Stomatal Traits Research Articles

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

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 of OsSTOMAGEN, 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 in stomagen and 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.

Stomatal development in the changing climate.

Stomata, microscopic pores flanked by symmetrical guard cells, are vital regulators of gas exchange that link plant processes with environmental dynamics. The formation of stomata involves the multi-step progression of a specialized cell lineage. Remarkably, this process is heavily influenced by environmental factors, allowing plants to adjust stomatal production to local conditions. With global warming set to alter our climate at an unprecedented pace, understanding how environmental factors impact stomatal development and plant fitness is becoming increasingly important. In this Review, we focus on the effects of carbon dioxide, high temperature and drought - three environmental factors tightly linked to global warming - on stomatal development. We summarize the stomatal response of a variety of plant species and highlight the existence of species-specific adaptations. Using the model plant Arabidopsis, we also provide an update on the molecular mechanisms involved in mediating the plasticity of stomatal development. Finally, we explore how knowledge on stomatal development is being applied to generate crop varieties with optimized stomatal traits that enhance their resilience against climate change and maintain agricultural productivity.

Machine learning-enabled computer vision for plant phenotyping: a primer on AI/ML and case study on stomatal patterning.

Artificial intelligence and machine learning (AI/ML) can be used to automatically analyze large image datasets. One valuable application of this approach is estimation of plant trait data contained within images. Here we review 39 papers that describe the development and/or application of such models for estimation of stomatal traits from epidermal micrographs. In doing so, we hope to provide plant biologists with a foundational understanding of AI/ML and summarize the current capabilities and limitations of published tools. While most models show human-level performance for stomatal density (SD) quantification at superhuman speed, they are often likely to be limited in how broadly they can be applied across phenotypic diversity associated with genetic, environmental or developmental variation. Other models can make predictions across greater phenotypic diversity and/or additional stomatal/epidermal traits, but require significantly greater time investment to generate ground-truth data. We discuss the challenges and opportunities presented by AI/ML-enabled computer vision analysis, and make recommendations for future work to advance accelerated stomatal phenotyping.

Effect of Colchicine and Bio-catharantin on the DNA Relative Content and Stomatal Structure of Black Rice (Oryza sativa L. var. Jeliteng)

<p>Black rice (<em>Oryza sativa</em> L. var. Jeliteng), known for its health benefits compared to white rice, faces challenges in productivity. Among varieties, this black rice is popular in Indonesia but shows low yield. Research on improving black rice through genetic manipulation with antimitotic substances is limited. Therefore, this study aims to compare the effects of colchicine and Bio-catharantin on the germination rate, DNA relative content, and stomatal structure of <em>O. sativa </em>L. var. Jeliteng. Seeds were treated with colchicine (0.1%, 0.2%, and 0.3%) and Bio-catharantin (0.1%, 0.2%, 0.3%, 0.4%, and 0.5%) at soaking durations of 12, 24, and 48 hours. Germination was assayed, ploidy was determined using flow cytometry, and stomatal traits, including size and density, were examined microscopically. The results showed that Bio-catharantin did not exhibit any toxic effects on germination rates, whereas colchicine reduced germination starting at 0.2% concentration. Both chemical agents modified the DNA relative content of Jeliteng black rice. Colchicine generally increased stomatal length and width while decreasing stomatal density, with significant changes at 0.3% concentration for 24 hours. Bio-catharantin also altered stomatal traits, enhancing length and width in most cases but significantly reducing density under certain conditions. Bio-catharantin emerged as a promising alternative to colchicine for inducing chromosomal mutations in plants, offering benefits in altered stomatal structures without the toxic effects on germination, compared to colchicine.</p>

Impacts of short-term rainfall and snowfall exclusions on hydraulic, economic and stomatal traits of Larix gmelinii in northeastern China

Impacts of short-term rainfall and snowfall exclusions on hydraulic, economic and stomatal traits of Larix gmelinii in northeastern China

Grass leaf structural and stomatal trait responses to climate gradients assessed over the 20th century and across the Great Plains, USA.

