Alteration in Light Spectra Causes Opposite Responses in Volatile Phenylpropanoids and Terpenoids Compared with Phenolic Acids in Sweet Basil (Ocimum basilicum) Leaves.

Basil (Ocimum basilicum, cv. Dolly) grew under three different light spectra (A, B, and C) created by light-emitting diode lamps. The proportions of UV-A, blue, and green-yellow wavelengths decreased linearly from A to C, and the proportions of red and far-red wavelengths increased from A to C. Photosynthetic photon flux density was 300 µmol m-2 s-1 in all spectra. The spectrum C plants had highest concentrations of phenolic acids (main compounds: rosmarinic acid and cichoric acid), lowest concentrations and emissions of phenylpropanoid eugenol and terpenoids (main compounds: linalool and 1,8-cineole), highest dry weight, and lowest water content. Conversely, spectra A and B caused higher terpenoid and eugenol concentrations and emissions and lower concentrations of phenolic acids. High density of peltate glandular trichomes explained high terpenoid and eugenol concentrations and emissions. Basil growth and secondary compounds affecting aroma and taste can be modified by altering light spectra; however, increasing terpenoids and phenylpropanoids decreases phenolic acids and growth and vice versa.

Effects of elevated ozone and warming on terpenoid emissions and concentrations of Norway spruce depend on needle phenology and age.

Norway spruce (Picea abies (L.) Karst) trees are affected by ongoing climate change, including warming and exposure to phytotoxic levels of ozone. Non-volatile terpenoids and volatile terpenoids (biogenic organic volatile compounds, BVOCs) protect spruce against biotic and abiotic stresses. BVOCs also affect the atmosphere's oxidative capacity. Four-year-old Norway spruce were exposed to elevated ozone (EO) (1.4 × ambient) and warming (1.1 °C + ambient air) alone and in combination on an open-field exposure site in Central Finland. Net photosynthesis, needle terpenoid concentrations and BVOC emissions were measured four times during the experiment's second growing season: after bud opening in May, during the mid-growing season in June, and after needle maturation in August and September. Warming increased terpene concentrations in May due to advanced phenology and decreased them at the end of the growing season in matured current-year needles. Ozone enhanced these effects of warming on several compounds. Warming decreased concentrations of oxygenated sesquiterpenes in previous-year needles. Decreased emissions of oxygenated monoterpenes by warming and ozone alone in May were less prominent when ozone and warming were combined. A similar interactive treatment response in isoprene, camphene, tricyclene and α-pinene was observed in August when the temperature and ozone concentration was high. The results suggest long-term warming may reduce the terpenoid-based defence capacity of young spruce, but the defence capacity can be increased during the most sensitive growth phase (after bud break), and when high temperatures or ozone concentrations co-occur. Reduced BVOC emissions from young spruce may decrease the atmosphere's oxidative capacity in the warmer future, but the effect of EO may be marginal because less reactive minor compounds are affected.

Cell structural changes in the mesophyll of Norway spruce needles by elevated ozone and elevated temperature in open-field exposure during cold acclimation.

The effects of elevated ozone (1.4× ambient) and temperature (ambient +1.3 °C) alone and in combination were studied on the needle cell structure of soil-grown Norway spruce seedlings in the late growing season and winter. Temperature treatment continued over winter and lengthened the snow-free period. Elevated temperature caused microscopic changes related to photosynthesis (decreased chloroplast size and increased number), carbon storage (reduced starch and increased cytoplasmic lipids) and defence (decreased mitochondrial size and proportion per cytoplasm, increased peroxisomes and plastoglobuli, altered appearance of tannins). The results suggest increased oxidative stress by elevated temperature and altered allocation of limited carbon reserve to defence. The number of peroxisomes and plastoglobuli remained high in the outer cells of needles of ozone-exposed seedlings but decreased in the inner cells. This may indicate defence allocation to cells close to the stomata and surface, which are experiencing more oxidative stress. Ozone reduced winter hardiness based on seasonal changes in chloroplast shape and location in the cells. The effects of ozone became evident at the end of the growing season, indicating the effect of cumulative ozone dose or that the seedlings were vulnerable to ozone at the later phases of winter hardening. Elevated temperature increased cellular damage in early winter and visible damage in spring, and the damage was enhanced by ozone. In conclusion, the study suggests that modest air temperature elevation increases stress at the cell structural level in spruce seedlings and is enhanced by low ozone elevation. Future climatic conditions where snow cover is formed later or is lacking but temperatures are low can increase the risk of severe seedling damage, and current and future predicted ozone concentrations increase this risk.

