Stomatal Pore Area Research Articles

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.

Partial root-zone drying irrigation improves intrinsic water-use efficiency and maintains high photosynthesis by uncoupling stomatal and mesophyll conductance in cotton leaves.

Partial root-zone drying irrigation (PRD) can improve water-use efficiency (WUE) without reductions in photosynthesis; however, the mechanism by which this is attained is unclear. To amend that, PRD conditions were simulated by polyethylene glycol 6000 in a root-splitting system and the effects of PRD on cotton growth were studied. Results showed that PRD decreased stomatal conductance (gs) but increased mesophyll conductance (gm). Due to the contrasting effects on gs and gm, net photosynthetic rate (AN) remained unaffected, while the enhanced gm/gs ratio facilitated a larger intrinsic WUE. Further analyses indicated that PRD-induced reduction of gs was related to decreased stomatal size and stomatal pore area in adaxial and abaxial surface which was ascribed to lower pore length and width. PRD-induced variation of gm was ascribed to the reduced liquid-phase resistance, due to increases in chloroplast area facing to intercellular airspaces and the ratio of chloroplast surface area to total mesophyll cell area exposed to intercellular airspaces, as well as to decreases in the distance between cell wall and chloroplast, and between adjacent chloroplasts. The above results demonstrate that PRD, through alterations to stomatal and mesophyll structures, decoupled gs and gm responses, which ultimately increased intrinsic WUE and maintained AN.

Drought response strategies are coupled with leaf habit in 35 evergreen and deciduous oak (Quercus) species across a climatic gradient in the Americas.

Distinct survival strategies can result from trade-offs in plant function under contrasting environments. Investment in drought resistance mechanisms can enhance survivorship but result in conservative growth. We tested the hypothesis that the widespread oaks (Quercus spp.) of the Americas exhibit an interspecific trade-off between drought resistance and growth capacity. Using experimental water treatments, we isolated adaptive trait associations among species in relation to their broad climates of origin and tested for correlated evolution between plant functional responses to water availability and habitat. Across all lineages, oaks displayed plastic drought responses - typically acclimating through osmolyte accumulation in leaves and/or employing conservative growth. Oaks from xeric climates had higher osmolytes and reduced stomatal pore area index, which allows for moderated gas exchange and limits tissue loss. Patterns suggest drought resistance strategies are convergent and under strong adaptive pressure. Leaf habit, however, mediates the growth and drought resistance strategies of oaks. Deciduous species, and evergreen species from xeric climates, have increased drought tolerance through osmoregulation, which allows for continuous, conservative growth. Evergreen mesic species show limited drought resistance but could enhance growth under well-watered conditions. Consequently, evergreen species from mesic environments are especially vulnerable to chronic drought and climate change.

Boosting biopesticide potential of Trichoderma harzianum for controlling the downy mildew and improving the growth and the productivity of King Ruby seedless grape

BackgroundPlasmopara viticola, the causal agent of grape downy mildew, is one of the most serious grape diseases. Application of Trichoderma harzianum grown on different potato dextrose supplemented broth media using some chemical inducers (TSDCIS), i.e., thiamine (T2), a mixture of micronutrients (T3) and potassium tartrate (T4), compared to Trichoderma without amendment (T1), was conducted under field conditions to evaluate the potential of T. harzianum against grape downy mildew, improving the growth, as well as the yield quality, and quantity.ResultsFoliar application of all TSDCIS significantly reduced the disease severity and increased the phenolic content, peroxidase, polyphenol oxidase enzyme activity, growth parameters, and yield parameters. Trichoderma growing on medium supplemented with potassium tartrate (T4) achieved the most significant reduction in the disease severity (78.9, 81.8%) than the control treatment in two growing seasons, respectively. In contrast, it decreased H2O2 content, lipid peroxidation, and cellular electrolyte leakage. Scanning electron microscopy observations revealed that the number of opened stomata, average stomatal area, and stomatal pore area decreased significantly in response to T4 treatment.ConclusionsIt can be concluded that using Trichoderma growing on medium supplemented with potassium tartrate (T4) to biosafety control the downy mildew disease of grape and improve its growth, yield, and fruit quality is recommended.

Stomatal pore width and area measurements in Zea mays.

