Effects of elevated ozone and low light on diurnal and seasonal carbon gain in sugar maple

The long‐term interactive effects of ozone and light on whole‐tree carbon balance of sugar maple (Acer saccharum Marsh.) seedlings were examined, with an emphasis on carbon acquisition, foliar partitioning into starch and soluble sugars, and allocation to growth. Sugar maple seedlings were fumigated with ambient, 1·7 × ambient and 3·0 × ambient ozone in open‐top chambers for 3 years under low and high light (15 and 35% full sunlight, respectively). Three years of ozone fumigation reduced the total biomass of seedlings in the low‐ and high‐light treatments by 64 and 41%, respectively, but had no effect on whole‐plant biomass allocation. Ozone had no effect on net photosynthesis until late in the growing season, with low‐light seedlings generally exhibiting more pronounced reductions in photosynthesis. The late‐season reduction in photosynthesis was not due to impaired stomatal function, but was associated more with accelerated senescence or senescence‐like injury. In contrast, the 3·0 × ambient ozone treatment immediately reduced diurnal starch accumulation in leaves by over 50% and increased partitioning of total non‐structural carbohydrates into soluble sugars, suggesting that injury repair processes may be maintaining photosynthesis in late spring and early summer at the expense of storage carbon. The results in the present study indicate that changes in leaf‐level photosynthesis may not accurately predict the growth response of sugar maple to ozone in different light environments. The larger reduction in seedling growth under low‐light conditions suggests that seedlings in gap or closed‐canopy environments are more susceptible to ozone than those in a clearing. Similarly, understanding the effects of tropospheric ozone on net carbon gain of a mature tree will require scaling of leaf‐level responses to heterogeneous light environments, where some leaves may be more susceptible than others.     

农田冬小麦生长和产量对臭氧动态暴露的响应

 评估臭氧(O₃)污染对农田冬小麦生长和产量的影响是污染生态学和生理生态学研究的重要内容之一。该研究运用开顶式气室(OTC),对冬小麦‘ 嘉403’(Triticum aestivum cv. Jia 403)进行了O₃动态暴露的田间原位试验。实验设置过滤空气组(CF)、自然大气组(NF)和两个不同浓度的 O₃动态暴露组(DO100和DO150)。结果表明：1) O₃浓度增加,一方面可以改变灌浆期冬小麦叶片气体交换参数的日变化规律;另一方面引起表观 光量子产额、光饱和点和光补偿点等光响应参数的显著降低。这表明灌浆期叶片光合能力的下降是气孔因素和非气孔因素共同作用的结果。2) O₃暴露可以改变小麦形态特征,如植株变矮、叶片衰老加速、 叶面积变小,并最终导致产量大幅下降。     

Ozone exposure over two growing seasons alters root-to-shoot ratio and chemical composition of birch (Betula pendula Roth)

Physiological and chemical responses of 17 birch (Betula pendula Roth) clones to 1.5–1.7 × ambient ozone were studied in an open‐field experiment over two growing seasons. The saplings were studied for growth, foliar visible injuries, net photosynthesis, stomatal conductance, and chlorophyll, carotenoid, Rubisco, total soluble protein, macronutrient and phenolic concentrations in leaves. Elevated ozone resulted in growth enhancement, changes in shoot‐to‐root (s/r) ratio, visible foliar injuries, reduced stomatal conductance, lower late‐season net photosynthesis, foliar nutrient imbalance, changes in phenolic composition, and reductions in pigment, Rubisco and soluble protein contents indicating accelerated leaf senescence. Majority of clones responded to ozone by changing C allocation towards roots, by stomatal closure (reduced ozone uptake), and by investment in low‐cost foliar antioxidants to avoid and tolerate ozone stress. A third of clones, showing increased s/r ratio, relied on inducible efficient high‐cost antioxidants, and enhanced leaf production to compensate ozone‐caused decline in leaf‐level net photosynthesis. However, the best ozone tolerance was found in two s/r ratio‐unaffected clones showing a high constitutive amount of total phenolics, investment in low‐cost antioxidants and N distribution to leaves, and lower stomatal conductance under ozone stress. The results highlight the importance of phenolic compounds in ozone defence mechanisms in the birch population. Depending on the genotype, ozone detoxification was improved by an increase in either efficient high‐cost or less efficient low‐cost antioxidative phenolics, with close connections to whole‐plant physiology.     

