和荧光显微镜比较研究几种植物细胞壁组织化学定位染色方法和技术，结果表明：（1）硫酸消化法和硫酸氢黄连素-苯胺兰对染法研究凯氏带，对取材时间和部位要求高，建议两种方法配合使用，可相互印证是否具凯氏带；（2）苏丹7B染色法，蓝色激发光下不染色和硫酸氢黄连素-苯胺兰对染研究细胞壁栓质层3种方法中，不染色蓝色激发光下结果比苏丹7B染色法敏感显色，但苏丹7B染色法在普通光学显微镜下观察较为便捷；（3）木质化细胞壁染色方法中硫酸氢黄连素-苯胺兰对染法比间苯三酚-盐酸染色法易显色观察；（4）甲苯胺兰快速染色细胞壁取代常规苏丹Ⅲ/Ⅳ法，细胞边界和层次更清楚。

关键词: 组织化学, 凯氏带, 栓质层, 木质化

Abstract: We carried out the comparative studies on several methods and techniques for cell wall histochemistry under light and epifluorescence microscope. The results showed that:(1) To study Casparian bands using staining methods of sulphuric acid digestion and berberine hemisulfate-aniline, the requirements of material collecting time and part were high, therefore, it was recommended to use the two methods to mutual confirmation; (2) To study suberin lamellae using Sudan red 7B staining, and none staining and berberine hemisulfate-aniline blue staining under blue excited, none staining was more sensitive than Sudan red 7B, but Sudan red 7B staining is convenient under light microscope; (3) To study lignified walls, berberine hemisulfate-aniline staining was more sensitive than phloroglucinol-HCl; (4) Using TBO fast wall staining to replace Sudan Ⅲ/Ⅳ conventional staining, the wall borders and layers were clearer.

Key words: walls histochemistry, Casparian bands, suberin lamellae, lignified walls

中图分类号:

Q93-333

张霞, 胡露洁, 周存宇, 杨朝东. 植物细胞壁组织化学定位染色方法和技术的比较研究[J]. 植物研究, 2017, 37(1): 147-154.

ZHANG Xia, HU Lu-Jie, ZHOU Cun-Yu, YANG Chao-Dong. Comparative Study on Staining Methods and Techniques of Cell Wall Histochemistry[J]. Bulletin of Botanical Research, 2017, 37(1): 147-154.

