Subcellular Compartmentation of Indole-3-acetic Acid in Mesophyll Cells of Spinacia oleracea

Chloroplasts, Compartmental Analysis, Indole-3-acetic Acid (LAA), IAA Conjugation, Permeability A compartmental analysis of the distribution of the plant hormone indole-3-acetic acid (IAA) was performed with mesophyll cells of spinach leaves. The results indicated that up to about 45% of the total amount of free IAA in the laminae of spinach is localized within the chloroplasts, although these organelles occupy only about 7% of the tissue volume. This distribution is due to the dissociation properties of IAA which has a pKa of 4,7. The chloroplast envelope is easily permeable for the undissociated acid (IAAH) and less permeable for the anion (IAA“). The rate of uptake of LAA by chloroplasts was linearly dependent on the IAAH concentration gradient between medium and stroma. Uptake was increased by lowering the extraplastidic pH or by alkalization of the stroma during illumination. In illuminated chloroplasts, IAA was accumulated up to 4 fold compared to the surrounding medium owing to the pH gradient between the medium (pH 7.6) and the stroma (pH 8.0). The unexpectedly high plastidic IAA concentration observed in the dark (concentration ratio stroma/medium was about 2) suggested binding of IAA to chloroplast components. Due to this binding, the IAA distribution between medium and chloroplasts must not be used for calculating the stroma pH, as is possible with ABA. However, from the different distribution of IAA in light and darkness the light-induced alkalization of the chloroplast stroma can be calculated.