Function of laccases in cell wall biosynthesis

Laccases are multicopper oxidases capable of polymerizing monolignols. Histochemical assays have shown temporal and spatial correlation with secondary cell wall formation in both herbs and woody perennials. However, in plants laccases constitutes a relatively large group of isoenzymes with unique substrate specificities and expression patterns. As part of the strategic research centre Bio4Bio, the present project deals with laccase functions in relation to cell wall formation in grasses based on a study of the model species Brachypodium distachyon. Thirty-one isozymes have been retrieved from the recently published genome. Histochemical assays show high correlation between lignification and laccase-like activity in this species. Temporal and spatial correlation between these processes will be further studied by analysis of gene expression based on quantitative PCR and fluorescent in situ hybridization. Specific isozymes that show high correlation with the process of secondary cell wall formation will be further studied in a reverse genetic study in which candidates will be knocked out using RNA interference. Phenotypes of knock-out mutants are to be described in relation to cell wall degradability in order to reveal possible gains for saccharification of grass-derived feedstocks in bioethanol production and industrial processing. Laccases are multicopper oxidases capable of polymerizing monolignols. Histochemical assays have shown temporal and spatial correlation with secondary cell wall formation in both herbs and woody perennials. However, in plants laccases constitutes a relatively large group of isoenzymes with unique substrate specificities and expression patterns. As part of the strategic research centre Bio4Bio, the present project deals with laccase functions in relation to cell wall formation in grasses based on a study of the model species Brachypodium distachyon. Thirty-one isozymes have been retrieved from the recently published genome. Histochemical assays show high correlation between lignification and laccase-like activity in this species. Temporal and spatial correlation between these processes will be further studied by analysis of gene expression based on quantitative PCR and fluorescent in situ hybridization. Specific isozymes that show high correlation with the process of secondary cell wall formation will be further studied in a reverse genetic study in which candidates will be knocked out using RNA interference. Phenotypes of knock-out mutants are to be described in relation to cell wall degradability in order to reveal possible gains for saccharification of grass-derived feedstocks in bioethanol production and industrial processing.