Climate change challenges, plant science solutions.

Climate change is a defining challenge of the 21st century, and this decade is a critical time for action to mitigate the worst effects on human populations and ecosystems. Plant science can play an important role in developing crops with enhanced resilience to harsh conditions (e.g. heat, drought, salt stress, flooding, disease outbreaks) and engineering efficient carbon-capturing and carbon-sequestering plants. Here, we present examples of research being conducted in these areas and discuss challenges and open questions as a call to action for the plant science community.

Similar photosynthetic but different yield responses of C3 and C4 crops to elevated O3.

The deleterious effects of ozone (O3) pollution on crop physiology, yield, and productivity are widely acknowledged. It has also been assumed that C4 crops with a carbon concentrating mechanism and greater water use efficiency are less sensitive to O3 pollution than C3 crops. This assumption has not been widely tested. Therefore, we compiled 46 journal articles and unpublished datasets that reported leaf photosynthetic and biochemical traits, plant biomass, and yield in five C3 crops (chickpea, rice, snap bean, soybean, and wheat) and four C4 crops (sorghum, maize, Miscanthus × giganteus, and switchgrass) grown under ambient and elevated O3 concentration ([O3]) in the field at free-air O3 concentration enrichment (O3-FACE) facilities over the past 20 y. When normalized by O3 exposure, C3 and C4 crops showed a similar response of leaf photosynthesis, but the reduction in chlorophyll content, fluorescence, and yield was greater in C3 crops compared with C4 crops. Additionally, inbred and hybrid lines of rice and maize showed different sensitivities to O3 exposure. This study quantitatively demonstrates that C4 crops respond less to elevated [O3] than C3 crops. This understanding could help maintain cropland productivity in an increasingly polluted atmosphere.

Plant biochemistry influences tropospheric ozone formation, destruction, deposition, and response.

Tropospheric ozone (O3) is among the most damaging air pollutant to plants. Plants alter the atmospheric O3 concentration in two distinct ways: (i) by the emission of volatile organic compounds (VOCs) that are precursors of O3; and (ii) by dry deposition, which includes diffusion of O3 into vegetation through stomata and destruction by nonstomatal pathways. Isoprene, monoterpenes, and higher terpenoids are emitted by plants in quantities that alter tropospheric O3. Deposition of O3 into vegetation is related to stomatal conductance, leaf structural traits, and the detoxification capacity of the apoplast. The biochemical fate of O3 once it enters leaves and reacts with aqueous surfaces is largely unknown, but new techniques for the tracking and identification of initial products have the potential to open the black box.

Genetic strategies for improving crop yields.

The current trajectory for crop yields is insufficient to nourish the world's population by 20501. Greater and more consistent crop production must be achieved against a backdrop of climatic stress that limits yields, owing to shifts in pests and pathogens, precipitation, heat-waves and other weather extremes. Here we consider the potential of plant sciences to address post-Green Revolution challenges in agriculture and explore emerging strategies for enhancing sustainable crop production and resilience in a changing climate. Accelerated crop improvement must leverage naturally evolved traits and transformative engineering driven by mechanistic understanding, to yield the resilient production systems that are needed to ensure future harvests.

A guide to photosynthetic gas exchange measurements: Fundamental principles, best practice and potential pitfalls.

Gas exchange measurements enable mechanistic insights into the processes that underpin carbon and water fluxes in plant leaves which in turn inform understanding of related processes at a range of scales from individual cells to entire ecosytems. Given the importance of photosynthesis for the global climate discussion it is important to (a) foster a basic understanding of the fundamental principles underpinning the experimental methods used by the broad community, and (b) ensure best practice and correct data interpretation within the research community. In this review, we outline the biochemical and biophysical parameters of photosynthesis that can be investigated with gas exchange measurements and we provide step-by-step guidance on how to reliably measure them. We advise on best practices for using gas exchange equipment and highlight potential pitfalls in experimental design and data interpretation. The Supporting Information contains exemplary data sets, experimental protocols and data-modelling routines. This review is a community effort to equip both the experimental researcher and the data modeller with a solid understanding of the theoretical basis of gas-exchange measurements, the rationale behind different experimental protocols and the approaches to data interpretation.