Carbohydrate concentrations and freezing stress resistance of silver birch buds grown under elevated temperature and ozone.

The effects of slightly elevated temperature (+0.8 °C), ozone (O3) concentration (1.3 × ambient O3 concentration) and their combination on over-wintering buds of Betula pendula Roth were studied after two growing seasons of exposure in the field. Carbohydrate concentrations, freezing stress resistance (FSR), bud dry weight to fresh weight ratio, and transcript levels of cytochrome oxidase (COX), alternative oxidase (AOX) and dehydrin (LTI36) genes were studied in two clones (clones 12 and 25) in December. Elevated temperature increased the bud dry weight to fresh weight ratio and the ratio of raffinose family oligosaccharides to sucrose and the transcript levels of the dehydrin (LTI36) gene (in clone 12 only), but did not alter the FSR of the buds. Genotype-specific alterations in carbohydrate metabolism were found in the buds grown under elevated O3. The treatments did not significantly affect the transcript level of the COX or AOX genes. No clear pattern of an interactive effect between elevated temperature and O3 concentration was found. According to these data, the increase in autumnal temperatures and slightly increasing O3 concentrations do not increase the risk for freeze-induced damage in winter in silver birch buds, although some alterations in bud physiology occur.

Needle metabolome, freezing tolerance and gas exchange in Norway spruce seedlings exposed to elevated temperature and ozone concentration.

Northern forests are currently experiencing increasing mean temperatures, especially during autumn and spring. Consequently, alterations in carbon sequestration, leaf biochemical quality and freezing tolerance (FT) are likely to occur. The interactive effects of elevated temperature and ozone (O(3)), the most harmful phytotoxic air pollutant, on Norway spruce (Picea abies (L.) Karst.) seedlings were studied by analysing phenology, metabolite concentrations in the needles, FT and gas exchange. Sampling was performed in September and May. The seedlings were exposed to a year-round elevated temperature (+1.3 °C), and to 1.4× ambient O(3) concentration during the growing season in the field. Elevated temperature increased the concentrations of amino acids, organic acids of the citric acid cycle and some carbohydrates, and reduced the concentrations of phenolic compounds, some organic acids of the shikimic acid pathway, sucrose, cyclitols and steroids, depending on the timing of the sampling. Although growth onset occurred earlier at elevated temperature, the temperature of 50% lethality (LT(50)) was similar in the treatments. Photosynthesis and the ratio of photosynthesis to dark respiration were reduced by elevated temperature. Elevated concentrations of O(3) reduced the total concentration of soluble sugars, and tended to reduce LT(50) of the needles in September. These results show that alterations in needle chemical quality can be expected at elevated temperatures, but the seedlings' sensitivity to autumn and spring frosts is not altered. Elevated O(3) has the potential to disturb cold hardening of Norway spruce seedlings in autumn, and to alter the water balance of the seedling through changes in stomatal conductance (g(s)), while elevated temperature is likely to reduce g(s) and consequently reduce the O(3)-flux inside the leaves.

Vertical profiles reveal impact of ozone and temperature on carbon assimilation of Betula pendula and Populus tremula.

Rising temperature and tropospheric ozone (O(3)) concentrations are likely to affect carbon assimilation processes and thus the carbon sink strength of trees. In this study, we investigated the joint action of elevated ozone and temperature on silver birch (Betula pendula) and European aspen (Populus tremula) saplings in field conditions by combining free-air ozone exposure (1.2 × ambient) and infrared heaters (ambient +1.2 °C). At leaf level measurements, elevated ozone decreased leaf net photosynthesis (P(n)), while the response to elevated temperature was dependent on leaf position within the foliage. This indicates that leaf position has to be taken into account when leaf level data are collected and applied. The ozone effect on P(n) was partly compensated for at elevated temperature, showing an interactive effect of the treatments. In addition, the ratio of photosynthesis to stomatal conductance (P(n)/g(s) ratio) was decreased by ozone, which suggests decreasing water use efficiency. At the plant level, the increasing leaf area at elevated temperature resulted in a considerable increase in photosynthesis and growth in both species.

Differential gene expression in senescing leaves of two silver birch genotypes in response to elevated CO2 and tropospheric ozone.