不同秧苗素质和移栽密度条件下臭氧胁迫对水稻光合作用、物质生产和产量的影响

近地层臭氧浓度升高使水稻生长受抑进而使产量下降,但这种影响是否因不同栽培条件而异尚不清楚。2011年依托先进的稻田臭氧FACE(Free Air gas Concentration Enrichment)技术平台,以汕优63为供试材料,臭氧设置大气臭氧浓度(Ambient)和高臭氧浓度(比Ambient高50%),秧苗素质设置弱苗(移栽时无分蘖)和壮苗(移栽时带两个分蘖),移栽密度设置低密度(16穴/m2)、中密度(24穴/m2)和高密度(32穴/m2),研究不同秧苗素质和移栽密度条件下臭氧胁迫对水稻生长和产量的影响。结果表明:高浓度臭氧使水稻结实期叶片SPAD值、净光合速率、气孔导度和蒸腾速率明显下降,但胞间CO2浓度和叶温无显著变化。高浓度臭氧对水稻拔节前物质生产量没有影响,但使拔节至抽穗期、抽穗至成熟期物质生产量平均分别降低13%和29%,进而使成熟期生物产量和籽粒产量均显著下降。方差分析表明,臭氧与秧苗素质间没有互作效应,但臭氧与移栽密度的互作对最终产量的影响达显著水平。以上结果表明,臭氧胁迫使水稻生长后期光合受阻,导致物质生产和产量显著下降;适当增加移栽密度可能会减少臭氧胁迫下水稻产量的损失。     

Physiological, stomatal and ultrastructural ozone responses in birch (Betula pendula Roth.) are modified by water stress

The physiological, stomatal and ultrastructural responses to ozone and drought of ozone-sensitive and more ozone-tolerant birch ( Betula pendula Roth.) clones were studied singly and in combination, in a high-stress chamber experiment and in a low-stress open-field experiment. In the chamber experiment, well watered (WW), moderately watered (MW) or drought-stressed (DS) saplings were exposed for 36 d to 0 or 130 nmol mol^∠1 ozone. In the open-field experiment, well watered or drought-stressed saplings were grown for one growing season in ambient air or exposed to 1·8 × ambient ozone. Drought stress reduced growth rate, stomatal conductance, stomatal density and the proportion of starch and thylakoids in chloroplasts, but stimulated net photosynthesis, Rubisco and chlorophyll quantity at the end of the growing season, and increased the size and density of plastoglobuli. Ozone fumigations caused more variable, clone- and exposure-dependent responses in growth, decreased stomatal conductance and net photosynthesis, an increased number of stomata, visible and ultrastructural chloroplast injuries, and enhanced autumn yellowing of the leaves. Ozone-induced changes in plastoglobuli, starch and thylakoids resembled drought responses. The two experiments revealed that, depending on the experimental conditions and the variable, the response to drought and ozone stress can be independent, additive or interactive. Drought protected the plants from ozone injuries under high-stress conditions in the chamber experiment. In the low-stress, open-field experiment, however, enhanced ozone damage was observed in birch saplings grown under restricted water supply.     

Photosynthesis responses to ozone in young trees of three species with different sensitivities, in a 2-year open-top chamber experiment (Curno, Italy)

This paper reports the findings of an open-top chamber experiment carried out in northern Italy (Forest nursery at Curno), during the 2004 and 2005 growth seasons, on Fagus sylvatica and Quercus robur seedlings and on Populus nigra cuttings, in order to test their photosynthesis response to ambient ozone. The experimental protocols were non-filtered air (NF), charcoal-filtered air (CF) and open air (OA). Tests performed included morphological features of leaves; development of foliar symptoms; chlorophyll content, determined by non-destructive means; chlorophyll fluorescence (direct fluorescence and JIP test) and gas exchanges and net photosynthesis (P_N). Main findings were as follows: (1) symptoms occurred early and were extensive in P. nigra , and they occurred later in F. sylvatica , whereas early degeneration of chlorophyll occurred in late summer in Q. robur ; (2) in conditions of ozone exposure, the three species all presented a decline in photosynthesis efficiency and a decrease in P_N, regardless of the symptomatology they displayed; (3) leaf traits are predictors of species-specific sensitivity to ozone—the high density of Q. robur foliar tissues prevents this species from developing visible symptoms and reduces the extent of physiological responses and (4) physiological responses varied from year to year in the same species—responses were lower in the second year of the experiment, when plants had become better acclimatized to plot conditions.     

