CDK8 of the mediator kinase module connects leaf development to the establishment of correct stomata patterning by regulating the levels of the transcription factor SPEECHLESS (SPCH).

The components of the mediator kinase module are highly conserved across all eukaryotic lineages, and cyclin-dependent kinase 8 (CDK8) is essential for correct cell proliferation and differentiation in diverse eukaryotic systems. We show that CDK8 couples leaf development with the establishment of correct stomata patterning for prevailing CO2 conditions. In Arabidopsis, the basic helix-loop-helix (bHLH) transcription factor SPEECHLESS (SPCH) controls cellular entry into the stomatal cell lineage, and CDK8 interacts with and phosphorylates SPCH, controlling SPCH protein levels and thereby also expression of the SPCH target genes encoding key regulators of cell fate and asymmetric cell divisions. The lack of the CDK8-mediated control of SPCH results in an increased number of meristemoid and guard mother cells, and increased stomata index in the cdk8 mutants. Increasing atmospheric CO2 concentrations trigger a developmental programme controlling cell entry into stomatal lineage by limiting the asymmetric divisions. In cdk8, the number of meristemoids and guard mother cells remains the same under ambient and high CO2 concentrations, as the accumulated levels of SPCH caused by the lack of CDK8 appear to override the negative regulation of increased CO2. Thus, our work provides novel mechanistic understanding of how plants alter critical leaf properties in response to increasing atmospheric CO2.

Response of stomatal conductance, transpiration, and photosynthesis to light and CO2 for rice leaves with different appearance days.

To investigate the dynamics of stomata, transpiration, and photosynthesis under varying light intensities and CO2 conditions during leaf development, the light response and CO2 response of stomatal conductance (g sw), transpiration rate (T r), and net photosynthetic rate (P n) were observed for rice leaves at different days after leaf emergence (DAE). The results showed that (1) as photosynthetically active radiation (PAR) increased, leaf g sw, T r, and P n initially increased rapidly and linearly, followed by a more gradual rise to maximum values, and then either stabilized or showed a declining trend. The maximum g sw, T r, and P n were smaller and occurred earlier for old leaves than for young leaves. The g sw, T r, and P n all exhibited a linear decreasing trend with increasing DAE, and the rate of decrease slowed down with the reduction in PAR; (2) as the CO2 concentration (C a) increased, g sw and T r decreased gradually to a stable minimum value, while P n increased linearly and slowly up to the maximum and then kept stable or decreased. The g sw, T r, and P n values initially kept high and then decreased with the increase of DAE. These results contribute to understanding the dynamics in g sw, T r, and P n during rice leaf growth and their response to varied light and CO2 concentration conditions and provide mechanistic support to estimate dynamic evapotranspiration and net ecosystem productivity at field-scale and a larger scale in paddy field ecosystems through the upscaling of leaf-level stomatal conductance, transpiration, and photosynthesis.

Preparation and Effect of CO2 Response Gel for Plugging Low-Permeability Reservoirs.

In order to solve the problem of gas channeling during CO2 flooding in low-permeability reservoirs, a novel CO2 responsive gel channeling system was prepared by using carrageenan, branched polyethylene imide and ethylenediamine under laboratory conditions. Based on the Box-Behnken response surface design method, the optimal synthesis concentration of the system was 0.5 wt% carrageenan, 2.5 wt% branchized polyethylenimide and 6.5 wt% ethylenediamine. The micromorphology of the system before and after response was characterized by scanning electron microscopy. The rheology and dehydration rate were tested under different conditions. The channeling performance and enhanced oil recovery effect of the gel system were simulated by a core displacement experiment. The experimental results show that the internal structure of the system changes from a disordered, smooth and loosely separated lamellae structure to a more uniform, complete and orderly three-dimensional network structure after exposure to CO2. The viscosity of the system was similar to aqueous solution before contact with CO2 and showed viscoelastic solid properties after contact with CO2. The experiment employing dehydration rates at different temperatures showed that the internal structure of the gel would change at a high temperature, but the gel system had a certain self-healing ability. The results of the displacement experiment show that the plugging rate of the gel system is stable at 85.32% after CO2 contact, and the recovery rate is increased by 17.06%, which provides an important guide for the development of low-permeability reservoirs.

