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32 records · Page 2

Testing unified theories for ozone response in C 4 species

Abstract There is tremendous interspecific variability in O 3 sensitivity among C 3 species, but variation among C 4 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 O 3 across species. In this study, we investigated leaf morphological, chemical, and photosynthetic responses of 22 genotypes of four C 4 bioenergy species (switchgrass, sorghum, maize, and miscanthus) to elevated O 3 in side‐by‐side field experiments using free‐air O 3 concentration enrichment (FACE). The C 4 species varied largely in leaf morphology, physiology, and nutrient composition. Elevated O 3 did not alter leaf morphology, nutrient content, stomatal conductance, chlorophyll fluorescence, and respiration in most genotypes but reduced net CO 2 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 O 3 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 O 3 were not affected by LMA directly but negatively influenced by LMA indirectly through stomatal conductance. These results demonstrate that there is significant variation in O 3 sensitivity among C 4 species with maize and sorghum showing greater sensitivity of photosynthesis to O 3 than switchgrass and miscanthus. Interspecific variation in O 3 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 O 3 sensitivity in C 4 bioenergy grasses. These findings advance understanding of O 3 tolerance in C 4 grasses and could aid in optimal placement of diverse C 4 bioenergy feedstock across a polluted landscape.

54 ENVIRONMENTAL SCIENCES↗

Increased activity of core photorespiratory enzymes and CO 2 transfer conductances are associated with higher and more optimal photosynthetic rates under elevated temperatures in the extremophile Rhazya stricta

Increase photorespiration and optimising intrinsic water use efficiency are unique challenges to photosynthetic carbon fixation at elevated temperatures. To determine how plants can adapt to facilitate high rates of photorespiration at elevated temperatures while also maintaining water-use efficiency, we performed in-depth gas exchange and biochemical assays of the C 3 extremophile, Rhazya stricta. These results demonstrate that R. stricta supports higher rates of photorespiration under elevated temperatures and that these higher rates of photorespiration correlate with increased activity of key photorespiratory enzymes; phosphoglycolate phosphatase and catalase. The increased photorespiratory enzyme activities may increase the overall capacity of photorespiration by reducing enzymatic bottlenecks and allowing minimal inhibitor accumulation under high photorespiratory rates. Additionally, we found the CO 2 transfer conductances (stomatal and mesophyll) are re-allocated to increase the water-use efficiency in R. stricta but not necessarily the photosynthetic response to temperature. These results suggest important adaptive strategies in R. stricta that maintain photosynthetic rates under elevated temperatures with optimal water loss. The strategies found in R. stricta may inform breeding and engineering efforts in other C 3 species to improve photosynthetic efficiency at high temperatures.

59 BASIC BIOLOGICAL SCIENCES↗

Improving Representation of Deforestation Effects on Evapotranspiration in the E3SM Land Model

Abstract Evapotranspiration (ET) plays an important role in land‐atmosphere coupling of energy, water, and carbon cycles. Following deforestation, ET is typically observed to decrease substantially as a consequence of decreases in leaf area and roots and increases in runoff. Changes in ET (latent heat flux) revise the surface energy and water budgets, which further affects large‐scale atmospheric dynamics and feeds back positively or negatively to long‐term forest sustainability. In this study, we used observations from a recent synthesis of 29 pairs of adjacent intact and deforested FLUXNET sites to improve model parameterization of stomatal characteristics, photosynthesis, and soil water dynamics in version 1 of the Energy Exascale Earth System Model (E3SM) Land Model (ELMv1). We found that default ELMv1 predicts an increase in ET after deforestation, likely leading to incorrect estimates of the effects of deforestation on land‐atmosphere coupling. The calibrated model accurately represented the FLUXNET observed deforestation effects on ET. Importantly, the search for global optimal parameters converged at values consistent with recent observational syntheses, confirming the reliability of the calibrated physical parameters. Applying this improved model parameterization to the globe scale reduced the bias of annual ET simulation by up to ~600 mm/year. Analysis on the roles of parameters suggested that future model development to improve ET simulation should focus on stomatal resistance and soil water‐related parameterizations. Finally, our predicted differences in seasonal ET changes from deforestation are large enough to substantially affect land‐atmosphere coupling and should be considered in such studies.