Long-term effects of elevated CO(2) and O(3) concentrations on gene expression in silver birch (Betula pendula Roth) leaves were studied during the end of the growing season. Two birch genotypes, clones 4 and 80, with different ozone growth responses, were exposed to 2x ambient CO(2) and/or O(3) in open-top chambers (OTCs). Microarray analyses were performed after 2 years of exposure, and the transcriptional profiles were compared to key physiological characteristics during leaf senescence. There were genotypic differences in the responses to CO(2) and O(3). Clone 80 exhibited greater transcriptional response and capacity to alter metabolism, resulting in better stress tolerance. The gene expression patterns of birch leaves indicated contrasting responses of senescence-related genes to elevated CO(2) and O(3). Elevated CO(2) delayed leaf senescence and reduced associated transcriptional changes, whereas elevated O(3) advanced leaf senescence because of increased oxidative stress. The combined treatment demonstrated that elevated CO(2) only temporarily alleviated the negative effects of O(3). Gene expression data alone were insufficient to explain the O(3) response in birch, and additional physiological and biochemical data were required to understand the true O(3) sensitivity of these clones.

Leaf size and surface characteristics of Betula papyrifera exposed to elevated CO2 and O3.

Betula papyrifera trees were exposed to elevated concentrations of CO(2) (1.4 x ambient), O(3) (1.2 x ambient) or CO(2) + O(3) at the Aspen Free-air CO(2) Enrichment Experiment. The treatment effects on leaf surface characteristics were studied after nine years of tree exposure. CO(2) and O(3) increased epidermal cell size and reduced epidermal cell density but leaf size was not altered. Stomatal density remained unaffected, but stomatal index increased under elevated CO(2). Cuticular ridges and epicuticular wax crystallites were less evident under CO(2) and CO(2) + O(3). The increase in amorphous deposits, particularly under CO(2) + O(3,) was associated with the appearance of elongated plate crystallites in stomatal chambers. Increased proportions of alkyl esters resulted from increased esterification of fatty acids and alcohols under elevated CO(2) + O(3). The combination of elevated CO(2) and O(3) resulted in different responses than expected under exposure to CO(2) or O(3) alone.

Diurnal changes in photosynthetic parameters of Populus tremuloides, modulated by elevated concentrations of CO2 and/or O3 and daily climatic variation.

The diurnal changes in light-saturated photosynthesis (Pn) under elevated CO(2) and/or O(3) in relation to stomatal conductance (g(s)), water potential, intercellular [CO(2)], leaf temperature and vapour-pressure difference between leaf and air (VPD(L)) were studied at the Aspen FACE site. Two aspen (Populus tremuloides Michx.) clones differing in their sensitivity to ozone were measured. The depression in Pn was found after 10:00 h. The midday decline in Pn corresponded with both decreased g(s) and decreased Rubisco carboxylation efficiency, Vc(max). As a result of increasing VPD(L), g(s) decreased. Elevated [CO(2)] resulted in more pronounced midday decline in Pn compared to ambient concentrations. Moreover, this decline was more pronounced under combined treatment compared to elevated CO(2) treatment. The positive impact of CO(2) on Pn was relatively more pronounced in days with environmental stress but relatively less pronounced during midday depression. The negative impact of ozone tended to decrease in both cases.

Will photosynthetic capacity of aspen trees acclimate after long-term exposure to elevated CO2 and O3?

Photosynthetic acclimation under elevated carbon dioxide (CO(2)) and/or ozone (O(3)) has been the topic of discussion in many papers recently. We examined whether or not aspen plants grown under elevated CO(2) and/or O(3) will acclimate after 11 years of exposure at the Aspen Face site in Rhinelander, WI, USA. We studied diurnal patterns of instantaneous photosynthetic measurements as well as A/C(i) measurements monthly during the 2004-2008 growing seasons. Our results suggest that the responses of two aspen clones differing in O(3) sensitivity showed no evidence of photosynthetic and stomatal acclimation under either elevated CO(2), O(3) or CO(2) + O(3). Both clones 42E and 271 did not show photosynthetic nor stomatal acclimation under elevated CO(2) and O(3) after a decade of exposure. We found that the degree of increase or decrease in the photosynthesis and stomatal conductance varied significantly from day to day and from one season to another.

Interactive effect of elevated temperature and O3 on antioxidant capacity and gas exchange in Betula pendula saplings.

