Isoprene emission rate and intercellular isoprene concentration as influenced by stomatal distribution and conductance

Isoprene emission in relation to stomatal distribution and conductance was determined for the hypostomatous species, aspen and white oak, and the amphistomatous species, cottonwood. For aspen and oak, isoprene emission from the adaxial (nonstomatal) surface was <2% of that from the abaxial (stomatal) surface, even when stomata were closed by addition of abscisic acid (ABA). When treated with ABA, the total flux rate of isoprene emission from leaves of these two hypostomatous species was unchanged, despite decreases in stomatal conductance of over 90%. The lack of control over isoprene emission rate by stomatal conductance, despite the apparent movement of isoprene through the stomatal pores, was due to increases in the intercellular isoprene concentration that compensated for the decreased stomatal conductance and restored the equilibrium between the isoprene synthesis rate and emission rate. This relationship was demonstrated by (a) an experiment in which the decrease in the internal isoprene pool following the imposition of darkness took longer in the presence of ABA than in its absence, and (b) direct measurements of the internal isoprene concentration through vacuum extraction, which revealed substantially higher values in the presence of ABA than in its absence. In the amphistomatous species, cottonwood, isoprene was emitted from both surfaces and addition of ABA caused an increase in isoprene emission from one surface coupled with a decrease from the other surface. The specific surface exhibiting an increase varied among leaves, with some leaves exhibiting an increase from the adaxial surface and other leaves from the abaxial surface. We interpret this as indicating nonuniform stomatal closure with concomitant emission of isoprene at the greatest rate from the surface with the highest stomatal conductance. We also observed an increase in the total isoprene emission rate from cottonwood leaves following treatment with ABA. We interpret this as indicating a stimulation of isoprene synthesis in response to ABA or stomatal closure, with unknown cause.     

Ozone‐induced stomatal sluggishness changes stomatal parameters of Jarvis‐type model in white birch and deciduous oak

• Stomatal ozone flux is closely related to ozone injury to plants. Jarvis‐type multiplicative model has been recommended for estimating stomatal ozone flux in forest trees. Ozone can change stomatal conductance by both stomatal closure and less efficient stomatal control (stomatal sluggishness). However, current Jarvis‐type models do not account for these ozone effects on stomatal conductance in forest trees.
• We examined seasonal course of stomatal conductance in two common deciduous tree species native to northern Japan (white birch: Betula platyphylla var. japonica ; deciduous oak: Quercus mongolica var. crispula ) grown under free‐air ozone exposure. We innovatively considered stomatal sluggishness in the Jarvis‐type model using a simple parameter, s , relating to cumulative ozone uptake (defined as POD : phytotoxic ozone dose).
• We found that ozone decreased stomatal conductance of white birch leaves after full expansion (?28%). However, such a reduction of stomatal conductance by ozone fell in late summer (?10%). At the same time, ozone reduced stomatal sensitivity of white birch to VPD and increased stomatal conductance under low light conditions. In contrast, in deciduous oak, ozone did not clearly change the model parameters.
• The consideration of both ozone‐induced stomatal closure and stomatal sluggishness improved the model performance to estimate stomatal conductance and to explain the dose–response relationship on ozone‐induced decline of photosynthesis of white birch. Our results indicate that ozone effects on stomatal conductance (i.e . stomatal closure and stomatal sluggishness) are crucial for modelling studies to determine stomatal response in deciduous trees, especially in species sensitive to ozone.

     

Model-based analysis of avoidance of ozone stress by stomatal closure in Siebold's beech (Fagus crenata)

Background and Aims

Resistance of plants to ozone stress can be classified as either avoidance or tolerance. Avoidance of ozone stress may be explained by decreased stomatal conductance during ozone exposure because stomata are the principal interface for entry of ozone into plants. In this study, a coupled photosynthesis–stomatal model was modified to test whether the presence of ozone can induce avoidance of ozone stress by stomatal closure.

Methods

The response of Siebold''s beech (Fagus crenata), a representative deciduous tree species, to ozone was studied in a free-air ozone exposure experiment in Japan. Photosynthesis and stomatal conductance were measured under ambient and elevated ozone. An optimization model of stomata involving water, CO₂ and ozone flux was tested using the leaf gas exchange data.

