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A fundamental objective of evolutionary biology is to understand the origin of independently evolving species. Phylogenetic studies of species radiations rarely are able to document ongoing speciation; instead, modes of speciation, entailing geographic separation and/or ecological differentiation, are posited retrospectively. The Oreinotinus clade of Viburnum has radiated recently from north to south through the cloud forests of Mexico and Central America to the Central Andes. Our analyses support a hypothesis of incipient speciation in Oreinotinus at the southern edge of its geographic range, from central Peru to northern Argentina. Although several species and infraspecific taxa have been recognized in this area, multiple lines of evidence and analytical approaches (including analyses of phylogenetic relationships, genetic structure, leaf morphology, and climatic envelopes) favor the recognition of just a single species, V. seemenii. We show that what has previously been recognized as V. seemenii f. minor has recently occupied the drier Tucuman-Bolivian forest region from Samaipata in Bolivia to Salta in northern Argentina. Plants in these populations form a well-supported clade with a distinctive genetic signature and they have evolved smaller, narrower leaves. We interpret this as the beginning of a within-species divergence process that has elsewhere in the neotropics resulted repeatedly in Viburnum species with a particular set of leaf ecomorphs. Specifically, the southern populations are in the process of evolving the small, glabrous, and entire leaf ecomorph that has evolved in four other montane areas of endemism. As predicted based on our studies of leaf ecomorphs in Chiapas, Mexico, these southern populations experience generally drier conditions, with large diurnal temperature fluctuations. In a central portion of the range of V. seemenii, characterized by wetter climatic conditions, we also document what may be the initial differentiation of the leaf ecomorph with larger, pubescent, and toothy leaves. The emergence of these ecomorphs thus appears to be driven by adaptation to subtly different climatic conditions in separate geographic regions, as opposed to parapatric differentiation along elevational gradients as suggested by Viburnum species distributions in other parts of the neotropics., (© The Author(s) 2024. Published by Oxford University Press, on behalf of the Society of Systematic Biologists. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

Understanding the global variation of plant strategies is essential for unravelling eco-evolutionary processes and ecosystem functions. Variation in ten fundamental aboveground and fine-root traits is summarised in four dimensions, the first of which relates to aboveground plant size. However, there is no consensus about how root size fits within this scheme. Here, we add rooting depth and lateral spread, compiling a set of twelve key traits that define the fundamental investments of plants in growth, reproduction, and survival. We examine whether the inclusion of root size alters the dimensionality and structure of trait correlations defining plant functional strategies. Our results show that including root size traits does not alter the fundamental structure and dimensionality of the plant functional space, regardless of trait completeness and phylogenetic relatedness. Plant size defines a single continuum of allometric investments at the global scale, independent from leaf and root economic strategies., (© 2024. The Author(s).)

Plants belonging to the genus Cotoneaster can be valuable sources of phytochemicals with potential therapeutic properties. The natural habitats of most of these species are situated in Asia, Africa and southern Europe. Introducing them into other climatic conditions could expose them to abiotic and biotic stresses, affecting their bioactive properties. The aim of this study was to assess and compare the performance of four non-native Cotoneaster species (C. roseus, C. hissaricus, C. hsingshangensis, C. nebrodensis) grown in eastern Poland in terms of leaf morphology, anatomy, and efficiency of the photosynthetic apparatus in relation to lipid peroxidation level, being an indicator of oxidative stress. Photosynthetic pigments concentration and chlorophyll a fluorescence parameters were used to evaluate the photosynthetic capacity. The morphological and anatomical analysis of leaves did not show any anomalies or stress symptoms. Cotoneaster roseus was characterized by the highest chlorophyll concentration, viability index (Rfd) and a moderate lipid peroxidation level. On the other hand, the lowest values of photochemical quenching (qP), maximum fluorescence (F m ), Rfd, and quantum yield of the photosystem II (QY) observed for C. nebrodensis might indicate an inferior efficiency of the photosynthetic apparatus in this species; however, it demonstrated the lowest lipid peroxidation level. Nevertheless, the content and proportion of the photosynthetic pigments as well as overall chlorophyll a fluorescence parameters and lipid peroxidation levels indicate a good physiological condition of all examined plants. The observed differences between the species may be rather species specific and genetically established and not indicate their sensitivity to non-native growth conditions. This is the first report about the physiological parameters of the four Cotoneaster species, proving they are well adapted to growth in the climatic conditions of central Europe providing thus a raw material with potential for pharmaceutical applications., (© 2024. The Author(s).)

Allagoptera campestris is an acaulescent, rhizomatous palm tree that occurs in grassland and savanna areas (Cerrado). In the Santa Bárbara Ecological Station (Águas de Santa Bárbara, São Paulo, Brazil) the species is found growing in three distinct conditions: 1) in the understory of Pinus species plantations introduced in the 1970s in formerly open savanna, 2) in an area where Pinus species cultivated in the 1970s were later removed and the remaining material burned, and 3) in an open, undisturbed savanna area without the interference of pines. Anatomical studies carried out with A. campestris leaves collected in the same three areas indicated leaf plasticity in response to growth conditions. To verify whether there are differences in vessel element morphology in belowground organs, light, and scanning electron microscopy analyses were conducted on portions just below the crown, in the middle of the rhizome, and the median portions of three longer adventitious roots sampled from three plants in each area. The study reveals significant variations in vessel element characteristics of A. campestris, with roots consistently displaying longer and larger elements than rhizomes, and environmental conditions, especially in pine understory, influence vessel dimensions, and hydraulic conductivity in a negative manner.

