Characterization of the Rate-Limiting Steps in the Dark-To-Light Transitions of Closed Photosystem II: Temperature Dependence and Invariance of Waiting Times during Multiple Light Reactions

Rate-limiting steps in the dark-to-light transition of Photosystem II (PSII) were discovered by measuring the variable chlorophyll-a fluorescence transients elicited by single-turnover saturating flashes (STSFs). It was shown that in diuron-treated samples: (i) the first STSF, despite fully reducing the Q(A) quinone acceptor molecule, generated only an F(1)(<F(m)) fluorescence level; (ii) to produce the maximum (F(m)) level, additional excitations were required, which, however, (iii) were effective only with sufficiently long Δτ waiting times between consecutive STSFs. Detailed studies revealed the gradual formation of the light-adapted charge-separated state, PSII(L). The data presented here substantiate this assignment: (i) the Δτ(1/2) half-increment rise (or half-waiting) times of the diuron-treated isolated PSII core complexes (CCs) of Thermostichus vulcanus and spinach thylakoid membranes displayed similar temperature dependences between 5 and –80 °C, with substantially increased values at low temperatures; (ii) the Δτ(1/2) values in PSII CC were essentially invariant on the F(k−)to-F(k+1) (k = 1–4) increments both at 5 and at −80 °C, indicating the involvement of the same physical mechanism during the light-adaptation process of PSII(L). These data are in harmony with the earlier proposed role of dielectric relaxation processes in the formation of the light-adapted charge-separated state and in the variable chlorophyll-a fluorescence of PSII.