Stomatal pores are adjustable microscopic holes on the surface of photosynthetic tissues that help regulate multiple aspects of plant physiology. Stomatal pores facilitate gas exchange necessary for photosynthesis, water transport, and temperature regulation. Pore size is influenced by many intertwined environmental, molecular, cellular, and physiological cues. Accurate and precise measurements of pore size is important for understanding the mechanisms that adjust pores and plant physiology. Here we investigate whether conventional pore measurements of width are appropriate for the economically important crop plant Zea mays . Our studies demonstrate that pore area is a more sensitive measurement than width in this plant.

Evaluating an Ultrasonic Magnetostrictive Transducer with Conical Nickel Core: Performance and Application

In recent years, ultrasonic machining has been developing rapidly, and it is used in areas such as abrasive machining, cleaning, and welding. In this research, a magnetostrictive device with pure nickel conical core with cone angle of 30° was constructed. The observed sound pressure level was used as a measure of transducer performance. Also, the ability of the device to reduce the residual pesticide on cucumber surface due to cavitation was evaluated. The results showed that the sound pressure from simulation by JMAG-Designer software is almost the same as the sound pressure produced in the constructed transducer. To assess the performance of the device in removing residual pesticide from cucumber surface and evaluate the changes in peel texture of cucumber, the gas chromatography (GC) and scanning electron microscope (SEM) methods were used, respectively. The GC results showed that with 20 min treatment, the removal of pesticide based on height and chromatogram area was 75% and 83%, respectively. The SEM results showed that by increasing the treatment time, the stomatal pore area reduced from 144.74 \({\mathrm{\mu m}}^{2}\) (reference) to 30.56 \({\mathrm{\mu m}}^{2}\) (20 min treatment). These results are promising; hence, further research is suggested towards enhancing the device for commercial use in biomaterials processing operations such as cleaning and removing pesticides from fruits and vegetables.Graphical Abstract

Maize stomatal responses against the climate change.

Drought and heat, in the context of climate change, are expected to increase in many agricultural areas across the globe. Among current abiotic stresses, they are the most limiting factors that influence crop growth and productivity. Maize is one of most widely produced crops of the world, being the first in grain production with a yield that exceeded 1.1 billion tons in 2021. Despite its wide distribution in semi-arid regions, it is highly vulnerable to climate change, which triggers important losses in its productivity. This article explores how maize yield may persevere through climate change by focusing on the stomatal regulation of gas exchange. The emerging picture unravels that maize copes with drought stress by reducing stomatal size and stomatal pore area, and increasing stomatal density, which, in turn, reduces transpiration and photosynthetic rate. When drought and heat co-occur, heat enhances stomatal response to drought stress. To avoid plant heat damage, the decline in stomatal aperture could trigger the expansion of the distance of action, from the longitudinal leaf veins, of ZmSHR1, which might act to positively regulate ZmSPCHs/ZmICE1 heterodimers, increasing the stomatal density. Only when drought is not very severe, elevated CO2 levels reduce yield losses. The knowledge of the upcoming climate changes together with the prediction of the developmental and physiological stomatal responses will allow not only to anticipate maize yield in the next years, but also to contribute to the correct decision-making in the management of this important crop.

Flexible drought deciduousness in a neotropical understory herb.

PremiseAdaptive divergence across environmental gradients is a key driver of speciation. Precipitation seasonality gradients are common in the tropics, yet drought adaptation is nearly unexplored in neotropical understory herbs. Here, we examined two recently diverged neotropical spiral gingers, one adapted to seasonal drought and one reliant on perennial water, to uncover the basis of drought adaptation.MethodsWe combined ecophysiological trait measurements in the field and greenhouse with experimental and observational assessments of real‐time drought response to determine how Costus villosissimus (Costaceae) differs from C. allenii to achieve drought adaptation.ResultsWe found that drought‐adapted C. villosissimus has several characteristics indicating flexible dehydration avoidance via semi‐drought‐deciduousness and a fast economic strategy. Although the two species do not differ in water‐use efficiency, C. villosissimus has a more rapid growth rate, lower leaf mass per area, lower stem density, higher leaf nitrogen, and a strong trend of greater light‐saturated photosynthetic rates. These fast economic strategy traits align with both field‐based observations and experimental dry‐down results. During drought, C. villosissimus displays facultative drought‐deciduousness, losing lower leaves during the dry season and rapidly growing new leaves in the wet season.ConclusionsWe revealed a drought adaptation strategy that has not, to our knowledge, previously been documented in tropical herbs. This divergent drought adaptation evolved recently and is an important component of reproductive isolation between C. villosissimus and C. allenii, indicating that adaptive shifts to survive seasonal drought may be an underappreciated axis of neotropical understory plant diversification.

