Sub-microsecond chlorophyll a delayed fluorescence from photosystem I. Magnetic field-induced increase of the emission yield.

(1) In photosystem I (PS I) particles in the presence of dithionite and intense background illumination at 290 K, an external magnetic field (0-0.22 T) induced an increase, delta F, of the low chlorophyll a emission yield, F (delta F/F approximately or equal to 1-1.5%). Half the effect was obtained at about 35-60 mT and saturation occurred for magnetic fields higher than about 0.15 T. In the absence of dithionite, no field-induced increase was observed. Cooling to 77 K decreased delta F at 685 nm, but not at 735 nm, to zero. Measuring the emission spectra of F and delta F, using continuous excitation light, at 82, 167 and 278 K indicated that the spectra of F and delta F have about the same maximum at about 730, 725 and 700 nm, respectively. However, the spectra of delta F show more long-wavelength emission than the corresponding spectra of F. (2) Only in the presence of dithionite and with (or after) background illumination, was a luminescence (delayed fluorescence) component observed at 735 nm, ater a 15 ns laser flash (530 nm), that decayed in about 0.1 microseconds at room temperature and in approx. 0.2 microseconds at 77 K. A magnetic field of 0.22 T caused an appreciable increase in luminescence intensity after 250 ns, probably mainly caused by an increase in decay time. The emission spectra of the magnetic field-induced increase of luminescence, delta L, at 82, 167 and 278 K coincided within experimental error with those of delta F mentioned above. The temperature dependence of delta F and delta L was found to be nearly the same, both at 685 and at 735 nm. (3) Analogously to the proposal concerning the 0.15 microseconds luminescence in photosystem II (Sonneveld, A., Duysens, L.N.M. and Moerdijk, A. (1980) Proc. Natl. Acad. Sci. U.S.A. 77, 5889-5893), we propose that recombination of the oxidized primary donor P-700+ and the reduced acceptor A-, probably A-1, of PS I causes the observed fast luminescence. The effect of an external magnetic field on this emission may be explained by the radical pair mechanism. The field-induced increase of the 0.1-0.2 microseconds luminescence seems to be at least in large part responsible for the observed increase of the total (prompt + delayed) emission measured during continuous illumination in the presence of a magnetic field.

Transfer and trapping of excitation energy in photosystem II as studied by chlorophyll alpha 2 fluorescence quenching by dinitrobenzene and carotenoid triplet. The matrix model.

1. The curves representing the reciprocal fluorescence yield of chlorophyll alpha of Photosystem II (PS II) in Chlorella vulgaris as a function of the concentration of m-dinitrobenzene in the states P Q and P Q-, are found to be straight parallel lines; P is the primary donor and Q the primary acceptor of PS II. In the weakly trapping state P Q- the half-quenching of dinitrobenzene is about 0.2 mM, in vitro it is of the order of 10 mM. The fluorescence yield as a function of the concentration of a quencher is described for three models for the energy transfer between the units, and the matrix model. If it is assumed that the rate constant of quenching by dinitrobenzene is high and thus the number of dinitrobenzene molecules per reaction center low, it can be concluded that the pigment system of PS II in C. vulgaris is a matrix of chlorophyll molecules in which the reaction centers are embedded. Theoretical and experimental evidence is consistent with such an assumption. For Cyanidium caldarium the zero fluorescence yield phi 0 and its quenching by dinitrobenzene were found to be much smaller than the corresponding quantities for C. vulgaris. Nevertheless, our measurements on C. caldarium could be interpreted by the assumption that the essential properties (rate constants, dinitrobenzene quenching) of PS II are the same for these two species belonging to such widely different groups. 2. The measured dinitrobenzene concentrations required for half-quenching in vivo and other observations are explained by (non-rate-limiting) energy transfer between the chlorophyll alpha molecules of PS II and by the assumptions that dinitrobenzene is approximately distributed at random in the membrane and does not diffuse during excitation. 3. The fluorescence kinetics of C. vulgaris during a 350 ns laser flash of variable intensity could be simulated on a computer using the matrix model. From the observed fluorescence quenching by the carotenoid triplet (CT) and the measurement of the the number of CT per reaction center via difference absorption spectroscopy, the rate constant for quenching of CT is calculated to be kT = 3.3 . 10(11)s-1 which is almost equal to the rate constant of trapping by an open reaction center (Duysens, L.N.M. (1979) CIBA Foundation Symposium 61 (New Series), pp. 323--340). 4. The fluorescence quenching by CT in non-treated spinach chloroplasts after a 500 ns laser flash (Breton, J., Geacintov, N.E. and Swenberg, C.E. (1979) Biochim, Biophys. Acta 548, 616--635) could be explained within the framework of the matrix model when the value for kT is used as given in point 3. 5. The observations mentioned under point 1 indicate that the fluorescence yield phi 0 for centers in trapping state P Q is probably for a fraction exceeding 0.8 emitted by PS II.

Magnetic field-induced increase in chlorophyll a delayed fluorescence of photosystem II: A 100- to 200-ns component between 4.2 and 300 K.

At room temperature the delayed fluorescence (luminescence) of spinach chloroplasts, in which the acceptor Q is prereduced, consists of a component with a lifetime of 0.7 mus and a more rapid component, presumably with a lifetime of 100-200 ns and about the same integrated intensity as the 0.7- mus component. Between 4.2 and 200 K only a 100- to 200-ns luminescence component was found, with an integrated intensity appreciably larger than that at room temperature. At 77 K the 150-ns component approached 63% of saturation at roughly the same energy as the variable fluorescence of photosystem II at room temperature. At 77 K the emission spectra of prompt fluorescence but not that of the 150-ns luminescence had a preponderant additional band at about 735 nm. The 150-ns emission also occurred in the photosystem I-lacking mutant FL5 of Chlamydomonas. These experiments indicate that the 150-ns component originates from photosystem II. At room temperature a magnetic field of 0.22 T stimulated the 0.7-mus delayed fluorescence by about 10%. At 77 K the field-induced increase of the 150-ns component amounted to 40-50%, being responsible for the observed approximately 2% increase of the total emission; the magnetic field increased the lifetime about 20%. In order to explain these phenomena a scheme for photosystem II is presented with an intermediary acceptor W between Q and the primary donor chlorophyll P-680; recombination of P-680(+) and W(-) causes the fast luminescence. The magnetic field effect on this emission is discussed in terms of the radical pair mechanism.

Chlorophyll a fluorescence as a monitor of nanosecond reduction of the photooxidized primary donor P-680 Of photosystem II.

1. Changes in the fluorescence yield of aerobic Chlorella vulgaris have been measured in laser flashes of 15 ns, 30 ns and 350 ns half time. The kinetics after the first flash given after a 3 min dark period could be simulated on a computer using the hypothesis that the oxidized acceptor Q and primary donor P+ are fluorescence quenchers, and Q- is a weak quencher, and that the reduction time for P+ is 20-35 ns. 2. The P+ reduction time for at least an appreciable part of the reaction centers was found to be longer after the second and subsequent flashes. In the first 5 flashes an oscillation was observed. Under steady state conditions, with a pulse separation of 3 s, a reduction time for P+ of about 400 ns for all reaction centers gave the best correspondence between computed and experimental fluorescence kinetics.

Novel method for determination of flow velocities with pulsed nuclear magnetic resonance.

A method for the determination of flow velocities with pulsed nuclear magnetic resonance is presented, based on a sequence of inhomogeneous 180 degrees pulses and a gradient in the stationary magnetic field. Results are shown for a capillary containing water with flow velocities in the range of 0.5 to 5 mm/s.