The Use of an Advanced Intelligent-Responsive Polymer for the Study of Dynamic Water-Carbon Dioxide Alternating Displacement.

Addressing the issue of inadequate temperature tolerance in traditional polymers, in this study, we successfully executed a one-step synthesis of intelligent-responsive polymers which have excellent adaptability in water-gas alternating displacement scenarios. Utilizing the fatty acid method, we produced OANND from oleic acid (OA) and N,N-dimethyl-1,3-propanediamine (NND). Upon testing the average particle size in the aqueous solution both prior and subsequent to CO2 passage, it became evident that OANND assumes the form of a small-molecule particle in the aqueous phase, minimizing damage during formation. Notably, upon CO2 exposure, it promptly organizes into stable micelles with an average size of 88 nm and a relatively uniform particle distribution. This unique characteristic endows it with a rapid CO2 response mechanism and the ability to form a highly resilient gel. In the exploration of viscoelastic fluids, we observed the remarkable behavior of the AONND aqueous solution when CO2/N2 was introduced. This system displayed repeatable transitions between aqueous and gel states, with the highest viscosity peaking at approximately 3895 mPa·s, highlighting its viscosity reversibility and reusability properties. The rheological property results that we obtained indicate that an elongated micellar structure is present in the solution system, with the optimal concentration ratio for its formation determined as 0.8, which is the molar ratio of the OANND-NaOA system. In the sealing performance tests, a 1.0 wt% concentration of the gel system exhibited excellent injectability properties. At 80 °C, this gel effectively reduced the permeability of a sand-filled model to 94.5% of its initial value, effectively sealing potential leakage paths or gas fluxes. This remarkable ability to block leakage paths and reduce seepage capacity highlights the material's superior blocking effect and erosion resistance properties. Furthermore, even at a temperature of 90 °C and an injection pore volume (PV) of 3, this plugging system could reduce the permeability of a high-permeability sand-filled model to over 90% of its initial value.

Effects of Rainfall Exclusion Treatment on Photosynthetic Characteristics of Black Locust in the Sub-Humid Region of the Loess Plateau, China.

The mesic-origin species Robinia pseudoacacia L. (black locust) is widely planted in the semiarid and sub-humid areas of the Loess Plateau for the reforestation of vegetation-degraded land. Under the scenario of changing precipitation patterns, exploring the response of photosynthesis to drought allows us to assess the risk to sustainable development of these plantations. In this study, paired plots were established including the control and a treatment of 30% exclusion of throughfall (since 2018). The photosynthetic characteristics were investigated using a portable photosynthesis system for four periods in the full-leaf growing season of 2021-2022, the fourth and fifth years, on both treated and controlled sampling trees. Leaf gas exchange parameters derived from diurnal changing patterns, light response curves, and CO2 response curves showed significant differences except for period II (9-11 September 2021) between the two plots. The photosynthetic midday depression was observed in 2022 in the treated plot. Meanwhile, the decline of net photosynthetic rate in the treated plot was converted from stomatal limitation to non-stomatal limitation. Furthermore, we observed that black locust adapted to long-term water deficiency by reducing stomatal conductance, increasing water use efficiency and intrinsic water use efficiency. The results demonstrate that reduction in precipitation would cause photosynthesis decrease, weaken the response sensitivity to light and CO2, and potentially impair photosynthetic resilience of the plantations. They also provide insights into the changes in photosynthetic functions under global climate change and a reference for management of plantations.

Contrasting leaf-scale photosynthetic low-light response and its temperature dependency are key to differences in crop-scale radiation use efficiency.