We studied the effects of slightly elevated temperature (T), O(3) concentration (O(3)) and their combination (T + O(3)) on the antioxidant defense, gas exchange and total leaf area of Betula pendula saplings in field conditions. During the second year of the experiment, T enhanced the total leaf area, net photosynthesis (P (n)) and maximum capacity of carboxylation, redox state of ascorbate and total antioxidant capacity in the apoplast. O(3) did not affect the total leaf area, but P (n) was slightly and g (s) significantly reduced. The saplings responded to elevated O(3) level by closing the stomata and by developing leaves with a lower leaf area per mass, rather than by accumulating ascorbate in the apoplast. The effects of T and O(3) on total leaf area and P (n) were counteractive. Elevated O(3) reduced the saplings' ability to utilize the warmer growth environment by increasing the stomatal limitation for photosynthesis and by reducing the redox state of ascorbate in the apoplast in the combination treatment as compared to T alone.

Rising atmospheric CO2 concentration partially masks the negative effects of elevated O3 in silver birch (Betula pendula Roth).

This review summarizes the main results from a 3-year open top chamber experiment, with two silver birch (Betula pendula Roth) clones (4 and 80) where impacts of 2x ambient [CO2] (EC) and [O3] (EO) and their combination (EC + EO) were examined. Growth, physiology of the foliage and root systems, crown structure, wood properties, and biological interactions were assessed to understand the effects of a future climate on the biology of silver birch. The clones displayed great differences in their reaction to EC and EO. Growth in clone 80 increased by 40% in EC and this clone also appeared O3-tolerant, showing no growth reduction. In contrast, growth in clone 4 was not enhanced by EC, and EO reduced growth with root growth being most affected. The physiological responses of the clones to EO were smaller than expected. We found no O3 effect on net photosynthesis in either of the clones, and many parameters indicated no change compared with chamber controls, suggesting active detoxification and defense in foliage. In EO, increased rhizospheric respiration over time and accelerated leaf senescence was common in both clones. We assumed that elevated O3 offsets the positive effects of elevated CO2 when plants were exposed to combined EC + EO treatment. In contrast, the responses to EC + EO mostly resembled the ones in EC, at least partly due to stomatal closure, which thus reduced O3 flux to the leaves. However, clear cellular level symptoms of oxidative stress were observed also in EC + EO treatment. Thus, we conclude that EC masked most of the negative O3 effects during long exposure of birch to EC + EO treatment. Biotic interactions were not heavily affected. Only some early season defoliators may suffer from faster maturation of leaves due to EO.

Stomatal characteristics and infection biology of Pyrenopeziza betulicola in Betula pendula trees grown under elevated CO2 and O3.

Two silver birch clones were exposed to ambient and elevated concentrations of CO(2) and O(3), and their combination for 3 years, using open-top chambers. We evaluated the effects of elevated CO(2) and O(3) on stomatal conductance (g(s)), density (SD) and index (SI), length of the guard cells, and epidermal cell size and number, with respect to crown position and leaf type. The relationship between the infection biology of the fungus (Pyrenopeziza betulicola) causing leaf spot disease and stomatal characteristics was also studied. Leaf type was an important determinant of O(3) response in silver birch, while crown position and clone played only a minor role. Elevated CO(2) reduced the g(s), but had otherwise no significant effect on the parameters studied. No significant interactions between elevated CO(2) and O(3) were found. The infection biology of P. betulicola was not correlated with SD or g(s), but it did occasionally correlate positively with the length of the guard cells.

Carbon gain and bud physiology in Populus tremuloides and Betula papyrifera grown under long-term exposure to elevated concentrations of CO2 and O3.

Paper birch (Betula papyrifera Marsh.) and three trembling aspen clones (Populus tremuloides Michx.) were studied to determine if alterations in carbon gain in response to an elevated concentration of CO(2) ([CO(2)]) or O(3) ([O(3)]) or a combination of both affected bud size and carbohydrate composition in autumn, and early leaf development in the following spring. The trees were measured for gas exchange, leaf size, date of leaf abscission, size and biochemical characteristics of the overwintering buds and early leaf development during the 8th-9th year of free-air CO(2) and O(3) exposure at the Aspen FACE site located near Rhinelander, WI. Net photosynthesis was enhanced 49-73% by elevated [CO(2)], and decreased 13-30% by elevated [O(3)]. Elevated [CO(2)] delayed, and elevated [O(3)] tended to accelerate, leaf abscission in autumn. Elevated [CO(2)] increased the ratio of monosaccharides to di- and oligosaccharides in aspen buds, which may indicate a lag in cold acclimation. The total carbon concentration in overwintering buds was unaffected by the treatments, although elevated [O(3)] decreased the amount of starch by 16% in birch buds, and reduced the size of aspen buds, which may be related to the delayed leaf development in aspen during the spring. Elevated [CO(2)] generally ameliorated the effects of elevated [O(3)]. Our results show that both elevated [CO(2)] and elevated [O(3)] have the potential to alter carbon metabolism of overwintering buds. These changes may cause carry-over effects during the next growing season.