Interactive effects of elevated ozone and springtime frost on growth and physiology of birch (<Emphasis Type="Italic">Betula pendula</Emphasis>) in field conditions

We studied the impact of ozone enrichment and late frost, singly and interactively, on four birch (Betula pendula Roth) families selected from a naturally regenerated birch stand in southeastern Finland. Seedlings were exposed to 1.5× ambient ozone over one and a half growing seasons using free-air ozone enrichment system. Simulated springtime frost was implemented at the beginning of the second study year, 4 weeks after the bud burst. Plants were measured for timing of bud burst, visible ozone injuries, chlorophyll fluorescence, net photosynthesis and concentrations of photosynthetic pigments, as well as for growth and carbon allocation. Frost treatment caused a rapid 60% decline in net photosynthesis. The recovery of net photosynthesis from acute frost treatment was not complete during the subsequent 3 weeks, which led to significant growth reductions, decreased shoot/root ratio and accumulation of excess nitrogen in the leaves. Photosynthetic responses to ozone were very variable and family-specific. Concentrations of photosynthetic pigments were sensitive to both stress factors, while the maximum quantum yield of PSII was unaffected. Ozone exacerbated the effect of frost only on diameter increment. However, ozone and frost affected different seedling characters, e.g., ozone reduced pigments and frost collapsed net photosynthesis, and these effect combined appear to damage birch seedlings more than a single stress situation.     

Shift in birch leaf metabolome and carbon allocation during long-term open-field ozone exposure

Current and future ozone concentrations have the potential to reduce plant growth and increase carbon demand for defence and repair processes, which may result in reduced carbon sink strength of forest trees in long‐term. Still, there is limited understanding regarding the alterations in plant metabolism and variation in ozone tolerance among tree species and genotypes. Therefore, this paper aims to study changes in birch leaf metabolome due to long‐term realistic ozone stress and to relate these shifts in the metabolism with growth responses. Two European white birch (Betula pendula Roth) genotypes showing different ozone sensitivity were growing under 1.4–1.7 × ambient ozone in open‐field conditions in Central Finland. After seven growing seasons, the trees were analysed for changes in leaf metabolite profiling, based on 339 low molecular weight compounds (including phenolics, polar and lipophilic compounds, and pigments) and related whole‐tree growth responses. Genotype caused most of the variance of metabolite concentrations, while ozone concentration was the second principal component explaining the metabolome profiling. The main ozone caused changes included increases in quercetin‐phenolic compounds and compounds related to leaf cuticular wax layer, whereas several compounds related to carbohydrate metabolism and function of chloroplast membranes and pigments (such as chlorophyll‐related phytol derivatives) were decreasing. Some candidate compounds such as surface wax‐related squalene, 1‐dotriacontanol, and dotriacontane, providing growth‐related tolerance against ozone were demonstrated. This study indicated that current growth‐based ozone risk assessment methods are inadequate, because they ignore ecophysiological impacts due to alterations in leaf chemistry.     

Photosynthesis, Dark Respiration, and Growth of Rumex patientia L. Exposed to Ultraviolet Irradiance (288 to 315 Nanometers) Simulating a Reduced Atmospheric Ozone Column

Net photosynthesis, dark respiration, and growth of Rumex patientia L. exposed to a ultraviolet irradiance (288-315 nanometers) simulating a 0.18 atm·cm stratospheric ozone column were determined. The ultraviolet irradiance corresponding to this 38% ozone decrease from normal was shown to be an effective inhibitor of photosynthesis and leaf growth. The repressive action on photosynthesis accumulated through time whereas leaf growth was retarded only during the initial few days of exposure. Small increases in dark respiration rates occurred but did not continue to increase with longer exposure periods. A reduction in total plant dry weight and leaf area of approximately 50% occurred after 22 days of treatment, whereas chlorophyll concentrations remained unaltered.     