54 ENVIRONMENTAL SCIENCES↗

Drivers of Natural Variation in Water-Use Efficiency Under Fluctuating Light Are Promising Targets for Improvement in Sorghum

Improving leaf intrinsic water-use efficiency ( iWUE ), the ratio of photosynthetic CO 2 assimilation to stomatal conductance, could decrease crop freshwater consumption. iWUE has primarily been studied under steady-state light, but light in crop stands rapidly fluctuates. Leaf responses to these fluctuations substantially affect overall plant performance. Notably, photosynthesis responds faster than stomata to decreases in light intensity: this desynchronization results in substantial loss of iWUE . Traits that could improve iWUE under fluctuating light, such as faster stomatal movement to better synchronize stomata with photosynthesis, show significant natural diversity in C 3 species. However, C 4 crops have been less closely investigated. Additionally, while modification of photosynthetic or stomatal traits independent of one another will theoretically have a proportionate effect on iWUE , in reality these traits are inter-dependent. It is unclear how interactions between photosynthesis and stomata affect natural diversity in iWUE , and whether some traits are more tractable drivers to improve iWUE . Here, measurements of photosynthesis, stomatal conductance and iWUE under steady-state and fluctuating light, along with stomatal patterning, were obtained in 18 field-grown accessions of the C 4 crop sorghum. These traits showed significant natural diversity but were highly correlated, with important implications for improvement of iWUE . Some features, such as gradual responses of photosynthesis to decreases in light, appeared promising for improvement of iWUE . Other traits showed tradeoffs that negated benefits to iWUE , e.g., accessions with faster stomatal responses to decreases in light, expected to benefit iWUE , also displayed more abrupt losses in photosynthesis, resulting in overall lower iWUE . Genetic engineering might be needed to break these natural tradeoffs and achieve optimal trait combinations, e.g., leaves with fewer, smaller stomata, more sensitive to changes in photosynthesis. Traits describing iWUE at steady-state, and the change in iWUE following decreases in light, were important contributors to overall iWUE under fluctuating light.

54 ENVIRONMENTAL SCIENCES↗

IRF1 Maintains Optimal Constitutive Expression of Antiviral Genes and Regulates the Early Antiviral Response

Viral defense at mucosal sites depends on interferons (IFN) and IFN stimulated genes (ISGs), either of which may be constitutively expressed to maintain an “antiviral state” (AVS). However, the mechanisms that govern the AVS are poorly defined. Using a BEAS-2B respiratory epithelial cell line deficient in IRF1, we demonstrate higher susceptibility to infection with vesicular stomatitis virus (VSV) and influenza virus. IRF1-mediated restriction of VSV is IFN-independent, as blockade of types I and III IFNs and JAK-STAT signaling before infection did not affect VSV infection of either parent or IRF1 KO cells. Transcriptome analysis revealed that IRF1 regulates constitutive expression of ~300 genes, including antiviral ISGs: OAS2, BST2, and RNASEL and knockdown of any of these IRF1-dependent genes increased VSV infection. Additionally, IRF1 enhances rapid expression of IFNβ and IFNλ after stimulation with poly I:C and also regulates ISG expression. Mechanistically, IRF1 enhances recruitment of BRD4 to promotor-enhancer regions of ISGs for rapid expression and maintains levels of histone H3K4me1 for optimal constitutive expression. Finally, IRF1 also regulates constitutive expression of TLR2 and TLR3 and promotes signaling through these pattern recognition receptors (PRR). These data reveal multiple roles for IRF1 toward effective anti-viral responses by maintaining IFN-independent constitutive expression of anti-viral ISGs and supporting early IFN-dependent responses to PRR stimulation.