Key Results

The data suggest that there are two phases in the avoidance of ozone stress via stomatal closure for Siebold''s beech: (1) in early summer ozone influx is efficiently limited by a reduction in stomatal conductance, without any clear effect on photosynthetic capacity; and (2) in late summer and autumn the efficiency of ozone stress avoidance was decreased because the decrease in stomatal conductance was small and accompanied by an ozone-induced decline of photosynthetic capacity.

Conclusions

Ozone-induced stomatal closure in Siebold''s beech during early summer reduces ozone influx and allows the maximum photosynthetic capacity to be reached, but is not sufficient in older leaves to protect the photosynthetic system.     

The capacity for thermal protection of photosynthetic electron transport varies for different monoterpenes in Quercus ilex

Heat stress resistance of foliar photosynthetic apparatus was investigated in the Mediterranean monoterpene-emitting evergreen sclerophyll species Quercus ilex. Leaf feeding with fosmidomycin, which is a specific inhibitor of the chloroplastic isoprenoid synthesis pathway, essentially stopped monoterpene emission and resulted in the decrease of the optimum temperature of photosynthetic electron transport from approximately 38 degrees C to approximately 30 degrees C. The heat stress resistance was partly restored by fumigation with 4 to 5 nmol mol(-1) air concentrations of monoterpene alpha-pinene but not with fumigations with monoterpene alcohol alpha-terpineol. Analyses of monoterpene physicochemical characteristics demonstrated that alpha-pinene was primarily distributed to leaf gas and lipid phases, while alpha-terpineol was primarily distributed to leaf aqueous phase. Thus, for a common monoterpene uptake rate, alpha-terpineol is less efficient in stabilizing membrane liquid-crystalline structure and as an antioxidant in plant membranes. Furthermore, alpha-terpineol uptake rate (U) strongly decreased with increasing temperature, while the uptake rates of alpha-pinene increased with increasing temperature, providing a further explanation of the lower efficiency of thermal protection by alpha-terpineol. The temperature-dependent decrease of alpha-terpineol uptake was both due to decreases in stomatal conductance, g(w), and increased volatility of alpha-terpineol at higher temperature that decreased the monoterpene diffusion gradient between the ambient air (F(A)) and leaf (F(I); U = g(w)[F(A) - F(I)]). Model analyses suggested that alpha-pinene reacted within the leaf at higher temperatures, possibly within the lipid phase, thereby avoiding the decrease in diffusion gradient, F(A) - F(I). Thus, these data contribute to the hypothesis of the antioxidative protection of leaf membranes during heat stress by monoterpenes. These data further suggest that fumigation with the relatively low atmospheric concentrations of monoterpenes that are occasionally observed during warm windless days in the Mediterranean canopies may significantly improve the heat tolerance of nonemitting vegetation that grows intermixed with emitting species.     

Constitutive and herbivore-induced monoterpenes emitted by Populus × euroamericana leaves are key volatiles that orient Chrysomela populi beetles

Chrysomela populi beetles feed on poplar leaves and extensively damage plantations. We investigated whether olfactory cues orientate landing and feeding. Young, unexpanded leaves of hybrid poplar emit constitutively a blend of monoterpenes, primarily ( E )- β -ocimene and linalool. This blend attracts inexperienced adults of C. populi that were not previously fed with poplar leaves. In mature leaves constitutively emitting isoprene, insect attack induces biosynthesis and emission of the same blend of monoterpenes, but in larger amount than in young leaves. The olfactometric test indicates that inexperienced beetles are more attracted by adult than by young attacked leaves, suggesting that attraction by induced monoterpenes is dose dependent. The blend does not attract adults that previously fed on poplar leaves. Insect-induced emission of monoterpenes peaks 4 d after the attack, and is also detected in non-attacked leaves. Induced monoterpene emission is associated in mature leaves with a larger decrease of isoprene emission. The reduction of isoprene emission is faster than photosynthesis reduction in attacked leaves, and also occurs in non-attacked leaves. Insect-induced monoterpenes are quickly and completely labelled by ¹³C. It is speculated that photosynthetic carbon preferentially allocated to constitutive isoprene in healthy leaves is in part diverted to induced monoterpenes after the insect attack.     

