Plastoquinone PQ is engaged in photosynthesis 1. Difference spectra in the UV-region indicate that PQ is an intermediate in the electron transport chain 2. PQ is located as a pool between the two light reactions I and II 3. PQ is reduced by hνII and oxidized by hνI3. In this paper the difference spectra which occur during light excitation of spinach chloroplasts and chlorella vulgaris were measured in the UV-region with high resolution by the high sensitive method of periodical flash photometry 6. On excitation with long flashes (10—1 sec) the difference spectra are similar to those obtained when plastoquinone is reduced to hydroquinone in vitro (see figs. 1, 3 and 6). Deviations in the case of chlorella (fig. 4) are caused by additional NADP-reduction. After extraction of plastoquinone from chloroplasts the difference spectra do not occur during light excitation but they can be produced in full after reconstitution with synthetic plastoquinone A (see fig. 2). In the presents of far red background light (718 nm) which excites only light reaction I the magnitude of the spectrum is doubled (see fig. 3). By excitation with short flashes (10—5 sec), two different spectra were found. The difference spectrum with a life-time of 5 · 10—4 sec (fig. 7) is new and does not correspond to that of plastoquinone in vitro. The difference spectrum with a life-time of about 2·10—2 sec (fig. 5) corresponds to the plastoquinone reduction. The magnitude of this spectrum is ten times smaller than that obtained by excitation with long flashes (fig. 6). The 1:10 ratio of the magnitude of the spectra in short and long flashes can be interpreted by a pool of plastoquinone between the two light reactions with a dynamic capacity of ten electrons. The doubled magnitude of the spectra in far red background light can be interpreted by an electron acceptor pool for plastoquinone with a capacity of five electrons (see also the following papers).