Plasmodesmata transport of GFP alone or fused to potato virus X TGBp1 is diffusion driven

Plasmodesmata (Pd) provide a pathway for exchanging various macromolecules between neighboring plant cells. Researchers routinely characterize the mobility of the green-fluorescent protein (GFP) and GFP fusions through Pd by calculating the proportion of sites in bombarded leaves which show fluorescence in multiple cell clusters (% movement). Here, the Arrhenius equation was used to describe the temperature dependence of GFP and GFP-TGBp1 (potato virus X triple gene block protein1) movement, using % movement values, and to calculate the activation energy for protein transport. The resulting low activation energy indicates GFP and GFP-TGBp1 movement are diffusion driven. Furthermore, GFP movement is inversely proportional to the leaf surface area of expanding leaves. The increase in leaf area results mainly from cell expansion during the sink–source transition. The increasing cell size results in lower Pd density, which decreases the probability that a GFP attains an open Pd by diffusion. The decline in GFP movement as leaf area expands indicates that, in addition to GFP diffusion through Pd, attaining an open Pd by undirected diffusion might be limiting for Pd transport. In summary, this report provides a new quantitative method for studying Pd conductivity.