Magnesium carbonate elicits defense-related genes in King Ruby grapevines against downy mildew and improves its growth, yield, and berries quality

Downy mildew, caused by Plasmopara viticola (Berk. and M. A. Curtis) Berl. and De Toni, is a serious disease of grapevines in general and King Ruby seedless cultivar in particular, affecting their growth and yield. Magnesium carbonate (MgCO3) is an antitranspirant, which induces stomatal closing and enhances plant growth and physiology. In this study, effect of foliar application of MgCO3 at 1 and 3% on plant resistance, growth, yield and physiology of grapevines (cv. King Ruby seedless) infected with downy mildew was investigated under field conditions. The obtained results showed that foliar application of MgCO3 at 3% led to upregulation of the transcription factor JERF3 (9.6-fold), and the defense-related genes GLU (6.3-fold), POD (8.7-fold), PR1 (9.6-fold), and CHI II (8.6-fold). In addition, this treatment led to a reduction in the disease severity (78%), and an increment in the yield per grapevine (20%). Furthermore, biochemical properties of berries, total contents of the photosynthetic pigments, phenolic compounds, and activities of the antioxidant enzymes peroxidase and polyphenol oxidase also enhanced. In contrast, lipid peroxidation, and H2O2 content in grapevines leaves reduced in response to MgCO3 spraying. Light microscope observations revealed that average number of closed stomata increased and the average stomatal pore area decreased in grapevines leaves as a result to MgCO3 spraying. Based on these results, we can conclude that spraying with MgCO3 at 3% has effective roles in inducing the plant resistance against downy mildew, and improving the growth and yield of grapevines.

Morpho-Anatomical Traits and Soluble Sugar Concentration Largely Explain the Responses of Three Deciduous Tree Species to Progressive Water Stress.

A better understanding of plant drought responses is essential to improve plant water use efficiency, productivity, and resilience to ever-changing climatic conditions. Here, we investigated the growth, morpho-anatomical, physiological, and biochemical responses of Quercus acutissima Carruth., Quercus serrata Murray, and Betula schmidtii Regel to progressive water-stress. Seedlings were subjected to well-watered (WW) and water-stressed (WS) conditions while regularly monitoring the soil volumetric water content, stem diameter (SD), height, biomass, stomatal conductance (gs), intercellular CO2 concentration (Ci), and leaf relative water content (RWC). We also investigated the variation in stomatal pore (SP) area, specific leaf area (SLA), root xylem vessel diameter (VD), and total soluble sugar (TSS) concentration between treatments. After 2 months, WS significantly suppressed SD growth of Q. acutissima and B. schmidtii but had no impact on Q. serrata. Total biomass significantly declined at WS-treated seedlings in all species. WS resulted in a smaller SLA than WW in all species. The SP of WS-treated seedlings of Q. acutissima and B. schmidtii significantly decreased, whereas it increased significantly with time in Q. serrata. Larger vessels (i.e., >100 to ≤ 130) were more frequent at WS for Q. acutissima and B. schmidtii, whereas smaller vessels (i.e., >40 to ≤ 90) were more frequent at WS than at WW for Q. serrata after 8 weeks. Tylosis was more frequent at WS than WW for Q. serrata and B. schmidtii at eighth week. WS seedlings showed lower gs, Ci, and RWC compared with WW-treated ones in Q. acutissima and B. schmidtii. TSS concentration was also higher at WS-treated seedlings in two Quercus species. Overall, principal component analysis (PCA) showed that SLA and SP are associated with WS seedlings of Q. serrata and B. schmidtii and the tylosis frequency, TSS, and VD are associated with WS seedlings of Q. acutissima. Therefore, water-stressed plants from all species responded positively to water stress with increasing experimental duration and stress intensity, and that is largely explained by morpho-anatomical traits and soluble sugar concentration. The present study should enhance our understanding of drought-induced tree growth and short-term tree-seedling responses to drought.

