Heritable variation in stomatal responses to elevated CO2 in wild radish, Raphanus raphanistrum (Brassicaceae)

Rising atmospheric carbon dioxide may affect plant populations in the short term through effects on photosynthesis and carbon allocation, and over the long term as an agent of natural selection. To test for heritable effects of elevated CO2 on stomatal responses and plant fecundity in Raphanus raphanistrum, we grew plants from 12 paternal families in outdoor open-top chambers at ambient (35 Pa) or elevated (67 Pa) CO2. Contrary to results from a previous study of this species, total flower and fruit production were marginally lower under elevated CO2. Across families, stomatal index and guard cell length showed little response to CO2 enrichment, but these characters varied significantly among paternal families in both the direction and magnitude of their response to changing CO2. Although these family-by-CO2 interactions suggest that natural selection might affect stomatal characters when ambient CO2 levels increase, we found no significant correlation between either character and flower or fruit production. Therefore, our data suggest that while heritable variation for stomatal index and guard cell length exists in this population, selection due to increasing CO2 is not likely to act on these traits because they had no detectable effect on lifetime fecundity.     

Elevated carbon dioxide concentrations indirectly affect plant fitness by altering plant tolerance to herbivory

Global environmental changes, such as rising atmospheric CO₂ concentrations, have a wide range of direct effects on plant physiology, growth, and fecundity. These environmental changes also can affect plants indirectly by altering interactions with other species. Therefore, the effects of global changes on a particular species may depend on the presence and abundance of other community members. We experimentally manipulated atmospheric CO₂ concentration and amounts of herbivore damage (natural insect folivory and clipping to simulate browsing) to examine: (1) how herbivores mediate the effects of elevated CO₂ (eCO₂) on the growth and fitness of Arabidopsis thaliana; and (2) how predicted changes in CO₂ concentration affect plant resistance to herbivores, which influences the amount of damage plants receive, and plant tolerance of herbivory, or the fitness consequences of damage. We found no evidence that CO₂ altered resistance, but plants grown in eCO₂ were less tolerant of herbivory—clipping reduced aboveground biomass and fruit production by 13 and 22%, respectively, when plants were reared under eCO₂, but plants fully compensated for clipping in ambient CO₂ (aCO₂) environments. Costs of tolerance in the form of reduced fitness of undamaged plants were detected in eCO₂ but not aCO₂ environments. Increased costs could reduce selection on tolerance in eCO₂ environments, potentially resulting in even larger fitness effects of clipping in predicted future eCO₂ conditions. Thus, environmental perturbations can indirectly affect both the ecology and evolution of plant populations by altering both the intensity of species interactions as well as the fitness consequences of those interactions.     

Pollen performance of Raphanus sativus (Brassicaceae) declines in response to elevated [CO2]

Although increases in atmospheric [CO₂] are known to affect plant physiology, growth and reproduction, understanding of these effects is limited because most studies of reproductive consequences focus solely on female function. Therefore, we examined the effects of CO₂ enrichment on male function in the annual Raphanus sativus. Pollen donors grown under elevated [CO₂] initially sired a higher proportion of seeds per fruit than ambient [CO₂]-grown plants when each was tested against two different standard competitors; however, by the end of the 5-month experiment, these pollen donors sired fewer seeds than ambient [CO₂]-grown plants and produced a lower proportion of viable pollen grains. The results of this experiment confirm that elevated [CO₂] can alter reproductive success. Additionally, the change in response to elevated [CO₂] over time varied among pollen donor families; thus, changes in [CO₂] could act as a selective force on this species.     

Elevated CO2 and tree fecundity: the role of tree size, interannual variability, and population heterogeneity