Radiation use efficiency (RUE) is a key crop adaptation trait that quantifies the potential amount of aboveground biomass produced by the crop per unit of solar energy intercepted. But it is unclear why elite maize and grain sorghum hybrids differ in their RUE at the crop level. Here, we used a non-traditional top-down approach via canopy photosynthesis modelling to identify leaf-level photosynthetic traits that are key to differences in crop-level RUE. A novel photosynthetic response measurement was developed and coupled with use of a Bayesian model fitting procedure, incorporating a C4 leaf photosynthesis model, to infer cohesive sets of photosynthetic parameters by simultaneously fitting responses to CO2 , light, and temperature. Statistically significant differences between leaf photosynthetic parameters of elite maize and grain sorghum hybrids were found across a range of leaf temperatures, in particular for effects on the quantum yield of photosynthesis, but also for the maximum enzymatic activity of Rubisco and PEPc. Simulation of diurnal canopy photosynthesis predicted that the leaf-level photosynthetic low-light response and its temperature dependency are key drivers of the performance of crop-level RUE, generating testable hypotheses for further physiological analysis and bioengineering applications.

Central CO2 chemosensitivity and CO2 controller gain independently contribute to daytime Pco2 in young subjects with congenital central hypoventilation syndrome.

Whether peripheral chemoreceptor response is altered in congenital central hypoventilation syndrome (CCHS) remains debated. Our aim was to prospectively evaluate both peripheral and central CO2 chemosensitivity and to evaluate their correlations with daytime Pco2 and arterial desaturation during exercise in CCHS. To this end, tidal breathing was recorded in patients with CCHS allowing the calculation of loop gain and its components {steady-state controller (assumed to mainly be peripheral chemosensitivity) and plant gains using a bivariate [end-tidal Pco2 ([Formula: see text]) and ventilation] constrained model}, a hyperoxic, hypercapnic ventilatory response test (central chemosensitivity), and a 6-min walk test (arterial desaturation). The results of loop gain were compared with those previously obtained in a healthy group of similar age. The study prospectively included 23 subjects with CCHS, without daytime ventilatory support; the subjects had a median age of 10 (5.6 to 27.4) yr (15 females) with moderate polyalanine repeat mutation (PARM: 20/25, 20/26, n = 11), severe PARM (20/27, 20/33, n = 8), or non-PARM (n = 4). As compared with 23 healthy subjects (4.9-27.0 yr), the subjects with CCHS had a decreased controller gain and an increased plant gain. Mean daytime [Formula: see text] level of subjects with CCHS correlated negatively to both Log(controller gain) and the slope of CO2 response. Genotype was not related to chemosensitivity. Arterial desaturation on exercise correlated negatively with Log(controller) gain but not with the slope of the CO2 response. In conclusion, we demonstrate that peripheral CO2 chemosensitivity is altered in some patients with CCHS and that the daytime [Formula: see text] depends on central and peripheral chemoreceptor responses.NEW & NOTEWORTHY Altered central CO2 chemosensitivity is a hallmark of congenital central hypoventilation syndrome (CCHS). Peripheral CO2 chemosensitivity can be partly assessed by controller gain measurement obtained from tidal breathing recording. In young subjects with CCHS, this study shows that both central and peripheral CO2 sensitivities independently contribute to daytime Pco2. Hypocapnia during nighttime-assisted ventilation is associated with higher peripheral chemosensitivity that is further associated with lesser arterial desaturation at walk.

Effects of 5-HT on the cold resistance of mangrove Kandelia obovata seedlings.

