Efficient Agrobacterium-Mediated Transformation of Green Arabidopsis Suspension Cells

Abstract

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.

FIGURE 1

Analysis of GFP‐positive Arabidopsis cells after co‐cultivation with Agrobacterium in liquid or on solid ABM‐MS medium for 2 days with or without 0.05% (w/v) PF68 and incubation on solid medium for 3 days. Images were then acquired with a microplate confocal imaging system. “Control” are cells that were incubated without Agrobacterium. The column on the right of the figures shows the percentage of GFP‐positive cells, as calculated by image analysis and data processing. Scale bar = 2000 µm.