Ambient ozone effects on forest trees of the eastern United States: a review

Tropospheric ozone can affect crop yield and has been reported to cause reductions in growth and biomass of forest tree species in laboratory and glasshouse studies. However, linkages between growth and ambient ozone concentrations in the field are not well established for forest trees. Ambient ozone concentrations have been shown to cause foliar injury on a number of tree species throughout much of the eastern USA. Symptom expression is influenced by endogenous and exogenous factors and, therefore, ozone-exposure/tree-response relationships have been difficult to confirm. Clearly defined, cause-effect relationships between visible injury and growth losses due to ozone have not been validated. Generalizations of sensitivity of forest trees to ozone are complicated by tree development stage, microclimate, leaf phenology, compensatory processes, within-species variation and other interacting stresses. In general, decreases in above-ground growth at ambient ozone levels in the eastern USA appear to be in the range of 0–10% per year. However, these conclusions are based on a small number of tree species, with the vast majority of studies involving individual tree seedlings in a non-competitive environment. Comparative studies of small and large trees indicate that seedlings are not suitable surrogates for predicting responses of mature trees to ozone. Process-level modelling is a promising methodology that has been recently utilized to assess ozone effects on a stand to regional scale, indicating that ozone is affecting forest growth in the eastern USA. The extent and magnitude of the response is variable and depends on many edaphic and climatic factors. It is imperative when conducting assessment exercises, however, that forest biologists constantly keep in mind the tremendous variability that exists within natural systems. Scaling of single site/physiological response phenomena from an individual tree to an ecosystem and/or region necessitates further research.     

不同密度下臭氧胁迫对Ⅱ优084水稻光合作用和物质生产的影响—FACE研究

依托中国稻田臭氧FACE(free air ozone concentration enrichment)技术平台,以超级稻Ⅱ优084为供试材料,臭氧设置当前大气臭氧浓度和高臭氧浓度(比前者高50%),移栽密度设置低密度(16穴·m^-2)、中密度(24穴·m^-2)和高密度(32穴·m^-2),研究不同移栽密度条件下近地层臭氧浓度升高对水稻光合作用、物质生产以及茎鞘非结构性碳水化合物浓度和含量的影响.结果表明: 臭氧浓度升高使水稻移栽后63 d、77 d和86 d剑叶SPAD值分别下降6%、11%和13%,均达显著或极显著水平.臭氧胁迫下结实期叶片净光合速率、气孔导度和蒸腾速率的降幅亦随时间推移而明显增加.高臭氧浓度使水稻抽穗至成熟期的物质生产量平均下降46%,从而使最终生物产量下降25%,均达显著水平.臭氧浓度升高使水稻拔节后茎鞘可溶性糖和淀粉的浓度和含量均显著降低,但使抽穗前茎鞘贮藏同化物的转运率大幅增加.方差分析表明,臭氧与密度间的互作对水稻所有测定参数均无显著影响.综上,近地层臭氧浓度升高使超级稻Ⅱ优084生育中后期的光合和生长均明显受抑,但这种抑制作用不受移栽密度的影响.
     

Whole-tree water use efficiency is decreased by ambient ozone and not affected by O3-induced stomatal sluggishness

Steady-state and dynamic gas exchange responses to ozone visible injury were investigated in an ozone-sensitive poplar clone under field conditions. The results were translated into whole tree water loss and carbon assimilation by comparing trees exposed to ambient ozone and trees treated with the ozone-protectant ethylenediurea (EDU). Steady-state stomatal conductance and photosynthesis linearly decreased with increasing ozone visible injury. Dynamic responses simulated by severing of a leaf revealed that stomatal sluggishness increased until a threshold of 5% injury and was then fairly constant. Sluggishness resulted from longer time to respond to the closing signal and slower rate of closing. Changes in photosynthesis were driven by the dynamics of stomata. Whole-tree carbon assimilation and water loss were lower in trees exposed to ambient O(3) than in trees protected by EDU, both under steady-state and dynamic conditions. Although stomatal sluggishness is expected to increase water loss, lower stomatal conductance and premature leaf shedding of injured leaves aggravated O(3) effects on whole tree carbon gain, while compensating for water loss. On average, WUE of trees exposed to ambient ozone was 2-4% lower than that of EDU-protected control trees in September and 6-8% lower in October.     

Responses of selected birch (Betula pendula Roth) clones to ozone change over time

A long-term free air ozone fumigation experiment was conducted to study changes in physiological ozone responses during tree ontogeny and exposure time in ozone sensitive and tolerant clones of European white birch (Betula pendula Roth), originated from south and central Finland. The trees were grown in soil in natural microclimatic conditions under ambient ozone (control) and 1.4-1.7 x ambient (elevated) ozone from May 1996 to October 2001, and were measured for stem and foliage growth, net photosynthesis, stomatal conductance, stomatal density, visible injuries, foliar starch content and bud formation. After 6 years of exposure, the magnitude of ozone-induced growth reductions in the sensitive clone was 12-48% (significant difference), levels similar or greater than those reported earlier for 2- and 3-year-old saplings undergoing shorter exposures. In the tolerant clone, growth of these larger trees was reduced by 1-38% (significant difference in stem volume), although the saplings had previously been unaffected. In both clones, ozone stress led to significantly reduced leaf-level net photosynthesis but significantly increased stomatal conductance rates during the late summer, resulting in a lower carbon gain for bud formation and the onset of visible foliar injuries. Increasing ozone sensitivity with duration of exposure was explained by a change in growth form (relatively reduced foliage mass), a lower photosynthesis to stomatal conductance ratio during the late summer, and deleterious carry-over effects arising from the reduced number of over-wintering buds.     

