Viscoelastic properties of cell walls of single living plant cells determined by dynamic nanoindentation

Plant development results from controlled cell divisions, structural modifications, and reorganizations of the cell wall. Thereby, regulation of cell wall behaviour takes place at multiple length scales involving compositional and architectural aspects in addition to various developmental and/or environmental factors. The physical properties of the primary wall are largely determined by the nature of the complex polymer network, which exhibits time-dependent behaviour representative of viscoelastic materials. Here, a dynamic nanoindentation technique is used to measure the time-dependent response and the viscoelastic behaviour of the cell wall in single living cells at a micron or sub-micron scale. With this approach, significant changes in storage (stiffness) and loss (loss of energy) moduli are captured among the tested cells. The results reveal hitherto unknown differences in the viscoelastic parameters of the walls of same-age similarly positioned cells of the Arabidopsis ecotypes (Col 0 and Ws 2). The technique is also shown to be sensitive enough to detect changes in cell wall properties in cells deficient in the activity of the chromatin modifier ATX1. Extensive computational modelling of the experimental measurements (i.e. modelling the cell as a viscoelastic pressure vessel) is used to analyse the influence of the wall thickness, as well as the turgor pressure, at the positions of our measurements. By combining the nanoDMA technique with finite element simulations quantifiable measurements of the viscoelastic properties of plant cell walls are achieved. Such techniques are expected to find broader applications in quantifying the influence of genetic, biological, and environmental factors on the nanoscale mechanical properties of the cell wall.

[1]  D. Cosgrove,et al.  © 2005 BioMed Central Ltd Summary , 2005 .

[2]  A. Jaffrin,et al.  Analysis of Pressure-Volume Curves of Leaves of Rosa hybrida cv. Sonia , 1993 .

[3]  K. Bowman.  Mechanical Behavior of Materials , 2003 .

[4]  G. Pharr,et al.  An improved technique for determining hardness and elastic modulus using load and displacement sensing indentation experiments , 1992 .

[5]  D. Cosgrove.  Relaxation in a high-stress environment: the molecular bases of extensible cell walls and cell enlargement. , 1997, The Plant cell.

[6]  Mark R Marten,et al.  Elastic Properties of the Cell Wall of Aspergillus nidulans Studied with Atomic Force Microscopy , 2008, Biotechnology progress.

[7]  P. Green.  Organogenesis-A Biophysical View , 1980 .

[8]  J. Müssig,et al.  Fibre matrix adhesion of natural fibres cotton, flax and hemp in polymeric matrices analyzed with the single fibre fragmentation test , 2008 .

[9]  Ingo Burgert,et al.  Exploring the micromechanical design of plant cell walls. , 2006, American journal of botany.

[10]  A. Shilatifard,et al.  Histone H3 lysine 4 (H3K4) methylation in development and differentiation. , 2010, Developmental biology.

[11]  J. Verbelen,et al.  Xyloglucan endotransglucosylase activity loosens a plant cell wall. , 2007, Annals of botany.

[12]  J. Rose,et al.  Characterization of a new xyloglucan endotransglucosylase/hydrolase (XTH) from ripening tomato fruit and implications for the diverse modes of enzymic action. , 2006, The Plant journal : for cell and molecular biology.

[13]  M. Jarvis.  Structure and properties of pectin gels in plant cell walls , 1984 .

[14]  W. Chiou,et al.  Causes and consequences of high osmotic potentials in epiphytic higher plants. , 2004, Journal of plant physiology.

[15]  F. Baluška,et al.  The Arabidopsis homolog of trithorax, ATX1, binds phosphatidylinositol 5-phosphate, and the two regulate a common set of target genes. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[16]  D. J. Montgomery,et al.  The physics of rubber elasticity , 1949 .

[17]  U. Grossniklaus,et al.  ATX-1, an Arabidopsis Homolog of Trithorax, Activates Flower Homeotic Genes , 2003, Current Biology.

[18]  F. Micheli.  Pectin methylesterases: cell wall enzymes with important roles in plant physiology. , 2001, Trends in plant science.

[19]  Y. Manetas,et al.  Optical Properties of Hairs During the Early Stages of Leaf Development in Platanus orientalis , 1996 .

[20]  Keiko Sugimoto-Shirasu,et al.  Tensile Properties of Arabidopsis Cell Walls Depend on Both a Xyloglucan Cross-Linked Microfibrillar Network and Rhamnogalacturonan II-Borate Complexes1 , 2003, Plant Physiology.

[21]  David Stuart Thompson,et al.  How do cell walls regulate plant growth? , 2005, Journal of experimental botany.

[22]  Kathryn J. Wahl,et al.  Nanoindentation and contact stiffness measurement using force modulation with a capacitive load-displacement transducer. , 1999 .

[23]  Markus Pauly,et al.  Glycosyltransferases and cell wall biosynthesis: novel players and insights. , 2004, Current opinion in plant biology.

[24]  R. Murphy,et al.  A New Pressure Probe Method to Determine the Average Volumetric Elastic Modulus of Cells in Plant Tissue , 1995, Plant physiology.

