Wang C X, Wang L, McQueen-Mason S J, Pritchard J, Thomas C R
School of Dentistry, The University of Birmingham, St Chad's Queensway, Birmingham, B4 6NN, UK.
J Plant Res. 2008 Sep;121(5):527-34. doi: 10.1007/s10265-008-0176-6. Epub 2008 Jul 10.
The aim of this study was to measure key material properties of the cell walls of single suspension-cultured plant cells and relate these to cell-wall biochemistry. To this end, micromanipulation was used to compress single tomato cells between two flat surfaces until they ruptured, and force-deformation data were obtained. In addition to measuring the bursting force, we also determined the elastic (Young's) modulus of the cell walls by matching low strain (< or = 20% deformation) experimental data with a cell compression model, assuming linear elastic cell walls. The walls were most elastic at pH 4.5, the pH optimum for expansin activity, with an elastic modulus of 2.0 +/- 0.1 GPa. Following the addition of exogenous expansins, cell walls became more elastic at all pH values. Western blot analysis of proteins from walls of cultured cells revealed the presence of expansin epitopes, suggesting that the inherent pH dependence of elasticity and other compression phenomena is related to the presence of endogenous expansin proteins and their wall-loosening ability. Although strict application of the linear-elastic model could not be applied to large deformations-for example, up to cell bursting-because of irreversible behaviour, the deviation of the data from the model was generally small enough to allow estimation of the strain in the cell wall at failure. This strain was greater at pH 4.5 and when expansins were added to the suspension. The changes in elasticity are consistent with suggestions about the mode of expansin action. The estimated strains at failure are compatible with data on the failure of Acetobacter-derived cellulose-xyloglucan composites and proposed mechanisms of such failure. Through the measurement of cell-wall material properties using micromanipulation, it may be possible to understand more fully how cell-wall composition, structure and biochemistry lead to cell mechanical behaviour.
本研究的目的是测量单个悬浮培养植物细胞细胞壁的关键材料特性,并将这些特性与细胞壁生物化学联系起来。为此,采用显微操作技术将单个番茄细胞压缩在两个平面之间,直至其破裂,并获取力-变形数据。除了测量破裂力外,我们还通过将低应变(≤20%变形)实验数据与细胞压缩模型相匹配,假定细胞壁为线性弹性,来确定细胞壁的弹性(杨氏)模量。细胞壁在pH 4.5时弹性最大,这是扩展蛋白活性的最适pH值,弹性模量为2.0±0.1 GPa。添加外源扩展蛋白后,细胞壁在所有pH值下都变得更具弹性。对培养细胞细胞壁蛋白进行的蛋白质印迹分析显示存在扩展蛋白表位,这表明弹性和其他压缩现象固有的pH依赖性与内源性扩展蛋白及其细胞壁松弛能力有关。尽管由于不可逆行为,线性弹性模型不能严格应用于大变形(例如直至细胞破裂),但数据与模型的偏差通常小到足以估计细胞壁在破裂时的应变。该应变在pH 4.5时以及向悬浮液中添加扩展蛋白时更大。弹性的变化与关于扩展蛋白作用模式的推测一致。估计的破裂应变与醋杆菌属来源的纤维素-木葡聚糖复合材料的破裂数据以及此类破裂的推测机制相符。通过使用显微操作测量细胞壁材料特性,有可能更全面地了解细胞壁组成、结构和生物化学如何导致细胞的力学行为。