Submitted to: BioMed Central (BMC)Biotechnology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 6/3/2013
Publication Date: 8/5/2013
Publication URL: http://handle.nal.usda.gov/10113/57429
Citation: Trabucco, G., Matos, D., Lee, S., Saathoff, A.J., Sarath, G., Hazen, S. 2013. Functional characterization of cinnamyl alcohol dehydrogenase and caffeic acid O-methyltransferase in Brachypodium distachyon. . BioMed Central (BMC)Biotechnology. doi: 10.1186/10.1186/1472-6750-13-61. Interpretive Summary: Lignin is a component of plant cell walls and biomass from plants containing reduced lignin concentration have been shown to have the potential to produce more ethanol in a biorefinery. The purpose of this work was to determine the effects of modifying two genes involved in lignin synthesis in the model grass plant Brachypodium distachyon. Brachypodium plants are easy to propagate in the laboratory and amenable to genetic engineering, making them an attractive model to assess the roles of genes that impact biofuel yields. Plants with reduced expression of the two modified genes had lower lignin amounts and altered growth characteristics. Potential ethanol yields were higher from the altered plants, which indicated that strategies targeting these genes in bioenergy grasses may help in developing better plants for use as bioenergy crops.
Technical Abstract: Lignin is a significant recalcitrant in the conversion of plant biomass to bioethanol. Cinnamyl alcohol dehydrogenase (CAD) and caffeic acid O-methyltransferase (COMT) catalyze key steps in the pathway of lignin monomer biosynthesis. Brown midrib mutants in Zea mays and Sorghum bicolor with impaired CAD or COMT activity have attracted considerable agronomic interest for their altered lignin composition and improved digestibility. We identified candidate genes encoding CAD and COMT enzymes in the grass model species Brachypodium distachyon and developed transgenic plants overexpressing artificial microRNA designed to silence BdCAD1 or BdCOMT4. Both transgenes caused altered flowering time and stem count and weight. Downregulation of BdCAD1 caused a leaf brown midrib phenotype, the first time this phenotype has been observed in a C3 plant. While acetyl bromide soluble lignin measurements were equivalent in BdCAD1 downregulated and wildtype plants, histochemical staining and thioacidolysis indicated a decrease in lignin syringyl units and reduced syringyl/guaiacyl ratio in the transgenic plants. BdCOMT4 downregulated plants exhibited a decrease in total lignin content, a significant reduction of guaiacyl lignin, and a modest reduction of syringyl lignin. Ethanol yield by microbial fermentation was enhanced in both BdCAD1- and BdCOMT4-downregulated plants. These results have elucidated two key genes in the lignin biosynthetic pathway in B. distachyon that, when perturbed, may result in greater biomass yield and bioconversion efficiency.