Taxonomic relevance of leaf surface micromorphology in Korean Clematis L. (Ranunculaceae).

Clematis, one of the largest genera of the family Ranunculaceae, has a wide array of morphological variation and is considered the most difficult group of taxa in terms of infrageneric discrimination. This study assessed the leaf micromorphological features of 19 Clematis taxa (16 species and three varieties) found in Korea. The leaf surface features were studied under scanning electron microscopy, and the stomatal counting and measurement were carried out under light microscopy. Clematis are hypostomatic, meaning the stomata are only found on the abaxial surface of the leaf. Observed taxa showed near uniformity in the epidermal cell type, structure, and morphology on both surfaces of the leaf. Differences were observed in the presence and absence and/or abundance of trichomes on both the adaxial and abaxial surfaces, the epidermal cell boundary, and the periclinal and anticlinal wall of the cells. Differences were also observed in the number of the epidermal cells connected with the stomata on the abaxial surface, with small differences noted in epidermal cell shapes. The ANOVA showed a significant variation in the stomata density in the studied taxa (P < 0.0001). The cluster analysis based on 13 leaf micromorphological features generated four major clusters. These results indicated similarities in certain key leaf micromorphological features among taxa from the Tubulosae, Clematis, and Virona sections. In the genus Clematis, as with other morphological characteristics, using leaf micromorphological characters alone, which possess limited taxonomic value, proves inadequate for resolving infrageneric relationships. However, incorporating certain features with other morphological characteristics offers a possible alternative means of determining the infrageneric relationships within the genus.

Growing at the arid edge: Anatomical variations in leaves are more extensive than in stems of five Mediterranean species across contrasting moisture regimes.

PREMISE: Increasing aridity in the Mediterranean region affects ecosystems and plant life. Various anatomical changes in plants help them cope with dry conditions. This study focused on anatomical differences in leaves and xylem of five co-occurring Mediterranean plant species namely Quercus calliprinos, Pistacia palaestina, Pistacia lentiscus, Rhamnus lycioides, and Phillyrea latifolia in wet and dry sites. METHODS: Stomatal density, stomatal length, leaf mass area, lamina composition, percentage of intercellular air spaces, and mesophyll cell area in leaves of plants in wet and dry sites were analyzed. Xylem anatomy was assessed through vessel length and area in branches. RESULTS: In the dry site, three species had increased stomatal density and decreased stomatal length. Four species had increased palisade mesophyll and reduced air space volume. In contrast, phenotypic changes in the xylem were less pronounced; vessel length was unaffected by site conditions, but vessel diameter decreased in two species. Intercellular air spaces proved to be the most dynamic anatomical feature. Quercus calliprinos had the most extensive anatomical changes; Rhamnus lycioides had only minor changes. All these changes were observed in comparison to the species in the wet site. CONCLUSIONS: This study elucidated variations in anatomical responses in leaves among co-occurring Mediterranean plant species and identified the most dynamic traits. Understanding these adaptations provides valuable insights into the ability of plants to thrive under changing climate conditions.

Rapid morphological change in UK populations of Impatiens glandulifera.

The highly invasive Impatiens glandulifera (Himalayan balsam) is one of the most prolific and widespread invasive plants in the British Isles. Introduced in the early nineteenth century, it has now been reported in almost every vice county across the UK and is a fierce competitor that has adverse effects on the local community structure. Despite the negative impacts that invaders like I. glandulifera have on local communities, there have been very few studies which address the morphological changes that invasive plant populations have undergone since their initial introduction. This is the first study of its kind to investigate the morphological changes that have occurred in I. glandulifera. 315 herbarium specimens dating from 1865 to 2017 were used to measure changes in morphological traits such as leaf size, flower length and stomatal characteristics. We found that since 1865, there has been a significant reduction in overall leaf size, a significant reduction in stomatal density and a significant increase in the overall flower length. These results highlight the importance of monitoring the evolutionary change in prolific alien species over the course of their invasion, providing useful insights into changes in competitive ability which may prove useful in managing dispersal and providing options for potential management.

Morpho-anatomical studies of family lamiaceae species of district Lahore, Punjab: a revision to flora of Pakistan.

