Mitochondrial Transformation of Mutant Arabidopsis thaliana to Rescue the Post-Transcriptional Modification and Assembly of F-ATP Synthase

The mitochondrion plays a fundamental role in cellular respiration, photosynthesis, and metabolism in plants, and these organelles possess an independent genome separate from the nuclear genome. Mitochondrial genomes in higher plants are often incredibly large and complex, offering a localized mechanism of independent genome regulation. Though nuclear genetic transformation in plants has been widely studied, genetic transformation of the plant mitochondrial genome in a vascular plant has yet to be achieved. If successful, an in vivo mitochondrial transformation would open a new avenue for the genetic engineering of plants and crops, and further the research and application of hybrid crop development via methods including cytoplasmic male sterility. Here, we attempt to achieve a mitochondrial transformation of a mutant seed line of Arabidopsis thaliana in which ATP synthesis has been significantly hindered. Two methods were tested to achieve such a transformation. The first, the use of peptide-nanoparticle DNA carriers, relies on the transport abilities of peptides to deliver genetic information. The second, Agrobacterium tumefaciens-mediated transformation, modifies previously existing methods in plant transformation to direct in vitro synthesized DNA to the mitochondria using the natural transfection mechanisms of Agrobacterium. The results of both methods have shown promise in infiltrating plant cells with exogenous DNA constructs but as of yet, we have been unsuccessful in integrating said genetic information into the mitochondrial genome. Regardless, the methodology remains a promising approach to mitochondrial genome transformation, and further research on the matter is recommended.