beta-1,3-Glucan in Developing Cotton Fibers: Structure, Localization, and Relationship of Synthesis to That of Secondary Wall Cellulose

Abstract

Evidence is presented for the existence of a noncellulosic beta-1,3-glucan in cotton fibers. The glucan can be isolated as distinct fractions of varying solubility. When fibers are homogenized rigorously in aqueous buffer, part of the total beta-1,3-glucan is found as a soluble polymer in homogenates freed of cell walls. The proportion of total beta-1,3-glucan which is found as the soluble polymer varies somewhat as a function of fiber age. The insoluble fraction of the beta-1,3-glucan remains associated with the cell wall fraction. Of this cell wall beta-1,3-glucan, a variable portion can be solubilized by treatment of walls with hot water, a further portion can be solubilized by alkaline extraction of the walls, and 17 to 29% of the glucan remains associated with cellulose even after alkaline extraction. A portion of this glucan can also be removed from the cell walls of intact cotton fibers by digestion with an endo-beta-1,3-glucanase. The glucan fraction which can be isolated as a soluble polymer in homogenates freed of cell walls is not associated with membranous material, and we propose that it represents glucan which is also extracellular but not tightly associated with the cell wall. Enzyme digestion studies indicate that all of the cotton fiber glucan is beta-linked, and methylation analyses and enzyme studies both show that the predominant linkage in the glucan is 1 --> 3. The possibility of some minor branching at C-6 can also be deduced from the methylation analyses. The timing of deposition of the beta-1,3-glucan during fiber development coincides closely with the onset of secondary wall cellulose synthesis. Kinetic studies performed with ovules and fibers cultured in vitro show that incorporation of radioactivity from [(14)C]glucose into beta-1,3-glucan is linear with respect to time almost from the start of the labeling period; however, a lag is observed before incorporation into cellulose becomes linear with time, suggesting that these two different glucans are not polymerized directly from the same substrate pool. Pulse-chase experiments indicate that neither the beta-1,3-glucan nor cellulose exhibits significant turnover after synthesis.