Variable Fluorescence Of Photosystem Research Articles

Sulfide and pH effects on variable fluorescence of photosystem II in two strains of the cyanobacterium Oscillatoria amphigranulata

Changes in fluorescence of photosystem II (PS II) chlorophyll were used to monitor the in vivo effects of sulfide and pH on photosynthesis by the cyanobacterium Oscillatoria amphigranulata. O. amphigranulata is capable of both oxygenic photosynthesis and sulfide dependent anoxygenic photosynthesis. A genetic variant of O. amphigranulata which photosynthesizes oxygenically at normal rates, but is incapable of anoxygenic photosynthesis and cannot tolerate sulfide, was also used to explore the mode of action of sulfide. In vivo fluorescence responses of PS II chlorophyll in the first few seconds of exposure to light (Kautsky transients) reflected the electrochemical states of PS II and associated electron donors and acceptors. Kautsky transients showed a distinct difference between PS II of the wild type and the variant, but sulfide lowered fluorescence in both. Kautsky transients with sulfide were similar to transients with addition of NH2OH, NH4 (+) or HCN, indicating sulfide interacts with a protein on the donor side of PS II. The fluorescence steady-state (after 2 min) was measured in the presence of sulfide, cyanide and ammonium with pH ranging from 7.2-8.7. Sulfide and cyanide had the most impact at pH 7.2, ammonium at pH 8.7. This suggests that the uncharged forms (HCN, NH3 and H2S) had the strongest effect on PS II, possibly because of increased membrane permeability.

Inhibition of photosynthesis by freezing temperatures and high light levels in cold‐acclimated seedlings of Scots pine (Pinus sylvestris). – II. Effects on chlorophyll fluorescence at room temperature and 77 K.

Shoots of cold‐acclimated seedlings of Pinus sylvestris L. were exposed to a temperature of –7°C for 4 h, in darkness or at a photon flux density of 1 300 μmol m‐2s‐1. Before and after freezing, fluorescence kinetics of intact needles and isolated chloroplast membranes were measured at both room temperature and 77 K. Maximum and variable fluorescence yield of photosystem II both at room temperature and 77 K decreased strongly after freezing in light, whereas the initial fluorescence yield was little affected. Quenching of maximum and variable fluorescence of photosystem I at 77 K also occurred. The results show that freezing in light damages photosystem II, thereby increasing the radiationless decay at the reaction centres of photosystem II. This is a typical symptom of photoinhibition of photosynthesis. Freezing in darkness did not significantly reduce fluorescence yield of photosystem II or photosystem I. Moreover, electron transport capacity was not significantly affected. We therefore suggest that the inhibition of the CO2 assimilation in pine seedlings by freezing alone does not involve thylakoid inactivation.

Variable fluorescence of photosystem I particles and its application to the study of the structure and function of photosystem I

Chlorophyll a fluorescence in Photosystem I (PSI) particles isolated according to the method of Bengis and Nelson [ J. Biol. Chem. 252, 4564–4569 (1977)]was found to be dependent on the redox state of both P700 and X (an acceptor on the reducing side of PSI). Addition of dithionite plus neutral red to PSI caused an increase in fluorescence intensity and a shift of the main fluorescence peak from 689 to 674 nm. Addition of electron acceptors such as ferredoxin and methyl viologen decreased the fluorescence yield when added to PSI incubated under anaerobic conditions in the presence of excess dichlorophenol indophenol (DCIPH 2). The K m for ferredoxin agreed with that determined from direct measurements of ferredoxin reduction, showing that X is a quencher of fluorescence. P700 was also found to be a quencher of fluorescence, since electron donors such as DCIPH 2, TMPD, and plastocyanin decreased fluorescence with K m 's nearly identical to those observed for P700 + reduction. Chemical modification of PSI (with ethylene diamine + a water-soluble carbodiimide) to make it positively charged increased the fluorescence yield and shifted the 689-nm peak to 674 nm. The K m 's for DCIPH 2 and ferredoxin were decreased. In contrast, modification of PSI with succinic anhydride, which increased the net negative charge, increased the K m for ferredoxin. Salts affected the interaction of methyl viologen with PSI. Both anion and cation selectivity were observed. Limited proteolysis increased the K m for both methyl viologen and ferredoxin, indicating that their binding site on PSI was altered. These results suggest that the binding site for ferredoxin is on either the 70- or the 20-kDa subunit of PSI.