Author Correction: Increasing CO2 threatens human nutrition.

An Amendment to this paper has been published and can be accessed via a link at the top of the paper.

Moderate heat stress prevented the observed biomass and yield stimulation caused by elevated CO2 in two well-watered wheat cultivars.

KEY MESSAGE: Heat stress (HS) under well-watered conditions was not detrimental to leaf photosynthesis or yield but modified the elevated CO2 response of photosynthesis and yield in two contrasting wheat cultivars. Climate change is increasing the frequency of extreme events such as heat waves, adversely affecting crop productivity. While positive impacts of elevated carbon dioxide (eCO2) on crop productivity are evident, the interactive effects of eCO2 and environmental stresses are still unclear. To investigate the interactive effects of elevated CO2 and heat stress (HS), we grew two contrasting wheat cultivars, early-maturing Scout and high-tillering Yitpi, under non-limiting water and nutrients at ambient (aCO2, 450 ppm) or elevated (eCO2, 650 ppm) CO2 and 22 °C in the glasshouse. Plants were exposed to two 3-day HS cycles at the vegetative (38.1 °C) and/or flowering (33.5 °C) stage. At aCO2, both wheat cultivars showed similar responses of photosynthesis and mesophyll conductance to temperature and produced similar grain yield. Relative to aCO2, eCO2 enhanced photosynthesis rate and reduced stomatal conductance and maximal carboxylation rate (Vcmax). During HS, high temperature stimulated photosynthesis at eCO2 in both cultivars, while eCO2 stimulated photosynthesis in Scout. Electron transport rate (Jmax) was unaffected by any treatment. eCO2 equally enhanced biomass and grain yield of both cultivars in control, but not HS, plants. HS reduced biomass and yield of Scout at eCO2. Yitpi, the cultivar with higher grain nitrogen, underwent a trade-off between grain yield and nitrogen. In conclusion, eCO2 improved photosynthesis of control and HS wheat, and improved biomass and grain yield of control plants only. Under well-watered conditions, HS was not detrimental to photosynthesis or growth but precluded a yield response to eCO2.

Carbon sink strength of nodules but not other organs modulates photosynthesis of faba bean (Vicia faba) grown under elevated [CO2 ] and different water supply.

Photosynthetic stimulation by elevated [CO2 ] (e[CO2 ]) may be limited by the capacity of sink organs to use photosynthates. In many legumes, N2 -fixing symbionts in root nodules provide an additional sink, so that legumes may be better able to profit from e[CO2 ]. However, drought not only constrains photosynthesis but also the size and activity of sinks, and little is known about the interaction of e[CO2 ] and drought on carbon sink strength of nodules and other organs. To compare carbon sink strength, faba bean was grown under ambient (400 ppm) or elevated (700 ppm) atmospheric [CO2 ] and subjected to well-watered or drought treatments, and then exposed to 13 C pulse-labelling using custom-built chambers to track the fate of new photosynthates. Drought decreased 13 C uptake and nodule sink strength, and this effect was even greater under e[CO2 ], and was associated with an accumulation of amino acids in nodules. This resulted in decreased N2 fixation, and increased accumulation of new photosynthates (13 C/sugars) in leaves, which in turn can feed back on photosynthesis. Our study suggests that nodule C sink activity is key to avoid sink limitation in legumes under e[CO2 ], and legumes may only be able to achieve greater C gain if nodule activity is maintained.

Early vigour in wheat: Could it lead to more severe terminal drought stress under elevated atmospheric [CO2 ] and semi-arid conditions?

Early vigour in wheat is a trait that has received attention for its benefits reducing evaporation from the soil surface early in the season. However, with the growth enhancement common to crops grown under elevated atmospheric CO2 concentrations (e[CO2 ]), there is a risk that too much early growth might deplete soil water and lead to more severe terminal drought stress in environments where production relies on stored soil water content. If this is the case, the incorporation of such a trait in wheat breeding programmes might have unintended negative consequences in the future, especially in dry years. We used selected data from cultivars with proven expression of high and low early vigour from the Australian Grains Free Air CO2 Enrichment (AGFACE) facility, and complemented this analysis with simulation results from two crop growth models which differ in the modelling of leaf area development and crop water use. Grain yield responses to e[CO2 ] were lower in the high early vigour group compared to the low early vigour group, and although these differences were not significant, they were corroborated by simulation model results. However, the simulated lower response with high early vigour lines was not caused by an earlier or greater depletion of soil water under e[CO2 ] and the mechanisms responsible appear to be related to an earlier saturation of the radiation intercepted. Whether this is the case in the field needs to be further investigated. In addition, there was some evidence that the timing of the drought stress during crop growth influenced the effect of e[CO2 ] regardless of the early vigour trait. There is a need for FACE investigations of the value of traits for drought adaptation to be conducted under more severe drought conditions and variable timing of drought stress, a risky but necessary endeavour.