Abstract. Using herbarium specimens spanning 133 years and field-collected measurements, we assessed intraspecific trait (leaf structural and stomatal) variability from grass species in the Great Plains of North America. We focused on two widespread, closely related grasses from the tribe Paniceae: Dichanthelium oligosanthes subsp. scribnerianum (C3) and Panicum virgatum (C4). Thirty-one specimens per taxon were sampled from local herbaria from the years 1887 to 2013 to assess trait responses across time to changes in atmospheric [CO2] and growing season precipitation and temperature. In 2021 and 2022, the species were measured from eight grasslands sites to explore how traits vary spatially across natural continental precipitation and temperature gradients. Δ13C increased with atmospheric [CO2] for D. oligosanthes but decreased for P. virgatum, likely linked to increases in precipitation in the study region over the past century. Notably, this is the first record of decreasing Δ13C over time for a C4 species illustrating 13C linkages to climate. As atmospheric [CO2] increased, C:N increased and δ15N decreased for both species and %N decreased for D. oligosanthes. Across a large precipitation gradient, D. oligosanthes leaf traits were more responsive to changes in precipitation than those of P. virgatum. In contrast, only two traits of P. virgatum responded to increases in temperature across a gradient: specific leaf area (increase) and leaf dry matter content (decrease). The only shared significant trend between species was increased C:N with precipitation. Our work demonstrates that these closely related grass species with different photosynthetic pathways exhibited various trait responses across temporal and spatial scales, illustrating the key role of scale of inquiry for forecasting leaf trait responses to future environmental change.

Stomata Are Driving the Direction of CO2-Induced Water-Use Efficiency Gain in Selected Tropical Trees in Fiji.

Understanding plant physiological response to a rising atmospheric CO2 concentration (ca) is key in predicting Earth system plant-climate feedbacks; however, the effects of long-term rising ca on plant gas-exchange characteristics in the tropics are largely unknown. Studying this long-term trend using herbarium records is challenging due to specimen trait variation. We assessed the impact of a ca rise of ~95 ppm (1927-2015) on the intrinsic water-use efficiency (iWUE) and maximum stomatal conductance (gsmax) of five tropical tree species in Fiji using the isotopic composition and stomatal traits of herbarium leaves. Empirical results were compared with simulated values using models that uniquely incorporated the variation in the empirical gsmax responses and species-specific parameterisation. The magnitude of the empirical iWUE and gsmax response was species-specific, ranging from strong to negligible. Stomatal density was more influential than the pore size in determining the gsmax response to ca. While our simulation results indicated that photosynthesis is the main factor contributing to the iWUE gain, stomata were driving the iWUE trend across the tree species. Generally, a stronger increase in the iWUE was accompanied by a stronger decline in stomatal response. This study demonstrates that the incorporation of variation in the gsmax in simulations is necessary for assessing an individual species' iWUE response to changing ca.

Efficient gene editing of a model fern species through gametophyte-based transformation.

The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated nuclease (Cas) system allows precise and easy editing of genes in many plant species. However, this system has not yet been applied to any fern species through gametophytes due to the complex characteristics of fern genomes, genetics, and physiology. Here, we established a protocol for gametophyte-based screening of single-guide RNAs (sgRNAs) with high efficiency for CRISPR/Cas9-mediated gene knockout in a model fern species, Ceratopteris richardii. We utilized the C. richardii ACTIN promoter to drive sgRNA expression and the enhanced CaMV 35S promoter to drive the expression of Streptococcus pyogenes Cas9 in this CRISPR-mediated editing system, which was employed to successfully edit a few genes, such as Nucleotidase/phosphatase 1 (CrSAL1) and Phytoene Desaturase (CrPDS), which resulted in an albino phenotype in C. richardii. Knockout of CrSAL1 resulted in significantly (P<0.05) reduced stomatal conductance (gs), leaf transpiration rate (E), guard cell length, and abscisic acid (ABA)-induced reactive oxygen species (ROS) accumulation in guard cells. Moreover, CrSAL1 overexpressing plants showed significantly increased net photosynthetic rate (A), gs, and E as well as most of the stomatal traits and ABA-induced ROS production in guard cells compared to the wild-type (WT) plants. Taken together, our optimized CRISPR/Cas9 system provides a useful tool for functional genomics in a model fern species, allowing the exploration of fern gene functions for evolutionary biology, herbal medicine discovery, and agricultural applications.

