Efficient Agrobacterium-Mediated Transformation of Green Arabidopsis Suspension Cells

Photosynthetic plant cell suspension cultures are a valuable experimental system for analyzing various physiological processes. This system bypasses the structural complexity of the whole plant organism and can be manipulated under uniform conditions. However, achieving a highly efficient and consistent transformation of plant suspension cells remains challenging. By using green fluorescent protein (GFP) and a microplate confocal imaging system for high-throughput analysis, we optimized the transformation of green Arabidopsis suspension cells to infect almost 100% of the cells. Key elements of our protocol included using the hypervirulent Agrobacterium tumefaciens strain AGL1, co-cultivating agrobacteria and suspension cells on solidified medium plates, and adding AB minimal salts and the surfactant Pluronic F68. The presented method can significantly increase the transformation rate of plant suspension cells, facilitating the introduction of genetic pathways for producing industrial, cosmetic, or pharmaceutical compounds in these systems.