60 APPLIED LIFE SCIENCES↗

The Community Land Model Version 5: Description of New Features, Benchmarking, and Impact of Forcing Uncertainty

The Community Land Model (CLM) is the land component of the Community Earth System Model (CESM) and is used in several global and regional modeling systems. In this paper, we introduce model developments included in CLM version 5 (CLM5), which is the default land component for CESM2. We assess an ensemble of simulations, including prescribed and prognostic vegetation state, multiple forcing data sets, and CLM4, CLM4.5, and CLM5, against a range of metrics including from the International Land Model Benchmarking (ILAMBv2) package. CLM5 includes new and updated processes and parameterizations: (1) dynamic land units, (2) updated parameterizations and structure for hydrology and snow (spatially explicit soil depth, dry surface layer, revised groundwater scheme, revised canopy interception and canopy snow processes, updated fresh snow density, simple firn model, and Model for Scale Adaptive River Transport), (3) plant hydraulics and hydraulic redistribution, (4) revised nitrogen cycling (flexible leaf stoichiometry, leaf N optimization for photosynthesis, and carbon costs for plant nitrogen uptake), (5) global crop model with six crop types and time-evolving irrigated areas and fertilization rates, (6) updated urban building energy, (7) carbon isotopes, and (8) updated stomatal physiology. New optional features include demographically structured dynamic vegetation model (Functionally Assembled Terrestrial Ecosystem Simulator), ozone damage to plants, and fire trace gas emissions coupling to the atmosphere. Conclusive establishment of improvement or degradation of individual variables or metrics is challenged by forcing uncertainty, parametric uncertainty, and model structural complexity, but the multivariate metrics presented here suggest a general broad improvement from CLM4 to CLM5.

58 GEOSCIENCES↗

Engineering quantitative stomatal trait variation and local adaptation potential by cis‐regulatory editing

Summary Cis‐regulatory element editing can generate quantitative trait variation that mitigates extreme phenotypes and harmful pleiotropy associated with coding sequence mutations. Here, we applied a multiplexed CRISPR/Cas9 approach, informed by bioinformatic datasets, to generate genotypic variation in the promoter ofOsSTOMAGEN, a positive regulator of rice stomatal density. Engineered genotypic variation corresponded to broad and continuous variation in stomatal density, ranging from 70% to 120% of wild‐type stomatal density. This panel of stomatal variants was leveraged in physiological assays to establish discrete relationships between stomatal morphological variation and stomatal conductance, carbon assimilation and intrinsic water use efficiency in steady‐state and fluctuating light conditions. Additionally, promoter alleles were subjected to vegetative drought regimes to assay the effects of the edited alleles on developmental response to drought. Notably, the capacity for drought‐responsive stomatal density reprogramming instomagenand two cis‐regulatory edited alleles was reduced. Collectively our data demonstrate that cis‐regulatory element editing can generate near‐isogenic trait variation that can be leveraged for establishing relationships between anatomy and physiology, providing a basis for optimizing traits across diverse environments.

Biotechnology & Applied Microbiology↗

When a cuvette is not a canopy: a caution about measuring leaf temperature during gas exchange measurements

Plant gas exchange systems are widely used to study leaf physiological processes and properties such as stomatal function and the maximum carboxylation rate of Rubisco. Increasingly, these systems are used to assess how leaf gas exchange varies with temperature in order to better understand how rising temperature will impact plant function. Examples of such studies include variation in optimal temperatures of photosynthesis as a function of species and growth environment, and respiratory acclimation to higher temperatures. Leaf thermoregulation leading to homeothermy has been reported based on leaf gas exchange measurements spanning a large (~25 °C) temperature range. However, as we show here, the design of a popular gas exchange system used for temperature-response measurements can lead to biased measurements of leaf temperature. Furthermore, we demonstrate this with an example showing that apparent leaf thermoregulatory behavior can arise even in empty cuvettes. More broadly, our results have implications for other temperature manipulations in similar gas exchange systems.

59 BASIC BIOLOGICAL SCIENCES↗

Eco-evolutionary optimality as a means to improve vegetation and land-surface models

Global vegetation and land-surface models embody interdisciplinary scientific understanding of the behaviour of plants and ecosystems, and are indispensable to project the impacts of environmental change on vegetation and the interactions between vegetation and climate. Furthermore, systematic errors and persistently large differences among carbon and water cycle projections by different models highlight the limitations of current process formulations. In this review, focusing on core plant functions in the terrestrial carbon and water cycles, we show how unifying hypotheses derived from eco-evolutionary optimality (EEO) principles can provide novel, parameter-sparse representations of plant and vegetation processes. We present case studies that demonstrate how EEO generates parsimonious representations of core, leaf-level processes that are individually testable and supported by evidence. EEO approaches to photosynthesis and primary production, dark respiration and stomatal behaviour are ripe for implementation in global models. EEO approaches to other important traits, including the leaf economics spectrum and applications of EEO at the community level are active research areas. Independently tested modules emerging from EEO studies could profitably be integrated into modelling frameworks that account for the multiple time scales on which plants and plant communities adjust to environmental change.