Leaf stomatal configuration and photosynthetic traits jointly affect leaf water use efficiency in forests along climate gradients.

Water use efficiency (WUE) represents the trade-off between carbon assimilation and water loss in plants. It remains unclear how leaf stomatal and photosynthetic traits regulate the spatial variation of leaf WUE in different natural forest ecosystems. We investigated 43 broad-leaf tree species spanning from cold-temperate to tropical forests in China. We quantified leaf WUE using leaf δ13C and measured stomatal traits, photosynthetic traits as well as maximum stomatal conductance ( ) and maximum carboxylation capacity ( ). We found that leaves in cold-temperate forests displayed 'fast' carbon economics, characterized by higher leaf nitrogen, Chl, specific leaf area, and , as an adaptation to the shorter growing season. However, these leaves exhibited 'slow' hydraulic traits, with larger but fewer stomata and similar , resulting in higher leaf WUE. By contrast, leaves in tropical forests had smaller and denser stomata, enabling swift response to heterogeneous light conditions. However, this stomatal configuration increased potential water loss, and coupled with their low photosynthetic capacity, led to lower WUE. Our findings contribute to understanding how plant photosynthetic and stomatal traits regulate carbon-water trade-offs across climatic gradients, advancing our ability to predict the impacts of climate changes on forest carbon and water cycles.

Integrated Effects of Soil Moisture on Wheat Hydraulic Properties and Stomatal Regulation.

The development of water-saving management relies on understanding the physiological response of crops to soil drought. The coordinated regulation of hydraulics and stomatal conductance in plant water relations has steadily received attention. However, research focusing on grain crops, such as winter wheat, remains limited. In this study, three soil water supply treatments, including high (H), moderate (M), and low (L) soil water contents, were conducted with potted winter wheat. Leaf water potential (Ψleaf), leaf hydraulic conductance (Kleaf), and stomatal conductance (gs), as well as leaf biochemical parameters and stomatal traits were measured. Results showed that, compared to H, predawn leaf water potential (ΨPD) significantly reduced by 48.10% and 47.91%, midday leaf water potential (ΨMD) reduced by 40.71% and 43.20%, Kleaf reduced by 64.80% and 65.61%, and gs reduced by 21.20% and 43.41%, respectively, under M and L conditions. Although gs showed a significant difference between M and L, Ψleaf and Kleaf did not show significant differences between these treatments. The maximum carboxylation rate (Vcmax) and maximum electron transfer rate (Jmax) under L significantly decreased by 23.11% and 28.10%, stomatal density (SD) and stomatal pore area index (SPI) under L on the abaxial side increased by 59.80% and 52.30%, respectively, compared to H. The leaf water potential at 50% hydraulic conduction loss (P50) under L was not significantly reduced. The gs was positively correlated with ΨMD and Kleaf, but it was negatively correlated with abscisic acid (ABA) and SD. A threshold relationship between gs and Kleaf was observed, with rapid and linear reduction in gs occurring only when Kleaf fell below 8.70 mmol m-2 s-1 MPa-1. Our findings demonstrate that wheat leaves adapt stomatal regulation strategies from anisohydric to isohydric in response to reduced soil water content. These results enrich the theory of trade-offs between the carbon assimilation and hydraulic safety in crops and also provide a theoretical basis for water management practices based on stomatal regulation strategies under varying soil water conditions.

A continental-scale analysis reveals the latitudinal gradient of stomatal density across amphistomatous species: Evolutionary history vs. present-day environment.

