Subscribe to RSS
DOI: 10.1055/s-0029-1185801
© Georg Thieme Verlag KG Stuttgart · New York
Characterization and Immunolocalization of Arabinogalactan-Proteins in Roots of Echinacea purpurea
Publication History
received January 12, 2009
revised April 16, 2009
accepted May 11, 2009
Publication Date:
26 June 2009 (online)
Abstract
From the high molecular weight fraction of an aqueous extract from roots of Echinacea purpurea L. Moench, arabinogalactan-proteins (AGPs), a class of proteoglycans proposed to be involved in cell differentiation and plant growth, were purified and characterized with regard to amino acid composition and structure of the polysaccharide moiety. The protein content of the AGP was 5.0 % (w/w) with the dominating amino acids Glx, Hyp, Asx, Ser, Thr and Ala. The highly branched polysaccharide moiety shows a linkage composition typical of AGPs with 1,3-, 1,6- and 1,3,6-linked galactopyranosyl residues and arabinofuranosyl residues predominantly as terminal and 1,5-linked residues. Terminal units of glucuronopyranose acid were also detected. Furthermore, a new method for the localization of AGPs in plant tissue has been developed. The synthetic (β-D-Glc)3 Yariv phenylgycoside (βGlcY) is known to specifically bind to AGPs. For immunolocalization, polyclonal βGlcY-antibodies have been generated and were used to label Yariv-treated thin sections of roots from E. purpurea. After addition of the FITC-conjugated secondary antibody, the sections were analyzed by confocal laser scanning microscopy. AGPs are detected mainly in the central cylinder in the area of the xylem. Cell walls of vessels and tracheids are strongly labelled, especially at the inner area of the wall. Furthermore, there is intense labelling of the pit canals.
Key words
Echinacea purpurea - Asteraceae - immunolocalization - arabinogalactan‐proteins - β‐D‐glucosyl Yariv reagent
References
- 1 Hoheisel O, Sandberg M, Bertram S, Bulitta M, Schäfer M. Echinagard treatment shortens the course of the common cold: a double-blind, placebo-controlled clinical trial. Eur J Clin Res. 1997; 9 261-268
-
2 Bauer R.
The Echinacea story – the scientific development of an herbal immunostimulant. Prendergast HD, Etkin NL, Harris DR, Houghton PJ Plants for food and medicine. London, Kiew; Royal Botanic Gardens 1998: 317-332 - 3 Classen B, Witthohn K, Blaschek W. Characterization of an arabinogalactan-protein isolated from pressed juice of Echinacea purpurea by precipitation with the β-glucosyl Yariv reagent. Carbohydr Res. 2000; 327 497-504
- 4 Thude S, Classen B. High molecular weight constituents from roots of Echinacea pallida: an arabinogalactan-protein and an arabinan. Phytochemistry. 2005; 66 1026-1032
- 5 Alban S, Classen B, Brunner G, Blaschek W. Differentiation between the complement modulating effects of an arabinogalactan-protein from Echinacea purpurea and heparin. Planta Med. 2002; 68 1118-1124
- 6 Classen B, Thude S, Blaschek W, Wack M, Bodinet C. Immunomodulatory effects of arabinogalactan-proteins from Baptisia and Echinacea. Phytomedicine. 2006; 13 688-694
- 7 Thude S, Classen B, Blaschek W, Barz D, Thude H. Binding studies of an arabinogalactan-protein from Echinacea purpurea to leucocytes. Phytomedicine. 2006; 13 425-427
- 8 Clarke A E, Anderson R L, Stone B A. Form and function of arabinogalactans and arabinogalactan-proteins. Phytochemistry. 1979; 18 521-540
- 9 Nothnagel E A. Proteoglycans and related components in plant cells. Int Rev Cytol. 1997; 174 195-291