5-hydroxytryptamine (5-HT) participates in plant growth and development, and can also delay senescence and cope with abiotic stress. To explore the role of 5-HT in regulating the abilities of mangrove in cold resis-tance, we examined the effects of cold acclimation and the spraying of p-chlorophenylalanine (p-CPA, 5-HT synthesis inhibitor) on leaf gas exchange parameters and CO2 response curves (A/Ca), as well as the endogenous phytohormone content levels in the mangrove species Kandelia obovata seedlings under low temperature stress. The results showed that low temperature stress significantly reduced the contents of 5-HT, chlorophyll, endogenous auxin (IAA), gibberellin (GA), and abscisic acid (ABA). It weakened the CO2 utilization abilities of plants and reduced net photosynthetic rate, which ultimately reduced carboxylation efficiency (CE). Under low temperature stress, exogenous p-CPA reduced the contents of photosynthetic pigments, endogenous hormones, and 5-HT in the leaves, which aggravated the damages caused by low temperature stress on photosynthesis. By enhancing cold acclimation abilities, the endogenous IAA content in the leaves could was reduced under low temperature stress, promoted the production of 5-HT, improved the contents of photosynthetic pigments, GA, and ABA, as well as enhanced photosynthetic carbon assimilation abilities, which would increase photosynthesis in the K. obovata seedlings. Under cold acclimation conditions, the spraying of p-CPA could significantly inhibit the synthesis of 5-HT, promote the production of IAA, and reduce the contents of photosynthetic pigments, GA, ABA, and CE, which would weaken the effects of cold acclimation by improving the cold resistance of mangroves. In conclusion, cold acclimation could improve the cold resistance abilities of K. obovata seedlings by regulating photosynthetic carbon assimilation capacity and the contents of endogenous phytohormone. 5-HT synthesis is one of the necessary conditions for improving the cold resistance abilities of mangroves.

A high efficiency CO2 concentration interval optimization method for lettuce growth.

The decline in carbon fertilization effects has shifted scientific focus toward the efficient and suitable regulation of CO2 concentration ([CO2]) for plant growth. In this study, the rapid A/CO2 response curve (RAC) data of lettuce were analyzed statistically under nine photosynthetic photon flux densities (PPFDs) and four temperatures. An efficient CO2 supplementation interval acquisition method based on the frequency distribution characteristics of RACs was proposed. The characteristic subsections of jumping were obtained depending on the frequency distribution of RACs. The cumulative contribution rate (CCR) of the characteristic subsections were >97 %, which showed the efficiency of the method. Additionally, U-chord curvature theory was used to simultaneously obtain the optimal regulated [CO2] for the same RAC curves, and the results showed that the [CO2] obtained by U-chord length were all within the interval obtained by the method, which proved the rationality of the method. The [CO2] interval supplement improved the daily CO2 exchange rate by 20.27 % and 21.64 % at 150 and 200 µmol·m-2·s-1, and increased the lettuce fresh biomass by 26.78 % at 150 µmol·m-2·s-1. Based on the interval of [CO2] efficient utilization regulation at various temperatures and PPFDs, a genetic algorithm-support vector regression model was built with R2 of the model was >0.84 and the root mean square error was <35.2256 µmol·mol-1. In conclusion, the [CO2] interval obtained by this method has a positive effect on lettuce growth. This work provides a new method for obtaining high-efficiency supplementary concentration of CO2 during the growth of lettuce.

Elevated [CO2] negatively impacts C4 photosynthesis under heat and water stress without penalizing biomass.

Elevated [CO2] (eCO2) and water stress reduce leaf stomatal conductance (gs), which may affect leaf thermoregulation during heat waves (heat stress). Two sorghum lines, with different leaf width were grown in a glasshouse at a mean day temperature of 30 °C, under different [CO2] and watering levels, and subjected to heat stress (43 °C) for 6 d at the start of the reproductive stage. We measured leaf photosynthetic and stomatal responses to light transients before harvesting the plants. Photosynthesis at growth conditions (Agrowth) and biomass accumulation were enhanced by eCO2 under control conditions. Heat stress increased gs, especially in wider leaves, and reduced the time constant of stomatal opening (kopen) at ambient [CO2] but not eCO2. However, heat stress reduced photosynthesis under water stress and eCO2 due to increased leaf temperature and reduced evaporative cooling. eCO2 prevented the reduction of biomass under both water and heat stress, possibly due to improved plant and soil water status as a result of reduced gs. Our results suggest that the response of the C4 crop sorghum to future climate conditions depends on the trade-off between low gs needed for high water use efficiency and drought tolerance, and the high gs needed for improved thermoregulation and heat tolerance under an eCO2 future.

