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CRISPR genome editing in Populus: Lignin biosynthesis revisited

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Monday 26 September 2016, 11:00 - 12:30

 
ABSTRACT

CRISPR/Cas9 genome editing in Populus was achieved with unprecedented efficiency, extending the species range of this powerful technology to outcrossing woody perennials.  For two 4-coumarate:CoA ligase (4CL) targets, all stably transformed plants harbored bi-allelic DNA mutations, resulting in strikingly uniform phenotypes across independent transgenic lines.  Sequence polymorphisms in outcrossing species pose an underappreciated obstacle to efficient genome editing.  Such data are often inaccessible via current genome portals, and are not considered in popular gRNA design programs. We developed variant-sensitive bioinformatics pipelines to address this limitation, but challenges remain for efficient genome editing of outcrossing, hybrid or polyploid species.  

 

With careful gRNA design, we achieved specific editing of the lignin-associated 4CL1 without affecting its genome duplicate 4CL5. The 4CL1-null poplars exhibited reduced lignin accrual and a reddish-brown discoloration in stem wood, as expected. However, the lignin S/G ratio decreased, which contrasts with increased S/G reported for herbaceous mutants.  RNA-Seq revealed distinct transcriptional responses of lignin pathway genes in Populus and Arabidopsis mutants.  The data suggest that fine-tuning of the monolignol branchways, perhaps via metabolic channeling, contributes to the plasticity and variation of lignin biosynthesis in herbaceous annuals and woody perennials.    

Location Jozef Schell seminar room
Contact Prof C-J Tsai
Warnell School of Forestry and Natural Resources, Department of Genetics
University of Georgia
USA