Fitzgerald L. Booker

Ecological issues related to ozone: agricultural issues

Research on the effects of ozone on agricultural crops and agro-ecosystems is needed for the development of regional emission reduction strategies, to underpin practical recommendations aiming to increase the sustainability of agricultural land management in a changing environment, and to secure food supply in regions with rapidly growing populations. Major limitations in current knowledge exist in several areas: (1) Modelling of ozone transfer and specifically stomatal ozone uptake under variable environmental conditions, using robust and well-validated dynamic models that can be linked to large-scale photochemical models lack coverage. (2) Processes involved in the initial reactions of ozone with extracellular and cellular components after entry through the stomata, and identification of key chemical species and their role in detoxification require additional study. (3) Scaling the effects from the level of individual cells to the whole-plant requires, for instance, a better understanding of the effects of ozone on carbon transport within the plant. (4) Implications of long-term ozone effects on community and whole-ecosystem level processes, with an emphasis on crop quality, element cycling and carbon sequestration, and biodiversity of pastures and rangelands require renewed efforts. The UNECE Convention on Long Range Trans-boundary Air Pollution shows, for example, that policy decisions may require the use of integrated assessment models. These models depend on quantitative exposure-response information to link quantitative effects at each level of organization to an effective ozone dose (i.e., the balance between the rate of ozone uptake by the foliage and the rate of ozone detoxification). In order to be effective in a policy, or technological context, results from future research must be funnelled into an appropriate knowledge transfer scheme. This requires data synthesis, up-scaling, and spatial aggregation. At the research level, interactions must be considered between the effects of ozone and factors that are either directly manipulated by man through crop management, or indirectly changed. The latter include elevated atmospheric CO(2), particulate matter, other pollutants such as nitrogen oxides, UV-B radiation, climate and associated soil moisture conditions.

Arbuscular Mycorrhizal Fungi Increase Organic Carbon Decomposition Under Elevated CO2

A Fungal Culprit to Carbon Loss In some ecosystems, such as in the layer of soil containing plant roots, fungi, and bacteria, increased levels of CO2 should stimulate more efficient aboveground photosynthesis, which in turn should promote increased sequestration of organic carbon in soil through the protective action of arbuscular mycorrhizal fungi. However, in a series of field and microcosm experiments performed under elevated levels of CO2 thought to be consistent with future emissions scenarios, Cheng et al. (p. 1084; see the Perspective by Kowalchuk) observed that these fungi actually promote degradation of soil organic carbon, releasing more CO2 in the process. Counter to expectations, fungi associated with plant roots diminish the carbon pool in soil ecosystems under elevated levels of carbon dioxide. The extent to which terrestrial ecosystems can sequester carbon to mitigate climate change is a matter of debate. The stimulation of arbuscular mycorrhizal fungi (AMF) by elevated atmospheric carbon dioxide (CO2) has been assumed to be a major mechanism facilitating soil carbon sequestration by increasing carbon inputs to soil and by protecting organic carbon from decomposition via aggregation. We present evidence from four independent microcosm and field experiments demonstrating that CO2 enhancement of AMF results in considerable soil carbon losses. Our findings challenge the assumption that AMF protect against degradation of organic carbon in soil and raise questions about the current prediction of terrestrial ecosystem carbon balance under future climate-change scenarios.

Is increased UV-B a threat to crop photosynthesis and productivity?

It has been suggested that increases in ground-level UV-B, as a result of stratospheric ozone depletion, may have major deleterious effects on crop photosynthesis and productivity. The direct consequences of such effects have been projected by some as a world-wide decrease in crop yields of 20–25%. Further losses, or unrealized gains, have also been suggested as a result of increased UV-B counteracting the beneficial effects of elevated atmospheric CO2. Deleterious UV-B effects may be largely partitioned between damage to the plant genome and damage to the photosynthetic machinery. Direct damage to DNA is a common result of absorption of high energy UV-B photons. However, most plants possess repair mechanisms adequate to deal with the levels of damage expected from projected increases in ground-level UV-B. In addition, most plants have the ability to increase production of UV-absorbing compounds in their leaves as a result of exposure to UV-B, UV-A and visible radiation. These compounds contribute substantially to reducing UV-B damage in situ. It has also been shown that in some plants, under the proper conditions, almost every facet of the photosynthetic machinery can be damaged directly by very high UV-B exposures. However, electron transport, mediated by Photosystem II (PS II) appears to be the most sensitive part of the system. Various laboratories have reported damage to virtually all parts of the PS II complex from the Mn binding site to the plastoquinone acceptor sites on the opposite surface of the thylakoid membrane. However, a critical review of the literature with emphasis on exposure protocols and characterization of the radiation environment, revealed that most growth chamber and greenhouse experiments and very many field experiments have been conducted at unrealistic or indeterminate UV-B exposure levels, especially with regard to the spectral balance of their normal radiation environment. Thus, these experiments have led directly to large overestimates of the potential for damage to crop photosynthesis and yield within the context of 100 year projections for stratospheric ozone depletion. Indeed, given the massive UV-B exposures necessary to produce many of these effects, we suggest it is unlikely that they would occur in a natural setting and urge reconsideration of the purported impacts of projected increases of UV-B on crop productivity.

