Abstract. Green barley leaves (Hordeum vulgaris) floated on the surface of 0.05 Mi ethylenediamine disodium tetraacetate, EDTA-ZNa, pH 7.0 and exposed to light (5000 lux) at 250 exhibited a marked bleaching (EDTA-bleaching) visible to the naked eye and paralleled by a striking reduction in content of chlorophylls a and b. This loss of color did not occur in controls which were treated with H,O instead of EDTA (water controls). In darkness the leaves in the water controls were bleached while EDTA-treated leaves retained all their color.EDTA bleaching was observed only in intact leaves. When leaves were boiled EDTA protected their pigment against photodecomposition. Without EDTA boiled leaves were bleached completely in light.When intact green leaves which had been floated on water and exposed to light for 48 hr were treated with boiling ethanol or acetone, the chlorophylls extracted by this procedure did not undergo bleaching if EDTA were present in solution. Under these conditions a green fine grain precipitate formed which was insoluble in ethanol or acetone and was stable in light or darkness.EDTA bleaching of green barley leaveE was inhibited by KCN, and by the addition of casein hydrolysate.During the past 8 years the authors have investigated the effect of gaseous ions on higher plants and have found that moderate concentrations of positive and negative ions markedly stimulate plant growth and increase oxygen consumption (2, 3, 6, 7). They increase both the rates of shrinking and swelling of isolated chloroplasts and the utilization of ATP in the process (4). They enhance iron uptake and in plants suffering from iron chlorosis increase the "residual" iron pool (including cytochromes and other Fe-containing enzymes) while the "active" iron fraction of the tissues decreases and the chlorophvll content is diminished (2, /7). Active iron, contained in the chloroplasts, participates in the production of chlorophyll while residual iron is not associated with chloroplasts nor is it involved in the biosynthesis of chlorophyll (1, 8).In studying the effect of air ions on chlorophyll biosynthesis attempts have been made to use etiolated seedlings for the preparation of isolated chloroplast suspensions which had retained their ability to produce chlorophyll. Unfortunately such isolates bleach when exposed to light; even green chloroplasts become decolorized during light treatment for 72 hr. In our experience retention of the green color of chloroplasts is closelv linked to their retention of catalytic activitv. Therefore it seemed reasonable that if one knew how to prevent the bleaching of chloroplasts, he might hope to obtain etiolated chloroplast suspensions capable of synthesizing chlorophyll in the light. These considerations led us to study the mechanism of decolorization of isolated green chloroplasts in a defined medium.In preliminary experiments it was noted that EDTA, a component of the isolation and suspension medium, was responsible for the decolorization of green chloroplasts. It appeared to us that an ...