Solar-induced chlorophyll fluorescence captures the effects of elevated ozone on canopy structure and acceleration of senescence in soybean.

Solar-induced chlorophyll fluorescence (SIF) provides an opportunity to rapidly and non-destructively investigate how plants respond to stress. Here, we explored the potential of SIF to detect the effects of elevated O3 on soybean in the field where soybean was subjected to ambient and elevated O3 throughout the growing season in 2021. Exposure to elevated O3 resulted in a significant decrease in canopy SIF at 760 nm (SIF760), with a larger decrease in the late growing season (36%) compared with the middle growing season (13%). Elevated O3 significantly decreased the fraction of absorbed photosynthetically active radiation by 8-15% in the middle growing season and by 35% in the late growing stage. SIF760 escape ratio (fesc) was significantly increased under elevated O3 by 5-12% in the late growth stage due to a decrease of leaf chlorophyll content and leaf area index. Fluorescence yield of the canopy was reduced by 5-11% in the late growing season depending on the fesc estimation method, during which leaf maximum carboxylation rate and maximum electron transport were significantly reduced by 29% and 20% under elevated O3. These results demonstrated that SIF could capture the elevated O3 effect on canopy structure and acceleration of senescence in soybean and provide empirical support for using SIF for soybean stress detection and phenotyping.

Does the response of Rubisco and photosynthesis to elevated [CO2] change with unfavourable environmental conditions?

Climate change due to anthropogenic CO2 emissions affects plant performance globally. To improve crop resilience, we need to understand the effects of elevated CO2 concentration (e[CO2]) on CO2 assimilation and Rubisco biochemistry. However, the interactive effects of e[CO2] and abiotic stress are especially unclear. This study analyses the CO2 effect on photosynthetic capacity under different water availability and temperature conditions in 42 different crop species, varying in functional group, photosynthetic pathway and phenological stage. We analysed close to 3000 data points extracted from 120 published manuscripts. For C3 species, e[CO2] increases net photosynthesis and intercellular [CO2], while reducing stomatal conductance and transpiration. Vmaxc, Rubisco in vitro extractable maximal activity and content also decrease with e[CO2] in C3 species, while C4 crops are less responsive to e[CO2]. The interaction with drought and/or heat stress does not significantly alter these photosynthetic responses, indicating that the photosynthetic capacity of stressed plants responds to e[CO2]. Moreover, e[CO2] has strong effect on the photosynthetic capacity of grasses mainly in the final stages of development. This study provides insight into the intricate interactions within the plant photosynthetic apparatus under the influence of climate change, enhancing the understanding of mechanisms governing plant responses to environmental parameters.

How do drought and heat affect the response of soybean seed yield to elevated O3? An analysis of 15 seasons of free-air O3 concentration enrichment studies.

The coincidence of rising ozone concentrations ([O3]), increasing global temperatures, and drought episodes is expected to become more intense and frequent in the future. A better understanding of the responses of crop yield to elevated [O3] under different levels of drought and high temperature stress is, therefore, critical for projecting future food production potential. Using a 15-year open-air field experiment in central Illinois, we assessed the impacts of elevated [O3] coupled with variation in growing season temperature and water availability on soybean seed yield. Thirteen soybean cultivars were exposed to a wide range of season-long elevated [O3] in the field using free-air O3 concentration enrichment. Elevated [O3] treatments reduced soybean seed yield from as little as 5.3% in 2005 to 35.2% in 2010. Although cultivars differed in yield response to elevated [O3] (R), ranging from 17.5% to -76.4%, there was a significant negative correlation between R and O3 dosage. Soybean cultivars showed greater seed yield losses to elevated [O3] when grown at drier or hotter conditions compared to wetter or cooler years, because the hotter and drier conditions were associated with greater O3 treatment. However, year-to-year variation in weather conditions did not influence the sensitivity of soybean seed yield to a given increase in [O3]. Collectively, this study quantitatively demonstrates that, although drought conditions or warmer temperatures led to greater O3 treatment concentrations and O3-induced seed yield reduction, drought and temperature stress did not alter soybean's sensitivity to O3. Our results have important implications for modeling the effects of rising O3 pollution on crops and suggest that altering irrigation practices to mitigate O3 stress may not be effective in reducing crop sensitivity to O3.