Leaf photosynthetic characteristics of silver birch during three years of exposure to elevated concentrations of CO2 and O3 in the field.

Effects of elevated concentrations of carbon dioxide ([CO2]) and ozone ([O3]) on photosynthesis and related biochemistry of two European silver birch (Betula pendula Roth) clones were studied under field conditions during 1999-2001. Seven-year-old trees of Clones 4 and 80 were exposed for 3 years to the following treatments in an open-top chamber experiment: outside control (OC), chamber control (CC), 2x ambient [CO2] (EC), 2x ambient [O3] (EO) and 2x ambient [CO2] + 2x ambient [O3] (EC+EO). During the experiment, gas exchange, chlorophyll fluorescence, amount and activity of Rubisco, concentrations of chlorophyll, soluble protein, soluble sugars, starch, nitrogen (N) and carbon:nitrogen (C:N) ratio were determined in short- and long-shoot leaves. Elevated [CO2] increased photosynthetic rate by around 30% when measurements were made at the growth [CO2]. When measured at ambient [CO2], photosynthesis was around 15% lower in EC trees than in CC trees. This was related to a approximately 10% decrease in total leaf N, to 26 and 20% decreases in the amount and activity of Rubisco, respectively, and to a 49% increase in starch concentration in elevated [CO2]. Elevated [O3] had no significant effect on gas exchange parameters and its effect on biochemistry was small in both clones. However, elevated [O3] decreased the proportion of Rubisco in total soluble proteins and the apparent quantum yield of photosystem II (PSII) photochemistry in light and increased non-photochemical quenching in 2000. The interactive effect of CO2 and O3 was variable. Elevated [O3] decreased chlorophyll concentration only in EO trees, and the EC+EO treatment decreased the total activity of Rubisco and increased the C:N ratio more than the EO treatment alone. The small effect of elevated [O3] on photosynthesis indicates that these young silver birches were fairly tolerant to annual [O3] exposures that were 2-3 times higher than the AOT40 value of 10 ppm.h. set as a critical dose for forest trees.

Silver birch and climate change: variable growth and carbon allocation responses to elevated concentrations of carbon dioxide and ozone.

We studied the effects of elevated concentrations of carbon dioxide ([CO2]) and ozone ([O3]) on growth, biomass allocation and leaf area of field-grown O3-tolerant (Clone 4) and O3-sensitive clones (Clone 80) of European silver birch (Betula pendula Roth) trees during 1999-2001. Seven-year-old trees of Clones 4 and 80 growing outside in open-top chambers were exposed for 3 years to the following treatments: outside control (OC); chamber control (CC); 2 x ambient [CO2] (EC); 2 x ambient [O3] (EO); and 2 x ambient [CO2] + 2 x ambient [O3] (EC+EO). When the results for the two clones were analyzed together, elevated [CO2] increased tree growth and biomass, but had no effect on biomass allocation. Total leaf area increased and leaf abscission was delayed in response to elevated [CO2]. Elevated [O3] decreased dry mass of roots and branches and mean leaf size and induced earlier leaf abscission in the autumn; otherwise, the effects of elevated [O3] were small across the clones. However, there were significant interactions between elevated [CO2] and elevated [O3]. When results for the clones were analyzed separately, stem diameter, volume growth and total biomass of Clone 80 were increased by elevated [CO2] and the stimulatory effects of elevated [CO2] on stem volume growth and total leaf area increased during the 3-year study. Clone 80 was unaffected by elevated [O3]. In Clone 4, elevated [O3] decreased root and branch biomass by 38 and 29%, respectively, whereas this clone showed few responses to elevated [CO2]. Elevated [CO2] significantly increased total leaf area in Clone 80 only, which may partly explain the smaller growth responses to elevated [CO2] of Clone 4 compared with Clone 80. Although we observed responses to elevated [O3], the responses to the EC+EO and EC treatments were similar, indicating that the trees only responded to elevated [O3] under ambient [CO2] conditions, perhaps reflecting a greater quantity of carbohydrates available for detoxification and repair in elevated [CO2].