Phenotypic plasticity and population differentiation in response to salinity in the invasive cordgrass <Emphasis Type="Italic">Spartina densiflora</Emphasis>

Salinity and tidal inundation induce physiological stress in vascular plant species and influence their distribution and productivity in estuarine wetlands. Climate change-induced sea level rise may magnify these abiotic stressors and the physiological stresses they can cause. Understanding the potential of invasive plants to respond to predicted salinity increases will elucidate their potential niche breadth. To examine potential phenotypic plasticity and functional trait responses to salinity stress in the invasive cordgrass Spartina densiflora, we collected rhizomes from four invasive populations occurring from California to Vancouver Island, British Columbia on the Pacific Coast of North America. In a glasshouse common garden experiment, we measured plant traits associated with growth and allocation, photosynthesis, leaf pigments, and leaf chemistry and calculated plasticity indices across imposed salinity treatments. Fifteen of 21 leaf chemistry, pigment, morphological and physiological traits expressed plastic responses to salinity. When averaged across all measured traits, degree of plasticity did not vary among sampled populations. However, differences in plasticity among populations in response to salinity were observed for 9 of 21 measured plant traits. Leaf chemistry and adaxial leaf rolling trait responses demonstrated the highest degree of plasticity, while growth and allocation measures were less plastic. Phenotypic plasticity of leaf functional traits to salinity indicates the potential of S. densiflora to maintain invasive growth in response to rising estuarine salinity with climate change.     

Anticipated yield loss in field-grown soybean under elevated ozone can be avoided at the expense of leaf growth during early reproductive growth stages in favourable environmental conditions

Ozone is a powerful oxidizing agent which is responsible for more damage to vegetation than any other air pollutant. In this study, leaf growth, photosynthesis, and carbohydrate content were analysed during the seed-filling growth stage of field-grown soybeans exposed to ambient air and 1.2 times ambient ozone concentration using a Free Air Concentration Enrichment (FACE) facility. By contrast to predictions based on controlled-environment and open-top chamber studies, final yield did not differ between treatments, although the cultivar used here was sensitive to ozone damage: growth and carbohydrate content of upper canopy leaves was reduced during the seed-filling stage in which an ozone-induced decrease of photosynthesis was present. However, 2004 was an ideal growing season in central Illinois and the cumulative ozone indices were lower than in previous years. Still, the results indicate that the anticipated yield loss under ozone concentrations was avoided at the expense of leaf growth, as reserves were diverted from vegetative to reproductive organs.     

Seasonal physiological responses and biomass growth in a bioenergy crop (<Emphasis Type="Italic">Phalaris arundinacea</Emphasis> L.) under elevated temperature and CO<Subscript>2</Subscript>, subjected to different water regimes in boreal conditions

We investigated the seasonal variability of effects of elevated temperature (+3.5°C), CO₂ elevation (700 μmol mol⁻¹) and varying water regimes (high to low water content) on physiological responses and biomass growth of reed canary grass (Phalaris arundinacea L., local field-grown cultivar) grown in a boreal environment. In controlled environment greenhouses, various physiological and growth parameters of grass, i.e., light-saturated net photosynthetic rates (P _nmax), water use efficiency (WUE) and optimal photochemical efficiency of photosystem II (F _v/F _m), and leaf area development and biomass of plant organs (leaf, stem, coarse, and fine root) were measured. During the early measurement periods, elevated temperature enhanced leaf photosynthesis and above-ground biomass of reed canary grass; however, this resulted in earlier senescence and lower biomass at the end of measurement period, compared to ambient temperature. This effect was more pronounced under water limitation. Elevated CO₂ enhanced P _nmax, WUE, and leaf area and total plant biomass (above- and below-ground) over growing season. The explanation for imbalance between stimulated photosynthesis and increase in above-ground biomass was that CO₂ enrichment causes a greater increase in the plant’s root system. The combination of elevated temperature and CO₂ slightly increases the growth of plant. Adequate water availability favored photosynthesis and biomass growth of reed canary grass. The temperature- and drought-induced stresses were partially mitigated by elevated CO₂. Other cultivars should be tested in order to identify those that are better adapted to elevated temperatures and CO₂ and variable water levels.     