A reduced-tillering trait shows small but important yield gains in dryland wheat production.

Reducing the number of tillers per plant using a tiller inhibition (tin) gene has been considered as an important trait for wheat production in dryland environments. We used a spatial analysis approach with a daily time-step coupled radiation and transpiration efficiency model to simulate the impact of the reduced-tillering trait on wheat yield under different climate change scenarios across Australia's arable land. Our results show a small but consistent yield advantage of the reduced-tillering trait in the most water-limited environments both under current and likely future conditions. Our climate scenarios show that whilst elevated [CO2 ] (e[CO2 ]) alone might limit the area where the reduced-tillering trait is advantageous, the most likely climate scenario of e[CO2 ] combined with increased temperature and reduced rainfall consistently increased the area where restricted tillering has an advantage. Whilst long-term average yield advantages were small (ranged from 31 to 51 kg ha-1  year-1 ), across large dryland areas the value is large (potential cost-benefits ranged from Australian dollar 23 to 60 MIL/year). It seems therefore worthwhile to further explore this reduced-tillering trait in relation to a range of different environments and climates, because its benefits are likely to grow in future dry environments where wheat is grown around the world.

Characteristics of free air carbon dioxide enrichment of a northern temperate mature forest.

In 2017, the Birmingham Institute of Forest Research (BIFoR) began to conduct Free Air Carbon Dioxide Enrichment (FACE) within a mature broadleaf deciduous forest situated in the United Kingdom. BIFoR FACE employs large-scale infrastructure, in the form of lattice towers, forming 'arrays' which encircle a forest plot of ~30 m diameter. BIFoR FACE consists of three treatment arrays to elevate local CO2 concentrations (e[CO2 ]) by +150 µmol/mol. In practice, acceptable operational enrichment (ambient [CO2 ] + e[CO2 ]) is ±20% of the set point 1-min average target. There are a further three arrays that replicate the infrastructure and deliver ambient air as paired controls for the treatment arrays. For the first growing season with e[CO2 ] (April to November 2017), [CO2 ] measurements in treatment and control arrays show that the target concentration was successfully delivered, that is: +147 ± 21 µmol/mol (mean ± SD) or 98 ± 14% of set point enrichment target. e[CO2 ] treatment was accomplished for 97.7% of the scheduled operation time, with the remaining time lost due to engineering faults (0.6% of the time), CO2 supply issues (0.6%) or adverse weather conditions (1.1%). CO2 demand in the facility was driven predominantly by wind speed and the formation of the deciduous canopy. Deviations greater than 10% from the ambient baseline CO2 occurred <1% of the time in control arrays. Incidences of cross-contamination >80 µmol/mol (i.e. >53% of the treatment increment) into control arrays accounted for <0.1% of the enrichment period. The median [CO2 ] values in reconstructed three-dimensional [CO2 ] fields show enrichment somewhat lower than the target but still well above ambient. The data presented here provide confidence in the facility setup and can be used to guide future next-generation forest FACE facilities built into tall and complex forest stands.

Increasing CO2 threatens human nutrition.

Dietary deficiencies of zinc and iron are a substantial global public health problem. An estimated two billion people suffer these deficiencies, causing a loss of 63 million life-years annually. Most of these people depend on C3 grains and legumes as their primary dietary source of zinc and iron. Here we report that C3 grains and legumes have lower concentrations of zinc and iron when grown under field conditions at the elevated atmospheric CO2 concentration predicted for the middle of this century. C3 crops other than legumes also have lower concentrations of protein, whereas C4 crops seem to be less affected. Differences between cultivars of a single crop suggest that breeding for decreased sensitivity to atmospheric CO2 concentration could partly address these new challenges to global health.

Elevated CO2 alleviates the negative impact of heat stress on wheat physiology but not on grain yield.

