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Structural analysis of wheat wax (Triticum aestivum, c.v. ‘Naturastar’ L.): from the molecular level to three dimensional crystals

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Abstract

In order to elucidate the self assembly process of plant epicuticular waxes, and the molecular arrangement within the crystals, re-crystallisation of wax platelets was studied on biological and non-biological surfaces. Wax platelets were extracted from the leaf blades of wheat (Triticum aestivum L., c.v. ‘Naturastar’, Poaceae). Waxes were analysed by gas chromatography (GC) and mass spectrometry (MS). Octacosan-1-ol was found to be the most abundant chemical component of the wax mixture (66 m%) and also the determining compound for the shape of the wax platelets. The electron diffraction pattern showed that both the wax mixture and pure octacosan-1-ol are crystalline. The re-crystallisation of the natural wax mixture and the pure octacosan-1-ol were studied by scanning tunnelling microscopy (STM), atomic force microscopy (AFM), scanning electron microscopy (SEM), and transmission electron microscopy (TEM). Crystallisation of wheat waxes and pure octacosano-1-ol on the non polar highly ordered pyrolytic graphite (HOPG) led to the formation of platelet structures similar to those found on the plant surface. In contrast, irregular wax morphologies and flat lying plates were formed on glass, silicon, salt crystals (NaCl) and mica surfaces. Movement of wheat wax through isolated Convallaria majalis cuticles led to typical wax platelets of wheat, arranged in the complex patterns typical for C. majalis. STM of pure octacosan-1-ol monolayers on HOPG showed that the arrangement of the molecules strictly followed the hexagonal structure of the substrate crystal. Re-crystallisation of wheat waxes on non-polar crystalline HOPG substrate showed that technical surfaces could be used to generate microstructured, biomimetic surfaces. AFM and SEM studies proved that a template effect of the substrate determined the orientation of the re-grown crystals. These effects of the structure and polarity of the substrate on the morphology of the epicuticular waxes are relevant for understanding interactions between biological as well as technical surfaces and waxes.

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Abbreviations

STM:

Scanning tunnelling microscopy

AFM:

Atomic force microscopy

SEM:

Scanning electron microscopy

TEM:

Transmission electron microscopy

TMS:

Trimethylsilyl

HOPG:

Highly ordered pyrolytic graphite

GC:

Gas chromatography

MS:

Mass spectrometry

References

  • Baker EA (1982) Chemistry and morphology of plant epicuticular waxes. In: Cutler DF, Alvin KL, Price CE (eds) The plant cuticle. Academic, London, pp 139–166

    Google Scholar 

  • Bargel H, Barthlott W, Koch K, Schreiber L, Neinhuis C (2004) Plant cuticles: multifunctional interfaces between plant and environment. In: Hemsley AR, Poole I (eds) Evolution of plant physiology. Academic, London, pp 171–194

    Google Scholar 

  • Barthlott W (1990) Scanning electron microscopy of the epidermal surface in plants. In: Claugher D (ed) Scanning electron microscopy in taxonomy and functional morphology. Syst. Assoc. Spec. Vol. No. 41, Clarendon Press, pp 69–94

  • Barthlott W, Fröhlich D (1983) Mikromorphologie und Orientierungsmuster epicuticularer Wachs-Kristalloide: Ein neues systematisches Merkmal bei Monokotylen. Plant Syst Evol 142:171–185

    Article  Google Scholar 

  • Barthlott W, Neinhuis C (1997) Purity of the sacred lotus or escape from contamination in biological surfaces. Planta 202:1–7

    Article  CAS  Google Scholar 

  • Barthlott W, Neinhuis C, Cutler D, Ditsch F, Meusel I, Theisen I, Wilhelmi H (1998) Classification and terminology of plant epicuticular waxes. Bot J Linnean Soc 126:237–260

    Article  Google Scholar 

  • Barthlott W, Theisen I, Borsch T, Neinhuis C (2003) Epicuticular waxes and vascular plant systematics: integrating micromorphological and chemical data. In: Stuessy TF, Mayer V, Hörandl E (eds) Deep morphology: toward a renaissance of morphology in plant systematics. Reg Veg Gantner Verlag Ruggell/Liechtenstein 141, pp 189–206

  • Batra IP, Garcian N, Rohrer H, Salmink H, Stoll E, Ciraci S (1987) A study of graphite surface with STM and electronic-structure calculations. Surf Sci 181:126–138

    Article  CAS  Google Scholar 

  • Bianchi G (1995) Plant waxes. In: Hamilton RJ (ed) Waxes: Chemistry, molecular biology and functions. The Oily Press, Glasgow, pp 175–222

    Google Scholar 

  • Bianchi G, Corbellini A (1977) Epicuticular wax of Triticum aestivum Demar 4. Phytochemistry 16:943–945

    Article  CAS  Google Scholar 

  • Bianchi G, Lupotto E, Corbellin M (1979) Composition of epicuticular waxes of Triticum aestivum demar 4 from different parts of the plant. Agrochimica 23:96–102

    CAS  Google Scholar 

  • Buchholz S, Rabe JM (1992) Molecular imaging of alkanol monolayers on graphite. Angew Chem 31:189–191