The challenge of making ozone risk assessment for forest trees more mechanistic.

Upcoming decades will experience increasing atmospheric CO2 and likely enhanced O3 exposure which represents a risk for the carbon sink strength of forests, so that the need for cause-effect related O3 risk assessment increases. Although assessment will gain in reliability on an O3 uptake basis, risk is co-determined by the effective dose, i.e. the plants sensitivity per O3 uptake. Recent progress in research on the molecular and metabolic control of the effective O3 dose is reported along with advances in empirically assessing O3 uptake at the whole-tree and stand level. Knowledge on both O3 uptake and effective dose (measures of stress avoidance and tolerance, respectively) needs to be understood mechanistically and linked as a pre-requisite before practical use of process-based O3 risk assessment can be implemented. To this end, perspectives are derived for validating and promoting new O3 flux-based modelling tools.

Catechin, proanthocyanidin and lignin contents of loblolly pine (Pinus taeda) needles after chronic exposure to ozone

Concentrations of soluble and bound phenolic compounds were measured in needles of 3-yr-old loblolly pine (Pinus taeda L.) trees exposed from May to November 1993 to a range of ozone (O3 ) concentrations in open-top field chambers, The treatments were charcoal-filtered air (CF). non-filtered air (NF), and NF air with O2 added at 1.5 times (NF 1.5) and 2(1 times (NF 2.0) the ambient O., concentration for 12 h daily. Average daily (0800-2000 hours) O3 concentrations in the CF. NF. NF 1.5 and NF 20 treatments were. 29. 47, 76 and 98 nl 1(-1) . respectively, for the 140 d treatment period. At the end of the treatment period, total phenolic and proanthocyanidin concentrations in the previous years needles were 25-29% higher in the NF 2.0 treatment compared with the lower O3 treatments. Catechin concentration increased in the previous years needles by as much as 81 % between the NF 2.0 treatment and the lower O3 treatments. Catechin is an effective antioxidant, and elevated levels might confer some protection against O3 injury. No significant differences in total phenolics and proanthocyanidins in the previous years needles were detected among the remaining treatments, or among any O3 treatment for the current years needles. Lignin content in needles of both years was not significantly affected by O3 exposure. Chances in the phenolic content of older needles in response to elevated O3 could alter plant-pathogen interactions and slow down microbiol decomposition, which could contribute to a decline in site soil quality.

Increased protein carbonylation in leaves of Arabidopsis and soybean in response to elevated [CO2]

While exposure of C3 plants to elevated [CO2] would be expected to reduce production of reactive oxygen species (ROS) in leaves because of reduced photorespiratory metabolism, results obtained in the present study suggest that exposure of plants to elevated [CO2] can result in increased oxidative stress. First, in Arabidopsis and soybean, leaf protein carbonylation, a marker of oxidative stress, was often increased when plants were exposed to elevated [CO2]. In soybean, increased carbonyl content was often associated with loss of leaf chlorophyll and reduced enhancement of leaf photosynthetic rate (Pn) by elevated [CO2]. Second, two-dimensional (2-DE) difference gel electrophoresis (DIGE) analysis of proteins extracted from leaves of soybean plants grown at elevated [CO2] or [O3] revealed that both treatments altered the abundance of a similar subset of proteins, consistent with the idea that both conditions may involve an oxidative stress. The 2-DE analysis of leaf proteins was facilitated by a novel and simple procedure to remove ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) from soluble soybean leaf extracts. Collectively, these findings add a new dimension to our understanding of global change biology and raise the possibility that oxidative signals can be an unexpected component of plant response to elevated [CO2].

Growth of Arabidopsis flavonoid mutants under solar radiation and UV filters

Abstract Growth of the chalcone isomerase defective tt-5 mutant of Arabidopsis thaliana and its Landsberg erecta progenitor were compared under a variety of full spectrum solar radiation conditions to determine if the tt-5 mutant could serve as an adequate subject for studies of the mechanisms of damage by UV-B radiation. An experiment was conducted in the fall of 1995 under open field filter frames using cellulose diacetate and Mylar filters to transmit and exclude natural UV-B irradiation, respectively. Even though growth under these conditions was slow and erratic owing to lack of temperature control, growth suppression as indicated by rosette diameter and harvest fresh weights provided a sensitive indicator of UV-B stress. This experience led to development of temperature-controlled Teflon-covered field chambers that admit up to 88% of the total daily PAR and about 85% of ambient UV-B, omit predators, and provide a generally stable environment for quantitative plant growth studies. The chambers were designed to facilitate the addition of optical filters and/or shade cloth and to accommodate control of the gaseous environment for pollutant and climate change studies and to provide clean air for other experiments. Three additional experiments were conducted in these chambers. Measurements of rosette diameter, weights of various aboveground plant parts, and plant height were evaluated as potential methods of comparing growth sensitivities of the tt-5 mutant to UV-B radiation. The weight of the reproductive parts (flowers and siliques) as a fraction of the total (e.g. harvest index) was consistently and negatively affected by solar UV-B, as was simple plant height. However, in no case, even in the virtual absence of UV-B, was growth of tt-5 comparable to that of Ler. We conclude that the disruption of secondary metabolism in tt-5 has growth implications far beyond the lack of UV-B protection, making it unsuitable as a surrogate for high UV-B experimentation.