Hot days are becoming hotter and more frequent, threatening wheat yields worldwide. Developing wheat varieties ready for future climates calls for improved understanding of how elevated CO2 (eCO2) and heat stress (HS) interactively impact wheat yields. We grew a modern, high-yielding wheat cultivar (Scout) at ambient CO2 (aCO2, 419 µl l -1) or eCO2 (654 µl l-1) in a glasshouse maintained at 22/15 °C (day/night). Half of the plants were exposed to HS (40/24 °C) for 5 d at anthesis. In non-HS plants, eCO2 enhanced (+36%) CO2 assimilation rates (Asat) measured at growth CO2 despite down-regulation of photosynthetic capacity. HS reduced Asat (-42%) in aCO2- but not in eCO2-grown plants because eCO2 protected photosynthesis by increasing ribulose bisphosphate regeneration capacity and reducing photochemical damage under HS. eCO2 stimulated biomass (+35%) of all plants and grain yield (+30%) of non-HS plants only. Plant biomass initially decreased following HS but recovered at maturity due to late tillering. HS equally reduced grain yield (-40%) in aCO2- and eCO2-grown plants due to grain abortion and reduced grain filling. While eCO2 mitigated the negative impacts of HS at anthesis on wheat photosynthesis and biomass, grain yield was reduced by HS in both CO2 treatments.

Water availability moderates N2 fixation benefit from elevated [CO2 ]: A 2-year free-air CO2 enrichment study on lentil (Lens culinaris MEDIK.) in a water limited agroecosystem.

Increased biomass and yield of plants grown under elevated [CO2 ] often corresponds to decreased grain N concentration ([N]), diminishing nutritional quality of crops. Legumes through their symbiotic N2 fixation may be better able to maintain biomass [N] and grain [N] under elevated [CO2 ], provided N2 fixation is stimulated by elevated [CO2 ] in line with growth and yield. In Mediterranean-type agroecosystems, N2 fixation may be impaired by drought, and it is unclear whether elevated [CO2 ] stimulation of N2 fixation can overcome this impact in dry years. To address this question, we grew lentil under two [CO2 ] (ambient ~400 ppm and elevated ~550 ppm) levels in a free-air CO2 enrichment facility over two growing seasons sharply contrasting in rainfall. Elevated [CO2 ] stimulated N2 fixation through greater nodule number (+27%), mass (+18%), and specific fixation activity (+17%), and this stimulation was greater in the high than in the low rainfall/dry season. Elevated [CO2 ] depressed grain [N] (-4%) in the dry season. In contrast, grain [N] increased (+3%) in the high rainfall season under elevated [CO2 ], as a consequence of greater post-flowering N2 fixation. Our results suggest that the benefit for N2 fixation from elevated [CO2 ] is high as long as there is enough soil water to continue N2 fixation during grain filling.

Benefits of increasing transpiration efficiency in wheat under elevated CO2 for rainfed regions.

Higher transpiration efficiency (TE) has been proposed as a mechanism to increase crop yields in dry environments where water availability usually limits yield. The application of a coupled radiation and TE simulation model shows wheat yield advantage of a high-TE cultivar (cv. Drysdale) over its almost identical low-TE parent line (Hartog), from about -7 to 558 kg/ha (mean 187 kg/ha) over the rainfed cropping region in Australia (221-1,351 mm annual rainfall), under the present-day climate. The smallest absolute yield response occurred in the more extreme drier and wetter areas of the wheat belt. However, under elevated CO2 conditions, the response of Drysdale was much greater overall, ranging from 51 to 886 kg/ha (mean 284 kg/ha) with the greatest response in the higher rainfall areas. Changes in simulated TE under elevated CO2 conditions are seen across Australia with notable increased areas of higher TE under a drier climate in Western Australia, Queensland and parts of New South Wales and Victoria. This improved efficiency is subtly deceptive, with highest yields not necessarily directly correlated with highest TE. Nevertheless, the advantage of Drysdale over Hartog is clear with the benefit of the trait advantage attributed to TE ranging from 102% to 118% (mean 109%). The potential annual cost-benefits of this increased genetic TE trait across the wheat growing areas of Australia (5 year average of area planted to wheat) totaled AUD 631 MIL (5-year average wheat price of AUD/260 t) with an average of 187 kg/ha under the present climate. The benefit to an individual farmer will depend on location but elevated CO2 raises this nation-wide benefit to AUD 796 MIL in a 2°C warmer climate, slightly lower (AUD 715 MIL) if rainfall is also reduced by 20%.