Silica Nanoparticles as a Probable Anti-Oomycete Compound Against Downy Mildew, and Yield and Quality Enhancer in Grapevines: Field Evaluation, Molecular, Physiological, Ultrastructural, and Toxicity Investigations.

Downy mildew is the most destructive disease of grapevines in the regions of relatively warm and humid climate causing up to 50% yield losses. Application of silicon- (Si-) based products have been extensively studied against various oomycete, fungal, bacterial, and viral plant diseases, but studies on Si application in their nanosize are limited. In this study, the field application of silica nanoparticles (SiNPs) on Thompson Seedless grapevines (H4 strain) infected with downy mildew was evaluated. In addition, molecular, physiological, ultrastructural, and toxicity investigations were also conducted. The obtained results revealed that spraying of grapevines with SiNPs at 150 ppm significantly overexpressed the transcription factor jasmonate and ethylene-responsive factor 3 recording 8.7-fold, and the defense-related genes β-1,3-glucanase (11-fold), peroxidase (10.7-fold) pathogenesis-related-protein 1 (10.6-fold), and chitinase (6.5-fold). Moreover, a reduction up to 81.5% in the disease severity was achieved in response to this treatment. Shoot length and yield per grapevine were considerably enhanced recording up to 26.3 and 23.7% increase, respectively. The berries quality was also improved. Furthermore, this treatment led to an enhancement in the photosynthetic pigments, induction of phenolic and ascorbic acid contents, an increase in the activity of peroxidase and polyphenol oxidase enzymes, and a reduction in the cellular electrolyte leakage, lipid peroxidation, and H2O2 content. Scanning electron microscopy observations showed an increase up to 86.6% in the number of closed stomata and a reduction up to 55% in the average stomatal pore area in response to this treatment. Observations of the transmission electron microscopy showed ultrastructural alterations in the cells of a grapevine leaf due to the infection with downy mildew, including plasmolysis and disruption of the cellular components, abnormal chloroplasts, and thickening of the cell wall and cell membrane. These abnormal alterations were reduced in response to SiNPs spray. In contrast, this study also showed that this treatment had considerable cytotoxic and genotoxic effects at this direct dose/concentration. So, additional investigations to determine the SiNPs residue in the produced edible plant parts are urgently needed. In addition, the pre-harvest interval, toxicity index, and risk assessment should be evaluated before any recommendation for use.

Leaf 13C and 15N composition shedding light on easing drought stress through partial K substitution by Na in eucalyptus species

This work aimed to investigate the partial K-replacement by Na supply to alleviate drought-induced stress in Eucalyptus species. Plant growth, leaf gas exchange parameters, water relations, oxidative stress (H2O2 and MDA content), chlorophyll concentration, carbon (C) and nitrogen (N) isotopic leaf composition (δ13C and δ15N) were analyzed. Drought tolerant E. urophylla and E. camaldulensis showed positive responses to the partial K substitution by Na, with similar dry mass yields, stomatal density and total stomatal pore area relative to the well K-supplied plants under both water conditions, suggesting that 50% of the K requirements is pressing for physiological functions that is poorly substituted by Na. Furthermore, E. urophylla and E. camaldulensis up-regulated leaf gas exchanges, leading to enhanced long-term water use efficiency (WUEL). Moreover, the partial K substitution by Na had no effects on plants H2O2, MDA, δ13C and δ15N, confirming that Na, to a certain extent, can effectively replace K in plants metabolism. Otherwise, the drought-sensitive E. saligna species was negatively affected by partial K replacement by Na, decreasing plants dry mass, even with up-regulated leaf gas exchange parameters. The exclusive Na-supplied plants showed K-deficient symptoms and lower growth, WUEL, and δ13C, besides higher Na accumulation, δ15N, H2O2 and MDA content.