Long‐term population effects of changes in atmospheric CO₂ will be largely determined by reproductive effort. Our research objectives were to quantify variability in seed production and rate of maturation among individual Pinus taeda L. (Pinaceae) trees growing in elevated CO₂ (ambient plus 200 μL L^?1) since 1996. Estimating tree fecundity in nature is frustrated by the difficulty of counting seeds from individual trees and the need for long‐term data. We have used a hierarchical Bayes approach to model individual tree fecundity, accounting for the complexity of experimentation in a natural setting over multiple years. The study presented here demonstrates large variability in natural fecundity rates and contributes to our understanding of how both interannual variation and population heterogeneity influence elevated CO₂ effects. We found that trees growing under elevated CO₂ matured earlier and produced more seeds and cones per unit basal area than ambient grown trees. By 2004, trees grown in high CO₂ had produced an average 300 more seeds per tree than ambient grown trees. Although there was a trend toward decreasing mean CO₂ effect (difference in fecundity between elevated and ambient treatments) over time, the hierarchical analysis indicates that this decrease comes from the emergence of a few highly fecund ambient grown trees by 2002, rather than acclimation or downregulation among the fumigated trees. The most important effect of increased CO₂ in forest ecosystems may be the increase in fecundity reported here. Although biomass responses can sometimes be large, the increase in fecundity can have long‐term impacts on forest dynamics that transcend the current generation.     

The impact of elevated CO2 on plant-herbivore interactions: experimental evidence of moderating effects at the community level

Surprisingly little research has been published on the responses to elevated [CO₂] at the community level, where herbivores can select their preferred food. We investigated the combined effects of atmospheric [CO₂] and herbivory on synthesised plant communities growing on soils of different fertility. Factorial combinations of two [CO₂] (350 or 700 l l⁻¹), two fertility (fertilised or non-fertilised), and two herbivory (herbivores present or absent) treatments were applied to a standard mixture of seven fast- and eight slow-growing plants in outdoor microcosms. The herbivores used were the grain aphid (Sitobion avenae) and the garden snail (Helix aspersa). We measured plant biomass, foliar nitrogen and soluble tannin concentration, aphid fecundity, and snail growth, fecundity, and feeding preferences over one growing season. Elevated [CO₂] did not have a significant impact on (1) the combined biomass of fast-growing or slow-growing plants, (2) herbivore feeding preferences, or (3) herbivore fitness. There was, however, a significant biomass increase of Carex flacca (which represented in all cases less than 5% of total live biomass), and some chemical changes in unpalatable plants under elevated [CO₂]. The herbivory treatment significantly increased the biomass of slow-growing plants over fast-growing plants, whereas fertilisation significantly increased the abundance of fast-growing plants over slow-growing plants. Predictions on the effects of elevated [CO₂] based on published single-species experiments were not supported by the results of this microcosm study. Received: 30 November 1997 / Accepted: 24 July 1998     

Genetic variation in plant below-ground response to elevated CO2 and two herbivore species

Aims

It is unclear how changing atmospheric conditions, including rising carbon dioxide concentration, influence interactions between above and below-ground systems and if intraspecific variation exists in this response.

Methods

We assessed interactive effects of atmospheric CO₂ concentration, above-ground herbivory, and plant genotype on root traits and mycorrhizal associations. Plants from five families of Asclepias syriaca, a perennial forb, were grown under ambient and elevated atmospheric CO₂ concentrations. Foliar herbivory by either lepidopteran caterpillars or phloem-feeding aphids was imposed. Mycorrhizal colonization, below-ground biomass, root biomass, and secondary defensive chemistry in roots were quantified.

Results

We observed substantial genetic variation among A. syriaca families in their mycorrhizal colonization levels in response to elevated CO₂ and herbivory treatments. Elevated CO₂ treatment increased root biomass in all genetic families, whereas foliar herbivory tended to decrease root biomass. Root cardenolide concentration and composition varied greatly among plant families, and elevated CO₂ treatment increased root cardenolides in two of the five plant families. Moreover, herbivores differentially affected the composition of cardenolides expressed below ground.

Conclusions

Increased atmospheric CO₂ has the potential to influence interactions among plants, herbivores and mycorrhizal fungi and intraspecific variation suggests that such interactions can evolve.     

Aphid suitability for the predatory hoverfly Episyrphus balteatus altered with elevating atmospheric CO2 and sinigrin

The fitness of natural enemies should be altered in response to changes in herbivore quality induced by the impact of increased atmospheric CO₂ levels on plants. We studied the effect of different CO₂ levels on the aphid predator Episyrphus balteatus DeGeer fed either specialist or generalist aphids reared on either of two host plants under laboratory conditions. In the host plant that contains sinigrin (black mustard), elevated CO₂ increased the sinigrin content of both host plant and the specialist aphid, but reduced the already very low levels in the generalist aphid. Predator development time increased with elevated CO₂, while fecundity decreased. Consequently, individual fitness decreased slightly with increasing atmospheric CO₂. Sinigrin significantly decreased fecundity and increased development time of the predator. As a result, fitness was significantly lower too. The consumption rate was influenced significantly by plant and prey solely and the interactions of host plant × prey type and CO₂ level × prey type. Further research on the effects of climate change parameters (e.g. greenhouse gases such as CO₂, ozone (O₃) and nitrogen dioxide (NO₂), etc.) separately and jointly under controlled environmental conditions will help to understand the nature and direction of their effects on natural enemies as part of the tritrophic system.     

