Ozone-induced accumulation of carbohydrates changes enzyme activities of carbohydrate metabolism in birch leaves

Birch cuttings (Betula pendula Roth) were grown in a sand-culture system with two concentrations (0.05, HF and 0.005%, LF) of fertilizer containing macronutrients and micronutrients, and were exposed to 90/40 nl l(-1) O(3) (day/night) and < 3 nl l(-1) O(3) (control) for one growing season in the field fumigation chambers at Birmensdorf (Switzerland). Leaves of different ages were analysed for gas exchange, contents of chlorophyll, protein, and for metabolites as well as enzyme activities of carbohydrate metabolism. Ozone reduced net photosynthesis and chlorophyll contents in mature leaves of both fertilization treatments, whereas that of protein was only reduced in high-fertilized plants (HF). However, net photosynthesis, chlorophyll, and protein increased in young leaves of low-fertilized plants (LF). The effects of ozone on enzyme activities of carbohydrate metabolism were most pronounced in leaves of LF plants. Specific activities of the sucrose-cleaving enzymes, sucrose synthase and alkaline invertase, were induced, whereas acid invertase was unchanged. Extractable activity of sucrose phosphate synthase, which is a key enzyme of sucrose synthesis, was reduced. Levels of fructose 2,6-bisphosphate, an inhibitor of sucrose synthesis, were increased in leaves of O(3)/LF plants, but reduced in O(3)/HF plants. In addition, activities of enzymes involved in starch metabolism, ADP-glucose pyrophosphorylase and starch phosphorylase, were lowered in ozone-treated samples and the ratio of ATP:ADP was increased. It is concluded that chronic ozone exposure leads to an inhibition of sucrose synthesis and favours sucrose degradation. This effect is modulated by the nutrient status of the plants, indicating higher O(3) tolerance in HF plants. Furthermore, as the metabolic responses in the ozone-treated samples resemble very closely those observed under end-product inhibition of photosynthesis, we assume that the O(3) effect is mainly due to reduced photosynthate export.