Redox changes of ferredoxin, P700, and plastocyanin measured simultaneously in intact leaves

Properties and performance of the recently introduced Dual/KLAS-NIR spectrophotometer for simultaneous measurements of ferredoxin (Fd), P700, and plastocyanin (PC) redox changes, together with whole leaf chlorophyll a (Chl) fluorescence (emission >760, 540 nm excitation) are outlined. Spectral information on in vivo Fd, P700, and PC in the near-infrared region (NIR, 780-1000 nm) is presented, on which the new approach is based. Examples of application focus on dark-light and light-dark transitions, where maximal redox changes of Fd occur. After dark-adaptation, Fd reduction induced by moderate light parallels the Kautsky effect of Chl fluorescence induction. Both signals are affected analogously by removal of O₂. A rapid type of Fd reoxidation, observed after a short pulse of light before light activation of linear electron transport (LET), is more pronounced in C4 compared to C3 leaves and interpreted to reflect cyclic PS I (CET). Light activation of LET, as assessed via the rate of Fd reoxidation after short light pulses, occurs at very low intensities and is slowly reversed (half-time ca. 20 min). Illumination with strong far-red light (FR, 740 nm) reveals two fractions of PS I, PS I (LET), and PS I (CET), differing in the rates of Fd reoxidation upon FR-off and the apparent equilibrium constants between P700 and PC. Parallel information on oxidation of Fd and reduction of P700 plus PC proves essential for identification of CET. Comparison of maize (C4) with sunflower and ivy (C3) responses leads to the conclusion that segregation of two types of PS I may not only exist in C4 (mesophyll and bundle sheath cells), but also in C3 photosynthesis (grana margins plus end membranes and stroma lamellae).