A feature of cell wall arabinoxylan in grasses is the presence of ferulic acid which upon oxidative coupling by the action of peroxidases forms diferuloyl bridges between formerly separated arabinoxylans. Ferulate cross-linking is suspected of playing various roles in different plant processes. Here we investigate the role of cell wall feruloyaltion in two major processes, that of leaf growth and the turnover of cell wall arabinoxylans on leaf senescence in tall fescue using plants in which the level of cell wall ferulates has been reduced by targeted expression of the Aspergillus niger ferulic acid esterase A (FAEA) to the apoplast or Golgi. Analysis of FAE expressing plants showed that all the lines had shorter and narrower leaves compared to control, which may be a consequence of the overall growth rate being lower and occurring earlier in FAE expressing leaves than in controls. Furthermore, the final length of epidermal cells was shorter than controls, indicating that their expansion was curtailed earlier than in control leaves. This may be due to the observations that the deposition of both ether and ester linked monomeric hydroxycinnamic acids and ferulate dimerization stopped earlier in FAE expressing leaves but at a lower level than controls, and hydroxycinnamic acid deposition started to slow down when peroxidase levels increased. It would appear therefore that one of the possible mechanisms for controlling overall leaf morphology such as leaf length and width in grasses, where leaf morphology is highly variable between species, may be the timing of hydroxycinnamic acid deposition in the expanding cell walls as they emerge from cell division into the elongation zone, controlled partially by the onset of peroxidase activity in this region.

While many aspects of the growth of maize are well understood, the role of cell wall feruloylation particularly during internode elongation has not been firmly established, but results so far indicate that it has significant implications for both biofuel feedstock conversion and for crop yield. The growth of the cell wall is achieved by synthesis, integration and cross-linking between wall polymers. As ferulate oxidative coupling of arabinoxylan side chains constitutes a significant type of cross-link in grass cell walls, it is expected to have a crucial role in plant growth. Making use of plants expressing an apoplast targeted Aspergillus niger FAEA under the control of either a constitutive or an inducible promoter, the role of cell wall feruloylation in maize internode expansion was investigated. Analysis of FAEA expressing plants showed that where FAEA was targeted to the apoplast under a constitutive promoter, plants varied in stature either from semi-dwarf plants with a 40-60% height reduction, to extreme dwarf mutants with over 90% reduction in plant heights compared to controls. Results indicate that disruption of cell wall feruloylation by FAEA occurs before the start of rapid internode expansion is initiated and affects the normal course of internode elongation, resulting in short internodes and dwarfed plants. In contrast, when under the inducible Lm See1 senescence promoter, FAEA activity was found to be low up to the VT stage of development but increased significantly at the VR stage as plants began to senesce, strongly suggesting that normal cell wall feruloylation is required for the process of internode expansion. In addition, with apoplast targeted expression of FAEA under control of the senescence enhanced promoter it was possible to demonstrate decreased cell wall feruloylation without affecting internode expansion or other aspects of plant development.

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