On the chlorophyll a fluorescence yield in chloroplasts upon excitation with twin turnover flashes (TTF) and high frequency flash trains.

Chlorophyll fluorescence is routinely taken as a quantifiable measure of the redox state of the primary quinone acceptor Q(A) of PSII. The variable fluorescence in thylakoids increases in a single turnover flash (STF) from its low dark level F (o) towards a maximum F (m) (STF) when Q(A) becomes reduced. We found, using twin single turnover flashes (TTFs) that the fluorescence increase induced by the first twin-partner is followed by a 20-30% increase when the second partner is applied within 20-100 micros after the first one. The amplitude of the twin response shows a period-of-four oscillation associated with the 4-step oxidation of water in the Kok cycle (S states) and originates from two different trapped states with a life time of 0.2-0.4 and 2-5 ms, respectively. The oscillation is supplemented with a binary oscillation associated with the two-electron gate mechanism at the PSII acceptor side. The F(t) response in high frequency flash trains (1-4 kHz) shows (i) in the first 3-4 flashes a transient overshoot 20-30% above the F (m) (STF) = 3*F (o) level reached in the 1st flash with a partial decline towards a dip D in the next 2-3 ms, independent of the flash frequency, and (ii) a frequency independent rise to F (m) = 5*F (o) in the 3-60 ms time range. The initial overshoot is interpreted to be due to electron trapping in the S(0) fraction with Q(B)-nonreducing centers and the dip to the subsequent recovery accompanying the reoxidation of the double reduced acceptor pair in these RCs after trapping. The rise after the overshoot is, in agreement with earlier findings, interpreted to indicate a photo-electrochemical control of the chlorophyll fluorescence yield of PSII. It is anticipated that the double exciton and electron trapping property of PSII is advantageous for the plant. It serves to alleviate the depression of electron transport in single reduced Q(B)-nonreducing RCs, associated with electrochemically coupled proton transport, by an increased electron trapping efficiency in these centers.