BACKGROUND: This study was aimed to determine the taxonomic position and delimitation of fifteen Lamiaceae taxa using leaf epidermal morpho-anatomical features in Lahore. A main objective of the study was also the revision and upgradation of Lamiaceae taxa in the flora of Pakistan, as no details of studied species are found in the flora of Pakistan. METHODS: The examination of significant anatomical parameters, such as epidermal cell shape and size, stomatal types, guard and subsidiary cells shape and size, stomatal cavity size, trichome size and shape, oil droplets, crystals, and secretory cavity characteristics were studied using light microscopic (LM) and scanning electron microscopic (SEM) techniques. Among all the studied Lamiaceae species, these anatomical features varied significantly. Principal component analysis and correlation were done to distinguish the species' similarities. RESULTS: Most species had pentagonal and hexagonal epidermal cells with straight anticlinal wall thickness. On the adaxial surface, paracytic stomata were found in Ocimum basilicum L. and Rosmarinus officinalis L. Diacytic stomata was observed in Ajuga reptans L. and anisocytic stomata in Galeopsis tetrahit L. In the abaxial surface, trichomes were present in five species, i.e., Mentha suaveolens Ehrh. A. reptans, Thymus vulgaris L., M. haplocalyx, and Salvia splendens Ewat. In S. splendens, peltate and glandular trichomes were seen whereas, in other species, trichomes were long, unbranched glandular and had tapering ends. In adaxial side trichomes were present only in M. suaveolens, A. reptans, S. bazyntina, O. basciculum, S. splendens, S. officinalis, S. rosemarinus. In other species, trichomes were absent on the adaxial surface. In abaxial view, M. suaveolens had the largest length of trichomes, and O. basciculum had the smallest. S. splendens L. had the largest trichome width, while T. vulgaris had the smallest. CONCLUSION: Hence, according to these findings, morpho-anatomical traits are useful for identifying Lamiaceae taxa. Also, there is a need of upgradation and addition of studied taxa in flora of Pakistan comprehensively.

Floral ontogeny reveals potential synapomorphies for Senegalia sect. Monacanthea p.p. (Leguminosae).

Senegalia was recently described as non-monophyletic; however, its sections exhibit robust monophyletic support, suggesting a potential reclassification into separate genera-Senegalia sect. Monocanthea p.p. is the largest section. It contains 164 species of pantropical distribution and includes all of the current 99 neotropical species of Senegalia; however, no morphological characteristics are available to differentiate this section. To characterize this section, we examined floral developmental traits in four species of Senegalia sect. Monocanthea p.p. These traits were previously considered as potentially distinguishing features within Acacia s.l. and include the onset patterns of the androecium, the timing of calyx union, the origin of the staminal disc, and the presence of stomata on the petals. Furthermore, we analyzed previously unexplored traits, such as corolla union types, inflorescence development, and micromorphological features related to the indumentum, as well as the presence and location of stomata. The characteristics proposed as potential synapomorphies of the group include the postgenital fusion of the corolla and the presence of a staminal disc formed at the base of the filaments. The other analyzed floral characteristics were not informative for the characterization of the group. Future studies of floral ontogeny will help to establish more precise patterns, mainly whether corolla union and staminal tube formation occur similarly in African and Asian sections of Senegalia.

Regional differences in leaf evolution facilitate photosynthesis following severe drought.

Characterizing physiological and anatomical changes that underlie rapid evolution following climatic perturbation can broaden our understanding of how climate change is affecting biodiversity. It can also provide evidence of cryptic adaptation despite stasis at higher levels of biological organization. Here, we compared evolutionary changes in populations of Mimulus cardinalis from historically different climates in the north and south of the species' range following an exceptional drought. We grew seeds produced from predrought ancestral plants alongside peak-drought descendants in a common glasshouse and exposed them to wet and dry conditions. Before the drought, northern ancestral populations expressed traits contributing to drought escape, while southern ancestral populations expressed drought avoidance. Following the drought, both regions evolved to reduce water loss and maintain photosynthesis in dry treatments (drought avoidance), but via different anatomical alterations in stomata, trichomes, and palisade mesophyll. Additionally, southern populations lost the ability to take advantage of wet conditions. These results reveal rapid evolution towards drought avoidance at an anatomical level following an exceptional drought, but suggest that differences in the mechanisms between regions incur different trade-offs. This sheds light on the importance of characterizing underlying mechanisms for downstream life-history and macromorphological traits.