A time-dependent mechanism mediated by SPL9 highlights how age-related growth reprogramming influences leaf shape modifications in simple- and complex-leaved, related plants. Extending cell proliferation and delaying differentiation drives the mechanism, offering insights into the shared developmental processes underlying leaf form diversification., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Fusing three-dimensional (3D) and multispectral (MS) imaging data holds promise for high-throughput and comprehensive plant phenotyping to decipher genome-to-phenome knowledge. Acquiring high-quality 3D MS point clouds (3DMPCs) of plants remains challenging because of poor 3D data quality and limited radiometric calibration methods for plants with a complex canopy structure. Here, we present a novel 3D spatial-spectral data fusion approach to collect high-quality 3DMPCs of plants by integrating the next-best-view planning for adaptive data acquisition and neural reference field (NeREF) for radiometric calibration. This approach was used to acquire 3DMPCs of perilla, tomato, and rapeseed plants with diverse plant architecture and leaf morphological features evaluated by the accuracy of chlorophyll content and equivalent water thickness (EWT) estimation. The results showed that the completeness of plant point clouds collected by this approach was improved by an average of 23.6% compared with the fixed viewpoints alone. The NeREF-based radiometric calibration with the hemispherical reference outperformed the conventional calibration method by reducing the root mean square error (RMSE) of 58.93% for extracted reflectance spectra. The RMSE for chlorophyll content and EWT predictions decreased by 21.25% and 14.13% using partial least squares regression with the generated 3DMPCs. Collectively, our study provides an effective and efficient way to collect high-quality 3DMPCs of plants under natural light conditions, which improves the accuracy and comprehensiveness of phenotyping plant morphological and physiological traits, and thus will facilitate plant biology and genetic studies as well as crop breeding., (Copyright © 2024 The Author. Published by Elsevier Inc. All rights reserved.)

Biological shape diversity is often manifested in modulation of organ symmetry and modification of the patterned elaboration of repeated shape elements. 1 , 2 , 3 , 4 , 5 Whether and how these two aspects of shape determination are coordinately regulated is unclear. 5 , 6 , 7 Plant leaves provide an attractive system to investigate this problem, because they often show asymmetries along the proximodistal (PD) axis of their blades, along which they can also produce repeated marginal outgrowths such as serrations or leaflets. 1 One aspect of leaf shape diversity is heteroblasty, where the leaf form in a single genotype is modified with progressive plant age. 8 , 9 , 10 , 11 In Arabidopsis thaliana, a plant with simple leaves, SQUAMOSA PROMOTER BINDING PROTEIN-LIKE 9 (SPL9) controls heteroblasty by activating CyclinD3 expression, thereby sustaining proliferative growth and retarding differentiation in adult leaves. 12 , 13 However, the precise significance of SPL9 action for leaf symmetry and marginal patterning is unknown. By combining genetics, quantitative shape analyses, and time-lapse imaging, we show that PD symmetry of the leaf blade in A. thaliana decreases in response to an age-dependent SPL9 expression gradient, and that SPL9 action coordinately regulates the distribution and shape of marginal serrations and overall leaf form. Using comparative analyses, we demonstrate that heteroblastic growth reprogramming in Cardamine hirsuta, a complex-leafed relative of A. thaliana, also involves prolonging the duration of cell proliferation and delaying differentiation. We further provide evidence that SPL9 enables species-specific action of homeobox genes that promote leaf complexity. In conclusion, we identified an age-dependent layer of organ PD growth regulation that modulates leaf symmetry and has enabled leaf shape diversification., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Premise: Leaf mass per area (LMA) is a widely used functional trait in both neobotanical and paleobotanical research that provides a window into how plants interact with their environment. Paleobotanists have used site-level measures of LMA as a proxy for climate, biome, deciduousness, and community-scale plant strategy, yet many of these relationships have not been grounded in modern data. In this study, we evaluated LMA from the paleobotanical perspective, seeking to add modern context to paleobotanical interpretations and discover what a combined modern and fossil data set can tell us about how LMA can be best applied toward interpreting plant communities., Methods: We built a modern data set by pulling plant trait data from the TRY database, and a fossil data set by compiling data from studies that have used the petiole-width proxy for LMA. We then investigated the relationships of species-mean, site-mean, and site-distribution LMA with different climatic, phylogenetic, and physiognomic variables., Results: We found that LMA distributions are correlated with climate, site taxonomic composition, and deciduousness. However, the relative contributions of these factors are not distinctive, and ultimately, LMA distributions cannot accurately reconstruct the biome or climate of an individual site., Conclusions: The correlations that make up the leaf economics spectrum are stronger than the correlations between LMA and climate, phylogeny, morphospace, or depositional environment. Fossil LMA should be understood as the culmination of the influences of these variables rather than as a predictor., (© 2024 Botanical Society of America.)