    Article  Google Scholar 

  • Canet D, Rohr R, Chamel A, Guillain F (1996) Atomic force microscopy study of isolated ivy leaf cuticles observed directly and after embedding in Epon. New Phytol 134:571–577

    Article  Google Scholar 

  • Chambers TC, Ritchie IM, Booth MA (1975) Chemical models for plant wax morphogenesis. New Phytol 77:43–49

    Article  Google Scholar 

  • Cyr DM, Venkataraman B, Flynn GW (1996) STM investigations of organic molecules physisorbed at the liquid–solid interface. Chem Mater 8:1600–1615

    Article  CAS  Google Scholar 

  • De Feyter S, De Schryver FC (2003) Two-dimensional supramolecular self-assembly probed by scanning tunnelling microscopy. Chem Soc Rev 32:139–150

    Article  PubMed  CAS  Google Scholar 

  • Dorset D (1999) Development of lamellar structures in natural waxes – an electron diffraction investigation. J Phys D-Appl Phys 32:1276–1280

    Article  CAS  Google Scholar 

  • Ensikat HJ, Barthlott W (1993) Liquid substitution: a versatile procedure for SEM specimen preparation of biological materials without drying or coating. J Microsc 172:195–203

    PubMed  CAS  Google Scholar 

  • Ensikat HJ, Neinhuis C, Barthlott W (2000) Direct access to plant epicuticular wax crystals by a new mechanical isolation method. Int J Plant Sci 161:143–148

    Article  PubMed  Google Scholar 

  • Jeffree CE (1974) A method for recrystallizing selected components of plant epicuticular waxes as surfaces for the growth of microorganisms. Trans Brit Mycol Soc 63:626–628

    Article  Google Scholar 

  • Jeffree CE (1996) Structure and ontogeny of plant cuticles. In: Kerstiens G (ed) Plant cuticles: an integrated functional approach. BIOS Scientific Pub, Oxford, pp 33–82

    Google Scholar 

  • Jeffree CE, Baker EA, Holloway PJ (1975) Ultrastructure and recrystallization of plant epicuticular waxes. New Phytol 75:539–549

    Article  Google Scholar 

  • Jeffree CE, Baker EA, Holloway PJ (1976) Origins of the fine structure of plant epicuticular waxes. In: Dickinson CH and Preece TF (eds) Microbiol aerial plant surf. Academic, London, pp 119–158

    Google Scholar 

  • Jetter R, Riederer M (1994) Epicuticular crystals of nonacosan-10-ol: In-vitro reconstitution and factors influencing crystal habits. Planta 195:257–270

    Article  CAS  Google Scholar 

  • Jetter R, Riederer M (1995) In vitro reconstitution of epicuticular wax crystals: formation of tubular aggregates by long chain secondary alkendiols. Bot Acta 108:111–120

    CAS  Google Scholar 

  • Jetter R, Schäffer S (2001) Chemical composition of the Prunus laurocerasus leaf surface. Dynamic changes of the epicuticular wax film during leaf development. Plant Physiol 126:1725–1737

    Article  PubMed  CAS  Google Scholar 

  • Koch K, Neinhuis C, Ensikat HJ, Barthlott W (2004) Self assembly of epicuticular waxes on plant surfaces investigated by Atomic Force Microscopy (AFM). J Exp Bot 55:711–718

    Article  PubMed  CAS  Google Scholar 

  • Kolattukudy PE (1980) Cutin, suberin, and waxes. In: Stumpf PK (ed) Lipids: structure and function. Academic, London, pp 571–646

    Google Scholar 

  • Kolattukudy PE (1996) Biosynthetic pathways of cutin and waxes. In: Kerstiens G (ed) Plant cuticles an integrated functional appraoch. Bios Scientific, Oxford, pp 83–108

    Google Scholar 

  • Kolattukudy PE (2001) Polyesters in higher plants. In Scheper T (ed) Biochemical engineering/biotechnology. Springer, Berlin Heidelberg New York, pp 4–49

    Google Scholar 

  • Kunst L, Samuels AS (2003) Biosynthesis and secretion of plant cuticular wax. Prog in Lipid Res 42:51–80

    Article  CAS  Google Scholar 

  • Le Poulennec C, Cousty J, Xie ZX, Miokowski C (2000) Self organisation of physisorbed secondary alcohol molecules on a graphite surface. Surface Sci 448:93–100

    Article  CAS  Google Scholar 

  • Matas AJ, MJ Sanz, Heredia A (2003) Studies on the structure of plant wax nonacosan-10-ol, the main component of epicuticular wax conifers. Int J Biol Macromol 33:31–35

    Article  PubMed  CAS  Google Scholar 

  • Meusel I, Neinhuis C, Markstädter K, Barthlott W (1999) Ultrastructure, chemical composition and recrystallization of epicuticular waxes: transversely ridged rodlets. Can J Bot 77:706–720

    Article  Google Scholar 

  • Meusel I, Barthlott W, Kutzke H, Barbier B (2000) Crystallographic studies of plant waxes. Powder Diffract 15:123–129