59 BASIC BIOLOGICAL SCIENCES↗

Water foraging with dynamic roots in E3SM; The role of roots in terrestrial ecosystem memory on intermediate timescales (Final Technical Report)

Terrestrial ecosystems can show sustained responses to stress events that may last for years after the initial perturbation. This phenomenon is referred to as legacy or memory and it emerges from a set of ecosystem processes that are poorly captured by Earth System and Land Surface Models. Consequently, these models struggle to predict the impact that extreme climate events have on surface energy, carbon and hydrological exchange once a stressor such as drought has relaxed or in response to repeated exposure to stress. In this project, we set out to test how the addition of dynamic root profiles in land surface models impact the capacity for Earth System Models, such as the Department of Energy’s E3SM, to capture realistic legacy effects. Observations have shown that vegetation shifts their root profiles during stress events to forage for water and these altered root profiles may take years to relax back to the initial state. We hypothesized that the alteration of the belowground root structures may be a source of terrestrial ecosystem legacy that is missing from models. To test this idea we undertook three core activities. (1) We developed a global-scale analysis of the impact of dynamic roots on ecosystem legacy building from a recently developed dynamic root module. (2) We developed new root dynamics in the DOE’s Energy Land Model (ELM) that allowed not only root profiles to shift their profile but also simultaneously alter carbon allocation to fine root pools. We then tested the impact of these new dynamics with intensive sensitivity analysis at four long-term AmeriFlux sites. (3) We undertook detailed isotopic analysis of tree rings from these AmeriFlux sites to assess ecophysiological and ecohydrological legacy to stress events that can be used to benchmark the sensitivity experiments with ELM. From these core activities, we report the following key findings. Firstly, on a global scale, the addition of dynamic roots led to chronically water-stressed ecosystems recovering faster to climate stress while wetter ecosystem showed enhanced legacy. This is because across ecosystems, stress events were almost universally associated with water shortages that led to the development of deeper root profiles. These deeper root profiles proved beneficial for recovery from drought stress. While the root dynamics did not universally improve the modeled representation of legacy it showed complex transient dynamics that emerge from the addition of dynamic roots. Secondly, the sensitivity analysis illustrated long term shifts in rooting depths away from the prescribed default profile suggesting that initializing of root profiles could benefit from spin-up simulations that converge on locally optimized root profiles. In addition, by enabling dynamic allocation some of the sites predicted unrealistically low allocation to roots (and high allocation to leaves). While these changes did not dramatically alter modeled gross primary productivity, they illustrated that without more sophisticated root processes in models, there is little penalty to dramatically disinvest in roots. Thirdly, the isotopic analysis of tree rings showed highly distinct legacy responses across species and sites. For example, T. canadensis showed reduced transpiration the year after stress events illustrated by sustained elevated $\delta^{18}$O. In contrast, A saccharum displayed elevated $\delta^{13}$C associated with reduced stomatal conductance in response to the previous years’ stress event. These geochemical signatures of legacy were present despite tree growth returning to normal the year after the stress. The results show the importance of species-level dynamics in legacy that are absent in modeling that assumes common traits within plant functional types. In summary, the work here established new avenues to explore root dynamics in models while illustrating how additional root processes are needed before dynamic carbon allocation can be implemented. Lastly, this project provided mentorship to a postdoctoral fellow, training for an early career scientist and multiple undergraduate students recruited from a minority serving institution.

