New Tools for Engineering the Arabidopsis Plastid Genome
International Conference on Plant Synthetic Biology and Bioengineering
2019
3rd International Conference on Plant Synthetic Biology, Bioengineering, and Biotechnology
Poster Session
Poster Session
Friday, October 4, 2019 - 5:00pm to 6:00pm
Plastid transformation in the spectinomycin-hypersensitive Col-0 acc2-1 and Sav-0 backgrounds was found to be 100-fold more efficient than the wild type (Yu et al., 2017). As a follow up to this study, we developed new tools to expand plastid transformation biotechnology in Arabidopsis. Newly constructed pMEK vectors harbor a left targeting region truncated at the HindIII site in the plastid rrn16 gene to enable utilization of the EcoRI-HindIII multiple cloning site. A one-step selective SPEED medium permits rapid evaluation of plastid transformation efficiency in 35 to 42 days (5 to 6 weeks) and transplastomic cells can be readily identified by GFP fluorescence. Mutations in the ACC2 N-terminal chloroplast transit peptide of RLD and Ws produced ACC2-knockout lines that are spectinomycin-hypersensitive and pliable for plastid transformation. We have advanced efforts to develop plastid transformation in Arabidopsis by developing a system for rapid evaluation of plastid transformation efficacy, complete with new vectors, culture media, and new ACC2-knockout lines that expand the range of accessions available for experimentation.