Pinpointing the causal influences of stomatal anatomy and behavior on minimum, operational, and maximum leaf surface conductance.

Leaf surface conductance to water vapor and CO2 across the epidermis (gleaf) strongly determines the rates of gas exchange. Thus, clarifying the drivers of gleaf has important implications for resolving the mechanisms of photosynthetic productivity and leaf and plant responses and tolerance to drought. It is well recognized that gleaf is a function of the conductances of the stomata (gs) and of the epidermis + cuticle (gec). Yet, controversies have arisen around the relative roles of stomatal density (d) and size (s), fractional stomatal opening (α; aperture relative to maximum), and gec in determining gleaf. Resolving the importance of these drivers is critical across the range of leaf surface conductances, from strong stomatal closure under drought (gleaf,min), to typical opening for photosynthesis (gleaf,op), to maximum achievable opening (gleaf,max). We derived equations and analyzed a compiled database of published and measured data for approximately 200 species and genotypes. On average, within and across species, higher gleaf,min was determined 10 times more strongly by α and gec than by d and negligibly by s; higher gleaf,op was determined approximately equally by α (47%) and by stomatal anatomy (45% by d and 8% by s), and negligibly by gec; and higher gleaf,max was determined entirely by d. These findings clarify how diversity in stomatal functioning arises from multiple structural and physiological causes with importance shifting with context. The rising importance of d relative to α, from gleaf,min to gleaf,op, enables even species with low gleaf,min, which can retain leaves through drought, to possess high d and thereby achieve rapid gas exchange in periods of high water availability.

Reponses of morphological and biochemical traits of bamboo trees under elevated atmospheric O3 enrichment.

Dwarf bamboo (Indocalamus decorus) is an O3-tolerant plant species. To identify the possible mechanism and response of leaf morphological, antioxidant, and anatomical characteristics to elevated atmospheric O3 (EO3) concentrations, we exposed three-year-old I. decorus seedlings to three O3 levels (low O3-LO: ambient air; medium O3-MO: Ambient air+70 ppb high O3-HO: Ambient air+140 ppb O3) over a growing season using open-top chambers. Leaf shape and stomatal characteristics, and leaf microscopic structure of I. decorus were examined. The results indicated that 1) the stomata O3 flux (Fst) of HO decreased more rapidly under EO3 as the exposure time increased. The foliar O3 injury of HO and MO occurred when AOT40 was 26.62 ppm h and 33.20 ppm h, respectively, 2) under EO3, leaf number, leaf mass per area, leaf area, and stomata length/width all decreased, while leaf thickness, stomatal density, width, and area increased compared to the control, 3) MDA and total soluble protein contents all showed significantly increase under HO (36.57% and 32.77%) and MO(31.91% and 19.52%) while proline contents only increased under HO(33.27%). 4) MO and HO increased bulliform cells numbers in the leaves by 6.28% and 23.01%, respectively. HO reduced the transverse area of bulliform cells by 13.73%, while MO treatments had no effect, and 5) the number of fusoid cells interspace, the transverse area of fusoid cells interspace, and mesophyll thickness of HO significantly increased by 11.16%, 28.58%, and 13.42%, respectively. In conclusion, I. decorus exhibits strong O3 tolerance characteristics, which stem from adaptions in the leaf's morphological, structural, antioxidant, and anatomical features. One critical attribute was the enlargement of the bulliform cell transverse area and the transverse area of fusoid cells interspace that drove this resistance to O3. Local bamboo species with high resistance to O3 pollution thus need to be promoted for sustained productivity and ecosystem services in areas with high O3 pollution.

Macromorphological and foliar epidermal anatomical characteristics of Lilium rosthornii (Liliaceae): Implications for morphological adaptations and taxonomic significance.