Volatile emissions and phenolic compound concentrations along a vertical profile of <Emphasis Type="Italic">Populus nigra</Emphasis> leaves exposed to realistic ozone concentrations

Plants are exposed to increasing levels of tropospheric ozone concentrations. This pollutant penetrates in leaves through stomata and quickly reacts inside leaves, thus making plants valuable ozone sinks, but at the same time triggers oxidation processes which lead to leaf injuries. To counteract these negative effects, plants produce an array of antioxidants which react with ozone and reactive molecules which ozone generates in the leaf tissues. In this study, we measured the effect of an ozone concentration which is likely to be attained in many areas of the world in the near future (80 ppb) on leaves of the vertical profile of the widespread agroforestry species Populus nigra. Changes in (1) physiological parameters (photosynthesis and stomatal conductance), (2) ozone uptake, (3) emission of volatile organic compounds (VOCs, i.e. isoprene, methanol and other oxygenated compounds), (4) concentration of antioxidant surface compounds, and (5) concentration of phenolic compounds were assessed. The aim was to assess whether the defensive pathways leading to isoprenoids and phenolics formation were induced when a moderate and chronic increment of ozone is not able to damage photosynthesis. No visual injuries and minor changes in physiology and ozone uptake were observed. The emission of isoprene and oxygenated six-carbon (C6) volatiles were inhibited by ozone, whereas methanol emission was increased, especially in developing leaves. We interpret these results as suggesting an ontogenetic shift in ozone-treated leaves, leading to a slower development and a faster senescence. Most surface and phenolic compounds showed a declining trend in concentration from the youngest to the fully expanded leaves. Ozone reduced the concentrations of chlorogenic acid derivatives at the leaf surface, whereas in total leaf extracts a metabolic shift towards few phenolics with higher antioxidant capacity was observed.     

Estimates of potential ozone stomatal uptake in mature trees of Quercus ilex in a Mediterranean climate

This paper is focused on evaluation of the potential ozone stomatal fluxes (FO₃) under seasonally varying microclimatic conditions at two levels of the canopy of an evergreen Mediterranean plant species, Holm oak (Quercus ilex L.), and on comparing them with ozone hourly averages, in order to assess what are the main environmental/physiological variables that most affect the definition of critical levels for Mediterranean vegetation. Microclimatic factors such as radiation, temperature and wind velocity greatly affected gas exchange rates and stomatal conductance to water vapour measured at different heights. O₃ concentration values highlight a daily cycle with higher nocturnal O₃ concentrations above the canopy than below it. Similar O₃ trends have been observed by using passive diffusive samplers. As a consequence, potential O₃ stomatal fluxes calculated for the upper layer of the canopy are higher than those below the canopy. Moreover, O₃ concentration values show an opposite seasonal trend compared to FO₃. These opposite trends are clearly due to the stomatal closure in drier months, as a protection against excessive water losses that yield low FO₃ values during high O₃ concentration. This paper highlights the different contribution of different Holm oak canopy portions to overall O₃ uptake impact, attributing important roles to microclimatic conditions and to physiological activity related to stomata opening, which in turn is affected by internal and external effectors (hormones, water availability, hydraulic conductance, etc.). We cannot exclude an O₃ effect on stomatal cell guards and on the carbon assimilation process working in the mesophyll cells. Further research needs to be considered to evaluate more clearly the risk of O₃ on Mediterranean vegetation taking into consideration microclimatic conditions, plant physiology and possible plant canopy defensive reactions to O₃, so as to define an air quality standard to protect the vegetation.     

Delayed Onset of Isoprene Emission in Developing Velvet Bean (Mucuna sp.) Leaves

Isoprene (2-methyl-1,3-butadiene) is one of the major volatile hydrocarbons emitted by plants, but its biosynthetic pathway and role in plant metabolism are unknown. Mucuna sp. (velvet bean) is an isoprene emitter, and leaf isoprene emission rate increased as much as 125-fold as leaves developed, and declined in older leaves. Net CO₂ assimilation and stomatal conductance, under different growth and environmental conditions, increased 3 to 5 days prior to an increase in isoprene emission rate, indicating that photosynthetic competence develops before significant isoprene emission occurs.     

