Summary
A fluorescent staining procedure to detect suberin, lignin and callose in plants has been developed. This procedure greatly improves on previous methods for visualizing Casparian bands in root exodermal and endodermal cells, and performs equally well on a variety of other plant tissues. Berberine was selected as the most suitable replacement forChelidonium majus root extract after comparing the staining properties of the extract with those of four of its constituent alkaloids. Aniline blue counterstaining efficiently quenched unwanted background fluorescence and nonspecific berberine staining, while providing a fluorochrome for callose. When used with multichambered holders which allow simultaneous processing of freehand sections, this efficient staining procedure facilitates morphological studies involving large numbers of samples.
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Abbreviations
- ISCC-NBS:
-
Inter-Society Color Council-National Bureau of Standards
- UV:
-
ultraviolet light
References
Combes RD, Havelant-Smith RB (1982) A review of the genotoxicity of food, drug and cosmetic colours and other azo, triphenylmethane and xanthene dyes. Mutat Res 98: 101–248
Currier HB (1957) Callose substance in plant cells. Am J Bot 44: 478–488
Elisei FG (1941) Richerche microfluoroscopiche sui punti de Caspary. Pavia Univ Inst Bot Atti 13: 1–64
Enerbach L (1974) Berberine sulphate binding to mast cell polyanions: a cytofluorometric method for the quantitation of heparin. Histochemistry 42: 301–313
Evans NA, Hoyne PA (1982) A fluorochrome from aniline blue: structure, synthesis and fluorescence properties. Aust J Chem 35: 2571–2575
— —,Stone BA (1984) Characteristics and specificity of the interaction of a fluorochrome from aniline blue (Sirofluor) with polysaccharides. Carbohydr Polymers 4: 215–230
French JC (1987) Systematic occurrence of a sclerotic hypodermis in roots ofAraceae. Am J Bot 74: 891–903
Frohlich MW (1984) Freehand sectioning with Parafilm. Stain Technol 59: 61–62
Jensen WA (1962) Botanical histochemistry: principles and practice. WH Freeman, San Francisco
Johansen DA (1940) Plant microtechnique. McGraw-Hill, New York
Johnson GD, Davidson RS, McNamee KC, Russell G, Goodwin D, Holborow E (1982) Fading of immunofluorescence during microscopy: a study of the phenomenon and its remedy. J Immunol Methods 55: 231–242
Karabestos JH, Pappelis AJ, Russo VM (1987) Visualization of halos in the epidermal cell wall ofAllium cepa caused byColletotrichum dematium f.circinans andBotrytis allii using fluorochromes. Mycopathologia 97: 137–141
Kelly KL (1965) ISCC-NBS color-name charts illustrated with centroid colors, standard sample #2106, suppl. Natl Bur Standards Circ 553. U.S. Government Printing Office, Washington, D.C.
O'Brien TP, McCully ME (1981) The study of plant structure principles and selected methods. Termarcarphi Pty, Melbourne
Peirson DR, Dumbroff EB (1969) Demonstration of a complete Casparian strip inAvena andIpomoea by a fluorescent staining technique. Can J Bot 47: 1869–1871
Peterson CA (1988) Exodermal Casparian bands: their significance for ion uptake by roots. Physiol Plant 72: 204–208
—,Emanuel ME, Wilson CW (1982) Identification of a Casparian band in the hypodermis of onion and corn roots. Can J Bot 60: 1529–1535
—,Peterson RL, Robards AW (1978) A correlated histochemical and ultrastructural study of the epidermis and hypodermis of onion roots. Protoplasma 96: 1–21
Philogene BJR, Arnason JT, Towers GHN, Abramowski Z, Campos F, Champagne D, McLachlan D (1984) Berberine: a naturally occurring phototoxic alkaloid. J Chem Ecol 10: 115–123
Stahl E, Schild W (1981) Pharmazeutische Biologie. Drogenanalyse, II Inhaltsstoffe und Isolierungen. Gustav Fischer, New York, pp 69–73
Tomlinson PB (1969) Anatomy of the monocotyledons, III Commelinales-Zingiberales. Clarendon Press, Oxford
Valnes K, Brandtzaeg P (1985) Retardation of immunofluorescence fading during microscopy. J Histochem Cytochem 33: 755–761
Weerdenburg CA, Peterson CA (1983) Structural changes in phi thickenings during primary and secondary growth in roots 1. Apple (Pyrus malus)Rosaceae. Can J Bot 61: 2570–2576
Wilcox W (1954) Primary organization of active and dormant roots of noble fir,Abies procera. Am J Bot 41: 812–821
Wilson CA, Peterson CA (1983) Chemical composition of the epidermal, hypodermal, endodermal and intervening cortical cell walls of various plant roots. Ann Bot 51: 759–769
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Brundrett, M.C., Enstone, D.E. & Peterson, C.A. A berberine-aniline blue fluorescent staining procedure for suberin, lignin, and callose in plant tissue. Protoplasma 146, 133–142 (1988). https://doi.org/10.1007/BF01405922
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DOI: https://doi.org/10.1007/BF01405922