Effects of continuous hypergravity stimuli on the amounts and composition of cell wall constituents were investigated in wheat shoots. Hypergravity (300 g) treatment for three days after germination increased the net amount of cell wall polysaccharides such as hemicellulose and cellulose, but reduced the shoot elongation. As a result, the amount of cell wall polysaccharides per unit length of shoot increased under hypergravity. The hemicellulose fraction contained polysaccharides in the middle and low molecular mass range (5 kDa-1 MDa) and increased in response to hypergravity. Also, the amounts of arabinose (Ara) and xylose (Xyl), the major sugar components of the hemicellulose fraction, increased under hypergravity conditions. In addition to wall polysaccharides, hypergravity increased the amounts of cell wall-bound phenolic acids, such as ferulic acid (FA) and diferulic acid (DFA). Furthermore, the activity of phenylalanine ammonia-lyase (PAL, EC 126.96.36.199) was enhanced under hypergravity conditions. These results suggest that continuous hypergravity stimulates the synthesis of cell wall constituents, especially hemicellulosic arabinoxylans and cell wall-bound FA and DFA in wheat shoots. The increased PAL activity may promote the formation of FA and DFA. These changes in cell wall architecture may be involved in making rigid and tough cell walls under hypergravity conditions and thereby contribute to the ability of plant to sustain their structures against gravitational stimuli.
Research Containing: ferulic acid
Increase in the level of arabinoxylan–hydroxycinnamate network in cell walls of wheat coleoptiles grown under continuous hypergravity conditions
Changes in the amount and composition of cell wall constituents in response to continuous hypergravity stimuli were studied in wheat (Triticum aestivum L.) coleoptiles. The lengths of coleoptiles grown under hypergravity (300 g) conditions for 2–4 days from germination stage were 60–70% of those of 1 g control. However, the net amounts of hemicellulosic polysaccharides and cellulose in hypergravity-treated coleoptiles increased progressively as much as those in the control coleoptiles. As a result, their contents per unit length of coleoptile largely increased under hypergravity conditions. In the hemicellulose fraction, the amounts of arabinose and xylose, the major components of the fraction, prominently increased in response to hypergravity. When hemicellulosic polysaccharides were separated into neutral and acidic polymers by an anion-exchange column, the amounts of the acidic fraction consisting of (glucurono)arabinoxylans were higher in hypergravity-treated coleoptiles than in control coleoptiles. The amounts of cell wall-bound ferulic acid and diferulic acid (DFA) increased dramatically in both 1 g control and hypergravity-treated coleoptiles. Particularly, the amounts of DFA in hypergravity-treated coleoptiles were significantly higher than those in control coleoptiles during the incubation period. These results suggest that continuous hypergravity increases the rigid network structures via arabinoxylan–hydroxycinnamate cross-links within cell wall architecture in wheat coleoptiles. These structures may have a load-bearing function and contribute to construct the stable cell wall against the gravitational force.
Oxalate oxidase (OXO) utilizes oxalate to generate hydrogen peroxide, and thereby acts as a source of hydrogen peroxide. The present study was carried out to investigate whether apoplastic OXO modifies the metabolism of cell wall-bound ferulates in wheat seedlings. Histochemical staining of OXO showed that cell walls were strongly stained, indicating the presence of OXO activity in shoot walls. When native cell walls prepared from shoots were incubated with oxalate or hydrogen peroxide, the levels of ester-linked diferulic acid (DFA) isomers were significantly increased. On the other hand, the level of ester-linked ferulic acid (FA) was substantially decreased. The decrease in FA level was accounted neither by the increases in DFA levels nor by the release of FA from cell walls during the incubation. After the extraction of ester-linked ferulates, considerable ultraviolet absorption remained in the hemicellulosic and cellulose fractions, which was increased by the treatment with oxalate or hydrogen peroxide. Therefore, a part of FA esters may form tight linkages within cell wall architecture. These results suggest that cell wall OXO is capable of modifying the metabolism of ester-linked ferulates in cell walls of wheat shoots by promoting the peroxidase action via supply of hydrogen peroxide.