Evolution of the isoprene biosynthetic pathway in kudzu

Isoprene synthase converts dimethylallyl diphosphate, derived from the methylerythritol 4-phosphate (MEP) pathway, to isoprene. Isoprene is made by some plants in substantial amounts, which affects atmospheric chemistry, while other plants make no isoprene. As part of our long-term study of isoprene synthesis, the genetics of the isoprene biosynthetic pathway of the isoprene emitter, kudzu (Pueraria montana), was compared with similar genes in Arabidopsis (Arabidopsis thaliana), which does not make isoprene. The MEP pathway genes in kudzu were similar to the corresponding Arabidopsis genes. Isoprene synthase genes of kudzu and aspen (Populus tremuloides) were cloned to compare their divergence with the divergence seen in MEP pathway genes. Phylogenetic analysis of the terpene synthase gene family indicated that isoprene synthases are either within the monoterpene synthase clade or sister to it. In Arabidopsis, the gene most similar to isoprene synthase is a myrcene/ocimene (acyclic monoterpenes) synthase. Two phenylalanine residues found exclusively in isoprene synthases make the active site smaller than other terpene synthase enzymes, possibly conferring specificity for the five-carbon substrate rather than precursors of the larger isoprenoids. Expression of the kudzu isoprene synthase gene in Arabidopsis caused Arabidopsis to emit isoprene, indicating that whether or not a plant emits isoprene depends on whether or not it has a terpene synthase capable of using dimethylallyl diphosphate.     

Does ozone increase ABA levels by non‐enzymatic synthesis causing stomata to close?

Reactive oxygen species (ROS) are widely recognized as important regulators of stomatal aperture and plant gas exchange. The pathways through which stomata perceive ROS share many common linkages with the well characterized signalling pathway for the hormone abscisic acid (ABA), a major driver of stomatal closure. Given reports that ABA receptor mutants have no stomatal response to ozone‐triggered ROS production, as well as evidence that all steps in the ABA biosynthetic pathway can be non‐enzymatically converted by ROS, here we investigated the possibility that ozone closes stomata by directly converting ABA precursors to ABA. In plants where stomata were responsive to ozone, we found that foliar ABA levels rapidly increased upon exposure to ozone. Recovery of gas exchange post‐exposure occurred only when ABA levels declined. Our data suggest that stomatal closure in response to ozone exposure occurs as a result of direct oxidation of ABA precursors leading to ABA production, but the importance of this ROS interaction remains uncertain under normal photosynthetic conditions.     

Isoprene Produced by Leaves Protects the Photosynthetic Apparatus against Ozone Damage, Quenches Ozone Products, and Reduces Lipid Peroxidation of Cellular Membranes

Many plants invest carbon to form isoprene. The role of isoprene in plants is unclear, but many experiments showed that isoprene may have a role in protecting plants from thermal damage. A more general antioxidant action has been recently hypothesized on the basis of the protection offered by exogenous isoprene in nonemitting plants exposed to acute ozone doses. We inhibited the synthesis of endogenous isoprene by feeding fosmidomycin and observed that Phragmites australis leaves became more sensitive to ozone than those leaves forming isoprene. Photosynthesis, stomatal conductance, and fluorescence parameters were significantly affected by ozone only in leaves on which isoprene was not formed. The protective effect of isoprene was more evident when the leaves were exposed for a long time (8 h) to relatively low (100 nL L(-1)) ozone levels than when the exposure was short and acute (3 h at 300 nL L(-1)). Isoprene quenched the amount of H(2)O(2) formed in leaves and reduced lipid peroxidation of cellular membranes caused by ozone. These results indicate that isoprene may exert its protective action at the membrane level, although a similar effect could be obtained if isoprene reacted with ozone before forming active oxygen species. Irrespective of the mechanism, our results suggest that endogenous isoprene has an important antioxidant role in plants.     

Ozone concentration in leaf intercellular air spaces is close to zero

Transpiration and ozone uptake rates were measured simultaneously in sunflower leaves at different stomatal openings and various ozone concentrations. Ozone uptake rates were proportional to the ozone concentration up to 1500 nanoliters per liter. The leaf gas phase diffusion resistance (stomatal plus boundary layer) to water vapor was calculated and converted to the resistance to ozone multiplying it by the theoretical ratio of diffusion coefficients for water vapor and ozone in air (1.67). The ozone concentration in intercellular air spaces calculated from the ozone uptake rate and diffusion resistance to ozone scattered around zero. The ozone concentration in intercellular air spaces was measured directly by supplying ozone to the leaf from one side and measuring the equilibrium concentration above the other side, and it was found to be zero. The total leaf resistance to ozone was proportional to the gas phase resistance to water vapor with a coefficient of 1.68. It is concluded that ozone enters the leaf by diffusion through the stomata, and is rapidly decomposed in cell walls and plasmalemma.     