The relationship between transpiration and nutrient uptake in wheat changes under elevated atmospheric CO2.

The impact of elevated [CO2 ] (e[CO2 ]) on crops often includes a decrease in their nutrient concentrations where reduced transpiration-driven mass flow of nutrients has been suggested to play a role. We used two independent approaches, a free-air CO2 enrichment (FACE) experiment in the South Eastern wheat belt of Australia and a simulation study employing the agricultural production systems simulator (APSIM), to show that transpiration (mm) and nutrient uptake (g m-2 ) of nitrogen (N), potassium (K), sulfur (S), calcium (Ca), magnesium (Mg) and manganese (Mn) in wheat are correlated under e[CO2 ], but that nutrient uptake per unit water transpired is higher under e[CO2 ] than under ambient [CO2 ] (a[CO2 ]). This result suggests that transpiration-driven mass flow of nutrients contributes to decreases in nutrient concentrations under e[CO2 ], but cannot solely explain the overall decline.

Elevated atmospheric [CO2 ] can dramatically increase wheat yields in semi-arid environments and buffer against heat waves.

Wheat production will be impacted by increasing concentration of atmospheric CO2 [CO2 ], which is expected to rise from about 400 µmol mol(-1) in 2015 to 550 µmol mol(-1) by 2050. Changes to plant physiology and crop responses from elevated [CO2 ] (e[CO2 ]) are well documented for some environments, but field-level responses in dryland Mediterranean environments with terminal drought and heat waves are scarce. The Australian Grains Free Air CO2 Enrichment facility was established to compare wheat (Triticum aestivum) growth and yield under ambient (~370 µmol(-1) in 2007) and e[CO2 ] (550 µmol(-1) ) in semi-arid environments. Experiments were undertaken at two dryland sites (Horsham and Walpeup) across three years with two cultivars, two sowing times and two irrigation treatments. Mean yield stimulation due to e[CO2 ] was 24% at Horsham and 53% at Walpeup, with some treatment responses greater than 70%, depending on environment. Under supplemental irrigation, e[CO2 ] stimulated yields at Horsham by 37% compared to 13% under rainfed conditions, showing that water limited growth and yield response to e[CO2 ]. Heat wave effects were ameliorated under e[CO2 ] as shown by reductions of 31% and 54% in screenings and 10% and 12% larger kernels (Horsham and Walpeup). Greatest yield stimulations occurred in the e[CO2 ] late sowing and heat stressed treatments, when supplied with more water. There were no clear differences in cultivar response due to e[CO2 ]. Multiple regression showed that yield response to e[CO2 ] depended on temperatures and water availability before and after anthesis. Thus, timing of temperature and water and the crop's ability to translocate carbohydrates to the grain postanthesis were all important in determining the e[CO2 ] response. The large responses to e[CO2 ] under dryland conditions have not been previously reported and underscore the need for field level research to provide mechanistic understanding for adapting crops to a changing climate.

Will intra-specific differences in transpiration efficiency in wheat be maintained in a high CO2 world? A FACE study.

This study evaluates whether the target breeding trait of superior leaf level transpiration efficiency is still appropriate under increasing carbon dioxide levels of a future climate using a semi-arid cropping system as a model. Specifically, we investigated whether physiological traits governing leaf level transpiration efficiency, such as net assimilation rates (A(net)), stomatal conductance (g(s)) or stomatal sensitivity were affected differently between two Triticum aestivum L. cultivars differing in transpiration efficiency (cv. Drysdale, superior; cv. Hartog, low). Plants were grown under Free Air Carbon dioxide Enrichment (FACE, approximately 550 µmol mol⁻¹ or ambient CO2 concentrations (approximately 390 µmol mol⁻¹). Mean A(net) (approximately 15% increase) and gs (approximately 25% decrease) were less affected by elevated [CO2] than previously found in FACE-grown wheat (approximately 25% increase and approximately 32% decrease, respectively), potentially reflecting growth in a dry-land cropping system. In contrast to previous FACE studies, analyses of the Ball et al. model revealed an elevated [CO2] effect on the slope of the linear regression by 12% indicating a decrease in stomatal sensitivity to the combination of [CO2], photosynthesis rate and humidity. Differences between cultivars indicated greater transpiration efficiency for Drysdale with growth under elevated [CO2] potentially increasing the response of this trait. This knowledge adds valuable information for crop germplasm improvement for future climates.