Effects of elevated CO2 associated with maize on multiple generations of the cotton bollworm,Helicoverpa armigera

Under elevated environmental carbon dioxide (CO₂), leaf chewers tend to compensate for decreased leaf nutritional quality with increased consumption; mortality and development times also increase and cause a reduction in the fitness of leaf chewers. However, the effect of elevated CO₂ on multiple successive generations of these and other insects is not well understood. Furthermore, information about the direct effects of increased environmental CO₂ on developmental time and consumption of herbivores is lacking. In this paper, we tested the hypothesis that cascade effects of elevated CO₂ through plants, rather than the direct effects of elevated CO₂, are the main factors decreasing the fitness of cotton bollworm, Helicoverpa armigera Hübner (Lepidoptera: Noctuidae). We used two series of experiments to quantify the growth, development, and consumption of H. armigera fed on an artificial diet or C₄ plants (maize) grown under two CO₂ levels (ambient vs. double ambient). In the first series of experiments, elevated CO₂ had no effect on the population abundance or individual consumption for three successive generations of cotton bollworms fed on an artificial diet. In the second series of experiments, elevated CO₂ reduced population abundance of cotton bollworm larvae for two successive generations when they were fed maize milky grains. The specific effects were longer larval duration, lower fecundity, and decreased r_m of cotton bollworms. Furthermore, elevated CO₂ increased individual consumption when cotton bollworm was fed maize milky grains for two successive generations and decreased the population’s total consumption in the first generation but increased it in the second generation. The results from this study indicate that: (1) The effects of elevated CO₂ on three successive generations of cotton bollworm fed on artificial diet were weak, or even non‐existent, and (2) elevated CO₂ increased the consumption when cotton bollworm were fed maize. Our study also suggests that the damage inflicted by cotton bollworm on maize (a C₄ plant) will be seriously affected by the increases in atmospheric CO₂, which is unlike our previous results for spring wheat (a C₃ plant).     

HERITABLE VARIATION FOR FECUNDITY IN FIELD-COLLECTED DROSOPHILA MELANOGASTER AND THEIR OFFSPRING REARED UNDER DIFFERENT ENVIRONMENTAL TEMPERATURES

Heritable variation for fitness components is normally measured under favorable laboratory conditions, but organisms in the field experience variable conditions that are often stressful and may affect the expression of heritable variation. We examined heritable variation for early fecundity in three samples of Drosophila melanogaster from the field. Flies were obtained from a rotting fruit pile in summer, autumn, and spring, and progeny were reared under laboratory conditions. Field parents were tested for fecundity at 14°C or 28°C depending on ambient temperatures. Wing/thorax length ratios measured on flies from the spring collection suggested that flies had developed at around 20°C. Progeny were reared and tested at 14°C, 25°C, and 28°C. In the summer collection, parent-offspring regression coefficients were high and significant, compared to nonsignificant values obtained in two of three autumn comparisons. In the spring collection, parent-offspring regressions were negative regardless of testing temperature, suggesting that field females with a high fecundity produced offspring with low scores. Comparisons of F₁ and F₂ laboratory generations indicated intermediate heritabilities for fecundity in the laboratory. The lower bound heritability estimate for fecundity in field individuals was 37% in summer and 59% in autumn. Estimates of field heritability and evolvability for wing length measured in the spring collection were lower than in the laboratory. The results indicate that heritabilities and additive genetic variances for fecundity can be high in field-reared flies, but that results may vary between field collections.     