Application of the rapid leaf A-Ci response (RACiR) technique: examples from evergreen broadleaved species.

Using steady-state photosynthesis-intercellular CO2 concentration (A-Ci) response curves to obtain the maximum rates of ribulose-1,5-bisphosphate carboxylase oxygenase carboxylation (Vcmax) and electron transport (Jmax) is time-consuming and labour-intensive. Instead, the rapid A-Ci response (RACiR) technique provides a potential, high-efficiency method. However, efficient parameter settings of RACiR technique for evergreen broadleaved species remain unclear. Here, we used Li-COR LI-6800 to obtain the optimum parameter settings of RACiR curves for evergreen broadleaved trees and shrubs. We set 11 groups of CO2 gradients ([CO2]), i.e. R1 (400-1500 ppm), R2 (400-200-800 ppm), R3 (420-20-620 ppm), R4 (420-20-820 ppm), R5 (420-20-1020 ppm), R6 (420-20-1220 ppm), R7 (420-20-1520 ppm), R8 (420-20-1820 ppm), R9 (450-50-650 ppm), R10 (650-50 ppm) and R11 (650-50-650 ppm), and then compared the differences between steady-state A-Ci and RACiR curves. We found that Vcmax and Jmax calculated by steady-state A-Ci and RACiR curves overall showed no significant differences across 11 [CO2] gradients (P > 0.05). For the studied evergreens, the efficiency and accuracy of R2, R3, R4, R9 and R10 were higher than the others. Hence, we recommend that the [CO2] gradients of R2, R3, R4, R9 and R10 could be applied preferentially for measurements when using the RACiR technique to obtain Vcmax and Jmax of evergreen broadleaved species.

Mesophyll conductance response to short-term changes in pCO2 is related to leaf anatomy and biochemistry in diverse C4 grasses.

Mesophyll CO2 conductance (gm ) in C3 species responds to short-term (minutes) changes in environment potentially due to changes in leaf anatomical and biochemical properties and measurement artefacts. Compared with C3 species, there is less information on gm responses to short-term changes in environmental conditions such as partial pressure of CO2 (pCO2 ) across diverse C4 species and the potential determinants of these responses. Using 16 C4 grasses we investigated the response of gm to short-term changes in pCO2 and its relationship with leaf anatomy and biochemistry. In general, gm increased as pCO2 decreased (statistically significant increase in 12 species), with percentage increases in gm ranging from +13% to +250%. Greater increase in gm at low pCO2 was observed in species exhibiting relatively thinner mesophyll cell walls along with greater mesophyll surface area exposed to intercellular air spaces, leaf N, photosynthetic capacity and activities of phosphoenolpyruvate carboxylase and Rubisco. Species with greater CO2 responses of gm were also able to maintain their leaf water-use efficiencies (TEi ) under low CO2 . Our study advances understanding of CO2 response of gm in diverse C4 species, identifies the key leaf traits related to this response and has implications for improving C4 photosynthetic models and TEi through modification of gm .

Fabrication of a CO2-responsive chitosan aerogel as an effective adsorbent for the adsorption and desorption of heavy metal ions.