Approaches to investigate crop responses to ozone pollution: from O3 -FACE to satellite-enabled modeling.

Ozone (O3 ) is a damaging air pollutant to crops. As one of the most reactive oxidants known, O3 rapidly forms other reactive oxygen species (ROS) once it enters leaves through stomata. Those ROS in turn can cause oxidative stress, reduce photosynthesis, accelerate senescence, and decrease crop yield. To improve and adapt our feed, fuel, and food supply to rising O3 pollution, a number of Free Air Concentration Enrichment (O3 -FACE) facilities have been developed around the world and have studied key staple crops. In this review, we provide an overview of the FACE facilities and highlight some of the lessons learned from the last two decades of research. We discuss the differences between C3 and C4 crop responses to elevated O3 , the possible trade-off between productivity and protection, genetic variation in O3 response within and across species, and how we might leverage this observed variation for crop improvement. We also highlight the need to improve understanding of the interaction between rising O3 pollution and other aspects of climate change, notably drought. Finally, we propose the use of globally modeled O3 data that are available at increasing spatial and temporal resolutions to expand upon the research conducted at the limited number of global O3 -FACE facilities.

Reductions in leaf area index, pod production, seed size, and harvest index drive yield loss to high temperatures in soybean.

Improvements in genetics, technology, and agricultural intensification have increased soybean yields; however, adverse climate conditions may prevent these gains from being fully realized in the future. Higher growing season temperatures reduce soybean yields in key production regions including the US Midwest, and better understanding of the developmental and physiological mechanisms that constrain soybean yield under high temperature conditions is needed. This study tested the response of two soybean cultivars to four elevated temperature treatments (+1.7, +2.6, +3.6, and +4.8 °C) in the field over three growing seasons and identified threshold temperatures for response and linear versus non-linear trait responses to temperature. Yield declined non-linearly to temperature, with decreases apparent when canopy temperature exceeded 20.9 °C for the locally adapted cultivar and 22.7°C for a cultivar adapted to more southern locations. While stem node number increased with increasing temperature, leaf area index decreased substantially. Pod production, seed size, and harvest index significantly decreased with increasing temperature. The seasonal average temperature of even the mildest treatment exceeded the threshold temperatures for yield loss, emphasizing the importance of improving temperature tolerance in soybean germplasm with intensifying climate change.

Anatomical determinants of gas exchange and hydraulics vary with leaf shape in soybean.

BACKGROUND AND AIMS: Leaf shape in crops can impact light distribution and carbon capture at the whole plant and canopy level. Given similar leaf inclination, narrow leaves can allow a greater fraction of incident light to pass through to lower canopy leaves by reducing leaf area index, which can potentially increase canopy-scale photosynthesis. Soybean has natural variation in leaf shape which can be utilized to optimize canopy architecture. However, the anatomical and physiological differences underlying variation in leaf shape remain largely unexplored. METHODS: In this study, we selected 28 diverse soybean lines with leaf length to width ratios (leaf ratio) ranging between 1.1 and 3.2. We made leaf cross-sectional, gas exchange, vein density and hydraulic measurements and studied their interrelationships among these lines. KEY RESULTS: Our study shows that narrow leaves tend to be thicker, with an ~30 µm increase in leaf thickness for every unit increase in leaf ratio. Interestingly, thicker leaves had a greater proportion of spongy mesophyll while the proportions of palisade and paraveinal mesophyll decreased. In addition, narrow and thicker leaves had greater photosynthesis and stomatal conductance per unit area along with greater leaf hydraulic conductance. CONCLUSIONS: Our results suggest that selecting for narrow leaves can improve photosynthetic performance and potentially provide a yield advantage in soybean.

Predicting biochemical acclimation of leaf photosynthesis in soybean under in-field canopy warming using hyperspectral reflectance.

