Proseus Timothy E, Boyer John S
College of Marine Studies and College of Agriculture and Natural Resources, 700 Pilottown Road, University of Delaware, Lewes, DE 19958, USA.
J Exp Bot. 2006;57(15):3989-4002. doi: 10.1093/jxb/erl166. Epub 2006 Nov 16.
Pectin, a normal constituent of cell walls, caused growth rates to accelerate to the rates in living cells when supplied externally to isolated cell walls of Chara corallina. Because this activity was not reported previously, the activity was investigated. Turgor pressure (P) was maintained in isolated walls or living cells using a pressure probe in culture medium. Pectin from various sources was supplied to the medium. Ca and Mg were the dominant inorganic elements in the wall. EGTA or pectin in the culture medium extracted moderate amounts of wall Ca and essentially all the wall Mg, and wall growth accelerated. Removing the external EGTA or pectin and replacing with fresh medium returned growth to the original rate. A high concentration of Ca2+ quenched the accelerating activity of EGTA or pectin and caused gelling of the pectin, physically inhibiting wall growth. Low pH had little effect. After the Mg had been removed, Ca-pectate in the wall bore the longitudinal load imposed by P. Removal of this Ca caused the wall to burst. Live cells and isolated walls reacted similarly. It was concluded that Ca cross-links between neighbouring pectin molecules were strong wall bonds that controlled wall growth rates. The central role of Ca-pectate chemistry was illustrated by removing Ca cross-links with new pectin (wall "loosening"), replacing vacated cross-links with new Ca2+ ("Ca2+-tightening"), or adding new cross-links with new Ca-pectate that gelled ("gel tightening"). These findings establish a molecular model for growth that includes wall deposition and assembly for sustained growth activity.
果胶是细胞壁的正常组成成分,当将其外源添加到轮藻分离的细胞壁时,可使生长速率加快至活细胞中的生长速率。由于此前未报道过这种活性,因此对该活性进行了研究。使用压力探针在培养基中维持分离细胞壁或活细胞中的膨压(P)。将来自各种来源的果胶添加到培养基中。钙和镁是细胞壁中的主要无机元素。培养基中的乙二醇双四乙酸(EGTA)或果胶可提取适量的细胞壁钙和几乎所有的细胞壁镁,细胞壁生长加速。去除外部的EGTA或果胶并用新鲜培养基替换后,生长恢复到原始速率。高浓度的Ca2+可抑制EGTA或果胶的促进活性,并导致果胶凝胶化,从而物理性抑制细胞壁生长。低pH值影响较小。去除镁后,细胞壁中的果胶酸钙承受膨压施加的纵向负荷。去除这种钙会导致细胞壁破裂。活细胞和分离的细胞壁反应相似。得出的结论是,相邻果胶分子之间的钙交联是控制细胞壁生长速率的强细胞壁键。通过用新果胶去除钙交联(细胞壁“松弛”)、用新的Ca2+替代空出的交联(“Ca2+收紧”)或添加新的果胶酸钙交联使其凝胶化(“凝胶收紧”),说明了果胶酸钙化学的核心作用。这些发现建立了一个生长的分子模型,该模型包括细胞壁沉积和组装以实现持续的生长活性。