In the traditional desorption method, strong acid is commonly used as an eluent for the regeneration of adsorbents. It is of critical economic and environmental significance to develop a chemical-free desorption method. In this study, a new CO2-responsive chitosan aerogel adsorbent was synthesized from CO2-responsive poly(acrylic acid-2-(dimethylamino)ethyl methacrylate) and chitosan by physicochemical double crosslinking for the adsorption of Cu2+. Compared with the chitosan aerogel, the adsorption capacity of Cu2+ and mechanical properties of CO2-responsive chitosan aerogel increased by 162% and 660%, respectively. Most importantly, after the adsorption of Cu2+ by CO2-responsive chitosan aerogel, the Cu2+ could be desorbed by CO2 bubbling, and the desorption rate of metal ions was more than 80%. The adsorption of Cu2+ by aerogel was attributed to chelation and complexation. The desorption of porous chitosan/P(AA-co-DMAEMA) aerogels (CPA) by CO2 mainly occurred through charge repulsion of protonated ‒NH2 and ‒N‒ groups. After 6 cycles, the adsorption capacity of CPA for metal ions still reached 70% of the initial adsorption capacity, and the desorption rate reached 75%. This novel CO2-responsive chitosan aerogel is a highly efficient and environmentally friendly adsorbent for the adsorption and recovery of metal ions.

Physiological and Molecular Analysis Reveals the Differences of Photosynthesis between Colored and Green Leaf Poplars.

Leaf coloration changes evoke different photosynthetic responses among different poplar cultivars. The aim of this study is to investigate the photosynthetic difference between a red leaf cultivar (ZHP) and a green leaf (L2025) cultivar of Populus deltoides. In this study, 'ZHP' exhibited wide ranges and huge potential for absorption and utilization of light energy and CO2 concentration which were similar to those in 'L2025' and even showed a stronger absorption for weak light. However, with the increasing light intensity and CO2 concentration, the photosynthetic capacity in both 'L2025' and 'ZHP' was gradually restricted, and the net photosynthetic rate (Pn) in 'ZHP' was significantly lower than that in 'L2025'under high light or high CO2 conditions, which was mainly attributed to stomatal regulation and different photosynthetic efficiency (including the light energy utilization efficiency and photosynthetic CO2 assimilation efficiency) in these two poplars. Moreover, the higher anthocyanin content in 'ZHP' than that in 'L2025' was considered to be closely related to the decreased photosynthetic efficiency in 'ZHP'. According to the results from the JIP-test, the capture efficiency of the reaction center for light energy in 'L2025' was significantly higher than that in 'ZHP'. Interestingly, the higher levels of light quantum caused relatively higher accumulation of QA- in 'L2025', which blocked the electron transport and weakened the photosystem II (PSII) performance as compared with 'ZHP'; however, the decreased capture of light quantum also could not promote the utilization of light energy, which was the key to the low photosynthetic efficiency in 'ZHP'. The differential expressions of a series of photosynthesis-related genes further promoted these specific photosynthetic processes between 'L2025' and 'ZHP'.

CO2 -responsiveness of leaf isoprene emission: Why do species differ?

Leaf isoprene emission rate, I, decreases with increasing atmospheric CO2 concentration with major implications for global change. There is a significant interspecific variability in [CO2 ]-responsiveness of I, but the extent of this variation is unknown and its reasons are not understood. We hypothesized that the magnitude of emission reduction reflects the size and changeability of precursor pools responsible for isoprene emission (dimethylallyl diphosphate, DMADP and 2-methyl-erythritol 2,4-cyclodiphosphate, MEcDP). Changes in I and intermediate pool sizes upon increase of [CO2 ] from 400 to 1500 µmol/mol were studied in nine woody species spanning boreal to tropical ecosystems. I varied 10-fold, total substrate pool size 37-fold and the ratio of DMADP/MEcDP pool sizes 57-fold. At higher [CO2 ], I was reduced on average by 65%, but [CO2 ]-responsiveness varied an order of magnitude across species. The increase in [CO2 ] resulted in concomitant reductions in both substrate pools. The variation in [CO2 ]-responsiveness across species scaled with the reduction in pool sizes, the substrate pool size supported and the share of DMADP in total substrate pool. This study highlights a major interspecific variation in [CO2 ]-responsiveness of isoprene emission and conclusively links this variation to interspecific variability in [CO2 ] effects on substrate availability and intermediate pool size.