This research is to examine the macromorphological and foliar epidermal anatomical features of Lilium rosthornii Diels and its ability to plastically adapt to environmental forces, which is crucial for its taxonomic classification. L. rosthornii has macromorphological characteristics such as linear to lanceolate leaves of up to 20 cm in length and 2-3 cm in breadth, grouped in a whorled pattern. The blooms are voluminous and conspicuous, measuring up to 15 cm in diameter and are supported by a towering stalk that grows up to 1 m in height. The foliar epidermal structure of L. rosthornii exhibits a stomatal length of 82.02 ± 5.77 µm and a width of 29.19 ± 1.39 µm. These measurements suggest that the plant's stomata are influenced by its ploidy levels and may serve as adaptive mechanisms to enhance water consumption efficiency. The leaf structure shows a significant thickness of 398.74 ± 97.96 µm, which might potentially contribute to its ability to withstand environmental challenges. Additionally, the presence of defensive adaptations in the top and lower epidermal layers further supports this observation. The palisade tissue measurement (58.87 ± 9.56 m) and spongy tissue measurement (32.42 ± 12.72 µm) indicate a potential for photosynthetic optimization. Furthermore, there is a possible correlation between the vascular bundle width (28.15 ± 6.52 °m) and the efficiency of nutrition delivery. The results of this study emphasize the notable diversity in the foliar structures of L. rosthornii, offering valuable understanding of its morphological adaptations that have ecological and taxonomic significance. The findings provide a deeper comprehension of the potential impact of anatomical characteristics on plant function and categorization, hence providing significant insights to the domain of plant morphology and systematics. RESEARCH HIGHLIGHTS: Examines Lilium rosthornii's anatomical features and environmental adaptability for taxonomic relevance. Leaf thickness and epidermal defenses indicate resilience to environmental stress. Highlights the diversity in L. rosthornii's foliar structures, with implications for ecological and taxonomic significance Offers insights into the impact of anatomical characteristics on plant function and classification.

Anatomical features of ecological importance and taxonomic value revealed by scanning electron microscopy and light microscopy in Camonea umbellata (L.) A.R. Simões & Staples (Convolvulaceae).

This article describes detailed and novel data on the anatomy and histochemistry of leaves, stems, and roots of Camonea umbellata (L.) A.R.Simões & Staples in different environments for the identification of characters with taxonomical value and of ecological importance, with provision of light and scanning electron microscopy images. To analyze the characters, we collected samples of the vegetative organs of three individuals in each of three populations, which were in a grazing area, an urban environment, and a biological reserve. The main diagnostic anatomical markers for the identification of C. umbellata include amphistomatic leaves, tetracytic and brachyparatetracytic stomata, peltate trichomes, long simple trichomes, epidermis with striated cuticle ornamentation, mesophyll with acute borders, presence of druses, secretory channels, angular collenchyma, fibrous pericycle in the stem, intraxylary phloem in the vegetative organs, oil bodies throughout the midrib, petiole, stem and root, and epicuticular waxes of the crust and coiled rodlet types. Since the characters above did not show variation in the environments evaluated, we consider these characters taxonomically useful for the identification of C. umbellata. RESEARCH HIGHLIGHTS: The anatomy of the aerial vegetative organs of Camonnea umbellata retains common Convolvulaceae characters. The sinuosity of the epidermal cell walls and the density of trichomes in the epidermis of the petiole were visually variable characters among the analyzed individuals. Amphistomatic leaves, tetracytic and brachyparatetracytic stomata, peltate trichomes, epidermis with striated cuticle ornamentation, dorsiventral mesophyll with border acute, presence of druses, secretory structures, angular collenchyma, fibrous pericycle in the stem, intraxillary phloem, presence of oil bodies in all organs, and epicuticular waxes of the crust type and coiled rods were considered important anatomical markers for the recognition and correct identification of Camonea umbellata.

Leaf morpho-anatomical adjustments in a Quercus pubescens forest after 10 years of partial rain exclusion in the field.