Amphistomy is a potential method for increasing photosynthetic rate; however, the latitudinal gradients of stomatal density across amphistomatous species and their drivers remain unknown. Here, the adaxial stomatal density (SDad) and abaxial stomatal density (SDab) of 486 amphistomatous species-site combinations, belonging to 32 plant families, were collected from China, and their total stomatal density (SDtotal) and stomatal ratio (SR) were calculated. Overall, these four stomatal traits did not show significant phylogenetic signals. There were no significant differences in SDab and SDtotal between woody and herbaceous species, but SDad and SR were higher in woody species than in herbaceous species. Besides, a significantly positive relationship between SDab and SDad was observed. We also found that stomatal density (including SDab, SDad, and SDtotal) decreased with latitude while SR increased with latitude, and temperature seasonality was the most important environmental factor driving it. Besides, evolutionary history (represented by both phylogeny and species) explained about 10-22 fold more of the variation in stomatal traits than the present-day environment (65.2%-71.1% vs. 2.9%-6.8%). Our study extended our knowledge of trait-environment relationships and highlighted the importance of evolutionary history in driving stomatal trait variability.

Variations in the leaf economics spectrum, anatomical, ultrastructural, and stomatal traits of five tree species in the urban-rural air pollution environment

Rapid urbanization has contributed to global increases in air pollution derived from urban areas. Unlike natural forests, urban forests are exposed to higher concentrations of airborne pollutants due to the strong urban-suburban-rural pollutant emission gradients. However, there remains a pressing lack of available information pertaining to the urban air pollution-related effects on the leaf economics spectrum, anatomical, ultrastructural, and stomatal traits of tree species along an urban-rural gradient. Here, the degree to which urban air pollution impacts the adaption of greening tree species and associated service functions was assessed by sampling five common tree species (Acer pictum, Fraxinus chinensis, Koelreuteria paniculata, Salix babylonica, Sophora japonica) along urban-rural-natural forests in the Beijing metropolitan region of China. These analyses revealed a significant reduction in leaf mass per unit area (–13.4 %), leaf thickness (–16.7 %), and stomatal area (–27.5 %) with increasing proximity to areas of greater air pollution that coincide with significant increases in leaf tissue density (+12.6 %), leaf nitrogen content (+10.1 %), relative chlorophyll content (+2.7 %), and stomatal density (+11.9 %). Higher air pollution levels were associated with organelle changes including gradual disintegration of chloroplasts, larger intercellular spaces and apparent starch and plastoglobuli deposition. Air pollution was conducive to the strengthening of the trade-off potential and adaptation strategies of trees in urban ecosystems, which are associated with trees with a rapid investment return strategy associated with thick leaves and strong photosynthetic capacity. These results provide strong empirical evidence of the profound air pollution-induced changes in leaf functional traits and adaption ability of urban forest tree species.

Decoding stomatal characteristics regulating water use efficiency at leaf and plant scales in rice genotypes.

Stomatal traits in rice genotypes affect water use efficiency. Low-frequency small-size stomata correlate with whole plant efficiency, while low-frequency large-size stomata show intrinsic efficiency and responsiveness to vapour pressure deficit. Leaf surface and the patterning of the epidermal layer play a vital role in determining plant growth. While the surface helps in determining radiation interception, epidermal pattern of stomatal factors strongly regulate gas exchange and water use efficiency (WUE). This study focuses on identifying distinct stomatal traits among rice genotypes to comprehend their influence on WUE. Stomatal frequency ranged from 353 to 687 per mm2 and the size varied between 128.31 and 339.01 μm2 among 150 rice germplasm with significant variability in abaxial and adaxial surfaces. The cumulative water transpired and WUE determined at the outdoor phenomics platform, over the entire crop growth period as well as during specific hours of a 24h-day did not correlate with stomatal frequency nor size. However, genotypes with low-frequency and large-size stomata recorded higher intrinsic water use efficiency (67.04μmol CO2 mol-1 H2O) and showed a quicker response to varying vapour pressure deficit that diurnally ranged between 0.03 and 2.17kPa. The study demonstrated the role of stomatal factors in determining physiological subcomponents of WUE both at single leaf and whole plant levels. Differential expression patterns of stomatal regulatory genes among the contrasting groups explained variations in the epidermal patterning. Increased expression of ERECTA, TMM and YODA genes appear to contribute to decreased stomatal frequency in low stomatal frequency genotypes. These findings underscore the significance of stomatal traits in breeding programs and strongly support the importance of these genes that govern variability in stomatal architecture in future crop improvement programs.