54 ENVIRONMENTAL SCIENCES↗

Derivation and Testing of Consumptive Water Use Fraction for Specialty Crops

The Crop Consumptive Use Fraction (CCUF) expresses beneficial water use in the form of seasonal evapotranspiration of applied water (ETAW), relative to total irrigation volume. The metric is an indicator of the efficiency of agricultural water use and is a recommended component for preparation of agricultural water management plans in California. An FAO-56 based web application has been developed to facilitate retrospective evaluation of ETAW, and hence CCUF, at field level. The current application is optimized for prevailing climate in four of the state’s main growing regions: San Joaquin Valley, Sacramento Valley, Central Coast, and North Coast. User inputs include crop type, soil texture, irrigation method, daily irrigation volume, daily rainfall, and seasonal start/stop dates to define the analysis period. Time series of fractional green canopy cover (Fc), based on Landsat and Sentinel-2 Earth-resource satellite observations, are imported from NASA’s Satellite Irrigation Management Support (SIMS) system. Grass reference evapotranspiration (ETo) time series are accessed from Spatial CIMIS (California Irrigation Management Information System). Daily crop height (h) is estimated as a simple function of typical maximum height for the given crop type (from FAO-56) and Fc. A vegetation density coefficient (Kd) is derived from Fc and h. Stomatal control factors during mid-and late-season are applied to tree and vine crops. Typical values for minimum daily relative humidity and mean daily windspeed, derived from historical CIMIS weather data, are used to correct for regional deviations from standard climate (defined as RHmin=45%, windspeed =2m/s). The resulting daily basal crop coefficient (Kcb) represents the ET of a well-watered crop with minimal soil evaporation, relative to ETo. A soil water balance sub-model for the top 0.1 m is used to calculate daily evaporation coefficients (Ke). A second sub-model applies to the root zone to calculate crop water stress coefficients (Ks) and effective precipitation (Pe), which is the fraction of rainfall that is available for crop use. Those coefficients are combined with Kcb and ETo to calculate daily ETc. ETAW is then derived as cumulative ETc less cumulative Pe. For verification purposes, sensor installations were used to measure seasonal ETc in commercial fields for several annual and perennial specialty crops by soil water balance and energy balance methods. Model estimates of seasonal ETc show mean absolute error of <10% compared to the ground measurements.

Derivation↗

Nitrogen nutrition and temporal effects of enhanced carbon dioxide on soybean growth

Plants grown on porous media at elevated CO2 levels generally have low concentrations of tissue N and often appear to require increased levels of external N to maximize growth response. This study determines if soybean [Glycine max (L.) Merr. Ransom'] grown hydroponically at elevated CO2 requires increases in external NO3- concentrations beyond levels that are optimal at ambient CO2 to maintain tissue N concentrations and maximize the growth response. This study also investigates temporal influences of elevated CO2 on growth responses by soybean. Plants were grown vegetatively for 34 d in hydroponic culture at atmospheric CO2 concentrations of 400, 650, and 900 microliters L-1 and during the final 18 d at NO3- concentrations of 0.5, 1.0, 5.0 and 10.0 mM in the culture solution. At 650 and 900 microliters L-1 CO2, plants had maximum increases of 31 and 45% in dry weight during the experimental period. Plant growth at 900 microliters L-1 CO2 was stimulated earlier than at 650 microliters L-1. During the final 18 d of the experiment, the relative growth rates (RGR) of plants grown at elevated CO2 declined. Elevated CO2 caused increases in total N and total NO3(-)-N content and leaf area but not leaf number. Enhancing CO2 levels also caused a decrease in root:shoot ratios. Stomatal resistance increased by 2.1- and 2.8-fold for plants at the 650 and 900 microliters L-1 CO2, respectively. Nitrate level in the culture solutions had no effect on growth or on C:N ratios of tissues, nor did increases in CO2 levels cause a decrease in N concentration of plant tissues. Hence, increases in NO3- concentration of the hydroponic solution were not necessary to maintain the N status of the plants or to maximize the growth response to elevated CO2.