Seedling density modifies the growth responses of yellow birch maternal families to elevated carbon dioxide

We studied seedling growth responses to ambient and elevated CO₂ (350 and 700 μL L^?1) of three maternal families of yellow birch (Betula alleghaniensis), raised both individually and in high-density stands. Seedlings in competitive, dense stands exhibited markedly lower average CO₂-induced growth enhancements than individually grown plants (16% vs. 49%). Maternal families differed in their growth responses to elevated CO₂. However, differences among families were contingent upon density; families which exhibited the greatest CO₂-induced growth at low density exhibited the least CO₂-responsiveness at high density. These data are discussed in two separate contexts; the reliability of estimates of the CO² fertilization potential of forest species based solely on individually grown plants, and the potential evolutionary consequences of rising CO₂ on regenerating forest tree populations.     

Towards an understanding of how phloem amino acid composition shapes elevated CO2‐induced changes in aphid population dynamics

1. The performance of foliage feeders tends to decrease under elevated CO₂, but the responses of phloem‐feeding insects have been much more equivocal. As phloem tissues are less accessible than whole‐plant tissues, much less is known about how phloem composition is altered under elevated CO₂ and the mechanisms driving changes in aphid performance. 2. In this study, the plant mechanisms underlying the performance of Rhopalosiphum padi aphids on Hordeum vulgare (barley) grown under ambient (390 ppm) and elevated (700 ppm) CO₂ were examined. We used aphid stylectomy to sample pure phloem from plants in CO₂‐controlled conditions and high‐performance liquid chromatography to analyse phloem samples for amino acid concentrations. 3. Aphid abundance significantly increased by 127% under elevated CO₂. Consequently, plant biomass decreased under elevated CO₂ in trials with herbivores present, possibly due to the increased herbivore load, but increased when aphids were absent. The intrinsic rate of population increase (r_m) was significantly higher under elevated CO₂; however, there were no statistically significant effects on aphid fecundity or development time. The concentration of individual amino acids tended to increase, although these increases were statistically significant in only a few cases. A principal components analysis revealed that the relative abundance (mol %) of those amino acids considered essential for aphids tended to increase under elevated CO₂. 4. These results indicate that CO₂ may affect nutrient translocation in plants in ways that are contrary to predictions about nitrogen metabolite responses to CO₂. Such plant biochemical responses may underlie observations of improved phloem feeder performance under elevated CO₂.     

Acorn mass and seedling growth in Quercus rubra in response to elevated CO2

Abstract. In order to explore whether seed size affects plant response to elevated CO₂, plants grown from red oak (Quercus rubra L.) acorns were studied for differences in their first year response to CO₂ concentrations of 350 and 700 μl/l. Overall, at final harvest, total biomass of plants grown in elevated CO₂ were 47 % larger than that of plants grown in ambient CO₂. There were significant interactions between CO₂ treatments and initial acorn mass for total biomass, as well as for root, leaf, and stem biomass. Although total biomass increased with increasing initial acorn mass for both high and ambient CO₂ plants, high CO₂ plants exhibited a greater increase than ambient CO₂ plants, as indicated by a steeper slope in high CO₂ plants. However, CO₂ levels did not affect biomass partitioning traits, such as root/shoot ratio, leaf, stem, and root weight ratios, and leaf area ratio. These results suggest that variation in seed size or initial plant size can cause intraspecific variation in response to elevated CO₂.     

Maternal and direct effects of elevated CO2 on seed provisioning, germination and seedling growth in Bromus erectus

Elevated CO₂ can affect plant fitness not only through its effects on seed production but also by altering the quality of seeds and therefore germination and seedling performance. We collected seeds from mother plants of Bromus erectus grown in field plots at ambient and elevated CO₂ (m-CO₂, maternal CO₂) and germinated them in the greenhouse in a reciprocal design under ambient and elevated CO₂ (o-CO₂, offspring CO₂). This design allowed us to examine both the direct effects of elevated CO₂ on germination and seedling growth and the indirect (maternal) effects via altered seed quality. Elevated m-CO₂ significantly increased seed mass and increased the C:N ratio of seeds from field-grown plants. Percentage and rate of germination were not affected by the m-CO₂ or o-CO₂ treatments. Similarly, elevated m-CO₂ had no significant effect on seedling size as estimated by the total leaf length. When differences in seed mass were adjusted by using seed mass as a covariate in ANOVA, a negative effect of m-CO₂ on seedling size appeared which increased with increasing seed mass (significant covariate×m-CO₂ interaction). This may indicate that the advantage of increased seed mass at elevated m-CO₂ was offset by the reduced concentration of nitrogen (and possibly other nutrients) in these seeds. In contrast to m-CO₂, elevated o-CO₂ greatly increased seedling size, and this stimulatory effect of elevated o-CO₂ was found to increase with increasing seed mass (significant covariate×o-CO₂ interaction). Taken together, these results suggest that in B. erectus transgenerational effects of elevated CO₂ are relatively small. However, other factors (genetic and environmental) that contribute to variation in seed provisioning can critically influence the responsiveness of seedlings to elevated CO₂. Received: 10 May 1999 / Accepted: 6 January 2000     