[25]  B. Petersen,et al.  Mechanical Properties of Plant Cell Walls Probed by Relaxation Spectra1[W][OA] , 2010, Plant Physiology.

[26]  C-E Aubin,et al.  Finite-Element Analysis of Geometrical Factors in Micro-Indentation of Pollen Tubes , 2006, Biomechanics and modeling in mechanobiology.

[27]  J. Pethica,et al.  Tip Surface Interactions in STM and AFM , 1987 .

[28]  M. Cannon,et al.  Self-assembly of the plant cell wall requires an extensin scaffold , 2008, Proceedings of the National Academy of Sciences.

[29]  R. Cleland.  The Instron technique as a measure of immediate-past wall extensibility , 1984, Planta.

[30]  G. Jeronimidis,et al.  Mechanical effects of plant cell wall enzymes on cellulose/xyloglucan composites. , 2004, The Plant journal : for cell and molecular biology.

[31]  G. Odegard,et al.  Characterization of viscoelastic properties of polymeric materials through nanoindentation , 2005 .

[32]  L Wang,et al.  Modelling the mechanical properties of single suspension-cultured tomato cells. , 2004, Annals of botany.

[33]  P B Green,et al.  Metabolic and physical control of cell elongation rate: in vivo studies in nitella. , 1971, Plant physiology.

[34]  D. Cosgrove.  Growth of the plant cell wall , 2005, Nature Reviews Molecular Cell Biology.

[35]  U. Zimmermann,et al.  Pressure probe technique for measuring water relations of cells in higher plants. , 1978, Plant physiology.

[36]  D. Reis,et al.  The helicoidal plant cell wall as a performing cellulose‐based composite , 1989 .

[37]  D. Wilman,et al.  Cell wall thickness and cell dimensions in plant parts of eight forage species , 1998, The Journal of Agricultural Science.

[38]  P. Fratzl,et al.  Cellulose microfibril orientation of Picea abies and its variability at the micron-level determined by Raman imaging , 2009, Journal of experimental botany.

[39]  M. C. Probine,et al.  Cell Growth and the Structure and Mechanical Properties of the Wall in Internodal Cells of Nitella opaca: II. MECHANICAL PROPERTIES OF THE WALLS , 1962 .

[40]  Z. Avramova,et al.  Wall-modifying genes regulated by the Arabidopsis homolog of trithorax, ATX1: repression of the XTH33 gene as a test case. , 2009, The Plant journal : for cell and molecular biology.

[41]  C. Nawrath,et al.  Unraveling the complex network of cuticular structure and function. , 2006, Current opinion in plant biology.

[42]  I. Ladunga,et al.  The Highly Similar Arabidopsis Homologs of Trithorax ATX1 and ATX2 Encode Proteins with Divergent Biochemical Functions[W] , 2008, The Plant Cell Online.

[43]  Arezki Boudaoud,et al.  In vivo analysis of local wall stiffness at the shoot apical meristem in Arabidopsis using atomic force microscopy. , 2011, The Plant journal : for cell and molecular biology.

[44]  T. Baskin.  Anisotropic expansion of the plant cell wall. , 2005, Annual review of cell and developmental biology.

[45]  Z. Avramova.  Evolution and pleiotropy of TRITHORAX function in Arabidopsis. , 2009, The International journal of developmental biology.

[46]  E. Steudle,et al.  Pressure probe and isopiestic psychrometer measure similar turgor. , 1987, Plant physiology.

[47]  U. Kutschera.  Cessation of cell elongation in rye coleoptiles is accompanied by a loss of cell-wall plasticity , 1996 .

[48]  P. A. Roelofsen.  The plant cell-wall , 1959 .

[49]  M. Cardoso.  Herbivore handling of a Plants trichome: the case of Heliconius charithonia (L.) (Lepidoptera: Nymphalidae) and Passiflora lobata (Killip) Hutch. (Passifloraceae). , 2008, Neotropical entomology.

[50]  K. Waldron,et al.  The mechanical properties and molecular dynamics of plant cell wall polysaccharides studied by Fourier-transform infrared spectroscopy. , 2000, Plant physiology.

[51]  P. Fratzl,et al.  Mechanical properties of spruce wood cell walls by nanoindentation , 2004 .

[52]  Park S. Nobel,et al.  Physicochemical and Environmental Plant Physiology , 1991 .

[53]  M. Shih,et al.  The GUS reporter-aided analysis of the promoter activities of Arabidopsis ACC synthase genes AtACS4, AtACS5, and AtACS7 induced by hormones and stresses. , 2005, Journal of experimental botany.

[54]  Anja Geitmann,et al.  Experimental approaches used to quantify physical parameters at cellular and subcellular levels. , 2006, American journal of botany.

[55]  M. Parker,et al.  Fracture mechanics of the cell wall of Chara corallina , 2001, Planta.

[56]  C. Dunand,et al.  Patterning of Arabidopsis epidermal cells: epigenetic factors regulate the complex epidermal cell fate pathway. , 2006, Trends in plant science.