In the Mediterranean region, a reduction of annual precipitation and a longer and drier summer season are expected with climate change by the end of the century, eventually endangering forest survival. To cope with such rapid changes, trees may modulate their morpho-anatomical and physiological traits. In the present study, we focused on the variation in leaf gas exchange and different leaf morpho-anatomical functional traits of Quercus pubescens Willd. in summer using a long-term drought experiment in natura consisting of a dynamic rainfall exclusion system where trees have been submitted to amplified drought (AD) (~-30% of annual precipitation) since April 2012 and compared them with trees under natural drought (ND) in a Mediterranean forest. During the study, we analyzed net CO2 assimilation (An), stomatal conductance (gs), transpiration (E), water-use efficiency (WUE), stomatal size and density, density of glandular trichomes and non-glandular trichomes, thickness of the different leaf tissues, specific leaf area and leaf surface. Under AD, tree functioning was slightly impacted, since only An exhibited a 49% drop, while gs, E and WUE remained stable. The decrease in An under AD was regulated by concomitant lower stomatal density and reduced leaf thickness. Trees under AD also featured leaves with a higher non-glandular trichome density and a lower glandular trichome density compared with ND, which simultaneously limits transpiration and production costs. This study points out that Q. pubescens exhibits adjustments of leaf morpho-anatomical traits which can help trees to acclimate to AD scenarios as those expected in the future in the Mediterranean region.

Endemic species analysis: Foliar epidermal anatomical characters of Aster glehnii F. Schmidt (Asteraceae).

The classification and identification of Aster glehnii F. Schmidt are determined from its foliar epidermal anatomical features. Scanning electronic microscopy has been used to determine the foliar epidermal anatomical characteristics of the species in detail. This study compared the qualitative and quantitative characteristics of the leaf epidermis of A. glehnii for taxonomic identification to be used as a reference for future studies on the species. A. glehnii has smooth, thin cuticles, depressed anomocytic stomata dispersed randomly throughout the leaf surface, polygonal epidermal cells with straight to slightly curved anticlinal walls, and no trichomes. There are obvious veins containing thick-walled bundle sheath cells. The stomatal density is between 100 and 150 stomata per millimeter. The vein density ranges from five to 10 veins per millimeter, and the epidermal cells are 10 to 20 µm long and 5 to 10 µm in width. Understanding the connections between the different A. glehnii species and categorizing and identifying them depend heavily on these foliar epidermal structural features. Taxonomy and conservation are closely intertwined because the former serves as the basis for comprehending and safeguarding biodiversity. RESEARCH HIGHLIGHTS: Optical microscopy of the A. glehnii leaf epidermis for taxonomic identification SEM was used to identify and authenticate endemic species Microscopic identification of endemic species can assist in the conservation.

Tolerance to mild shading levels in cattail as related to increased photosynthesis and changes in its leaf area and anatomy.

Shading is an environmental factor that has been little investigated regarding its effects on emergent aquatic plants. Typha domingensis Pers. is an emergent macrophyte that demonstrates some plasticity for self-shading, and as it can shade other species in the same area, the effect of shading on its traits deserves further investigation. The objective of the present study was to evaluate the gas exchange, leaf anatomy, and growth of T. domingensis cultivated under increasing shading intensities. The plants were collected and propagated in a greenhouse, and the clones were subjected to four shading intensities: 0% (unshaded), 35%, 73%, and 83% shading created by black nets. Growth traits, clonal production, photosynthesis, transpiration, and leaf anatomy were evaluated. The 73% and 83% shading promoted the death of all plants, but all plants survived in the 35% and unshaded treatments. Compared with the unshaded treatment, the 35% shading treatment promoted a higher photosynthetic rate and greater transpiration, supporting increased growth and production of clones. The increase in the photosynthetic rate in the 35% shading was related to the increase in leaf area which increased the photosynthesis of the whole plant. The 73% and 83% treatments inhibited the development of photosynthetic parenchyma and stomata in T. domingensis, leading to a drastic reduction in photosynthesis and energy depletion. Therefore, T. domingensis does not tolerate intense shading, but its photosynthetic characteristics and growth are favored by mild shading, a factor that may be of great importance for its competitiveness and invasive behavior.

How Important Are Functional and Developmental Constraints on Phenotypic Evolution? An Empirical Test with the Stomatal Anatomy of Flowering Plants.

