Visualization of the effect of a surfactant on the uptake of xenobiotics into plant foliage by confocal laser scanning microscopy

A radiolabelling method is generally used to determine the foliar uptake of xenobiotics. This technique cannot provide any information on the localization of chemicals inside leaf tissues. The influence of an alcohol ethoxylate surfactant on the uptake of three fluorescent dyes, difluorofluorescein (hydrophilic), rhodamine B (moderately lipophilic) and a naphthalimide dye (lipophilic), into the leaves of three contrasting species, bean (Vicia faba), wheat (Triticum aestivum) and cabbage (Brassica oleracea), at 16 h after treatment was measured using a conventional wash‐off method and also visualized in vivo by confocal laser scanning microscopy (CLSM). Whereas the lipophilic dyes showed greater intrinsic uptake than the hydrophilic one, the enhancing effect of the surfactant on uptake was more pronounced for the latter. CLSM revealed that the presence of the surfactant increased the transport of difluorofluorescein into the epidermal cells of bean and wheat leaves, but not cabbage leaves. Rhodamine B showed greatest transcuticular diffusion in all three species, but most of the dye moved into the vacuole of the epidermal cells. The naphthalimide dye was strongly retained by the wax–cuticle layer of all species, even in the presence of the surfactant. CLSM has proven to be an attractive tool for studying xenobiotic diffusion. The results obtained using fluorescent dyes are believed to be applicable to the foliar uptake of herbicides.