Does elevated ozone have differing effects in flowering and deblossomed strawberry?

Fruiting and deblossomed plants of strawberry ( Fragaria × ananassa ) were exposed to 92 ppb ozone or filtered air in open-top chambers for 69 d. Flower and fruit production, relative growth rate of leaf area, leaf gas exchange and plant biomass were investigated. Ozone caused an initial acceleration in inflorescence production, which was followed by a reduction in inflorescence production, fruit set, and, later, individual fruit weight, although total fruit yield was not affected before the end of the fumigation period. Ozone accelerated leaf senescence and had a greater negative effect on the rate of photosynthesis in older than in younger leaves in fruiting and deblossomed plants, but the response of net photosynthesis to ozone did not differ between the two groups of plants. Relative growth rate of leaf area was the first parameter to be reduced by ozone fumigation, with the effect being significant in fruiting, but not in deblossomed, plants. Final above-ground biomass was also significantly decreased by ozone in fruiting plants, but not in deblossomed plants. Root and crown biomass were not significantly affected by ozone fumigation in either fruiting or deblossomed plants.     

Herbivory alters plant carbon assimilation,patterns of biomass allocation and nitrogen use efficiency

Herbivory can trigger physiological processes resulting in leaf and whole plant functional changes. The effects of chronic infestation by an insect on leaf traits related to carbon and nitrogen economy in three Prunus avium cultivars were assessed. Leaves from non-infested trees (control) and damaged leaves from infested trees were selected. The insect larvae produce skeletonization of the leaves leaving relatively intact the vein network of the eaten leaves and the abaxial epidermal tissue. At the leaf level, nitrogen content per mass (N_mass) and per area (N_area), net photosynthesis per mass (A_mass) and per area (A_area), photosynthetic nitrogen-use efficiency (PNUE), leaf mass per area (LMA) and total leaf phenols content were measured in the three cultivars. All cultivars responded to herbivory in a similar fashion. The N_mass, A_mass, and PNUE decreased, while LMA and total content of phenols increased in partially damaged leaves. Increases in herbivore pressure resulted in lower leaf size and total leaf area per plant across cultivars. Despite this, stem cumulative growth tended to increase in infected plants suggesting a change in the patterns of biomass allocation and in resources sequestration elicited by herbivory. A larger N investment in defenses instead of photosynthetic structures may explain the lower PNUE and A_mass observed in damaged leaves. Some physiological changes due to herbivory partially compensate for the cost of leaf removal buffering the carbon economy at the whole plant level.     

Photosynthetic parameters as early indicators of ozone injury in apple leaves

Ozone may affect leaf photosynthesis even before visible symptoms become apparent. This study had the objective to test several parameters of chlorophyll fluorescence and leaf gas exchange for their usefulness as indicators of latent ozone injury in the field. Container-grown apple trees (Malus domestica Borkh. cv. Golden Delicious) were exposed to four different ozone levels in open-top chambers. Identical leaves were analyzed in fixed-time intervals for the characteristics of fast fluorescence induction kinetics in vivo. By using high-time resolution, characteristic parameters describing the early photochemical events could be calculated according to the JIP-test. Parameters responsive to the different ozone treatments showed clear dependence on the accumulated ozone dose. Ozone exposure immediately preceding the measurements was more important for the extent of the physiological effects than the total accumulated ozone dose. The most sensitive parameters were the turnover number N (indicating how many times Q_A has to be reduced for full reduction of all acceptors; positively correlated to ozone dose) and D₀, the density of reaction centres per leaf area (negatively correlated to ozone dose). Most parameters analyzed showed clearer responses to ozone on the adaxial than on the abaxial surface of the leaf. Changes in the parameter N were better correlated to ozone doses with low cut-offs (AOT00 and AOT20), whereas changes in D₀ and in the specific electron fluxes per reaction centre were mainly influenced by ozone doses with high cut-offs (AOT80 and AOT100). Leaf gas exchange analyses revealed a higher ozone sensitivity in carboxylation efficiency than in light utilization efficiency and in the rate of light-saturated net photosynthesis. All ozone-induced photosynthetic effects were observed in leaves showing no sign of visible leaf injury. This study identified fluorescence parameters that could be useful for rapid monitoring and early detection of latent leaf injury by ozone.