Coping with extremes: Responses of Quercus robur L. and Fagus sylvatica L. to soil drought and elevated vapour pressure deficit

Climate change, particularly droughts and heat waves, significantly impacts global photosynthesis and forest ecosystem sustainability. To understand how trees respond to and recover from hydrological stress, we investigated the combined effects of soil moisture and atmospheric vapour pressure deficit (VPD) on seedlings of the two major European broadleaved tree species Fagus sylvatica (FS) and Quercus robur (QR). The experiment was conducted under natural forest gap conditions, while soil water availability was strictly manipulated. We monitored gas exchange (net photosynthesis, stomatal conductance and transpiration rates), nonstructural carbohydrates (NSC) concentration in roots and stomatal morphometry (size and density) during a drought period and recovery. Our comparative empirical study allowed us to distinguish and quantify the effects of soil drought and VPD on stomatal behavior, going beyond theoretical models. We found that QR conserved water more conservatively than FS by reducing transpiration and regulating stomatal conductance under drought. FS maintained higher stomatal conductance and transpiration at elevated VPD until soil moisture became critically low. QR showed higher intrinsic water use efficiency than FS. Stomata density and size also likely played a role in photosynthetic rate and speed of recovery, especially since QR with its seasonal adjustments in stomatal traits (smaller, more numerous stomata in summer leaves) responded and recovered faster compared to FS. Our focal species showed different responses in NSC content under drought stress and recovery, suggesting possible different evolutionary pathways in coping with stress. QR mobilized soluble sugars, while FS relied on starch mobilization to resist drought. Although our focal species often co-occur in mixed forests, our study showed that they have evolved different physiological, morphological and biochemical strategies to cope with drought stress. This suggests that ongoing climate change may alter their competitive ability and adaptive potential in favor of one of the species studied.

Response strategies of leaf vein traits and stomatal traits of Aegiceras corniculatum to shrimp aquaculture wastewater discharge

Response strategies of leaf vein traits and stomatal traits of Aegiceras corniculatum to shrimp aquaculture wastewater discharge

PHYSIO-ANATOMICAL EVALUATION OF SOME TREE SPECIES FOR AFFORESTATION IN DRY REGION OF KOGI STATE, NIGERIA

The stomatal features of plant species have ability to release water vapour into the air. Hence, correlations between the stomatal features and transpiration rate of five tree species namely Danielli oliveri, Delonix regia, Piliostigma thonningii, Azadirachta indica and Tectona grandis was studied to evaluate their capacity for afforestation. The leaf epidermal layers were isolated using nail polish; they were observed under the light microscope to examine their stomatal features. The transpiratiom rate was evaluated using the cobalt chloride method. The results revealed that Delonix regia and Piliostigma thonningii are amphistomatic; while the remaining three species are hypostomatic. The stomatal complex types observed are anomocytic, brachyparacytic and paracytic, The stomatal density ranged from 14.41 mm-2 to 93.61 mm-2; the stomatal index ranged from 7.15% to 28.23%; while the stomatal size ranged from 11.19 µm2 to 29.36 µm2. The study revealed that stomatal traits such as hypostomatic leaf nature, stomatal complex types (i.e. paracytic, brachyparacytic), low stomatal index, small stomatal size possessed by the plant species may be responsible for their lower rate of transpiration; which in turn might be suitable for their afforestation in dry areas. Therefore, Tectona grandis which released the lowest amount of water (2.49 × 10-6 mol m-2s-1) into the atmosphere might be the most suitable for afforestation, followed by Piliostigma thonningii, Daniellia oliveri, Delonix regia and lastly Azadirachta indica (2.97 × 10-6 mol. m-2s-1). Conclusively, the stomatal features showed positive correlations with transpiration rates; thereby enhancing the potentials of the studied species for afforestation in dry region.