Drought-induced stress represents one of the most economically detrimental natural phenomena impacting grapevine (Vitis vinifera ) development, yield, and fruit characteristics. Also, auxin is one of the most important plant growth regulators that can reduce damage caused by stress in plants. In this study, the impact of exogenously sprayed auxin (0, 50, and 200mgL-1 ) on growth, biochemical, and anatomical parameters was investigated in two grapevine varieties (cvs. 'Rashe' and 'Fakhri') under water deficit. According to our findings, water deficit led to a notable decrease in growth, protein content, and anatomical parameters; but significantly enhanced electrolyte leakage. Grapevines exposed to water deficit exhibited substantial increases in total phenolic compounds and antioxidant activity. Applying 50mgL-1 napthalene acetic acid (NAA) reduced the effects of water deficit in both grapevine cultivars by decreasing electrolyte leakage (15% in 'Rashe' and 20% in 'Fakhri'), and accumulating protein content (22% 'Rashe' and 32% 'Fakhri'), total phenolic compounds (33%'Rashe' and 40% 'Fakhri'), and antioxidant capacity (11% 'Rashe' and 39% 'Fakhri'); anantomical parameters were also improved. However, application of 200mgL-1 NAA had adverse effects on growth and biochemical traits of grapevines, with a more pronounced impact on root growth and anatomical parameters compared to other NAA concentrations. In conclusion, the application of 50mgL-1 NAA enhanced grapevine growth, enabling them to better thrive under water deficit.

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., (© 2024 Botanical Society of America.)

Integrating traits across above- and belowground organs offers comprehensive insights into plant ecology, but their various functions also increase model complexity. This study aimed to illuminate the interspecific pattern of whole-plant trait correlations through a network lens, including a detailed analysis of the root system. Using a network algorithm that allows individual traits to belong to multiple modules, we characterize interrelations among 19 traits, spanning both shoot and root phenology, architecture, morphology, and tissue properties of 44 species, mostly herbaceous monocots from Northern Ontario wetlands, grown in a common garden. The resulting trait network shows three distinct yet partially overlapping modules. Two major trait modules indicate constraints of plant size and form, and resource economics, respectively. These modules highlight the interdependence between shoot size, root architecture and porosity, and a shoot-root coordination in phenology and dry-matter content. A third module depicts leaf biomechanical adaptations specific to wetland graminoids. All three modules overlap on shoot height, suggesting multifaceted constraints of plant stature. In the network, individual-level traits showed significantly higher centrality than tissue-level traits do, demonstrating a hierarchical trait integration. The presented whole-plant, integrated network suggests that trait covariation is essentially function-driven rather than organ-specific., (© 2024 The Author(s). New Phytologist © 2024 New Phytologist Foundation.)

Nicotiana benthamiana is predominantly distributed in arid habitats across northern Australia. However, none of six geographically isolated accessions shows obvious xerophytic morphological features. To investigate how these tender-looking plants withstand drought, we examined their responses to water deprivation, assessed phenotypic, physiological, and cellular responses, and analysed cuticular wax composition and wax biosynthesis gene expression profiles. Results showed that the Central Australia (CA) accession, globally known as a research tool, has evolved a drought escape strategy with early vigour, short life cycle, and weak, water loss-limiting responses. By contrast, a northern Queensland (NQ) accession responded to drought by slowing growth, inhibiting flowering, increasing leaf cuticle thickness, and altering cuticular wax composition. Under water stress, NQ increased the heat stability and water impermeability of its cuticle by extending the carbon backbone of cuticular long-chain alkanes from c. 25 to 33. This correlated with rapid upregulation of at least five wax biosynthesis genes. In CA, the alkane chain lengths (c. 25) and gene expression profiles remained largely unaltered. This study highlights complex genetic and environmental control over cuticle composition and provides evidence for divergence into at least two fundamentally different drought response strategies within the N. benthamiana species in < 1 million years., (© 2024 The Author(s). New Phytologist © 2024 New Phytologist Foundation.)

Leaf size is a crucial agronomic trait directly affecting crop yield, which is mainly determined by coordinated cell proliferation, growth, and differentiation. Although endoreduplication is known to be correlated with the onset of cell differentiation and leaf size, the underlying molecular mechanisms are largely unclear. The DnaJ-like zinc finger domain-containing protein ORANGE (OR) was initially demonstrated to confer the massive accumulation of carotenoids in cauliflower curds. However, the cauliflower or mutant also possesses other phenotypes such as smaller curds, smaller leaves with elongated petioles, and delayed flowering. Here, we demonstrated that OR physically interacts with the transcription factor TCP7, which promotes endoreduplication by inducing the expression of the cell cycle gene CYCLIN D 1;1 (CYCD1;1). Overexpression of OR resulted in smaller rosette leaves, whereas the OR-silencing plants had larger rosette leaves than wild-type plants. Our microscopic observations and flow cytometry analysis revealed that the variation in leaf size was a result of different endoreduplication levels. Genetic analyses showed that OR functions antagonistically with TCP7 in regulating the endoreduplication levels in leaf cells. While the expression of OR is induced by TCP7, OR represses the transactivation activity of TCP7 by affecting its binding capability to the TCP-binding motif in the promoter region of CYCD1;1. Through this interaction, OR negatively regulates the expression of CYCD1;1 and reduces the nuclear ploidy level in rosette leaf cells. Our findings provide new insights into the regulatory network of leaf size and also reveal a regulatory circuit controlling endoreduplication in leaf cells., (© 2024 Society for Experimental Biology and John Wiley & Sons Ltd.)

