Two electrogenic mechanisms contributing to the 560 nm absorption changes in intact Bryopsis chloroplasts

Light -induced absorbance changes at 560 nm in dark-adapted intact chloroplasts of the green alga, Bryopsis maxima were studied in the time range of 200 ms. The initial rise of the 560 nm signals constists of two major components which are both electrochromic absorbance changes of the carotenoids, sipnonein and/or siphonaxanthin, but different in mechanisms of the field formation.The first component (component S) is related to electron transport since it was sensitive to 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) and 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone (DBMIB) and showed a light-intensity dependence similar to that of electron transport in chloroplasts. In the presence of DCMU, component S could be restored on addition of proton-transporting electron donors such as reduced 2,6-dichlorophenol indophenol and phenazine methosulfate, but not on addition of N,N,N′,N′-tetramethyl-p-phenylenediamine which does not carry protons with electrons (Trebst, A. (1974) Annu. Rev. Plant Physiol. 25, 423–458). We propose that component S is due to the electric field set up by the proton translocation across the thylakoid membrane.The second component (component R) was resistant to DCMU and DBMIB. The light-intensity dependency of component R was similar to that of cytochrome f photooxidation which showed saturation at a relatively low light intensity. The magnitude of component R was markedly reduced by phenylmercuric acetate, suggesting the participation of ferredoxin and ferredoxin-NADP oxidoreductase in the mechanism of the field formation responsible for this component. In the presence of DCMU and phenylmercuric acetate, time courses of the 560 nm changes paralleled those of cytochrome f changes. These results indicate that component R is due to the electric field formed between oxidized cytochrome f and other intersystem electron carriers located in the inner part of the thylakoid membrane and reduced electron acceptors of Photosystem I situated on the membrane surface.The complex natures of the 560 nm changes, as well as the contributions of Photosystems I and II to the absorbance changes, are explained in terms of the two electrogenic mechanisms.