Elevated Carbon Dioxide and Ozone Effects on Peanut: I. Gas-Exchange, Biomass, and Leaf Chemistry

The effects of elevated CO2 and ozone (O3) on net photosynthetic rate (A) and growth are generally antagonistic although plant responses are highly dependent on crop sensitivity to the individual gases and their concentrations. In this experiment, we evaluated the effects of various CO2 and O3 mixtures on leaf gas-exchange, harvest biomass, and leaf chemistry in peanut (Arachis hypogaea L.), an O3-sensitive species, using open-top field chambers. Treatments included ambient CO2 (about 375 micromol mol-1) and CO2 enrichment of approximately 173 and 355 micromol mol-1 in combination with charcoal-filtered air (22 nmol O3 mol-1), nonfiltered air (46 nmol O3 mol-1), and nonfiltered air plus O3 (75 nmol O3 mol-1). Twice-ambient CO2 in charcoal-filtered air increased A by 23% while decreasing seasonal stomatal conductance (gs) by 42%. Harvest biomass was increased 12 to 15% by elevated CO2. In ambient CO2, nonfiltered air and added O3 lowered A by 21% and 48%, respectively, while added O3 reduced gs by 18%. Biomass was not significantly affected by nonfiltered air, but was 40% lower in the added O3 treatment. Elevated CO2 generally suppressed inhibitory effects of O3 on A and harvest biomass. Leaf starch concentration was increased by elevated CO2 and decreased by O3. Treatment effects on foliar N and total phenolic concentrations were minor. Increasing atmospheric CO2 concentrations should attenuate detrimental effects of ambient O3 and promote growth in peanut but its effectiveness declines with increasing O3 concentrations.

Re-evaluating the role of ascorbic acid and phenolic glycosides in ozone scavenging in the leaf apoplast of Arabidopsis thaliana L

Phenolic glycosides are effective reactive oxygen scavengers and peroxidase substrates, suggesting that compounds in addition to ascorbate may have functional importance in defence responses against ozone (O(3)), especially in the leaf apoplast. The apoplastic concentrations of ascorbic acid (AA) and phenolic glycosides in Arabidopsis thaliana L. Col-0 wild-type plants were determined following exposure to a range of O(3) concentrations (5, 125 or 175 nL L(-1)) in controlled environment chambers. AA in leaf apoplast extracts was almost entirely oxidized in all treatments, suggesting that O(3) scavenging by direct reactions with reduced AA was very limited. In regard to phenolics, O(3) stimulated transcription of numerous phenylpropanoid pathway genes and increased the apoplastic concentration of sinapoyl malate. However, modelling of O(3) scavenging in the apoplast indicated that sinapoyl malate concentrations were too low to be effective protectants. Furthermore, null mutants for sinapoyl esters (fah1-7), kaempferol glycosides (tt4-1) and the double mutant (tt4-1/fah1-7) were equally sensitive to chronic O(3) as Ler-0 wild-type plants. These results indicate that current understanding of O(3) defence schemes deserves reassessment as mechanisms other than direct scavenging of O(3) by extracellular AA and antioxidant activity of some phenolics may predominate in some plant species.

Minimal influence of G-protein null mutations on ozone-induced changes in gene expression, foliar injury, gas exchange and peroxidase activity in Arabidopsis thaliana L.

Ozone (O(3)) uptake by plants leads to an increase in reactive oxygen species (ROS) in the intercellular space of leaves and induces signalling processes reported to involve the membrane-bound heterotrimeric G-protein complex. Therefore, potential G-protein-mediated response mechanisms to O(3) were compared between Arabidopsis thaliana L. lines with null mutations in the α- and β-subunits (gpa1-4, agb1-2 and gpa1-4/agb1-2) and Col-0 wild-type plants. Plants were treated with a range of O(3) concentrations (5, 125, 175 and 300 nL L(-1)) for 1 and 2 d in controlled environment chambers. Transcript levels of GPA1, AGB1 and RGS1 transiently increased in Col-0 exposed to 125 nL L(-1) O(3) compared with the 5 nL L(-1) control treatment. However, silencing of α and β G-protein genes resulted in little alteration of many processes associated with O(3) injury, including the induction of ROS-signalling genes, increased leaf tissue ion leakage, decreased net photosynthesis and stomatal conductance, and increased peroxidase activity, especially in the leaf apoplast. These results indicated that many responses to O(3) stress at physiological levels were not detectably influenced by α and β G-proteins.