Project Number: 5325-21000-002-03
Project Type: Trust

Start Date: Jul 01, 2008
End Date: Sep 30, 2009

Objective:
Plant expression cassettes containing codon optimized cellulase genes will be constructed and transformed into Arabidopsis thaliana and Brachypodium distachyon plants. Several different assemblies of plant transformation vectors containing different promoters and localization signals will be tested for each of the two candidate genes. The strategies best suited for producing high levels of localized protein which are enzymatically active on the plant cell wall will be compared.

Approach:
Two dicot-expression constructs (one for each cellulase gene) will be constructed. Agrobacterium-mediated floral dip transformation will be performed to generate 20-30 independent transgenic Arabidopsis plants per construct. The Arabidopsis transgenic lines produced will be molecularly characterized and plant tissue will be collected and provided to the Sandia collaborators for analysis. Six monocot-expression constructs (three per cellulase gene) will be assembled and used to generate 5-15 independent transgenic Brachypodium distachyon plants per construct. The tissue culture based Agrobacterium-mediated transformation will produce the first transgenic lines (for two constructs initiated at the start of the project) in approximately 8-10 months, with the other constructs following in the next 6-8 months. Source material for Brachypodium transformation and the regenerated transgenic plants and their progeny will require approximately 2 benches (200 sq. ft.) of greenhouse space for two years. Transgenic Brachypodium lines will be characterized and tissue of confirmed lines will be provided to the Sandia collaborators for analysis. Documents Reimbursable with Sandia Labs. Log 36945.