-
10 Johnson K, Jones B, Schultz C J, Bacic A.
Non-enzymic cell wall (glyco)proteins. Rose J The plant cell wall. UK; Blackwell Publishing 2003: 111-154 - 11 Seifert G J, Roberts K. The biology of arabinogalactan proteins. Annu Rev Plant Biol. 2007; 58 137-161
- 12 Kreuger M, van Holst G J. Arabinogalactan-protein epitopes in somatic embryogenesis of Daucus carota L. Planta. 1995; 197 135-141
- 13 Hu Y, Qin Y, Zhao J. Localization of an arabinogalactan protein epitope and the effects of Yariv phenylglycoside during zygotic embryo development of Arabidopsis thaliana. Protoplasma. 2006; 229 21-31
- 14 Serpe M D, Nothnagel E A. Effects of Yariv phenylglycosides on Rosa cell suspensions: evidence for the involvement of arabinogalactan-proteins in cell proliferation. Planta. 1994; 193 542-550
- 15 Gao M, Showalter A M. Yariv reagent treatment induces programmed cell death in Arabidopsis cell cultures and implicates arabinogalactan protein involvement. Plant J. 1999; 19 321-331
- 16 Guan Y, Nothnagel E A. Binding of arabinogalactan proteins by Yariv phenylglycoside triggers wound-like responses in Arabidopsis cell cultures. Plant Physiol. 2004; 135 1346-1366
- 17 Majewska-Sawka A, Nothnagel E A. The multiple roles of arabinogalactan proteins in plant development. Plant Physiol. 2000; 122 3-9
- 18 Youl J J, Bacic A, Oxley D. Arabinogalactan-proteins from Nicotiana alata and Pyrus communis contain glycosylphosphatidylinositol membrane anchors. Proc Natl Acad Sci USA. 1998; 95 7921-7926
- 19 Knox J P. The use of antibodies to study the architecture and developmental regulation of plant cell walls. Int Rev Cytol. 1997; 171 79-120
- 20 Knox J P. Revealing the structural and functional diversity of plant cell walls. Curr Oppin Plant Biol. 2008; 11 308-313
- 21 Blake A W, Marcus S E, Copeland J E, Blackburn R S, Knox J P. In situ analysis of cell wall polymers associated with phloem fibre cells in stems of hemp, Cannabis sativa L. Planta. 2008; 228 1-13
- 22 Yariv J, Rapport M M, Graf L. Interaction of glycosides and saccharides with antibody to the corresponding phenylazo glycosides. Biochem J. 1962; 85 383-388
- 23 Putoczki T L, Pettolino F, Griffin M DW, Moeller R, Gerrard J A, Bacic A, Jackson S L. Characterization of the structure, expression and function of Pinus radiata D. Don arabinogalactan-proteins. Planta. 2007; 226 1131-1142
- 24 Classen B, Mau S L, Bacic A. The arabinogalactan-proteins from pressed juice of Echinacea purpurea belong to the “hybrid” class of hydroxyproline-rich glycoproteins. Planta Med. 2005; 71 59-66
- 25 Taylor R L, Conrad H E. Stoichiometric depolymerization of polyuronides and glycosaminoglycuronans to monosaccharides following reduction of their carbodiimide-activated carboxyl group. Biochemistry. 1972; 11 1383-1388
- 26 Gleeson P A, Clarke A E. Structural studies on the major component of Gladiolus style mucilage, an arabinogalactan-protein. Biochem J. 1979; 181 607-621
- 27 Blakeney A B, Harris P J, Henry R J, Stone B A. A simple and rapid preparation of alditol acetates for monosaccharide analysis. Carbohydr Res. 1983; 113 291-299
- 28 Harris P J, Henry R J, Blakeney A B, Stone B A. An improved procedure for the methylation analysis of oligosaccharides and polysaccharides. Carbohydr Res. 1984; 127 59-73
- 29 Woo K L, Ahan Y K. Determination of protein amino acids as benzylthiocarbamyl derivatives compared with phenylthiocarbamyl derivatives by reversed-phase high-performance liquid chromatography, ultraviolet detection and precolumn derivatization. J Chromatogr. 1996; 740 41-50
- 30 Stegemann H, Stalder K H. Determination of hydroxyproline. Clin Chim Acta. 1967; 18 267-273
- 31 Valnes K, Brandtzaeg P. Retardation of immunofluorescence fading during microscopy. J Histochem Cytochem. 1985; 33 755-761
- 32 Aspinall G O. Classification of polysaccharides. The polysaccharides, Vol. 1. New York; Academic Press 1982: 340
- 33 Willats W, McCartney L, Mackie W, Knox P. Pectin: cell biology and prospects for functional analysis. Plant Mol Biol. 2001; 47 9-27
- 34 Mollard M, Basile D. Acacia senegal cells cultured in suspension secrete a hydroxyproline-deficient arabinogalactan protein. Plant Physiol Biochem. 2000; 32 703-709
- 35 Lee K, Sakata Y, Mau S, Pettolino F, Bacic A, Quatrano R, Knight C, Knox P. Arabinogalactan proteins are required for apical cell extension in the moss Physcomitrella patens. Plant Cell. 2005; 17 3051-3065
- 36 Showalter A. Arabinogalactan-proteins: structure, expression and function. Cell Mol Life Sci. 2001; 58 1399-1417