NASA Program CELSS↗

Reducing Long‐Standing Surface Ozone Overestimation in Earth System Modeling by High‐Resolution Simulation and Dry Deposition Improvement

The overestimation of surface ozone concentration in low‐resolution global atmospheric chemistry and climate models has been a long‐standing issue. We first update the ozone dry deposition scheme in both high‐ (0.25°) and low‐resolution (1°) Community Earth System Model (CESM) version 1.3 runs, by adding the effects of leaf area index and correcting the sunlit and shaded fractions of stomatal resistances. With this update, 5‐year‐long summer simulations (2015–2019) using the low‐resolution CESM still exhibit substantial ozone overestimation (by 6.0–16.2 ppbv) over the U.S., Europe, eastern China, and ozone pollution hotspots. The ozone dry deposition scheme is further improved by adjusting the leaf cuticle conductance, reducing the mean ozone bias by 19%, and increasing the model resolution further reduces the ozone overestimation by 43%. We elucidate the mechanism by which model grid spacing influences simulated ozone, revealing distinctive pathways in urban versus rural areas. In rural areas, grid spacing mainly affects daytime ozone levels, where additional NO x emissions from nearby urban areas result in an ozone boost and overestimation in low‐resolution simulations. In contrast, over urban areas, daytime ozone overestimation follows a similar mechanism due to the influence of volatile organic compounds from surrounding rural areas. However, nighttime ozone overestimation is closely linked to weakened NO titration owing to the redistribution of urban NO x to rural areas. Additionally, stratosphere‐troposphere exchange may also contribute to reducing ozone bias in high‐resolution simulations, warranting further investigation. This optimized high‐resolution CESM may enhance understanding of ozone formation mechanisms, sources, and changes in a warming climate.

54 ENVIRONMENTAL SCIENCES↗

Engineering CAM Photosynthetic Machinery into Bioenergy Crops for Biofuels Production in Marginal Environments

CAM provides an excellent opportunity to engineer enhanced photosynthetic performance and water-use efficiency (WUE) into bioenergy crops. Our vision was to understand the enzymatic and regulatory pathways required for CAM in order to engineer CAM photosynthetic machinery into Arabidopsis and Populus. Major advances were made in understanding the molecular “parts list” necessary for engineering CAM through the sequencing the genomes and transcriptomes of several model and agronomically important CAM species including Agave, Ananas, Clusia, Kalanchoe, Mesembryanthemum, and Opuntia. Comparison of diel co-expression network analyses in Agave, Ananas, and Kalanchoe with expression patterns in C 3 photosynthesis species revealed the rescheduling of gene expression associated with stomatal opening and closing and led to the identification of many candidate CAM-related genes for further functional testing. Both RNAi-mediated gene silencing and CRISPR/Cas genome editing for gene knockout were used to investigate over 70 CAM-related enzymes, regulatory kinases/phosphatases, and transporters and 25 transcription factors implicated in playing key regulatory roles in CAM. Knockdown of key C 4 cycle enzymes within the carboxylation and decarboxylation modules of CAM not only perturbed CAM itself, but also altered the expression of core circadian clock components demonstrating that loss of CAM function also perturbs the central circadian clock and alters the regulatory patterns of key guard cell signaling genes that are linked with the characteristic inverse pattern of stomatal opening and closing during CAM. An advanced system for joining D.AN fragments from a universal library was developed that automatically maintains open reading frames (ORFs) and does not require linkers, adaptors, regions of sequence homology, amplification (or mutation for domestication) of fragments to function systems for the construction of gene circuits. Alternatively, Gibson assembly was used to create gene circuits consisting of up to 15 unique gene expression cassettes expressing either the core C 4 cycle of CAM comprising carboxylation or decarboxylation modules or both modules together. The expression of each gene cassette was driven by a drought-stress inducible 5’ regulatory region or promoter (and cognate 3’ terminator region) from A. thaliana so that CAM gene expression would be enhanced under conditions of water-deficit stress. These gene circuits were introduced into the C 3 photosynthesis model species A. thaliana. Expression of the nocturnal carboxylation module resulted in increased biomass and seed yield arising from net CO 2 fixation at night and malate accumulation typical of a CAM plant along with daytime CO 2 fixation typical of a C 3 photosynthesis plant. To create a leaf anatomy suited for the optimal performance of CAM in a C 3 photosynthesis plant, increased tissue succulence was engineered in A. thaliana (and Nicotiana sylvestris), which also improved biomass and seed yield and drought attenuation and improved salinity tolerance. The basic CAM Biodesign principles described in this report can be extended to increase the productivity and WUE of food and bioenergy crops and are expected to have broad applicability and ensure sustainable biofuel feedstock production in the face of the global climate crisis.

59 BASIC BIOLOGICAL SCIENCES↗