Multivariate analysis of physiological parameters reveals a consistent O3 response pattern in leaves of adult European beech (Fagus sylvatica).

• Increasing atmospheric concentrations of phytotoxic ozone (O(3) ) can constrain growth and carbon sink strength of forest trees, potentially exacerbating global radiative forcing. Despite progress in the conceptual understanding of the impact of O(3) on plants, it is still difficult to detect response patterns at the leaf level. • Here, we employed principal component analysis (PCA) to analyse a database containing physiological leaf-level parameters of 60-yr-old Fagus sylvatica (European beech) trees. Data were collected over two climatically contrasting years under ambient and twice-ambient O(3) regimes in a free-air forest environment. • The first principal component (PC1) of the PCA was consistently responsive to O(3) and crown position within the trees over both years. Only a few of the original parameters showed an O(3) effect. PC1 was related to parameters indicative of oxidative stress signalling and changes in carbohydrate metabolism. PC1 correlated with cumulative O(3) uptake over preceding days. • PC1 represents an O(3) -responsive multivariate pattern detectable in the absence of consistently measurable O(3) effects on individual leaf-level parameters. An underlying effect of O(3) on physiological processes is indicated, providing experimental confirmation of theoretical O(3) response patterns suggested previously.

Does Elevated [CO2] Only Increase Root Growth in the Topsoil? A FACE Study with Lentil in a Semi-Arid Environment.

Atmospheric carbon dioxide concentrations [CO2] are increasing steadily. Some reports have shown that root growth in grain crops is mostly stimulated in the topsoil rather than evenly throughout the soil profile by e[CO2], which is not optimal for crops grown in semi-arid environments with strong reliance on stored water. An experiment was conducted during the 2014 and 2015 growing seasons with two lentil (Lens culinaris) genotypes grown under Free Air CO2 Enrichment (FACE) in which root growth was observed non-destructively with mini-rhizotrons approximately every 2-3 weeks. Root growth was not always statistically increased by e[CO2] and not consistently between depths and genotypes. In 2014, root growth in the top 15 cm of the soil profile (topsoil) was indeed increased by e[CO2], but increases at lower depths (30-45 cm) later in the season were greater than in the topsoil. In 2015, e[CO2] only increased root length in the topsoil for one genotype, potentially reflecting the lack of plant available soil water between 30-60 cm until recharged by irrigation during grain filling. Our limited data to compare responses to e[CO2] showed that root length increases in the topsoil were correlated with a lower yield response to e[CO2]. The increase in yield response was rather correlated with increases in root growth below 30 cm depth.

Relations of sugar composition and delta 13C in phloem sap to growth and physiological performance of Eucalyptus globulus (Labill).

We characterized differences in carbon isotopic content (delta(13)C) and sugar concentrations in phloem exudates from Eucalyptus globulus (Labill) plantations across a rainfall gradient in south-western Australia. Phloem sap delta(13)C and sugar concentrations varied with season and annual rainfall. Annual bole growth was negatively related to phloem sap delta(13)C during summer, suggesting a water limitation, yet was positively related in winter. We conclude that when water is abundant, variations in carboxylation rates become significant to overall growth. Concentrations of sucrose in phloem sap varied across sites by up to 600 mm, and raffinose by 300 mm. These compounds play significant roles in maintaining osmotic balance and facilitating carbon movement into the phloem, and their relative abundances contribute strongly to overall delta(13)C of phloem sap. Taken together, the delta(13)C and concentrations of specific sugars in phloem sap provide significant insights to functions supporting growth at the tree, site and landscape scale.

Lack of genetic variation in tree ring delta13C suggests a uniform, stomatally-driven response to drought stress across Pinus radiata genotypes.