Phytofabricated gold nanoparticles as modulators of salt stress responses in spinach: implications for redox homeostasis, biochemical and physiological adaptation.

The utilization of plant material for synthesizing nanoparticles effectively triggers physiological and biochemical responses in plants to combat abiotic stresses. Salt stress, particularly caused by NaCl, significantly affects plant morphology and physiology, leading to reduced crop yields. Understanding the mechanisms of salt tolerance is crucial for maintaining crop productivity. In this study, we examined the effects of 150 μM spinach-assisted gold nanoparticles (S-AuNPs) on various parameters related to seed germination, growth attributes, photosynthetic pigments, stomatal traits, ion concentrations, stress markers, antioxidants, metabolites, and nutritional contents of spinach plants irrigated with 50 mM NaCl. Results showed that S-AuNPs enhanced chlorophyll levels, leading to improved light absorption, increased photosynthates production, higher sugar content, and stimulated plant growth under NaCl stress. Stomatal traits were improved, and partially closed stomata were reopened with S-AuNPs treatment, possibly due to K+/Na+ modulation, resulting in enhanced relative water content and stomatal conductance. ABA content decreased under S-AuNPs application, possibly due to K+ ion accumulation. S-AuNPs supplementation increased proline and flavonoid contents while reducing ROS accumulation and lipid peroxidation via activation of both non-enzymatic and enzymatic antioxidants. S-AuNPs also regulated the ionic ratio of K+/Na+, leading to decreased Na+ accumulation and increased levels of essential ions in spinach plants under NaCl irrigation. Overall, these findings suggest that S-AuNPs significantly contributeto salt stress endurance in spinach plants by modulating various physiological attributes.

Exploring natural genetic diversity in a bread wheat multi-founder population: Dual imaging of photosynthesis and stomatal kinetics.

A better understanding of crop phenotype under dynamic environmental conditions will help inform the development of new cultivars with superior adaptation to constantly changing field conditions. Recent research has shown that optimising photosynthetic and stomatal conductance traits holds promise for improved crop performance. However, standard phenotyping tools such as gas-exchange systems are limited by their throughput. In this work, a novel approach based on a bespoke gas-exchange chamber allowing combined measurement of the quantum yield of photosystem II (PSII) with an estimation of stomatal conductance via thermal imaging, was used to phenotype a range of bread wheat (Triticum aestivum L.) genotypes, that were a sub-set of a multi-founder experimental population. Datasets were further supplemented by measurement of photosynthetic capacity and stomatal density. First, we showed that measurement of stomatal traits using our dual imaging system compared to standard IRGA methods showed good agreement between the two methods (R2=0.86) for the rapidity of stomatal opening (Ki), with the dual-imager method resulting in less intra-genotype variation. Using the dual-imaging methods, and traditional approaches we found broad and significant variation in key traits, including photosynthetic CO2 uptake at saturating light and ambient CO2 concentration (Asat), photosynthetic CO2 uptake at saturating light and elevated CO2 concentration (Amax), the maximum velocity of Rubisco for carboxylation (Vcmax), time for stomatal opening (Ki), and leaf evaporative cooling. Anatomical analysis revealed significant variation in flag leaf adaxial stomatal density. Associations between traits highlighted significant relationships between leaf evaporative cooling, leaf stomatal conductance under low (gsmin) and high (gsmax) light intensity, and the operating efficiency of PSII (Fq'/Fm'), highlighting the importance of stomatal conductance and stomatal rapidity in maintaining optimal leaf temperature for photosynthesis in wheat. Additionally, gsmin and gsmax were positively associated, indicating that potential combination of preferable traits (i.e. inherently high gsmax, low Ki and maintained leaf evaporative cooling) are present in wheat. This work highlights for the first time the effectiveness of thermal imaging in screening dynamic stomatal conductance in a large panel of wheat genotypes. The wide phenotypic variation observed suggested the presence of exploitable genetic variability in bread wheat for dynamic stomatal conductance traits and photosynthetic capacity for targeted optimisation within future breeding programs.