Incorporating cultivar-specific stomatal traits into stomatal conductance models improves the estimation of evapotranspiration enhancing greenhouse climate management

The effect of considering cultivar differences in stomatal conductance (g s ) on relative air humidity (RH)-related energy demand was addressed. We conducted six experiments in order to study the variation in evapotranspiration (ET c ) of six pot rose cultivars, investigate the underlying processes and parameterise a g s -based ET c model. Several levels of crop ET c were realised by adjusting the growth environment. The commonly applied Ball–Woodrow–Berry g s -sub-model (BWB-model) in ET c models was validated under greenhouse conditions, and showed a close agreement between simulated and measured ET c . The validated model was incorporated into a greenhouse simulator. A scenario simulation study showed that selecting low-g s cultivars reduces energy demand (≤5.75%), depending on the RH set point. However, the BWB-model showed poor prediction quality at RH lower than 60% and a good fit at higher RH. Therefore, an attempt was made to improve model prediction: the in situ -obtained data were employed to adapt and extend either the BWB-model, or the Liu-extension with substrate water potential (Ψ; BWB-Liu-model). Both models were extended with stomatal density (D s ) or pore area. Although the modified BWB-Liu-model (considering D s ) allowed higher accuracy (R 2 = 0.59), as compared to the basic version (R 2 = 0.31), the typical lack of Ψ prediction in greenhouse models may be problematic for implementation into real-time climate control. The current study lays the basis for the development of cultivar specific cultivation strategies as well as improving the g s sub-model for dynamic climate conditions under low RH using model-based control systems. • Relative air humidity control is a key but costly aspect of greenhouse climate. • Accurate estimation of evapotranspiration (ET c ) is essential for relative air humidity regulation. • Six pot rose cultivars and several cultivation regimes were used. • ET c estimation was improved using cultivar-specific stomatal conductance parameters. • The use of low-stomatal conductance cultivars secures energy saving (2.5–5.75%).

Ozone Response of Leaf Physiological and Stomatal Characteristics in Brassica juncea L. at Supraoptimal Temperatures

Plants are affected by the features of their surrounding environment, such as climate change and air pollution caused by anthropogenic activities. In particular, agricultural production is highly sensitive to environmental characteristics. Since no environmental factor is independent, the interactive effects of these factors on plants are essential for agricultural production. In this context, the interactive effects of ozone (O3) and supraoptimal temperatures remain unclear. Here, we investigated the physiological and stomatal characteristics of leaf mustard (Brassica juncea L.) in the presence of charcoal-filtered (target concentration, 10 ppb) and elevated (target concentration, 120 ppb) O3 concentrations and/or optimal (22/20 °C day/night) and supraoptimal temperatures (27/25 °C). Regarding physiological characteristics, the maximum rate of electron transport and triose phosphate use significantly decreased in the presence of elevated O3 at a supraoptimal temperature (OT conditions) compared with those in the presence of elevated O3 at an optimal temperature (O conditions). Total chlorophyll content was also significantly affected by supraoptimal temperature and elevated O3. The chlorophyll a/b ratio significantly reduced under OT conditions compared to C condition at 7 days after the beginning of exposure (DAE). Regarding stomatal characteristics, there was no significant difference in stomatal pore area between O and OT conditions, but stomatal density under OT conditions was significantly increased compared with that under O conditions. At 14 DAE, the levels of superoxide (O2-), which is a reactive oxygen species, were significantly increased under OT conditions compared with those under O conditions. Furthermore, leaf weight was significantly reduced under OT conditions compared with that under O conditions. Collectively, these results indicate that temperature is a key driver of the O3 response of B. juncea via changes in leaf physiological and stomatal characteristics.

Is the Seasonal Variation in Frost Resistance and Plant Performance in Four Oak Species Affected by Changing Temperatures?