CO2 -responsive CCT protein interacts with 14-3-3 proteins and controls the expression of starch synthesis-related genes.

CO2 -responsive CCT protein (CRCT) is a positive regulator of starch synthesis-related genes such as ADP-glucose pyrophosphorylase large subunit 1 and starch branching enzyme I particularly in the leaf sheath of rice (Oryza sativa L.). The promoter GUS analysis revealed that CRCT expressed exclusively in the vascular bundle, whereas starch synthesis-related genes were expressed in different sites such as mesophyll cell and starch storage parenchyma cell. However, the chromatin immunoprecipitation (ChIP) using a FLAG-CRCT overexpression line and subsequent qPCR analyses showed that the 5'-flanking regions of these starch synthesis-related genes tended to be enriched by ChIP, suggesting that CRCT can bind to the promoter regions of these genes. The monomer of CRCT is 34.2 kDa; however, CRCT was detected at 270 kDa via gel filtration chromatography, suggesting that CRCT forms a complex in vivo. Immunoprecipitation and subsequent MS analysis pulled down several 14-3-3-like proteins. A yeast two-hybrid analysis and bimolecular fluorescence complementation assays confirmed the interaction between CRCT and 14-3-3-like proteins. Although there is an inconsistency in the place of expression, this study provides important findings regarding the molecular function of CRCT to control the expression of key starch synthesis-related genes.

The methylome is altered for plants in a high CO2 world: Insights into the response of a wild plant population to multigenerational exposure to elevated atmospheric [CO2 ].

Unravelling plant responses to rising atmospheric CO2 concentration ([CO2 ]) has largely focussed on plastic functional attributes to single generation [CO2 ] exposure. Quantifying the consequences of long-term, decadal multigenerational exposure to elevated [CO2 ] and the genetic changes that may underpin evolutionary mechanisms with [CO2 ] as a driver remain largely unexplored. Here, we investigated both plastic and evolutionary plant responses to elevated [CO2 ] by applying multi-omic technologies using populations of Plantago lanceolata L., grown in naturally high [CO2 ] for many generations in a CO2 spring. Seed from populations at the CO2 spring and an adjacent control site (ambient [CO2 ]) were grown in a common environment for one generation, and then offspring were grown in ambient or elevated [CO2 ] growth chambers. Low overall genetic differentiation between the CO2 spring and control site populations was found, with evidence of weak selection in exons. We identified evolutionary divergence in the DNA methylation profiles of populations derived from the spring relative to the control population, providing the first evidence that plant methylomes may respond to elevated [CO2 ] over multiple generations. In contrast, growth at elevated [CO2 ] for a single generation induced limited methylome remodelling (an order of magnitude fewer differential methylation events than observed between populations), although some of this appeared to be stably transgenerationally inherited. In all, 59 regions of the genome were identified where transcripts exhibiting differential expression (associated with single generation or long-term natural exposure to elevated [CO2 ]) co-located with sites of differential methylation or with single nucleotide polymorphisms exhibiting significant inter-population divergence. This included genes in pathways known to respond to elevated [CO2 ], such as nitrogen use efficiency and stomatal patterning. This study provides the first indication that DNA methylation may contribute to plant adaptation to future atmospheric [CO2 ] and identifies several areas of the genome that are targets for future study.

During photosynthetic induction, biochemical and stomatal limitations differ between Brassica crops.