A hypothesis on the evolution of isoprenoid emission by oaks based on the correlation between emission type and Quercus taxonomy

We show that Mediterranean oaks that emit isoprene, monoterpenes or no isoprenoids belong to different subgenera as indicated by morpho-taxonomy and molecular genetics. On the other hand, oaks from North America and Asia that are taxonomically similar to the Mediterranean monoterpene emitter Q. ilex emit isoprene only. We surmise that isoprene emission is a genetic character which evolved ancestrally in the oak genus since this is the prevalent emission type in oaks widespread around the world and adapted to different environments. This ancestral character may have been either lost or modified in more recent clades such as those originating the Mediterranean oaks. If our hypothesis is correct then the taxonomy of European oaks is validated by this independent trait. Isoprenoid emission could serve as a chemo-taxonomical marker and could be used to reconstruct the phylogeny of oaks in association with molecular markers. Received: 30 November 1997  / Accepted: 28 February 1998     

Distinct responses to ozone of abaxial and adaxial stomata in three Euramerican poplar genotypes

Ozone induces stomatal sluggishness, which impacts photosynthesis and transpiration. Stomatal responses to variation of environmental parameters are slowed and reduced by ozone and may be linked to difference of ozone sensitivity. Here we determine the ozone effects on stomatal conductance of each leaf surface. Potential causes of this sluggish movement, such as ultrastructural or ionic fluxes modification, were studied independently on both leaf surfaces of three Euramerican poplar genotypes differing in ozone sensitivity and in stomatal behaviour. The element contents in guard cells were linked to the gene expression of ion channels and transporters involved in stomatal movements, directly in microdissected stomata. In response to ozone, we found a decrease in the stomatal conductance of the leaf adaxial surface correlated with high calcium content in guard cells compared with a slight decrease on the abaxial surface. No ultrastructural modifications of stomata were shown except an increase in the number of mitochondria. The expression of vacuolar H⁺/Ca²⁺‐antiports (CAX1 and CAX3 homologs), β‐carbonic anhydrases (βCA1 and βCA4) and proton H⁺‐ATPase (AHA11) genes was strongly decreased under ozone treatment. The sensitive genotype characterized by constitutive slow stomatal response was also characterized by constitutive low expression of genes encoding vacuolar H⁺/Ca²⁺‐antiports.     

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.     

Monoterpene synthase activities in leaves of Picea abies (L.) Karst. and Quercus ilex L

In addition to direct ecological functions in the interaction of plants with the environment, the emission of monoterpenes, especially from the foliage of evergreen trees, is of great importance for the production of ozone and photochemical oxidants in the troposphere. In the present work, we established a reproducible non-radioactive standard enzyme assay and characterized monoterpene synthase activities in needles of Norway spruce (Picea abies (L.) Karst.) and in leaves of holm oak (Quercus ilex L.). In Norway spruce, the dominant monoterpenes formed were alpha-pinene, camphene, and to a lesser extent beta-pinene and limonene. In holm oak, alpha-pinene, sabinene, and beta-pinene were the main products, while limonene was a minor component. Under optimum conditions, in both Norway spruce and holm oak, monoterpene formation remained constant up to 180 min and 90 min, respectively, and varied with the buffer and Mg2+ and Mn2+ concentrations used. Optimum temperature for monoterpene synthase activity was 40 degrees C in both species; optimal pH ranged between 6.5 and 7.5 in both species. Apparent Michaelis-constants for the substrate GDP were ca. 17.9 +/- 5.1 microM for Norway spruce and ca. 69.4 +/- 22.1 microM for holm oak. Molecular weight determination by FPLC indicated that the monoterpene synthases in Norway spruce and holm oak have native molecular weights of ca. 59 and 50 kDa, respectively.     

Disruption of a gene encoding C4-dicarboxylate transporter-like protein increases ozone sensitivity through deregulation of the stomatal response in Arabidopsis thaliana

To understand better the plant response to ozone, we isolated and characterized an ozone-sensitive (ozs1) mutant strain from a set of T-DNA-tagged Arabidopsis thaliana ecotype Columbia. The mutant plants show enhanced sensitivity to ozone, desiccation and sulfur dioxide, but have normal sensitivity to hydrogen peroxide, low temperature and high light levels. The T-DNA was inserted at a single locus which is linked to ozone sensitivity. Identification of the genomic sequences flanking the T-DNA insertion revealed disruption of a gene encoding a transporter-like protein of the tellurite resistance/C(4)-dicarboxylate transporter family. Plants with either of two different T-DNA insertions in this gene were also sensitive to ozone, and these plants failed to complement ozs1. Transpiration levels, stomatal conductance levels and the size of stomatal apertures were greater in ozs1 mutant plants than in the wild type. The stomatal apertures of ozs1 mutant plants responded to light fluctuations but were always larger than those of the wild-type plants under the same conditions. The stomata of the mutant and wild-type plants responded similarly to stimuli such as light, abscisic acid, high concentrations of carbon dioxide and ozone. These results suggest that OZS1 helps to close stomata, being not involved in the responses to these signals.     