Variation in leaf venation network architecture may reflect trade-offs among multiple functions including efficiency, resilience, support, cost, and resistance to drought and herbivory. However, our knowledge about architecture-function trade-offs is mostly based on studies examining a small number of functional axes, so we still lack a more integrative picture of multidimensional trade-offs. Here, we measured architecture and functional traits on 122 ferns and angiosperms species to describe how trade-offs vary across phylogenetic groups and vein spatial scales (small, medium, and large vein width) and determine whether architecture traits at each scale have independent or integrated effects on each function. We found that generalized architecture-function trade-offs are weak. Architecture strongly predicts leaf support and damage resistance axes but weakly predicts efficiency and resilience axes. Architecture traits at different spatial scales contribute to different functional axes, allowing plants to independently modulate different functions by varying network properties at each scale. This independence of vein architecture traits within and across spatial scales may enable evolution of multiple alternative leaf network designs with similar functioning., (© 2024 The Author(s). New Phytologist © 2024 New Phytologist Foundation.)

Key Message: High-throughput next-generation sequencing of 161 olive germplas. 33 samples were selected as core olive germplasm and Fingerprints were constructed. After GWAS analysis of olive leaf shape, 14 candidate genes were localized. Olive (Olea europaea L.) has been introduced to China since the 1960s. After a prolonged period of variation and domestication, there is a lack of comprehensive research on its genetics. The olive oil directly extracted from Olea europaea L. is recognized as 'liquid gold', nevertheless, people constantly overlook the valuable wealth of olive leaves. High-throughput next-generation sequencing was performed on 161 olive germplasm to analyze the kinship, genetic structure and diversity of olives, and the core germplasm of olives were selected and fingerprints were constructed. Meanwhile, Genome-wide association analysis (GWAS) was performed to locate the gene for regulating olive leaf shape. Herein, the results parsed that most of the Chinese olive germplasm was more closely related to the Italian germplasm. A wealth of hybridized germplasm possessed high genetic diversity and had the potential to be used as superior parental material for olive germplasm. A total of 33 samples were selected and characterized as core germplasm of olive and Fingerprints were also constructed. A total of 14 candidate genes were localized after GWAS analysis of four olive leaf shape phenotypes, including leaf shape, leaf curvature shape, leaf tip and leaf base shape. Collectively, this study revealed the genetic basis of olives in China and also succeeded in constructing the core germplasm that stands for the genetic diversity of olives, which can contribute to the scientific and effective collection and preservation of olive germplasm resources, and provide a scientific basis for the in-depth excavation and utilization of genes regulating olive leaf shape., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Main Conclusion: The leaf color asymmetry found in the reciprocal hybrids C. hystrix × C. sativus (HC) and C. sativus × C. hystrix (CH) could be influenced by the CsPPR gene (CsaV3&#95;1G038250.1). Most angiosperm organelles are maternally inherited; thus, the reciprocal hybrids usually exhibit asymmetric phenotypes that are associated with the maternal parent. However, there are two sets of organelle genomes in the plant cytoplasm, and the mechanism of reciprocal differences are more complex and largely unknown, because the chloroplast genes are involved besides mitochondrial genes. Cucumis spp. contains the species, i.e., cucumber and melon, which chloroplasts and mitochondria are maternally inherited and paternally inherited, respectively, serving as good materials for the study of reciprocal differences. In this study, leaf color asymmetry was observed in the reciprocal hybrids (HC and CH) derived from C. sativus (2n = 14, CC) and C. hystrix (2n = 24, HH), where the leaves of HC were found to have reduced chlorophyll content, abnormal chloroplast structure and lower photosynthetic capacity. Transcriptomic analysis revealed that the chloroplast development-related genes were differentially expressed in leaf color asymmetry. Genetic analysis showed that leaf color asymmetry was caused by the maternal chloroplast genome. Comparative analysis of chloroplast genomes revealed that there was no mutation in the chloroplast genome during interspecific hybridization. Moreover, a PPR gene (CsaV3&#95;1G038250.1) with RNA-editing function was found to be involved in the regulation of leaf color asymmetry. These findings provide new insights into the regulatory mechanisms of asymmetric phenotypes in plant reciprocal crosses., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Withania (Solanaceae, Solanoideae) is a widespread genus. Comparative macro-, micro-morphological, anatomical, and molecular features of this genus in Egypt were examined using light and scanning electron microscopy to reassess the conflicted taxonomic relationships between the two studied species. The most significant morphological differences that have been found were: the shape of the lamina, apex, anther, and stigma, and the ratio of calyx tube/lobe; anatomical examination of taxonomic interest are as follows: number of vascular bundles, presence of ears and distribution of accessory vascular bundles in petiole and shape of spongy cells, and number of lower parenchyma in the midrib region of the leaf; trichomes of both species showed no significant differences; pollen, and seed characters are of taxonomic significance in differentiation and characterization between them. Protein profiling revealed that W. somnifera has only conserved proteins, while W. obtusifolia possessed both conserved and additional proteins in their SDS-PAGE banding patterns. Eleven starts codon-targeted (ScoT) primers were applied and produced 96 amplicons with an average of 70.83% polymorphism/primer. W. obtusifolia generated more polymorphic bands and maintained monomorphic ones. SDS-PAGE disclosed that both Withania species were 50% related. While Scot-Dendrogram revealed that both Withania species were poorly related. So, protein and molecular analyses showed considerable genetic variations between these two species., (© 2024. The Author(s).)

