Photosystem II function and herbicide binding sites during photoinhibition of spinach chloroplasts in-vivo and in-vitro

The time courses of some Photosystem II (PS II) parameters have been monitored during in-vivo and in-vitro photoinhibition of spinach chloroplasts, at room temperature and at 10 °C or 0 °C. Exposing leaf discs of low-light grown spinach at 25 °C to high light led to photoinhibition of chloroplasts in-vivo as manifested by a parallel decrease in the number of functional PS II centres, the variable chlorophyll fluorescence at 77K (F_v/F_m), and the number of atrazine-binding sites. When the photoinhibitory treatment was given at 10 °C, the former two parameters declined in parallel but the loss of atrazine-binding sites occurred more slowly and to a lesser extent. During in-vitro photoinhibition of chloroplast thylakoids at 25 °C, the loss of functional PS II centres proceeded slightly more rapidly than the loss of atrazine-binding sites, and this difference in rate was further increased when the thylakoids were photoinhibited at 0 °C. During the recovery phase of leaf discs (up to 9 h) the increases in F_v/F_m preceded that of the number of functional PS II centres, while only a further decline in the number of atrazine-binding sites was observed. The recovery of variable chlorophyll fluorescence and the concentration of functional PS II centres occurred more rapidly at 25 °C than at 10 °C. These results suggest that the photoinhibition of PS II function is a relatively temperature-independent early photochemical event, whereas the changes in the concentration of herbicide-binding sites appear to be a more complex biochemical process which can occur with a delayed time course.Abbreviations BSA bovine serum albumin - Chl chlorophyll - D1 32kDa herbicide-binding polypeptide in photosystem II and product of the psbA gene - D2 34kDa polypeptide in photosystem II which is the product of the psbD gene - DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea - DCPIP 2,6-dichlorophenolin-dophenol - F₀, F_v, F_m chlorophyll fluorescence with reaction centres open, variable and maximum fluorescence, respectively - LDS lithium dodecyl sulfate - MES 2-(N-morpholino) ethanesulfonic acid - PSII photosystem II - Q_A, Q_B first and second quinone-type PS II acceptor, respectively