Research Highlights: We found seasonal variation in frost resistance (FR) and plant performance which were affected by growth temperature. This helps to better understand ecophysiological processes in the light of climate change. Background and Objectives: FR and photosynthesis are important plant characteristics that vary with the season. The aim of this study was to find out whether there is a seasonal variation in FR, photosynthetic CO2 assimilation rates and leaf functional traits associated with performance such as specific leaf area (SLA), leaf dry matter content (LDMC), chlorophyll content, stomatal characteristics and leaf thickness in two evergreen and two deciduous species, and whether this is influenced by different temperature treatments. Additionally, the trade-off between FR and photosynthetic performance, and the influence of leaf functional traits was analyzed. By understanding these processes better, predicting species behavior concerning plant performance and its changes under varying climate regimes can be improved. Materials and Methods: 40 individuals of four oak species were measured weekly over the course of ten months with one half of the trees exposed to frost in winter and the other half protected in the green house. Two of these species were evergreen (Quercus ilex L., Quercus rhysophylla Weath.), and two were deciduous (Quercus palustris L., Quercus rubra L.). We measured FR, the maximum assimilation rate at light saturation under ambient CO2 concentrations (Amax), chlorophyll fluorescence and the leaf functional traits SLA, LDMC, stomatal pore area index (SPI), chlorophyll content (Chl) and leaf thickness. Results: All parameters showed a significant species-specific seasonal variation. There was a difference in all traits investigated between evergreen and deciduous species and between the two temperature treatments. Individuals that were protected from frost in winter showed higher photosynthesis values as well as SLA and Chl, whereas individuals exposed to frost had overall higher FR, LDMC, SPI and leaf thickness. A trade-off between FR and SLA, rather than FR and photosynthetic performance was found.

The timing of leaf senescence relates to flowering phenology and functional traits in 17 herbaceous species along elevational gradients

Abstract Leaf senescence is a major event in a plant's life history as autumn marks the end of the growing season. The optimal timing of leaf senescence is crucial to both minimise risks of low temperature events and to maximise carbon gain during the growing season. As abiotic conditions are currently changing at unprecedented rates, it is important to study how leaf senescence of different species is responding to these changes in order to forecast future growing season length and carbon sequestration potentials. In contrast to flowering phenology, data on autumn events are scarce and even more so for herbaceous than for woody plants, thus more information on this phenological stage is urgently needed. We studied leaf senescence in 632 populations from 17 herbaceous species located along elevational gradients. We focussed on the beginning (5% of the population senesce, LS5) and peak (50% senesce, LS50) of leaf senescence. To see whether we can predict species‐specific changes, we studied the link between LS5 and LS50 and flowering phenology as well as leaf functional traits related to plant performance. We looked at first and last flowering day and flowering duration as well as the traits specific leaf area (SLA), leaf dry matter content (LDMC), area‐based leaf nitrogen and carbon content, carbon isotope discrimination (Δ13C) and the stomatal pore area index. We found species‐specific slopes of the beginning of leaf senescence along the elevational gradient. The peak of leaf senescence was uniformly delayed with increasing elevation across all species. Flowering phenology as well as leaf functional traits had a close relationship with leaf senescence and thus can be used to forecast species‐specific responses to changes in abiotic conditions. High SLA and high leaf nitrogen were related to earlier senescence while high LDMC, high Δ13C and high stomatal pore area index were related to later senescence. Synthesis. The link between senescence, flowering phenology and plant functional traits will help to fine‐tune predictions of future growing season length and ecosystem function. To date, most analyses are based on spring phenology and traits, for which data are more abundant than data on autumn senescence.

Accelerating Automated Stomata Analysis Through Simplified Sample Collection and Imaging Techniques.

Digital image processing is commonly used in plant health and growth analysis, aiming to improve research efficiency and repeatability. One focus is analysing the morphology of stomata, with the aim to better understand the regulation of gas exchange, its link to photosynthesis and water use and how they are influenced by climatic conditions. Despite the key role played by these cells, their microscopic analysis is largely manual, requiring intricate sample collection, laborious microscope application and the manual operation of a graphical user interface to identify and measure stomata. This research proposes a simple, end-to-end solution which enables automatic analysis of stomata by introducing key changes to imaging techniques, stomata detection as well as stomatal pore area calculation. An optimal procedure was developed for sample collection and imaging by investigating the suitability of using an automatic microscope slide scanner to image nail polish imprints. The use of the slide scanner allows the rapid collection of high-quality images from entire samples with minimal manual effort. A convolutional neural network was used to automatically detect stomata in the input image, achieving average precision, recall and F-score values of 0.79, 0.85, and 0.82 across four plant species. A novel binary segmentation and stomatal cross section analysis method is developed to estimate the pore boundary and calculate the associated area. The pore estimation algorithm correctly identifies stomata pores 73.72% of the time. Ultimately, this research presents a fast and simplified method of stomatal assay generation requiring minimal human intervention, enhancing the speed of acquiring plant health information.

Heterogeneity of Stomatal Pore Area Is Suppressed by Ambient Aerosol in the Homobaric Species, Vicia faba.