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., Competing Interests: The authors declare that they have no competing interests., (© 2024 Ghimire et al.)

Traditional pastures in temperate regions face limitations such as reduced growth and nutritional quality during the summer season. Plantain (P. lanceolata L.) offers advantages like increased yield and decreased nitrogen losses from grazing ruminants. Effective grazing management is essential for pasture health, and defoliation frequency and intensity play a pivotal role. This study aimed to evaluate plantain's regrowth, yield, and morpho-physiological and chemical responses under different defoliation frequencies and intensities, with the goal of enhancing its management in pastures. The study was conducted in pots within a controlled-environment growth chamber, examining the impact of three defoliation frequencies (based on extended leaf length: 15, 25 and 35 cm) and two defoliation intensities (5 and 8 cm of residual heights) with four replicates (24 pots as experimental units). The variables of interest were morphological characteristics, dry matter (DM) accumulation, herbage chemical composition, growth rate traits, and photosynthetic parameters. Defoliation frequency affected plantain's growth and nutritional composition. More frequent cuts (15 cm) resulted in lower DM yield per cut and lower stem content, while less frequent cuts (35 cm) produced higher values. Defoliation intensity influenced the proportion of leaves and stems in the total DM, with 5 cm cuts favoring leaves. Nutrient content was also affected by defoliation frequency, with less frequent cuts (35 cm) showing lower crude protein concentration and metabolizable energy content but higher neutral detergent fiber and water-soluble carbohydrate concentration. Plantain's growth rate variables were mainly influenced by defoliation frequency, with less frequent cuts promoting faster leaf appearance and growth of new leaves. The basal fluorescence variables and chlorophyll content were affected by cutting frequency, being highest when cut less frequently (35 cm), while no differences were found in the actual quantum efficiency among different defoliation frequencies and intensities. The fraction of light dedicated to non-photochemical quenching was highest when cut less frequently and more intensively. Overall, defoliation at 25 cm of extended leaf length balanced plantain forage quality and regrowth capacity., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Merino et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Grain and flag leaf size are two important agronomic traits that influence grain yield in rice (Oryza sativa). Many quantitative trait loci (QTLs) and genes that regulate these traits individually have been identified, however, few QTLs and genes that simultaneously control these two traits have been identified. In this study, we conducted a genome-wide association analysis in rice and detected a major locus, WIDTH OF LEAF AND GRAIN (WLG), that was associated with both grain and flag leaf width. WLG encodes a RING-domain E3 ubiquitin ligase. WLGhap.B, which possesses five single nucleotide polymophysim (SNP) variations compared to WLGhap.A, encodes a protein with enhanced ubiquitination activity that confers increased rice leaf width and grain size, whereas mutation of WLG leads to narrower leaves and smaller grains. Both WLGhap.A and WLGhap.B interact with LARGE2, a HETC-type E3 ligase, however, WLGhap.B exhibits stronger interaction with LARGE2, thus higher ubiquitination activity toward LARGE2 compared with WLGhap.A. Lysine1021 is crucial for the ubiquitination of LARGE2 by WLG. Loss-of-function of LARGE2 in wlg-1 phenocopies large2-c in grain and leaf width, suggesting that WLG acts upstream of LARGE2. These findings reveal the genetic and molecular mechanism by which the WLG-LARGE2 module mediates grain and leaf size in rice and suggest the potential of WLGhap.B in improving rice yield., Competing Interests: Conflict of interest statement. None declared., (© The Author(s) 2024. Published by Oxford University Press on behalf of American Society of Plant Biologists. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

The leaf is a determinate organ with a final size under genetic control. Numerous factors that regulate the final leaf size have been identified in Arabidopsis thaliana; although most of these factors play their roles during the growth of leaf primordia, much less is known about leaf initiation and its effects on the final leaf size. In this study, we characterized oligocellula6-D (oli6-D), a semidominant mutant of A. thaliana with smaller leaves than the wild type (WT) due to its reduced leaf cell numbers. A time-course analysis showed that oli6-D had approximately 50% fewer leaf cells even immediately after leaf initiation; this difference was maintained throughout leaf development. Next-generation sequencing showed that oli6-D had chromosomal duplications involving 2-kb and 3-Mb regions of chromosomes 2 and 4, respectively. Several duplicated genes examined had approximately 2-fold higher expression levels, and at least one gene acquired a new intron/exon structure due to a chromosome fusion event. oli6-D showed reduced auxin responses in leaf primordia, primary roots and embryos, as well as reduced apical dominance and partial auxin-resistant root growth. CRISPR-associated protein-9-mediated genome editing enabled the removal of a 3-Mb duplicated segment, the largest targeted deletion in plants thus far. As a result, oli6-D restored the WT leaf phenotypes, demonstrating that oli6-D is a gain-of-function mutant. Our results suggest a new regulatory point of leaf size determination that functions at a very early stage of leaf development and is negatively regulated by one or more genes located in the duplicated chromosomal segments., (© The Author(s) 2024. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site–for further information please contact journals.permissions@oup.com.)