-
37 Bacic A, Currie G, Gilson P, Mau S L, Oxley D, Schultz C J, Sommer-Knudsen J, Clarke A E.
Structural classes of arabinogalactan-proteins. Nothnagel EA, Bacic A, Clarke AE Cell and developmental biology of arabinogalactan-proteins. Dordrecht; Kluwer 2000: 11-23 - 38 Osman M E, Menzies A R, Albo Martin B, Williams P A, Phillips G O, Baldwin T C. Characterization of gum arabic fractions obtained by anion exchange chromatography. Phytochemistry. 1995; 38 409-417
- 39 Girault R, His I, Andeme-Onzighi C, Driouich A, Morvan C. Identification and partial characterization of proteins and proteoglycans encrusting the secondary cell walls of flax fibres. Planta. 2000; 211 256-264
- 40 Classen B. Characterization of an arabinogalactan-protein from suspension culture of Echinacea purpurea. Plant Cell Tissue Organ Cult. 2007; 88 267-275
- 41 Motose H, Sugiyama M, Fukuda H. A proteoglycan mediates inductive interaction during plant vascular development. Nature. 2004; 429 873-878
- 42 Willats W, Knox J P. A role for arabinogalactan-proteins in plant cell expansion: evidence from studies on the interaction of β-glucosyl Yariv reagent with seedlings of Arabidopsis thaliana. Plant J. 1996; 9 919-925
- 43 Thompson H JM, Knox J P. Stage-specific responses of embryogenic carrot cell suspension cultures to arabinogalactan protein-binding β-glucosyl Yariv reagent. Planta. 1998; 205 32-38
- 44 Chaves I, Regalado A P, Chen M, Ricardo C P, Showalter A M. Programmed cell death induced by (β‐D-galactosyl)3 Yariv reagent in Nicotiana tabacum BY‐2 suspension-cultured cells. Physiol Plant. 2002; 116 548-553
- 45 Serpe M D, Nothnagel E A. Arabinogalactan-proteins in the multiple domains of the plant cell surface. Adv Bot Res. 1999; 30 207-289
- 46 Pennell R I, Knox J P, Scofield G N, Selvendran R R, Roberts K. A family of abundant plasma membrane-associated glycoproteins related to the arabinogalactan proteins is unique to flowering plants. J Cell Biol. 1989; 108 1967-1977
- 47 Basile D V, Basile M R. The occurrence of cell wall-associated arabinogalactan proteins in the Hepaticae. Bryologist. 1988; 90 401-404
- 48 Battaglia M, Solorzano R M, Hernandez M, Cuellar-Ortiz S, Garcia-Gomez B, Marquez J, Covarrubias A A. Proline-rich cell wall proteins accumulate in growing regions and phloem tissue in response to water deficit in common bean seedlings. Planta. 2007; 225 1121-1133
- 49 Samson M R, Jongeneel R, Klis F M. Arabinogalactan protein in the extracellular space of Phaseolus vulgaris hypocotyls. Phytochemistry. 1984; 23 493-496
- 50 Knox J P, Linstead P J, Peart J, Cooper C, Roberts K. Developmentally regulated epitopes of cell-surface arabinogalactan proteins and their relation to root-tissue pattern-formation. Plant J. 1991; 1 317-326
- 51 Dolan L, Roberts K. Secondary thickening in roots of Arabidopsis thaliana – anatomy and cell-surface changes. New Phytol. 1995; 131 121-128
- 52 Dolan L, Linstead P, Roberts K. An AGP epitope distinguishes a central metaxylem initial from other vascular initials in the Arabidopsis root. Protoplasma. 1995; 189 149-155
