Effects of ozone on apoplast/cytoplasm partitioning of ascorbic acid in snap bean

Apoplast/cytoplasm partitioning of ascorbic acid (AA) was examined in four genotypes of snap bean ( Phaseolus vulgaris L.) known to differ in ozone sensitivity. Plants were grown in pots under field conditions using open-top chambers to establish charcoal-filtered (CF) air (36 nmol mol^?1 ozone) or elevated ozone (77 nmol mol^?1 ozone) treatments. AA in fully expanded leaves of 36-day-old plants was separated into apoplast and cytoplasm fractions by vacuum infiltration methods using glucose 6-phosphate as a marker for cytoplasm contamination. Apoplast ascorbate levels ranged from 30 to 150 nmol g^?1 fresh weight. Ozone-sensitive genotypes partitioned 1–2% of total AA into the apoplast under CF conditions and up to 7% following a 7-day ozone exposure. In contrast, an ozone-tolerant genotype partitioned 3–4% of total leaf AA into the leaf apoplast in both CF and ozone-treated plants. The results suggest that genetic background and ozone stress are factors that affect AA levels in the extracellular space. For all genotypes, the fraction of AA in the oxidized form was higher in the apoplast compared to the cytoplasm, indicative of a more oxidizing environment within the cell wall.