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., Competing Interests: Conflict of interest statement. None declared., (© The Author(s) 2024. Published by Oxford University Press on behalf of American Society of Plant Biologists. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

The water relation strategy is a key issue in climate change. Given the difficulty of determining water relations strategy, there is a need for simple traits with a solid theoretical basis to estimate it. Traits associated with resource allocation patterns along a 'fast-slow' plant economics spectrum are particularly compelling, reflecting trade-offs between growth rate and carbon allocation. Avocado (Persea americana ), fig tree (Ficus carica ), mandarin (Citrus reticulata ), olive (Olea europaea ), pomegranate (Punica granatum ), and grapevine (Vitis vinifera ) were characterised in terms of iso-anisohydric strategy through stomatal behaviour, water potential at the turgor loss point (TLP), and hydroscape area. Additionally, the association of these metrics with leaf mass per area (LMA) and wood density (WDen) was explored. We observed high coordination between LMA and WDen, and both traits were related to metrics of water relation strategy. More anisohydric species tended to invest more carbon per unit leaf area or unit stem volume, which has implications for hydraulic efficiency and water stress tolerance. WDen and TLP were the most powerful traits in estimating the water relation strategy for six fruit species. These traits are easy to measure, time-cost efficient, and appear central to coordinating multiple traits and behaviours along the water relations strategies.

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., (© 2024 The Author(s). New Phytologist © 2024 New Phytologist Foundation.)

C 4 photosynthesis is a complex trait requiring multiple developmental and metabolic alterations. Despite this complexity, it has independently evolved over 60 times. However, our understanding of the transition to C 4 is complicated by the fact that variation in photosynthetic type is usually segregated between species that diverged a long time ago. Here, we perform a genome-wide association study (GWAS) using the grass Alloteropsis semialata, the only known species to have C 3 , intermediate, and C 4 accessions that recently diverged. We aimed to identify genomic regions associated with the strength of the C 4 cycle (measured using δ 13 C), and the development of C 4 leaf anatomy. Genomic regions correlated with δ 13 C include regulators of C 4 decarboxylation enzymes (RIPK), nonphotochemical quenching (SOQ1), and the development of Kranz anatomy (SCARECROW-LIKE). Regions associated with the development of C 4 leaf anatomy in the intermediate individuals contain additional leaf anatomy regulators, including those responsible for vein patterning (GSL8) and meristem determinacy (GIF1). The parallel recruitment of paralogous leaf anatomy regulators between A. semialata and other C 4 lineages implies the co-option of these genes is context-dependent, which likely has implications for the engineering of the C 4 trait into C 3 species., (© 2024 The Author(s). New Phytologist © 2024 New Phytologist Foundation.)

Plants express diverse nutrient use and acquisition traits, but it is unclear how trait combinations at the species level are constrained by phylogeny, trait coordination, or trade-offs in resource investment. One trait - nitrogen (N) fixation - is assumed to correlate with other traits and used to define plant functional groups, despite potential confounding effects of phylogeny. We quantified growth, carbon metabolism, fixation rate, root phosphatase activity (RPA), mycorrhizal colonization, and leaf and root morphology/chemistry across 22 species of fixing and nonfixing tropical Fabaceae trees under common conditions. Belowground trait variation was high even among closely related species, and most traits displayed a phylogenetic signal, including N-fixation rate and nodule biomass. Across species, we observed strong positive correlations between physiological traits such as RPA and root respiration. RPA increased ~ fourfold per unit increase in fixation, supporting the debated hypothesis that N-fixers 'trade' N for phosphatases to enhance phosphorus acquisition. Specific root length and root N differed between functional groups, though for other traits, apparent differences became nonsignificant after accounting for phylogenetic nonindependence. We conclude that evolutionary history, trait coordination, and fixation ability contribute to nutrient trait expression at the species level, and recommend explicitly considering phylogeny in analyses of functional groupings., (© 2024 The Author(s). New Phytologist © 2024 New Phytologist Foundation.)

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., (© 2024 Wiley Periodicals LLC.)

Leaves play a crucial role as ornamental organs in Spathiphyllum , exhibiting distinct differences across various Spathiphyllum varieties. Leaf development is intricately linked to processes of cell proliferation and expansion, with cell morphology often regulated by plant cell walls, primarily composed of cellulose. Alterations in cellulose content can impact cell morphology, subsequently influencing the overall shape of plant organs. Although cellulases have been shown to affect cellulose levels in plant cells, genetic evidence linking them to the regulation of leaf shape remains limited. This study took the leaves of Spathiphyllum 'Mojo' and its somatic variants as the research objects. We screened four cellulase gene family members from the transcriptome and then measured the leaf cellulose content, cellulase activity, and expression levels of cellulase-related genes. Correlation analysis pinpointed the gene SpGH9A3 as closely associated with leaf shape variations in the mutant. Green fluorescent fusion protein assays revealed that the SpGH9A3 protein was localized to the cell membrane. Notably, the expression of the SpGH9A3 gene in mutant leaves peaked during the early spread stage, resulting in smaller overall leaf size and reduced cellulose content upon overexpression in Arabidopsis .