The effects of ozone on growth and stomatal response in the F<Subscript>2</Subscript> generation of hybrid poplar (<Emphasis Type="Italic">Populus trichocarpa × Populus deltoides</Emphasis>)

Thirty-six F₂ hybrid poplar (Populus trichocarpa × P. deltoides) clones were fumigated with ozone to record its effects on growth, correlate them with stomatal response and screen for ozone sensitivity. Fumigation was applied for 6 to 9 h each day for approximately 3 months at ozone concentrations of 85 to 128 μg g⁻¹ using open-top chambers. Height, diameter, number of leaves, stomatal conductance, transpiration rate, total biomass, biomass components and root/shoot ratios were reduced by ozone stress. Percent of leaf fall in ozone-treated plants was nearly three times higher than in control plants exposed to charcoal-filtered air. Leaf senescence, because of ozone exposure, did not appear to be associated with reduced biomass production. Some clones had a high percentage of leaf-fall with ozone exposure, but were able to maintain total biomass production near that of the control. Their response may be an example of an ability to adjust or compensate for ozone damage. There was no significant or consistent relationship between stomatal conductance and total biomass or the change in stomatal conductance as a result of ozone exposure and the change in total biomass. Taken together, these results suggest that effects of ozone on poplar growth cannot be solely correlated to changes in stomatal conductance, more physiological and biochemical parameters should be examined.     

Defining hybrid poplar (Populus deltoides × Populus trichocarpa) tolerance to ozone: identifying key parameters

This study examined whether two genotypes of hybrid poplar ( Populus deltoides  ×  Populus trichocarpa ), previously classified as ozone tolerant and ozone sensitive, had differing physiological and biochemical responses when fumigated with 120 nL L⁻¹ ozone for 6 h per day for eight consecutive days. Isoprene emission rate, ozone uptake and a number of physiological and biochemical parameters were investigated before, during and after fumigation with ozone. Previous studies have shown that isoprene protects plants against oxidative stress. Therefore, it was hypothesized that these two genotypes would differ in either their basal isoprene emission rates or in the response of isoprene to fumigation by ozone.
Our results showed that the basal emission rates of isoprene, physiological responses and ozone uptake rates were all similar. However, significant differences were found in visible damage, carotenoids, hydrogen peroxide (H₂O₂), thiobarbituric acid reactions (TBARS) and post-fumigation isoprene emission rates. It is shown that, although the classification of ozone tolerance or sensitivity had been previously clearly and carefully defined using one particular set of parameters, assessment of other key variables does not necessarily lead to the same conclusions. Thus, it may be necessary to reconsider the way in which plants are classified as ozone tolerant or sensitive.     

On the monoterpene emission under heat stress and on the increased thermotolerance of leaves of Quercus ilex L. fumigated with selected monoterpenes

Leaves of the monoterpene emitter Quercus ilex were exposed to a temperature ramp with 5 °C steps from 30 to 55 °C while maintained under conditions in which endogenous emission of monoterpenes was allowed or suppressed, or under fumigation with selected exogenous monoterpenes. Fumigation with monoterpenes reduced the decline of photosynthesis, photorespiration and monoterpene emission found in non-fumigated leaves exposed to high temperatures. It also substantially increased respiration when photosynthesis and photorespiration were inhibited by low O₂ and CO₂-free air. These results indicate that, as previously reported for isoprene, monoterpenes may help plants cope with heat stress. Monoterpenes may enhance membrane stability, thus providing a rather non-specific protection of photosynthetic and respiratory processes. Monoterpene emission was maximal at a temperature of 35 °C and was inhibited at higher temperatures. This is likely to be the result of the temperature dependency of the enzymes involved in monoterpene synthesis. In contrast to other monoterpenes, cis- and trans- β -ocimene did not respond to exposure to high temperatures. Cis- β -ocimene also did not respond to low O₂ or to fumigation. These results indicate that cis and trans- β -ocimene may have a different pathway of formation that probably does not involve enzymatic synthesis.