Inter‐ and intraspecific stomatal morphological traits vary in response to topographic habitat changes

AbstractQuestionsStomata can reflect the plant's adaptation to environmental changes. However, the variation patterns of stomatal traits across different habitats and their relationships with environmental drivers are still poorly understood. Here, we assessed the extent of interspecific and intraspecific variation in stomatal traits in two typical riparian forests, and investigated how stomatal traits adapt to habitat change as well as trait–environment relationships.LocationThe Irtysh River Basin in China.MethodsWe measured the stomatal density (SD), stomatal size (SS), and stomatal relative area (SRA) of dominant plant species from two riparian forests: (1) a riparian forest in the valley and (2) a riparian forest in the pediment plain. We analyzed the stomatal trait variation patterns of dominant plant species in different habitats. We then quantified the magnitude of intraspecific and interspecific stomatal trait variability and evaluated the relationships between stomatal traits and environmental factors.ResultsWe found that SD and SRA were significantly higher in pediment plain forests than in valley forests, whereas SS was not significantly different between these habitats. The SD and SRA of trees in pediment plain forests were significantly higher than those in valley forests, whereas there were no significant differences in understory plants between habitats. The interspecific variation in each stomatal trait was significantly higher than the intraspecific variation. Stomatal traits were more related to the soil environment than to climatic factors.ConclusionsResponse strategies of stomatal traits to habitat changes differed between species and functional groups, and trait–environment relationships depended on the habitat type of the riparian forest. Our analysis of stomata trait variation implies adaptive strategies in species of natural riparian forest. Insights into trait–environment relationships could be used to predict carbon and water cycling, and vegetation changes in riparian forests of arid regions, especially in the context of climate change.

Exogenous melatonin enhances heat stress tolerance in sweetpotato by modulating antioxidant defense system, osmotic homeostasis and stomatal traits

Heat stress hinders the growth and productivity of sweetpotato plants, predominantly through oxidative damage to cellular membranes. Therefore, the development of efficient approaches for mitigating heat-related impairments is essential for the long-term production of sweetpotatoes. Melatonin has been recognised for its capacity to assist plants in dealing with abiotic stress conditions. This research aimed to investigate how different doses of exogenous melatonin influence heat damage in sweetpotato plants. Heat stress drastically affected shoot and root fresh weight by 31.8 and 44.5%, respectively. This reduction resulted in oxidative stress characterised by increased formation of hydrogen peroxide (H2O2) by 804.4%, superoxide ion (O2•−) by 211.5% and malondialdehyde (MDA) by 234.2%. Heat stress also reduced chlorophyll concentration, photosystem II efficiency (Fv/Fm) by 15.3% and gaseous exchange. However, pre-treatment with 100 μmol L−1 melatonin increased growth and reduced oxidative damage to sweetpotato plants under heat stress. In particular, melatonin decreased H2O2, O2•− and MDA by 64.8%, 42.7% and 38.2%, respectively. Melatonin also mitigated the decline in chlorophyll levels and improved stomatal traits, gaseous exchange and Fv/Fm (13%). Results suggested that the favorable outcomes of melatonin treatment can be associated with elevated antioxidant enzyme activity and an increase in non-enzymatic antioxidants and osmo-protectants. Overall, these findings indicate that exogenous melatonin can improve heat stress tolerance in sweetpotatoes. This study will assist researchers in further investigating how melatonin makes sweetpotatoes more resistant to heat stress.