Stomatal pore area is heterogeneous across leaf surfaces. This has been considered as “patchy stomatal conductance,” and may have substantial implications for photosynthetic efficiency. Aerosols have always been important elements of plant environments, but their effects on stomatal control of plant water relations, and stomatal heterogeneity specifically, have not been considered. Here we evaluate the spatial coordination of pore area in the glabrous and homobaric leaves of Vicia faba grown under two aerosol treatments and measured at four levels of VPD. We construct a large dataset (n > 88,000 discrete comparisons) of paired pore areas and distances between the pores. Plants were grown in ambient urban air and in filtered air (FA) to determine the effect of ambient aerosol on stomatal properties. Pore area exhibited spatial organization, as well as considerable variability among closely co-located pores. The difference between pore areas was positively correlated with the distance between the pores, in both aerosol treatments and at all VPDs. However, aerosol deposition reduced both the magnitude of variability between pores and the rate at which this variability increased with pore separation distance. These data support previous conclusions that deposition of hygroscopic aerosol may create a thin aqueous film across the leaf surface that connects neighboring stomata to each other and to the leaf interior. Aerosol impacts on stomatal heterogeneity and gas exchange are not adequately considered in current assessments of stomatal control.

Special issue in honour of Prof. Reto J. Strasser - Seasonal variation and trade-off between frost resistance and photosynthetic performance in woody species

We tested the seasonal variability of frost resistance and photosynthesis characteristics, namely net photosynthetic rate at saturating irradiance and ambient atmospheric CO2 concentrations, maximum carboxylation rate, and measurements based on the JIP-test, namely maximum quantum yield of PSII and the performance index. Additionally, we measured leaf functional traits, namely specific leaf area, leaf dry matter content, stomatal pore area index, and chlorophyll content. We assessed all traits weekly on two evergreen species Berberis vulgaris and Quercus × hispanica as well as the two summer green species Betula pendula and Quercus pubescens. We found that there was a high seasonal variability in all traits except stomatal pore area index. In a multivariate analysis, we could show trade-offs between growth rates and leaf thickness on the first axis, as well as resistance and performance on the second axis. The findings help understand ecophysiological processes and fine-tune biosphere models.

Plant intraspecific functional trait variation is related to within-habitat heterogeneity and genetic diversity in Trifolium montanum L.

Intraspecific trait variation (ITV), based on available genetic diversity, is one of the major means plant populations can respond to environmental variability. The study of functional trait variation and diversity has become popular in ecological research, for example, as a proxy for plant performance influencing fitness. Up to now, it is unclear which aspects of intraspecific functional trait variation (iFDCV) can be attributed to the environment or genetics under natural conditions. Here, we examined 260 individuals from 13 locations of the rare (semi‐)dry calcareous grassland species Trifolium montanum L. in terms of iFDCV, within‐habitat heterogeneity, and genetic diversity. The iFDCV was assessed by measuring functional traits (releasing height, biomass, leaf area, specific leaf area, leaf dry matter content, Fv/Fm, performance index, stomatal pore surface, and stomatal pore area index). Abiotic within‐habitat heterogeneity was derived from altitude, slope exposure, slope, leaf area index, soil depth, and further soil factors. Based on microsatellites, we calculated expected heterozygosity (He) because it best‐explained, among other indices, iFDCV. We performed multiple linear regression models quantifying relationships among iFDCV, abiotic within‐habitat heterogeneity and genetic diversity, and also between separate functional traits and abiotic within‐habitat heterogeneity or genetic diversity. We found that abiotic within‐habitat heterogeneity influenced iFDCV twice as strong compared to genetic diversity. Both aspects together explained 77% of variation in iFDCV (Radj2 = .77, F 2, 10 = 21.66, p < .001). The majority of functional traits (releasing height, biomass, specific leaf area, leaf dry matter content, Fv/Fm, and performance index) were related to abiotic habitat conditions indicating responses to environmental heterogeneity. In contrast, only morphology‐related functional traits (releasing height, biomass, and leaf area) were related to genetics. Our results suggest that both within‐habitat heterogeneity and genetic diversity affect iFDCV and are thus crucial to consider when aiming to understand or predict changes of plant species performance under changing environmental conditions.