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Between 2004 and 2005 a combined open plot and open-top chamber (OTC) experiment was carried out at Curno (Northern Italy) with cuttings of the poplar clone Oxford (Populus maximowiczii Henry x Populus berolinensis Dippel) grown in open plots (OPs, ambient air), charcoal-filtered OTCs (CF, ozone concentration reduced to 50% of ambient) or non-filtered OTCs (NF, ozone concentration reduced to 95% of ambient). Plants in half of the chambers were kept well-watered (WET), and plants in the remaining chambers were not watered (DRY). The onset and development of visible foliar injury and the stomatal conductance to water vapor (g(w)) were assessed during each growing season. A stomatal conductance model was parameterized by the Jarvis approach, allowing the calculation of ozone stomatal fluxes of plants in each treatment. The pattern of visible symptoms was analyzed in relation to ozone exposure (AOT40, accumulated ozone over a threshold of 40 ppb) and accumulated ozone stomatal fluxes (AF(ST)). Symptoms became visible at an AOT40 between 9584 and 13,110 ppb h and an AF(ST) between 27.85 and 30.40 mmol O(3) m(-2). The development of symptoms was more widespread and faster in plants in WET plots than in DRY plots. A slightly higher dose of ozone was required to cause visible symptoms in plants in DRY plots than in WET plots. By the end of each growing season, plants in the CF OTCs had absorbed a high dose of ozone (31.60 mmol O(3) m(-2) in 2004 and 32.83 mmol O(3) m(-2) in 2005, for WET plots), without developing any visible symptoms. A reliable dose-response relationship was defined by a sigmoidal curve model. The shape of this curve expresses the change in leaf sensitivity and physiologic state over a prolonged ozone exposure. After the appearance of the first symptoms, foliar injury increased more rapidly than the increases in ozone exposure and ozone absorbed dose; however, when the injury incidence reached 75%, the plant response declined.

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Studies on intact cells, membrane vesicles, and reconstituted proteoliposomes have demonstrated in higher plants the existence of an ATP-driven electrogenic proton pump operating at the plasmalemma. There is also evidence of a second ATP-driven H+ pump localized at the tonoplast. The characteristics of both these ATP-driven pumps closely correspond to those of the plasmalemma and tonoplast proton pumps of Neurospora and yeasts.