Manipulating the Phenolic Acid Content and Digestibility of Forage Grasses by Targeted Expression of Fungal Cell Wall Degrading Enzymes

Grass cell walls constitute 30-80% of forage dry matter, representing a major source of energy for ruminants. Ferulic acid (4-hydroxy-3-methoxy-cinnamic acid) and other hydroxycinnamic acids are ester linked to arabinosyl residues in arabinoxylans of grass cell walls and undergo oxidative coupling reactions resulting in the formation of a variety of dehydrodiferulate dimers which cross-link cell wall polymers. Although such cross-links have a number of important roles in the cell wall, they also hinder the rate and extent of cell wall degradation by ruminant microbial and fungal enzymes. We have shown previously the expression of a ferulic acid esterase gene from Aspergillus niger in Festuca arundinacea and the potential of the expressed FAE to break phenolic cross-links and release monomeric and dimeric ferulic acids on cell death in vacuole targeted FAE plants. This was enhanced several fold by the addition of exogenous recombinant xylanase (Buanafina et al., 2002). We propose to decrease the level of phenolic cross-linking of cell wall carbohydrate by inducible expression of FAE to the apoplast, ER and golgi and by co-expressing FAE and endo-ß-1,4-xylanase from Trichoderma reesei to the apoplast and vacuole