AbstractQuantifying the relative contribution of functional and developmental constraints on phenotypic variation is a long-standing goal of macroevolution, but it is often difficult to distinguish different types of constraints. Alternatively, selection can limit phenotypic (co)variation if some trait combinations are generally maladaptive. The anatomy of leaves with stomata on both surfaces (amphistomatous) present a unique opportunity to test the importance of functional and developmental constraints on phenotypic evolution. The key insight is that stomata on each leaf surface encounter the same functional and developmental constraints but potentially different selective pressures because of leaf asymmetry in light capture, gas exchange, and other features. Independent evolution of stomatal traits on each surface imply that functional and developmental constraints alone likely do not explain trait covariance. Packing limits on how many stomata can fit into a finite epidermis and cell size-mediated developmental integration are hypothesized to constrain variation in stomatal anatomy. The simple geometry of the planar leaf surface and knowledge of stomatal development make it possible to derive equations for phenotypic (co)variance caused by these constraints and compare them with data. We analyzed evolutionary covariance between stomatal density and length in amphistomatous leaves from 236 phylogenetically independent contrasts using a robust Bayesian model. Stomatal anatomy on each surface diverges partially independently, meaning that packing limits and developmental integration are not sufficient to explain phenotypic (co)variation. Hence, (co)variation in ecologically important traits like stomata arises in part because there is a limited range of evolutionary optima. We show how it is possible to evaluate the contribution of different constraints by deriving expected patterns of (co)variance and testing them using similar but separate tissues, organs, or sexes.

Anatomical drivers of stomatal conductance in sorghum lines with different leaf widths grown under different temperatures.

Sustaining crop productivity and resilience in water-limited environments and under rising temperatures are matters of concern worldwide. We investigated the leaf anatomical traits that underpin our recently identified link between leaf width (LW) and intrinsic water use efficiency (iWUE), as traits of interest in plant breeding. Ten sorghum lines with varying LW were grown under three temperatures to expand the range of variation of both LW and gas exchange rates. Leaf gas exchange, surface morphology and cross-sectional anatomy were measured and analysed using structural equations modelling. Narrower leaves had lower stomatal conductance (gs ) and higher iWUE across growth temperatures. They also had smaller intercellular airspaces, stomatal size, percentage of open stomatal aperture relative to maximum, hydraulic pathway, mesophyll thickness, and leaf mass per area. Structural modelling revealed a developmental association among leaf anatomical traits that underpinned gs variation in sorghum. Growing temperature and LW both impacted leaf gas exchange rates, but only LW directly impacted leaf anatomy. Wider leaves may be more productive under well-watered conditions, but consume more water for growth and development, which is detrimental under water stress.

Implications of foliar epidermal micromorphology using light and scanning electron microscopy: A useful tool in taxonomy of Korean irises.

The genus Iris L., comprising approximately 210 species, is one of the most species-rich genera in the family Iridaceae. In this study, the first comprehensive leaf micromorphological characters of Korean irises were studied using light and scanning electron microscopy. Our objective was to evaluate the foliar micromorphological characteristics (namely epidermal cells, stomata types, and guard cell size) of Korean Iris taxa in a systematic context. All the investigated Korean Iris taxa had amphistomatic or hypostomatic leaves with anomocytic stomatal complexes. Guard cell length varied among species, ranging from 24.8 µm (I. rossii) to 56.0 µm (I. domestica). Although the presence of papillae on the outer periclinal wall is not of taxonomic significance, leaf margin pattern, guard cell size, and sunken stomata type were useful for species-level identification of Korean Iris species. The occurrence of polymorphic stomatal types was reported here for the first time, and the correlation between genome size and epidermal guard cell length was discussed.

Stomatal density in Pinus sylvestris as an indicator of temperature rather than CO2 : Evidence from a pan-European transect.