Traditional gas exchange measurements are cumbersome, which makes it difficult to capture variation in biochemical parameters, namely the maximum rate of carboxylation measured at a reference temperature (Vcmax25 ) and the maximum electron transport at a reference temperature (Jmax25 ), in response to growth temperature over time from days to weeks. Hyperspectral reflectance provides reliable measures of Vcmax25 and Jmax25 ; however, the capability of this method to capture biochemical acclimations of the two parameters to high growth temperature over time has not been demonstrated. In this study, Vcmax25 and Jmax25 were measured over multiple growth stages during two growing seasons for field-grown soybeans using both gas exchange techniques and leaf spectral reflectance under ambient and four elevated canopy temperature treatments (ambient+1.5, +3, +4.5, and +6°C). Spectral vegetation indices and machine learning methods were used to build predictive models for Vcmax25 and Jmax25 , based on the leaf reflectance. Results showed that these models yielded an R2 of 0.57-0.65 and 0.48-0.58 for Vcmax25 and Jmax25 , respectively. Hyperspectral reflectance captured biochemical acclimation of leaf photosynthesis to high temperature in the field, improving spatial and temporal resolution in the ability to assess the impact of future warming on crop productivity.

The leaf economics spectrum of triploid and tetraploid C4 grass Miscanthus x giganteus.

The leaf economics spectrum (LES) describes multivariate correlations in leaf structural, physiological and chemical traits, originally based on diverse C3 species grown under natural ecosystems. However, the specific contribution of C4 species to the global LES is studied less widely. C4 species have a CO2 concentrating mechanism which drives high rates of photosynthesis and improves resource use efficiency, thus potentially pushing them towards the edge of the LES. Here, we measured foliage morphology, structure, photosynthesis, and nutrient content for hundreds of genotypes of the C4 grass Miscanthus× giganteus grown in two common gardens over two seasons. We show substantial trait variations across M.× giganteus genotypes and robust genotypic trait relationships. Compared to the global LES, M.× giganteus genotypes had higher photosynthetic rates, lower stomatal conductance, and less nitrogen content, indicating greater water and photosynthetic nitrogen use efficiency in the C4 species. Additionally, tetraploid genotypes produced thicker leaves with greater leaf mass per area and lower leaf density than triploid genotypes. By expanding the LES relationships across C3 species to include C4 crops, these findings highlight that M.× giganteus occupies the boundary of the global LES and suggest the potential for ploidy to alter LES traits.

Physiological trait networks enhance understanding of crop growth and water use in contrasting environments.

Plant function arises from a complex network of structural and physiological traits. Explicit representation of these traits, as well as their connections with other biophysical processes, is required to advance our understanding of plant-soil-climate interactions. We used the Terrestrial Regional Ecosystem Exchange Simulator (TREES) to evaluate physiological trait networks in maize. Net primary productivity (NPP) and grain yield were simulated across five contrasting climate scenarios. Simulations achieving high NPP and grain yield in high precipitation environments featured trait networks conferring high water use strategies: deep roots, high stomatal conductance at low water potential ("risky" stomatal regulation), high xylem hydraulic conductivity and high maximal leaf area index. In contrast, high NPP and grain yield was achieved in dry environments with low late-season precipitation via water conserving trait networks: deep roots, high embolism resistance and low stomatal conductance at low leaf water potential ("conservative" stomatal regulation). We suggest that our approach, which allows for the simultaneous evaluation of physiological traits, soil characteristics and their interactions (i.e., networks), has potential to improve our understanding of crop performance in different environments. In contrast, evaluating single traits in isolation of other coordinated traits does not appear to be an effective strategy for predicting plant performance.

Advances in field-based high-throughput photosynthetic phenotyping.