    CAS  Google Scholar 

  • Neinhuis C, Koch K, Barthlott W (2001) Movement and regeneration of epicuticular waxes through plant cuticles. Planta 213:427–434

    Article  PubMed  CAS  Google Scholar 

  • Rabe JP, Buchholz S (1991) Commensurability and mobility in 2-dimensional molecular patterns on graphite. Science 253:424–427

    Article  PubMed  CAS  Google Scholar 

  • Reynhardt EC, Riederer M (1994) Structures and molecular dynamics of plant waxes. II. Cuticular waxes from leaves of Fagus sylvatica L. and Hordeum vulgare L. Eur Biophys J 23:59–70

    Article  CAS  Google Scholar 

  • Schönherr J, Riederer M (1986) Plant cuticles sorb lipophilic compounds during enzymatic isolation. Plant Cell Environ 9:456–466

    Article  Google Scholar 

  • Schreiber L, Kirsch T, Riederer M (1996) Diffusion through cuticles: principles and models. In: Kerstiens G (ed) Plant cuticles—an integrated functional approach. BIOS Sci Pub Ltd, Oxford, pp 109–119

    Google Scholar 

  • Stevens JF, Hart H, Wollenweber E (1995) The systematic and evolutionary significance of exudate flavonoids in Aeonium. Phytochemistry 39:805–813

    Article  CAS  Google Scholar 

  • Truskett N van, Stebe KJ (2003) Influence of surfactants on an evaporating drop: fluorescence images and particle deposition paterns. Langmuir 19:8271–8279

    Article  CAS  Google Scholar 

  • Tulloch AP (1973) Composition of leaf surface waxes of Triticum species: variation with age and tissue. Phytochemistry 12:2225–2232

    Article  CAS  Google Scholar 

  • Tulloch AP, Bernard R, Hoffmann L (1980) A survey of epicuticular waxes among genera of Triticeae. 2. Chemistry. Can J Bot 58:2602–2615

    Article  CAS  Google Scholar 

  • Venkataraman B, Breen JJ, Flynn GW (1995) Scanning-tunneling-microscopy studies of solvent effects on the adsorbtion and mobility of triacontane triacontanol molecules adsorbed on graphite. J Phys Chem 99:6608–6619

    Article  CAS  Google Scholar 

  • Vogg G, Fischer S, Leide J, Emmanuel E, Jetter R, Levy AA, Riederer M (2004) Tomato fruit cuticular waxes and their effects on transpiration barrier properties: functional characterization of a mutant deficient in a very-long-chain fatty acid beta-ketoacyl-CoA synthase. J Exp Bot 55:1401–1410

    Article  PubMed  CAS  Google Scholar 

  • Walton TJ (1990) Waxes, cutin and suberin. In: Harwood JL, Bowyer JR (eds) Lipids, membranes and aspects of photobiology. Academic, London, pp 105–158

    Google Scholar 

  • von Wettstein-Knowles P (1971) The molecular phenotypes of the eceriferum mutants. In: Nilan RA (ed) Barley genetics. 2nd edn. Washington State Univ Press, Pullman, WA, pp 146–193

    Google Scholar 

  • Wilms M, Kruft M, Bermes G, Wandelt K (1999) A new and sophisticated electrochemical STM design for the investigation of potentiodynamic processes. Rev Sci Instrum 70:36–41

    Article  Google Scholar 

  • Zeier J, Schreiber L (1997) Chemical composition of hypodermal and endodermal cell walls and xylem vessels isolated from Clivia miniata: identification of the biopolymers lignin and suberin. Plant Phys 113:1223–1231

    CAS  Google Scholar 

Download references

Acknowledgements

We thank the Deutsche Forschungs Gemeinschaft, the Scientific Research-Flanders and the University of Bonn for the financial support of our research and the Physikalisch-Technische Bundesanstalt Braunschweig (Germany) for providing the standard normal for the AFM calibration, L. Schreiber and A. Dommisse (University of Bonn) for assistance in wax analysis, H.J. Ensikat for TEM analysis, the Institute for Organic Agriculture (University of Bonn) for providing the wheat seeds, and the reviewers for their helpful comments.

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Authors and Affiliations

  1. Nees-Institut für Biodiversität der Pflanzen, Meckenheimer Allee 170, 53115, Bonn, Germany

    K. Koch, W. Barthlott & S. Koch

  2. Institut für Theoretische und Physikalische Chemie, Wegeler Str. 12, 53115, Bonn, Germany

    A. Hommes, K. Wandelt & P. Broekmann

  3. Department of Chemistry, Katholieke Universiteit Leuven, Celestijnenlaan 200 F, 3001, Leuven, Belgium

    W. Mamdouh & S. De-Feyter

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  1. K. Koch
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  2. W. Barthlott
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  3. S. Koch
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Correspondence to K. Koch.

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Koch, K., Barthlott, W., Koch, S. et al. Structural analysis of wheat wax (Triticum aestivum, c.v. ‘Naturastar’ L.): from the molecular level to three dimensional crystals. Planta 223, 258–270 (2006). https://doi.org/10.1007/s00425-005-0081-3

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