We assessed the variation in delta(13)C signatures of Pinus radiata D. Don stemwood taken from three genetic trials in southern Australia. We sought to determine the potential of using delta(13)C signatures as selection criteria for drought tolerance. Increment cores were taken from P. radiata and were used to determine the basal area increment and the delta(13)C signature of extracted cellulose. Both growth increment and cellulose delta(13)C were affected by water availability. Growth increment and delta(13)C were negatively correlated suggesting that growth was water-limited. While there was significant genetic variation in growth, there was no significant genetic variation in cellulose delta(13)C of tree rings. This suggests that different genotypes of P. radiata display significant differences in growth and yet respond similarly to drought stress. The delta(13)C response to drought stress was more due to changes in stomatal conductance than to the variation in photosynthetic capacity, and this may explain the lack of genetic variation in delta(13)C. The lack of genetic variation in cellulose delta(13)C of tree rings precludes its use as a selection criterion for drought tolerance among P. radiata genotypes.

Effects of drought and canopy ozone exposure on antioxidants in fine roots of mature European beech (Fagus sylvatica).

We quantified ascorbate, glutathione and alpha-tocopherol in fine roots of mature Fagus sylvatica L. under free-air canopy ozone (O(3)) exposure (twice ambient O(3) concentration, 2x[O(3)]) during two growing seasons that differed in the extent of summer drought (exceptional drought year 2003, average year 2004). This design allowed us to test whether O(3) exposure or drought, or both, affected root antioxidants during the growing season. In both years, root ascorbate and alpha-tocopherol showed a similar relationship with volumetric soil water content (SWC): ascorbate concentrations on a root dry mass basis increased from about 6 to 12 micromol g(-1) when SWC dropped from 25 to 20%, and a-tocopherol increased from 100 to 150 nmol g(-1) at SWC values below 20%. Root glutathione showed no relationship with SWC or differences between the dry and the average year, but it was significantly and consistently diminished by 2x[O(3)]. Our results were inconclusive as to whether shoot-root translocation of glutathione or glutathione production in the roots was diminished. Phloem glutathione concentrations in the canopy remained constant, but reduced transport velocity in the phloem and, as a consequence, reduced mass flow of glutathione cannot be ruled out.

Prescreening in large populations as a tool for identifying elevated CO2-responsive genotypes in plants.

Elevated atmospheric CO2 concentration (e[CO2]) can stimulate the photosynthesis and productivity of C3 species including food and forest crops. Intraspecific variation in responsiveness to e[CO2] can be exploited to increase productivity under e[CO2]. However, active selection of genotypes to increase productivity under e[CO2] is rarely performed across a wide range of germplasm, because of constraints of space and the cost of CO2 fumigation facilities. If we are to capitalise on recent advances in whole genome sequencing, approaches are required to help overcome these issues of space and cost. Here, we discuss the advantage of applying prescreening as a tool in large genome×e[CO2] experiments, where a surrogate for e[CO2] was used to select cultivars for more detailed analysis under e[CO2] conditions. We discuss why phenotypic prescreening in population-wide screening for e[CO2] responsiveness is necessary, what approaches could be used for prescreening for e[CO2] responsiveness, and how the data can be used to improve genetic selection of high-performing cultivars. We do this within the framework of understanding the strengths and limitations of genotype-phenotype mapping.

The water use dynamics of canola cultivars grown under elevated CO2 are linked to their leaf area development.

The 'CO2 fertilisation effect' is often predicted to be greater under drier than wetter conditions, mainly due to hypothesised early season water savings under elevated [CO2] (e[CO2]). However, water savings largely depend on the balance between CO2-induced improvement of leaf-level water use efficiency and CO2-stimulation of transpiring leaf area. The dynamics of water use during the growing season can therefore vary depending on leaf area development. Two canola (Brassica napus L.) cultivars of contrasting growth and vigour (vigorous hybrid cv. Hyola 50 and non-hybrid cv. Thumper) were grown under ambient [CO2] (a[CO2], ∼400 µmol mol-1) or e[CO2] (∼700 µmol mol-1) with two water treatments (well-watered and mild drought) in a glasshouse to investigate the interdependence of leaf area development and water use. Dynamics of water use during the growing season varied depending on [CO2] and cultivars. Early stimulation of leaf growth under e[CO2], which also depended on cultivar, overcompensated for the effect of increased leaf-level water use efficiency, so that weekly water use was greater and water depletion from soil greater under e[CO2] than a[CO2]. This result shows that the balance between leaf area and water use efficiency stimulation by e[CO2] can tip towards early depletion of available soil water, so that e[CO2] does not lead to water savings, and the 'CO2 fertilisation effect' is not greater under drier conditions.