Effects of age and ontogeny on photosynthetic responses of a determinate annual plant to elevated CO2 concentrations

Plant responses to elevated CO₂ concentrations ([CO₂]) may be regulated by both accelerated ontogeny and allocational changes as plants grow. However, isolating ontogeny‐related effects from age‐related effects are difficult because these factors are often confounded. In this study, the roles of age and ontogeny in photosynthetic responses to elevated [CO₂] were examined on Xanthium strumarium L. grown at ambient (365 µmol mol^?1) and elevated (730 µmol mol^?1) [CO₂]. To examine age‐related effects, six cohorts were planted at 5‐day intervals. To examine ontogeny‐related effects, all plants were induced to flower at the same time; ontogeny in Xanthium is relatively unaffected by growth in elevated [CO₂]. Growth in elevated [CO₂] increased net photosynthetic rates by approximately 30% throughout vegetative growth (i.e. active carbohydrate sinks), approximately 10% during flowering (i.e. minimal sink activity), and approximately 20% during fruit production (i.e. active sinks). At the harvest, the ratio of source to sink tissue significantly decreased with increasing plant age and was correlated with leaf soluble sugar concentration. Leaf soluble sugar concentration was negatively correlated with the relative photosynthetic response to elevated [CO₂]. These results suggest that age and ontogeny independently affect photosynthetic responses to elevated [CO₂] and the effects are mediated by reversible changes in source : sink balance.     

Effects of Elevated CO2 and Defoliation on Compensatory Growth and Photosynthesis of Seedlings in a Tropical Tree,Copaifera aromatica1

After defoliation by herbivores, some plants exhibit enhanced rates of photosynthesis and growth that enable them to compensate for lost tissue, thus maintaining their fitness relative to competing, undefoliated plants. Our aim was to determine whether compensatory photosynthesis and growth would be altered by increasing concentrations of atmospheric CO₂. Defoliation of developing leaflets on seedlings of a tropical tree, Copaifera aromatica, caused increases in photosynthesis under ambient CO₂, but not under elevated CO₂. An enhancement in the development of buds in the leaf axils followed defoliation at ambient levels of CO₂. In contrast, under elevated CO₂, enhanced development of buds occurred in undefoliated plants with no further enhancement in bud development due to exposure to elevated CO₂. Growth of leaf area after defoliation was increased, particularly under elevated CO₂. Despite this increase, defoliated plants grown under elevated CO₂ were further from compensating for tissue lost during defoliation after 5¹/₂ weeks than those grown under ambient CO₂ concentrations.     

Population parameters and growth of Myzus persicae Sulzer (Hemiptera: Aphididae) under elevated CO2 concentrations in the air

The effects of three different CO₂ concentrations (400, 600, and 1000 ppm) on the population parameters and growth of the green peach aphid, Myzus persicae, were examined. Raw life history data from M. persicae were analyzed using an age-stage, two-sex life table to take into account the viable development rate among individuals. The population projections of M. persicae indicate the stage structure and variability of the population growth under different CO₂ concentrations based on an age-stage, two-sex life table analysis. Significantly longer oviposition duration and higher fecundity were observed under elevated CO₂ (600 and 1000 ppm) than those under ambient CO₂ (400 ppm). Furthermore, the M. persicae population reared under elevated CO₂ concentrations showed higher intrinsic and finite rates of population increase than under ambient CO₂ concentrations. These results indicate that the population parameters and growth of M. persicae were positively influenced in the fecundity by elevated CO₂ concentrations relative to the ambient CO₂. These findings indicate that it is basically remained to understand the direct effects of CO₂ elevation on the host plants, and the interaction between the host plants and M. persicae in the same CO₂ concentration for establishing more realistic population growth model systems for M. persicae in the aerial environment rising CO₂ concentration level.     