参考文献

1. Schreiber L,Franke R B. Endodermis and exodermis in roots[R]. Chichester:John Wiley and Sons Ltd,2011,doi:10. 1002/9780470015902. a0002086. pub2.
2. Abiko T,Kotula L,Shiono K,et al. Enhanced formation of aerenchyma and induction of a barrier to radial oxygen loss in adventitious roots of Zea nicaraguensis contribute to its waterlogging tolerance as compared with maize(Zea mays ssp. mays)[J]. Plant,Cell & Environment,2012,35(9):1618-1630.
3. Yang C D,Zhang X,Li J K,et al. Anatomy and histochemistry of roots and shoots in wild Rice(Zizania latifolia Griseb.)[J]. Journal of Botany,2014:181727.
4. 杨朝东,张霞. 双穗雀稗(Paspalum distichum)通透性生理和茎解剖结构补充研究[J]. 植物研究,2013,33(5):564-568.Yang C D,Zhang X. Permeability and supplement structures of stems of Paspalum distichum[J]. Bulletin of Botanical Research,2013,33(5):564-568.
5. 张霞,杨朝东,宁国贵. 狗牙根和双穗雀稗根中质外体屏障结构发育过程的比较研究[J]. 湖北农业科学,2013,52(20):4991-4994.Zhang X,Yang C D,Ning G G. The developmental comparison of apoplastic barriers in Cynodon dactylon and Paspalum distichum roots[J]. Hubei Agricultural Sciences,2013,52(20):4991-4994.
6. Seago J L Jr.,Peterson C A,Enstone D E,et al. Development of the endodermis and hypodermis of Typha glauca Godr. and T. angustifolia L. roots[J]. Canadian Journal of Botany,1999,77(1):122-134.
7. Soukup A,Armstrong W,Schreiber L,et al. Apoplastic barriers to radial oxygen loss and solute penetration:a chemical and functional comparison of the exodermis of two wetland species,Phragmites australis and Glyceria maxima[J]. New Phytologist,2007,173(2):264-278.
8. Yang C D,Zhang X,Zhou C Y,et al. Root and stem anatomy and histochemistry of four grasses from the Jianghan floodplain along the Yangtze River,China[J]. Flora,2011,206(7):653-661.
9. 杨朝东,李守峰,邓仕明,等. 白茅解剖结构和屏障结构特征研究[J]. 草业学报,2015,24(3):213-218.Yang C D,Li S F,Deng S M,et al. Study of the anatomy and apoplastic barrier characteristics of Imperata cylindrica[J]. Acta Prataculturae Sinica,2015,24(3):213-218.
10. 杨朝东,李守峰,姚兰,等. 天胡荽(Hydrocotyle sibthorpioides)的解剖和屏障结构特征研究[J]. 草业学报,2015,24(7):139-145.Yang C D,Li S F,Yao L,et al. A study of anatomical structure and apoplastic barrier characteristics of Hydrocotyle sibthorpioides[J]. Acta Prataculturae Sinica,2015,24(7):139-145.
11. López-pérez L,Fernández-garcía N,Olmos E,et al. The phi thickening in roots of broccoli plants:an acclimation mechanism to salinity[J]. International Journal of Plant Sciences,2007,168(8):1141-1149.
12. Fernández-garcía N,López-pérez L,Hernandez M,et al. Role of phi cells and the endodermis under salt stress in Brassica oleracea[J]. New Phytologist,2009,181(2):347-360.
13. Meyer C J,Peterson C A. Casparian bands occur in the periderm of Pelargonium hortorum stem and root[J]. Annals of Botany,2011,107(4):591-598.
14. Mcmanus H A,Seago J L,Jr.,Marsh L C. Epifluorescent and histochemical aspects of shoot anatomy of Typha latifolia L.,Typha angustifolia L. and Typha glauca Godr[J]. Annals of Botany,2002,90(4):489-493.
15. Vecchia F D,Cuccato F,Rocca N L,et al. Endodermis-like sheaths in the submerged freshwater macrophyte Ranunculus trichophyllus Chaix[J]. Annals of Botany,1999,83(1):93-97.
16. Watanabe H,Saigusa M,Morita S. Identification of Casparian bands in the mesocotyl and lower internodes of rice(Oryza sativa L.) seedlings using fluorescence microscopy[J]. Plant Production Science,2006,9(4):390-394.
17. Mertz R A,Brutnell T P. Bundle sheath suberization in grass leaves:multiple barriers to characterization[J]. Journal of Experimental Botany,2014,65(13):3371-3380.
18. Enstone D E,Peterson C A,Ma F S. Root endodermis and exodermis:structure,function,and responses to the environment[J]. Journal of Plant Growth Regulation,2002,21(4):335-351.
19. 杨朝东,张霞,刘国锋,等. 植物根中质外体屏障结构和生理功能研究进展[J]. 植物研究,2013,33(1):114-119.Yang C D,Zhang X,Liu G F,et al. Progress on the structure and physiological functions of apoplastic barriers in root[J]. Bulletin of Botanical Research,2013,33(1):114-119.
20. Fich E A,Segerson N A,Rose J K C. The plant polyester cutin:biosynthesis,structure,and biological roles[J]. Annual Review of Plant Biology,2016,67(1):207-233.
21. Brundrett M C,Enstone D E,Peterson C A. A berberine-aniline blue fluorescent staining procedure for suberin,lignin,and callose in plant tissue[J]. Protoplasma,1988,146(2):133-142.
22. Brundrett M C,Kendrick B,Peterson C A. Efficient lipid staining in plant material with Sudan red 7B or Fluoral yellow 088 in polyethylene glycol-glycerol[J]. Biotechnic & Histochemistry,1991,66(3):111-116.
23. Pauluzzi G,Divol F,Puig J,et al. Surfing along the root ground tissue gene network[J]. Developmental Biology,2012,365(1):14-22.
24. Naseer S,Lee Y,Lapierre C,et al. Casparian strip diffusion barrier in Arabidopsis is made of a lignin polymer without suberin[J]. Proceedings of the National Academy of Science of the United States of America,2012,109(25):10101-10106.
25. Alassimone J,Roppolo D,Geldner N,et al. The endodermis-development and differentiation of the plant's inner skin[J]. Protoplasma,2012,249(3):433-443.
26. Geldner N. The endodermis[J]. Annual Review of Plant Biology,2013,64(1):531-558.
27. Roppolo D,De Rybel B,Tendon V D,et al. A novel protein family mediates Casparian strip formation in the endodermis[J]. Nature,2011,473(7347):380-383.
28. Lee Y,Rubio M C,Alassimone J,et al. A mechanism for localized lignin deposition in the endodermis[J]. Cell,2013,153(2):402-412.
29. Jensen W A. Botanical histochemistry:principles and practice[M]. San Francisco,CA:W. H. Freeman,1962.
30. Meyer C J,Seago J L Jr.,Peterson C A. Environmental effects on the maturation of the endodermis and multiseriate exodermis of Iris germanica roots[J]. Annals of Botany,2009,103(5):687-702.
31. Roppolo D,Boeckmann B,Pfister A,et al. Functional and evolutionary analysis of the Casparian strip membrane domain protein family[J]. Plant Physiology,2014,165(4):1709-1722.
32. Pfister A,Barberon M,Alassimone J,et al. A receptor-like kinase mutant with absent endodermal diffusion barrier displays selective nutrient homeostasis defects[J]. eLIFE,2014,3:e03115.
33. Shiono K,Ogawa S,Yamazaki S,et al. Contrasting dynamics of radial O₂-loss barrier induction and aerenchyma formation in rice roots of two lengths[J]. Annals of Botany,2011,107(1):89-99.
34. Buschhaus C,Jetter R. Composition differences between epicuticular and intracuticular wax substructures:how do plants seal their epidermal surfaces[J]?Journal of Experimental Botany,2011,62(3):841-853.
35. Beisson F,Li-Beisson Y,Pollard M. Solving the puzzles of cutin and suberin polymer biosynthesis[J]. Current Opinion in Plant Biology,2012,15(3):329-337.
36. Schreiber L. Transport barriers made of cutin,suberin and associated waxes[J]. Trends in Plant Science,2010,15(10):546-553.
37. Watanabe K,Nishiuchi S,Kulichikhin K,et al. Does suberin accumulation in plant roots contribute to waterlogging tolerance[J]?Frontiers in Plant Science,2013,4(2):57-60.
38. 张晓民. 禾本科植物细胞壁的木质化和阿魏酰化[J]. 中国野生植物资源,2011,30(6):7-13.Zhang X M. Lignification and feruloylation in the cell wall of grass family[J]. Chinese Wild Plant Resources,2011,30(6):7-13.

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