Interventions to increase crop radiation use efficiency rely on understanding of how biochemical and stomatal limitations affect photosynthesis. When leaves transition from shade to high light, slow increases in maximum Rubisco carboxylation rate and stomatal conductance limit net CO2 assimilation for several minutes. However, as stomata open intercellular [CO2 ] increases, so electron transport rate could also become limiting. Photosynthetic limitations were evaluated in three important Brassica crops: Brassica rapa, Brassica oleracea and Brassica napus. Measurements of induction after a period of shade showed that net CO2 assimilation by B. rapa and B. napus saturated by 10 min. A new method of analyzing limitations to induction by varying intercellular [CO2 ] showed this was due to co-limitation by Rubisco and electron transport. By contrast, in B. oleracea persistent Rubisco limitation meant that CO2 assimilation was still recovering 15 min after induction. Correspondingly, B. oleracea had the lowest Rubisco total activity. The methodology developed, and its application here, shows a means to identify the basis of variation in photosynthetic efficiency in fluctuating light, which could be exploited in breeding and bioengineering to improve crop productivity.

Precipitation-drainage cycles lead to hot moments in soil carbon dioxide dynamics in a Neotropical wet forest.

Soil CO2 concentrations and emissions from tropical forests are modulated seasonally by precipitation. However, subseasonal responses to meteorological events (e.g., storms, drought) are less well known. Here, we present the effects of meteorological variability on short-term (hours to months) dynamics of soil CO2 concentrations and emissions in a Neotropical wet forest. We continuously monitored soil temperature, moisture, and CO2 for a three-year period (2015-2017), encompassing normal conditions, floods, a dry El Niño period, and a hurricane. We used a coupled model (Hydrus-1D) for soil water propagation, heat transfer, and diffusive gas transport to explain observed soil moisture, soil temperature, and soil CO2 concentration responses to meteorology, and we estimated soil CO2 efflux with a gradient-flux model. Then, we predicted changes in soil CO2 concentrations and emissions under different warming climate change scenarios. Observed short-term (hourly to daily) soil CO2 concentration responded more to precipitation than to other meteorological variables (including lower pressure during the hurricane). Observed soil CO2 failed to exhibit diel patterns (associated with diel temperature fluctuations in drier climates), except during the drier El Niño period. Climate change scenarios showed enhanced soil CO2 due to warmer conditions, while precipitation played a critical role in moderating the balance between concentrations and emissions. The scenario with increased precipitation (based on a regional model projection) led to increases of +11% in soil CO2 concentrations and +4% in soil CO2 emissions. The scenario with decreased precipitation (based on global circulation model projections) resulted in increases of +4% in soil CO2 concentrations and +18% in soil CO2 emissions, and presented more prominent hot moments in soil CO2 outgassing. These findings suggest that soil CO2 will increase under warmer climate in tropical wet forests, and precipitation patterns will define the intensity of CO2 outgassing hot moments.

Elevated CO2 promotes satellite stomata production in young cotyledons of Arabidopsis thaliana.

Stomata are tiny pores on plant leaves and stems surrounded by a pair of differentiated epidermal cells known as guard cells. Plants undergo guard cell differentiation in response to environmental cues, including atmospheric CO2 . To quantitatively evaluate stomatal development in response to elevated CO2 , imaging analysis of stomata was conducted using young cotyledons of Arabidopsis thaliana grown under ambient (380 ppm) and elevated (1,000 ppm) CO2 conditions. Our analysis revealed that treatment with 1,000 ppm CO2 did not affect stomatal numbers on abaxial sides of cotyledons but increased cotyledon area, resulting in decreased stomatal density, 7 days after germination. Interestingly, this treatment also perturbed the uniform distribution of stomata via excess satellite stomata and stomatal precursor cells. We used overexpression lines of the DNA replication licensing factor gene CDC6, a reported positive regulator of satellite stomata production. CDC6 overexpression decreased the speed of cotyledon expansion, even under treatment with 1,000 ppm CO2 , possibly by suppressing pavement cell maturation. In contrast, treatment with 1,000 ppm CO2 induced stomatal distribution changes in the overexpressor. These results suggest that treatment with 1,000 ppm CO2 enhances both cotyledon expansion and satellite stomata production via independent pathways, at least in young cotyledons of A. thaliana.