The unrolling of etiolated barley leaves, H. vulgare cv. Pallas, is phytochromemediated. After brief illumination with red light unrolling begins after 8 hr. The possibility that illumination results in the fall in content of an endogenous inhibitor led to the work reported.Inhibitory activity in an "acidic fraction" from etiolated barley leaves, as measured by leaf unrolling and wheat seed germination bioassays, declined in the 6-hr period in the dark following exposure to red light for 10 min.No abscisic acid (ABA) could be detected, either by paper chromatography and wheat coleoptile bioassay or by gas chromatography, in the inhibitory acidic fractions. The inhibitor is therefore not ABA. The inhibitory region on paper chromatograms did not exhibit a strong phenol-like reaction.Linoleic and linolenic acids, identified by gas chromatography and mass spectroscopy, were present in a 1 : 1 ratio in acidic fractions and linoleic acid inhibited unrolling. These fatty acids are chromatographically separable from the leaf-unrolling inhibitor(s). As they also increased nearly threefold in the leaves in the 6 hr following exposure to red light, they are not of interest to the present investigation. They possibly originate from etioplasts.