Gas exchange techniques revolutionized plant research and advanced understanding, including associated fluxes and efficiencies, of photosynthesis, photorespiration, and respiration of plants from cellular to ecosystem scales. These techniques remain the gold standard for inferring photosynthetic rates and underlying physiology/biochemistry, although their utility for high-throughput phenotyping (HTP) of photosynthesis is limited both by the number of gas exchange systems available and the number of personnel available to operate the equipment. Remote sensing techniques have long been used to assess ecosystem productivity at coarse spatial and temporal resolutions, and advances in sensor technology coupled with advanced statistical techniques are expanding remote sensing tools to finer spatial scales and increasing the number and complexity of phenotypes that can be extracted. In this review, we outline the photosynthetic phenotypes of interest to the plant science community and describe the advances in high-throughput techniques to characterize photosynthesis at spatial scales useful to infer treatment or genotypic variation in field-based experiments or breeding trials. We will accomplish this objective by presenting six lessons learned thus far through the development and application of proximal/remote sensing-based measurements and the accompanying statistical analyses. We will conclude by outlining what we perceive as the current limitations, bottlenecks, and opportunities facing HTP of photosynthesis.

Testing unified theories for ozone response in C4 species.

There is tremendous interspecific variability in O3  sensitivity among C3  species, but variation among C4  species has been less clearly documented. It is also unclear whether stomatal conductance and leaf structure such as leaf mass per area (LMA) determine the variation in sensitivity to O3 across species. In this study, we investigated leaf morphological, chemical, and photosynthetic responses of 22 genotypes of four C4 bioenergy species (switchgrass, sorghum, maize, and miscanthus) to elevated O3 in side-by-side field experiments using free-air O3 concentration enrichment (FACE). The C4  species varied largely in leaf morphology, physiology, and nutrient composition. Elevated O3 did not alter leaf morphology, nutrient content, stomatal conductance, chlorophyll fluorescence, and respiration in most genotypes but reduced net CO2 assimilation in maize and photosynthetic capacity in sorghum and maize. Species with lower LMA and higher stomatal conductance tended to show greater losses in photosynthetic rate and capacity in elevated O3 compared with species with higher LMA and lower stomatal conductance. Stomatal conductance was the strongest determinant of leaf photosynthetic rate and capacity. The response of both area- and mass-based leaf photosynthetic rate and capacity to elevated O3 were not affected by LMA directly but negatively influenced by LMA indirectly through stomatal conductance. These results demonstrate that there is significant variation in O3  sensitivity among C4  species with maize and sorghum showing greater sensitivity of photosynthesis to O3 than switchgrass and miscanthus. Interspecific variation in O3  sensitivity was determined by direct effects of stomatal conductance and indirect effects of LMA. This is the first study to provide a test of unifying theories explaining variation in O3  sensitivity in C4 bioenergy grasses. These findings advance understanding of O3 tolerance in C4  grasses and could aid in optimal placement of diverse C4 bioenergy feedstock across a polluted landscape.

Interannual variability of ecosystem iso/anisohydry is regulated by environmental dryness.

●Plants are characterized by the iso/anisohydry continuum depending on how they regulate leaf water potential (ΨL ). However, how iso/anisohydry changes over time in response to year-to-year variations in environmental dryness and how such responses vary across different regions remains poorly characterized. ●We investigated how dryness, represented by aridity index, affects the interannual variability of ecosystem iso/anisohydry at the regional scale, estimated using satellite microwave vegetation optical depth (VOD) observations. This ecosystem-level analysis was further complemented with published field observations of species-level ΨL . ●We found different behaviors in the directionality and sensitivity of isohydricity (σ) with respect to the interannual variation of dryness in different ecosystems. These behaviors can largely be differentiated by the average dryness of the ecosystem itself: in mesic ecosystems, σ decreases in drier years with a higher sensitivity to dryness; in xeric ecosystems, σ increases in drier years with a lower sensitivity to dryness. These results were supported by the species-level synthesis. ●Our study suggests that how plants adjust their water use across years - as revealed by their interannual variability in isohydricity - depends on the dryness of plants' living environment. This finding advances our understanding of plant responses to drought at regional scales.

Bioenergy sorghum maintains photosynthetic capacity in elevated ozone concentrations.