Corn, as one of the three major grain crops in China, plays a crucial role in ensuring national food security through its yield and quality. With the advancement of agricultural intelligence, agricultural robot technology has gained significant attention. High-precision navigation is the basis for realizing various operations of agricultural robots in corn fields and is closely related to the quality of operations. Corn leaf and stalk recognition and ranging are the prerequisites for achieving high-precision navigation and have attracted much attention. This paper proposes a corn leaf and stalk recognition and ranging algorithm based on multi-sensor fusion. First, YOLOv8 is used to identify corn leaves and stalks. Considering the large differences in leaf morphology and the large changes in field illumination that lead to discontinuous identification, an equidistant expansion polygon algorithm is proposed to post-process the leaves, thereby increasing the average recognition completeness of the leaves to 86.4%. Secondly, after eliminating redundant point clouds, the IMU data are used to calculate the confidence of the LiDAR and depth camera ranging point clouds, and point cloud fusion is performed based on this to achieve high-precision ranging of corn leaves. The average ranging error is 2.9 cm, which is lower than the measurement error of a single sensor. Finally, the stalk point cloud is processed and clustered using the FILL-DBSCAN algorithm to identify and measure the distance of the same corn stalk. The algorithm combines recognition accuracy and ranging accuracy to meet the needs of robot navigation or phenotypic measurement in corn fields, ensuring the stable and efficient operation of the robot in the corn field.

Background and Scope: Plant functional traits are the result of natural selection to optimize carbon gain, leading to a broad spectrum of traits across environmental gradients. Among plant traits, leaf water storage capacity is paramount for plant drought resistance. We explored whether leaf-succulent taxa follow trait correlations similar to those of non-leaf-succulent taxa to evaluate whether both are similarly constrained by relationships between leaf water storage and climate., Methods: We tested the relationships among three leaf traits related to water storage capacity and resource use strategies in 132 species comprising three primary leaf types: succulent, sclerophyllous, and leaves with rapid returns on water investment, referred to as fast return. Correlation coefficients among specific leaf area (SLA), water mass per unit of area (WMA), and saturated water content (SWC) were tested, along with relationships between leaf trait spectra and aridity determined from species occurrence records., Results: Both SWC and WMA at a given SLA were ~10-fold higher in succulent leaves than in non-succulent leaves. While SWC actually increased with SLA in non-succulent leaves, no relationship was detected between SWC and SLA in succulent leaves, although WMA decreased with SLA in all leaf types. A principal component analysis (PCA) revealed that succulent taxa occupied a widely different mean trait space than either fast-return (P < 0.0001) or sclerophyllous (P < 0.0001) taxa along the first PCA axis, which explained 63 % of mean trait expression among species. However, aridity only explained 12 % of the variation in PCA1 values. This study is among the first to establish a structural leaf trait spectrum in succulent leaf taxa and quantify contrasts in leaf water storage among leaf types relative to specific leaf area., Conclusions: Trait coordination in succulent leaf taxa may not follow patterns similar to those of widely studied non-succulent taxa., (© The Author(s) 2024. Published by Oxford University Press on behalf of the Annals of Botany Company. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

Background and Aims: Leaf area (A) is a crucial indicator of the photosynthetic capacity of plants. The Montgomery equation (ME), which hypothesizes that A is proportional to the product of leaf length (L) and width (W), is a valid tool for non-destructively measuring A for many broadleaved plants. At present, the methods used to compute L and W for the ME can be broadly divided into two kinds: using computer recognition and measuring manually. However, the potential difference in the prediction accuracy using either method has not been thoroughly examined in previous studies., Methods: In the present study, we measured 540 Alangium chinense leaves, 489 Liquidambar formosana leaves and 215 Liriodendron × sinoamericanum leaves, utilizing computer recognition and manual measurement methods to determine L and W. The ME was used to fit the data determined by the two methods, and the goodness of fits were compared. The prediction errors of A were analysed by examining the correlations with two leaf symmetry indices (areal ratio of the left side to the right side, and standardized index for bilateral asymmetry), as well as the leaf shape complexity index (the leaf dissection index)., Key Results: The results indicate that there is a neglectable difference in the estimation of A between the two methods. This further validates that the ME is an effective method for estimating A in broadleaved tree species, including those with lobes. Additionally, leaf shape complexity significantly influenced the estimation of A., Conclusions: These results show that the use of computer recognition and manual measurement in the field are both effective and feasible, although the influence of leaf shape complexity should be considered when applying the ME to estimate A in the future., (© The Author(s) 2024. Published by Oxford University Press on behalf of the Annals of Botany Company. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

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., (© 2024. Crown.)