- 53 Casero P J, Casimiro I, Knox J P. Occurence of cell surface arabinogalactan-protein and extensin epitopes in relation to pericycle and vascular tissue development in the root apex of four species. Planta. 1998; 204 252-259
- 54 Van Hengel A J, Roberts K. AtAGP30, an arabinogalactan-protein in the cell walls of the primary root, plays a role in root regeneration and seed germination. Plant J. 2003; 36 256-270
- 55 Zhang Y, Brown G, Whetten R, Loopstra C A, Neale D, Kieliszewski M J, Sederoff R R. An arabinogalactan protein associated with secondary cell wall formation in differentiating xylem of loblolly pine. Plant Mol Biol. 2003; 52 91-102
- 56 Kieliszewski M J. The latest hype on Hyp-O-glycosylation codes. Phytochemistry. 2001; 57 319-323
- 57 Ito S, Suzuki Y, Miyamoto K, Ueda J, Yamaguchi I. AtFLA11, a fasciclin-like arabinogalactan-protein, specifically localized in sclerenchym cells. Biosci Biotechnol Biochem. 2005; 69 1963-1969
- 58 Roach M J, Deyholos M K. Microarray analysis of flax (Linum usitatissimum L.) stems identifies transcripts enriched in fibre-bearing phloem tissues. Mol Genet Genomics. 2007; 278 149-165
- 59 Schindler T, Bergfeld R, Schopfer P. Arabinogalactan proteins in maize coleoptiles: developmental relationship to cell death during xylem differentiation but not to extensin growth. Plant J. 1995; 7 25-36
- 60 Stacey N J, Roberts K, Carpita N C, Wells B, McCann M C. Dynamic changes in cell surface molecules are very early events in the differentiation of mesophyll cells from Zinnia elegans into tracheary elements. Plant J. 1995; 8 891-906
- 61 Gao M, Showalter A M. Immunolocalization of LeAGP‐1, a modular arabinogalactan-protein, reveals its developmentally regulated expression in tomato. Planta. 2000; 210 865-874
- 62 Loopstra C A, Puryear J D, No E G. Purification and cloning of an arabinogalactan-protein from xylem of loblolly pine. Planta. 2000; 210 686-689
- 63 Yang S H, Wang H, Sathyan P, Stasolla C, Loopstra C A. Real-time RT‐PCR analysis of loblolly pine (Pinus taeda) arabinogalactan protein and arabinogalactan protein-like genes. Physiol Plant. 2005; 124 91-106
- 64 Liu C, Mehdy M C. A nonclassical arabinogalactan protein gene highly expressed in vascular tissues, AGP31, is transcriptionally repressed by methyl jasmonic acid in Arabidopsis. Plant Physiol. 2007; 145 863-874
- 65 Dahiya P, Findlay K, Roberts K, McCann M C. A fsciclin-domain containing gene, ZeFLA11, is expressed exclusively in xylem elements that have reticulate wall thickenings in the stem vascular system of Zinnia elegans cv Envy. Planta. 2006; 223 1281-1291
- 66 Lafarguette F, Leple J C, Dejardin A, Laurans F, Costa G, Lesage-Descauses M C, Pilate G. Poplar genes encoding fasciclin-like arabinogalactan proteins are highly expressed in tension wood. New Phytol. 2004; 164 107-121
- 67 Orfila C, Knox J P. Spatial regulation of pectic polysaccharides in relation to pit fields in cell walls of tomato fruit pericarp. Plant Physiol. 2000; 122 775-781
PD Dr. Birgit Classen
Department of Pharmaceutical Biology
Pharmaceutical Institute
Christian-Albrechts-University of Kiel
Gutenbergstraße 76
24118 Kiel
Germany
Phone: + 49 43 18 80 11 30
Fax: + 49 43 18 80 11 02
Email: bclassen@pharmazie.uni-kiel.de