Elevated tropospheric ozone concentration (O3 ) significantly reduces photosynthesis and productivity in several C4 crops including maize, switchgrass and sugarcane. However, it is unknown how O3 affects plant growth, development and productivity in sorghum (Sorghum bicolor L.), an emerging C4 bioenergy crop. Here, we investigated the effects of elevated O3 on photosynthesis, biomass and nutrient composition of a number of sorghum genotypes over two seasons in the field using free-air concentration enrichment (FACE), and in growth chambers. We also tested if elevated O3 altered the relationship between stomatal conductance and environmental conditions using two common stomatal conductance models. Sorghum genotypes showed significant variability in plant functional traits, including photosynthetic capacity, leaf N content and specific leaf area, but responded similarly to O3 . At the FACE experiment, elevated O3 did not alter net CO2 assimilation (A), stomatal conductance (gs ), stomatal sensitivity to the environment, chlorophyll fluorescence and plant biomass, but led to reductions in the maximum carboxylation capacity of phosphoenolpyruvate and increased stomatal limitation to A in both years. These findings suggest that bioenergy sorghum is tolerant to O3 and could be used to enhance biomass productivity in O3 polluted regions.

Unique contributions of chlorophyll and nitrogen to predict crop photosynthetic capacity from leaf spectroscopy.

The photosynthetic capacity or the CO2-saturated photosynthetic rate (Vmax), chlorophyll, and nitrogen are closely linked leaf traits that determine C4 crop photosynthesis and yield. Accurate, timely, rapid, and non-destructive approaches to predict leaf photosynthetic traits from hyperspectral reflectance are urgently needed for high-throughput crop monitoring to ensure food and bioenergy security. Therefore, this study thoroughly evaluated the state-of-the-art physically based radiative transfer models (RTMs), data-driven partial least squares regression (PLSR), and generalized PLSR (gPLSR) models to estimate leaf traits from leaf-clip hyperspectral reflectance, which was collected from maize (Zea mays L.) bioenergy plots with diverse genotypes, growth stages, treatments with nitrogen fertilizers, and ozone stresses in three growing seasons. The results show that leaf RTMs considering bidirectional effects can give accurate estimates of chlorophyll content (Pearson correlation r=0.95), while gPLSR enabled retrieval of leaf nitrogen concentration (r=0.85). Using PLSR with field measurements for training, the cross-validation indicates that Vmax can be well predicted from spectra (r=0.81). The integration of chlorophyll content (strongly related to visible spectra) and nitrogen concentration (linked to shortwave infrared signals) can provide better predictions of Vmax (r=0.71) than only using either chlorophyll or nitrogen individually. This study highlights that leaf chlorophyll content and nitrogen concentration have key and unique contributions to Vmax prediction.

30 years of free-air carbon dioxide enrichment (FACE): What have we learned about future crop productivity and its potential for adaptation?

Free-air CO2 enrichment (FACE) allows open-air elevation of [CO2 ] without altering the microclimate. Its scale uniquely supports simultaneous study from physiology and yield to soil processes and disease. In 2005 we summarized results of then 28 published observations by meta-analysis. Subsequent studies have combined FACE with temperature, drought, ozone, and nitrogen treatments. Here, we summarize the results of now almost 250 observations, spanning 14 sites and five continents. Across 186 independent studies of 18 C3 crops, elevation of [CO2 ] by ca. 200 ppm caused a ca. 18% increase in yield under non-stress conditions. Legumes and root crops showed a greater increase and cereals less. Nitrogen deficiency reduced the average increase to 10%, as did warming by ca. 2°C. Two conclusions of the 2005 analysis were that C4 crops would not be more productive in elevated [CO2 ], except under drought, and that yield responses of C3 crops were diminished by nitrogen deficiency and wet conditions. Both stand the test of time. Further studies of maize and sorghum showed no yield increase, except in drought, while soybean productivity was negatively affected by early growing season wet conditions. Subsequent study showed reduced levels of nutrients, notably Zn and Fe in most crops, and lower nitrogen and protein in the seeds of non-leguminous crops. Testing across crop germplasm revealed sufficient variation to maintain nutrient content under rising [CO2 ]. A strong correlation of yield response under elevated [CO2 ] to genetic yield potential in both rice and soybean was observed. Rice cultivars with the highest yield potential showed a 35% yield increase in elevated [CO2 ] compared to an average of 14%. Future FACE experiments have the potential to develop cultivars and management strategies for co-promoting sustainability and productivity under future elevated [CO2 ].