Thylakoid membrane stability to heat stress studied by flash spectroscopic measurements of the electrochromic shift in intact potato leaves: influence of the xanthophyll content

A flash-induced transthylakoid electric field was measured at 515 nm as an electrochromic absorbance shift in intact potato leaves using a double flash differential spectrophotometer. The decay rate of the electrochromic shift in dark-adapted samples was used to examine the conductance to ions of thylakoid membranes. Heat stress (39.5 degrees C for 15 min) was found to accelerate drastically the electric field decay, with the half decay time falling from more than 200 ms to less than 45 ms. Heat-induced acceleration of the electric field breakdown was insensitive to the PSII electron donor Hydroxylamine and to the ATPase inhibitor dicyclohexylcarbodiimide (DCCD), thus indicating that it reflects an increase in thylakoid membrane permeability after heat stress. This phenomenon did not involve peroxidative damage of membrane lipids. Acceleration of the electric field relaxation exhibited the same temperature dependence as that of PSII deactivation, suggesting that the ionic permeability of thylakoid membranes is one of the most heat-sensitive components of the photosynthetic apparatus. When potato leaves were infiltrated with 100 mol m-3 ascorbate (in a buffer of pH 5), there was massive conversion of the carotenoid violaxanthin to zeaxanthin. This change in carotenoid composition protected thylakoid membranes against heat-induced changes in permeability, as revealed by the maintenance of a slow decay of the 515 nm absorbance change after heat stress. No such effect was observed after treatments which did not induce the violaxanthin-to-zeaxanthin conversion: leaf infiltration with 0 mol m-3 ascorbate (at pH 5 or 8), 100 mol m-3 ascorbate at pH 8 or 100 mol m-3 ascorbate + 5 mol m-3 dithiothreitol at pH 5. Increased stability of the permeability properties of thylakoid membranes was also observed after a mild heat treatment (2 h at 35 degrees C). The data presented suggest that de-epoxidized xanthophylls in vivo stabilize thylakoid membranes and protect thylakoids against heat induced disorganization.