Field performance of sugarcane with TALEN mediated targeted mutagenesis of COMT

Transcription activator-like effector nuclease (TALEN) is a recently developed tool enabling precise genome modifications, such as targeted mutagenesis, gene replacement, or insertion. Sugarcane is a prime feedstock for bioethanol production, and utilizing both sucrose and cell wall bound sugars for fermentation will enhance the biofuel yield. We recently demonstrated that RNAi mediated downregulation of lignin biosynthetic gene Caffeic acid O-methyltransferase (COMT) is a successful strategy to improve bioethanol production from lignocellulosic sugarcane biomass. In this study, COMT was targeted for the TALEN induced multi-allelic mutagenesis to modify lignin biosynthesis in sugarcane. Targeted mutations following TALEN delivery were identified by capillary electrophoresis of the COMT amplicon. Events were confirmed by sequencing of the COMT amplicon which revealed the presence of insertions and deletions at the target site. Data comparing the total lignin content in the stem biomass of COMT mutant and RNAi suppressed sugarcane events and their field performance will be presented.