Sunflower seedlings subjected to increasing stress by water deficit: Changes in superoxide production related to the composition of thylakoid membranes

Free radicals, such as the superoxide anion, can damage the photosynthetic apparatus. At the same time they are increasingly formed as by-products of electron transport during stress conditions. For this reason, we studied the kinetics of formation and decay of superoxide anions in relation to changes, induced by increasing water deficit, in the composition of thylakoids. Seedlings of sunflower (Helianthus annuus L. cv. Licia Stella) were subjected to either a mild, a moderate, or a severe water deficit. Upon illumination of thylakoid membranes containing Tiron, we found first order kinetics of formation and decay of the Tiron semiquinone radical. At each stress intensity a higher rate of superoxide formation than in the control was observed. The different decay rates suggested a changed chemical environment around the radical. In comparison with the control, the levels of carotenoids and hydrophobic proteins in the thylakoids decreased, and the lipid/protein ratio increased after a mild water deficit. After moderate drought, the increased levels of hydrophilic proteins and carotenoids seem unsufficient to limit the improved capacity to leak electrons from the thylakoids. After a severe water stress, a decrease in unsaturation and in linolenic acid as well as a loss of lipolytic antioxidants took place, which may have rendered the thylakoids more sensiive to attack by activated forms of oxygen. The results suggest that during water deficit, the interactions among membrane components change in relation to a changed chemical composition. Such changes are probably among the causes for changes in the conformation of proteins and in the position of various molecular species in he lipid bilayer, so that they become more exposed to molecular oxygen.