The commonly observed negative relationship between stomatal density (SD) and atmospheric CO2 has led to SD being proposed as an indicator of atmospheric CO2 concentration. The use of SD as a proxy for CO2 , however, has been hampered by an insufficient understanding of the intraspecific variation of this trait. We hypothesized that SD in Pinus sylvestris, a widely distributed conifer, varies geographically and that this variation is determined by major climatic variables. By sampling needles from naturally growing trees along a latitudinal range of 32.25°, equivalent to 13.7°C gradient of mean annual temperature (MAT) across Europe, we found that SD decreased from the warmest southern sites to the coldest sites in the north at a rate of 4 stomata per mm2 for each 1°C, with MAT explaining 44% of the variation. Additionally, samples from a provenance trial exhibited a positive relationship between SD and the MAT of the original localities, suggesting that high SD is an adaptation to warm temperature. Our study revealed one of the strongest intraspecific relationships between SD and climate in any woody species, supporting the utility of SD as a temperature, rather than direct CO2 , proxy. In addition, our results predict the response of SD to climate warming.

A stomata classification and detection system in microscope images of maize cultivars.

Plant stomata are essential structures (pores) that control the exchange of gases between plant leaves and the atmosphere, and also they influence plant adaptation to climate through photosynthesis and transpiration stream. Many works in literature aim for a better understanding of these structures and their role in the evolution process and the behavior of plants. Although stomata studies in dicots species have advanced considerably in the past years, even there is not much knowledge about the stomata of cereal grasses. Due to the high morphological variation of stomata traits intra- and inter-species, detecting and classifying stomata automatically becomes challenging. For this reason, in this work, we propose a new system for automatic stomata classification and detection in microscope images for maize cultivars based on transfer learning strategy of different deep convolution neural netwoks (DCNN). Our performed experiments show that our system achieves an approximated accuracy of 97.1% in identifying stomata regions using classifiers based on deep learning features, which figures out as a nearly perfect classification system. As the stomata are responsible for several plant functionalities, this work represents an important advance for maize research, providing an accurate system in replacing the current manual task of categorizing these pores on microscope images. Furthermore, this system can also be a reference for studies using images from different cereal grasses.

Macroscopic variation in Arabidopsis mutants despite stomatal uniformity across soil nutrient environments.

Stomata are essential pores flanked by guard cells that control gas exchange in plants. We can utilize stomatal size and density measurements as a proxy for a plant's capacity for gas exchange. While stomatal responses to stressful environments are well studied; data are lacking in the responses across mutant genotypes of the same species in these trait and treatment interactions or genetic variation in phenotypic plasticity. We evaluated the effects of soil nutrient variation on macroscopic and stomatal traits of Arabidopsis thaliana T-DNA insertion mutants for which prior performance in a single benign growing condition were available. Nutrient-induced stress significantly impacted traits including plant biomass, height, fruit number, and leaf number which we denote as macroscopic traits. We found evidence that genotype by environment effects exist for macroscopic traits, yet total stomatal area variation, or "microscopic variation" across environments was modest. Divergence from the wildtype line varied by mutant background and these responses were variable among traits. These findings suggest that Arabidopsis employs a strategy of physiological compensation, sacrificing morphological traits to maintain stomatal production.

Callitriche as a potential model system for evolutionary studies on the dorsiventral distribution of stomata.

Controlling the distribution of stomata is crucial for the adaptation of plants to new, or changing environments. While many plant species produce stomata predominantly on the abaxial leaf surface (hypostomy), some produce stomata on both surfaces (amphistomy), and the remaining few produce them only on the adaxial surface (hyperstomy). Various selective pressures have driven the evolution of these three modes of stomatal distribution. Despite recent advances in our understanding of stomatal development and dorsiventral leaf polarity, the genetic basis for the evolution of different stomatal distributions is still unclear. Here, we propose the genus Callitriche as a new model system to investigate patterns in the evolution of stomatal distribution. Callitriche comprises species with diverse lifestyles, including terrestrial, amphibious, and obligately aquatic plants. We found that species in this genus cover all three modes of dorsiventral stomatal distribution, making it a desirable model for comparative and evolutionary analyses on distribution modes. We further characterized the genetic basis of the different distribution modes, focusing on the stomatal key transcription factor SPEECHLESS. Future research using the promising model system Callitriche would open a new direction for evolutionary developmental biology studies on stomata.