Infection with the parasitic angiosperm Striga hermonthica influences the response of the C3 cereal Oryza sativa to elevated CO2

Upland rice (Oryza sativa L.) was grown at both ambient (350 μmol mol^?1) and elevated (700 μmol mol^?1) CO₂ in either the presence or absence of the root hemi‐parasitic angiosperm Striga hermonthica (Del) Benth. Elevated CO₂ alleviated the impact of the parasite on host growth: biomass of infected rice grown at ambient CO₂ was 35% that of uninfected, control plants, while at elevated CO₂, biomass of infected plants was 73% that of controls. This amelioration occurred despite the fact that O. sativa grown at elevated CO₂ supported both greater numbers and a higher biomass of parasites per host than plants grown at ambient CO₂. The impact of infection on host leaf area, leaf mass, root mass and reproductive tissue mass was significantly lower in plants grown at elevated as compared with ambient CO₂. There were significant CO₂ and Striga effects on photosynthetic metabolism and instantaneous water‐use efficiency of O. sativa. The response of photosynthesis to internal [CO₂] (A/C_i curves) indicated that, at 45 days after sowing (DAS), prior to emergence of the parasites, uninfected plants grown at elevated CO₂ had significantly lower CO₂ saturated rates of photosynthesis, carboxylation efficiencies and ribulose‐1,5‐bisphosphate carboxylase/oxygenase (Rubisco; EC 4.1.1.39) contents than uninfected, ambient CO₂‐grown O. sativa. In contrast, infection with S. hermonthica prevented down‐regulation of photosynthesis in O. sativa grown at elevated CO₂, but had no impact on photosynthesis of hosts grown at ambient CO₂. At 76 DAS (after parasites had emerged), however, infected plants grown at both elevated and ambient CO₂ had lower carboxylation efficiencies and Rubisco contents than uninfected O. sativa grown at ambient CO₂. The reductions in carboxylation efficiency (and Rubisco content) were accompanied by similar reductions in nitrogen concentration of O. sativa leaves, both before and after parasite emergence. There were no significant CO₂ or infection effects on the concentrations of soluble sugars in leaves of O. sativa, but starch concentration was significantly lower in infected plants at both CO₂ concentrations. These results demonstrate that elevated CO₂ concentrations can alleviate the impact of infection with Striga on the growth of C₃ hosts such as rice and also that infection can delay the onset of photosynthetic down‐regulation in rice grown at elevated CO₂.     

QUANTITATIVE GENETICS OF FITNESS TRAITS IN A WILD POPULATION OF PHLOX

To determine if the evolution of fitness traits in the annual plant, Phlox drummondii, is constrained by lack of genetic variation, we calculated the heritability and genetic correlation of 16 traits in a field population. Full- and half-sib families of seeds were generated in the greenhouse and planted back into the study population. Of 855 seeds that germinated, 609 survived to produce fruit. For each plant we measured several aspects of plant size and three components of female fecundity: total number of fruits produced, number of seeds per fruit, and mass of individual seeds. Heritability of traits ranged from 0.00 to 0.15. Several traits showed significant levels of additive genetic variance, but we found no evidence of additive genetic variance in components of female fecundity and no evidence of negative genetic correlation between fitness traits. These results suggest that evolution in this population would be constrained by lack of heritable variation in fitness traits.     

Poa annua shows inter-generational differences in response to elevated CO2

Inter-generational effects on the growth of Poa annua (L.) in ambient and elevated atmospheric CO₂ conditions (350 and 550 μl l^–1, respectively) were studied in two different experiments. Both experiments showed similar results. In a greenhouse experiment growth, measured as the numbers of tillers produced per week, was compared for plants grown from first and second generation seeds. Second generation seeds were obtained from plants grown for one whole generation in either ambient or elevated atmospheric CO₂ (‘ambient’ and ‘elevated’ seeds, respectively). First generation plants and second generation ‘ambient’ plants did not respond to elevated CO₂. Second generation ‘elevated’ plants produced significantly more tillers in elevated CO₂. In the second experiment model terrestrial ecosystems growing in the Ecotron and which included Poa annua were used. Above-ground biomass after one and two generations of growth were compared. At the end of Generation 1 no difference was found in biomass production while at the end of Generation 2 biomass increased in elevated CO₂ by 50%. The implications for climate change research are discussed.