Poster Presentation 47th Lorne Genome Conference 2026

Construction of the first plant artificial chromosome (PAC) (135437)

James Lloyd 1
  1. The University of Western Australia, Crawley, WA, Australia

Re-writing genomes in complex eukaryotes such as plants and humans have been challenging. While bacterial artificial chromosomes (BACs) and yeast artificial chromosomes (YACs) were developed decades ago, limited progress in the construction of a functioning plant artificial chromosome (PAC) has been made. This is in part due to the complex natural genomes of plants and long generation times of plants. Unlike in yeast, plant centromeres and origins of DNA replication are not encoded purely at the sequence level and are instead controlled by epigenetic mechanisms. The centromeres of plants are defined not by their underlying sequence identify but by the self-reinforcing presence of the cenH3 variant of H3. Furthermore, most plants lack the molecular genetic toolkit that was critical for such developments in yeast and bacteria, limiting progress. Here, we propose to make the first PAC using the model moss Physcomitrium patens (Physco). Physco, unlike other land plants, is able to edit DNA via homologous recombination much like yeast can, thus allowing for in planta alterations to the PAC. Furthermore, episomes without any known origin of DNA replication can be maintained in Physco. Thus, we aim to construct a synthetic centromere (synCEN), synthetic telomeres (synTEL), and reporter genes as a genetic payload into a PAC and show that it is more faithfully inherited than a regular moss episome. This will then be expanded in planta to a 1 Mb PAC and transferred to our crop target, potato, to demonstrate cross-species compatibility of our PAC. Ultimately, we aim to create the first modular orthogonal synthetic scaffold for trait augmentation in plants, providing a new platform for delivery of large and complex genetic payloads to improve plant performance, productivity, and robustness.