Semi‐automated workflow for high‐throughput Agrobacterium‐mediated plant transformation

• DOI: 10.1111/tpj.70118
• Publication Date: 2025-04-12T21:11:50Z
• Authors: Davide Annese; Facundo Romani; Carolina Grandellis; Lesley Ives; Eftychios Franged...; Felipe X. Buson; Jennifer C. Molloy; Jim Haseloff

High-throughput experiments in plants are hindered by long generation times and high costs. To address these challenges, we present an optimized pipeline for Agrobacterium tumefaciens transformation a simplified protocol to obtain stable transgenic lines of the model liverwort Marchantia polymorpha, paving way efficient high-throughput plant synthetic biology other applications. Our involves freeze-thaw method six-well plates that can be adapted robotic automation. Using Opentrons open-source platform, implemented semi-automated showing similar efficiency compared manual manipulation. Additionally, have streamlined process selection M. reducing cost, time, labor without compromising efficiency. The addition sucrose media significantly enhances production gemmae, accelerating isogenic plants. We believe protocols potential facilitate screenings diverse species represent significant step towards full automation pipelines. This approach allows testing ~100 constructs per month, using conventional tissue culture facilities. recently demonstrated successful implementation this screening hundreds fluorescent reporters gemmae.