Induced Plant Cell Wall Modifications: Use of Plant Cells with Altered Walls to Study Wall Structure, Growth and Potential for Genetic Modification

Our previous work indicated that suspension-cultured plant cells show remarkable flexibility in altering cell wall structure in response either to growth on saline medium or in the presence of the cellulose synthesis inhibitor 2,-6-dichlorobenzonitrile (DCB). We have continued to analyze the structure of these modified cell walls to understand how the changes modify wall strength, porosity, and ability to expand. The major load-bearing network in the walls of DCB-adapted dicot cells that lack a substantial cellulose-xyloglucan network is comprised of Ca2+-bridged pectates; these cells also have an unusual and abundant soluble pectic fraction. By contrast, DCB-adapted barley, a graminaceous monocot achieves extra wall strength by enhanced cross-linking of its non-cellulosic polysaccharide network via phenolic residues. Our results have also shed new light on normal wall stucture: 1) the cellulose-xyloglucan network may be independent of other wall networks in dicot primary walls and accounts for about 70% of the total wall strength; 2) the pectic network in dicot walls is the primary determinant of wall porosity; 3) both wall strength and porosity in graminaceous monocot primary walls is greatly influenced by the degree of phenolic cross-linking between non-cellulosic polysaccharides; and 4) the fact that the monocot cells do not secrete excess glucuronoarabinoxylan and mixed-linked glucan in response to growth on DCB, suggests that these two non-cellulosic polymers do not normally interact with cellulose in a manner similar to xyloglucan. We also attempted to understand the factors which limit cell expansion during growth of cells in saline medium. Analyses of hydrolytic enzyme activities suggest that xyloglucan metabolism is not repressed during growth on NaCl. Unlike non-adapted cells, salt-adapted cells were found to lack pectin methyl esterase, but it is not clear how this difference could relate to alterations in wall expansibility. Salt-adaped cell walls contain reduced hyp and secrete two unique PRPP-related proteins suggesting that high NaCl inhibits the cross-linking of these proteins into the walls, a finding that might relate to their altered expansibility.