Various attributes are hypothesized to facilitate the dominance of an invasive species in non-native geographical and ecological regimes. To explore the characteristic invasive attributes of the family Asteraceae, a comparative study was conducted among nine species of this family, co-occurring in the western Himalayan region. Based on their nativity and invasion status, the species were categorized as "Invasive", "Naturalized", and "Native". Fifteen plant functional traits, strongly linked with invasion, were examined in the test species. The analyses revealed a strong dissimilarity between all the plant functional traits (except leaf carbon [Leaf C]) represented by "Invasive" and "Native" categories and most of the traits (except leaf area [LA], leaf nitrogen [Leaf N], Leaf C, and leaf carbon-nitrogen ratio [C: N]) represented by the "Naturalized" and "Native" categories. Similarly, "Invasive" and "Naturalized" categories also varied significantly for most of the traits (except Leaf N, Leaf C, capitula per m² population [C m² ], seeds per capitula [S capitula ], and seed mass). Invasive species are characterized by high LA, specific leaf area [SLA] and germination, and low C:N and leaf construction costs [LCC]. Most of the traits represented by native species justify their non-invasive behavior; whereas the naturalized species, despite having better size metrics (plant height), resource investment strategy (aboveground non-reproductive biomass [BNR], and aboveground reproductive biomass [BR]), and reproductive output (capitula per individual plant [C plant ], and seeds per individual plant [S plant ]) failed to invade, which implies that the role of these functional aspects in imparting invasion potential to a species is not consistent in all the ecosystems and/or phylogenetic groups. Results of PCA revealed that trait divergence plays a more imperative role in invasion success than naturalization in the species of the family Asteraceae. The present study is intended to refine the pre-generalized invasion concepts associated with family Asteraceae to ensure more accurate identification of the potential invaders and better management of the existing ones., (© 2024. The Author(s).)

Key Message: Multi-environmental characterization of flag leaf morphology traits in the US winter wheat revealed nine stable genomic regions for different flag leaf-related traits including a major region governing flag leaf angle. Flag leaf in wheat is the primary contributor to accumulating photosynthetic assimilates. Flag leaf morphology (FLM) traits determine the overall canopy structure and capacity to intercept the light, thus influencing photosynthetic efficiency. Hence, understanding the genetic control of these traits could be useful for breeding desirable ideotypes in wheat. We used a panel of 272 accessions from the hard winter wheat (HWW) region of the USA to investigate the genetic architecture of five FLM traits including flag leaf length (FLL), width (FLW), angle (FLANG), length-width ratio, and area using multilocation field experiments. Multi-environment GWAS using 14,537 single-nucleotide polymorphisms identified 36 marker-trait associations for different traits, with nine being stable across environments. A novel and major stable region for FLANG (qFLANG.1A) was identified on chromosome 1A accounting for 9-13% variation. Analysis of spatial distribution for qFLANG.1A in a set of 2354 breeding lines from the HWW region showed a higher frequency of allele associated with narrow leaf angle. A KASP assay was developed for allelic discrimination of qFLANG.1A and was used for its independent validation in a diverse set of spring wheat accessions. Furthermore, candidate gene analysis for two regions associated with FLANG identified seven putative genes of interest for each of the two regions. The present study enhances our understanding of the genetic control of FLM in wheat, particularly FLANG, and these results will be useful for dissecting the genes underlying canopy architecture in wheat facilitating the development of climate-resilient wheat varieties., (© 2024. The Author(s).)

The grass family (Poaceae) dominates ~43 % of Earth's land area and contributes 33 % of terrestrial primary productivity that is critical to naturally regulating atmosphere CO 2 concentration and global climate change. Currently grasses comprise ~11,780 species and ~50 % of them (~6000 species) utilize C 4 photosynthetic pathway. Generally, grass species have smaller leaves under colder and drier environments, but it is unclear whether the primary drivers of leaf size differ between C 3 and C 4 grasses on a global scale. Here, we analyzed 34 environmental variables, such as latitude, elevation, mean annual temperature, mean annual precipitation, and solar radiation etc., through a comparatively comprehensive database of ~3.0 million occurrence records from 1380 C 3 and 978 C 4 grass species (2358 species in total). Results from this study confirm that C 4 grasses have occupied habitats with lower latitudes and elevations, characterized by warmer, sunnier, drier and less fertile environmental conditions. Grass leaf size correlates positively with mean annual temperature and precipitation as expected. Our results also demonstrate that the mean temperature of the wettest quarter of the year is the primary control for C 3 leaf size, whereas C 4 leaf size is negatively correlated with the difference between summer and winter temperatures. For C 4 grasses, phylogeny exerts a significant effect on leaf size but is less important than environmental factors. Our findings highlight the importance of evolutionarily contrasting variations in leaf size between C 3 and C 4 grasses for shaping their geographical distribution and habitat suitability at the global scale., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

For the Caatinga vegetation, it has not yet been definitively established which adaptive strategies best define the functional groups of woody plants and which syndromes emerge from the relationships between functional traits to achieve success in a semi-arid tropical region. To fill some of these gaps, we analyzed a specific set of characteristics that make up the LHS scheme of the plant ecological strategy (leaf-heigh-seed). The LHS scheme captures the functional niche of plants regarding the functional traits specific leaf area (SLA), plant height (HE), and seed mass (SM). We associate wood density (WD) to this scheme because this trait is a key feature for the identification of functional strategies in seasonally dry vegetation such as the Caatinga. We measured these characteristics in eight woody species and classified them according to their leaf phenology into deciduous and evergreen. The experiment was carried out between December 2017 and November 2018 in Caatinga areas located in the municipality of Barra de Santana, PB, a semiarid region of Brazil. Using cluster analysis, principal component analysis (PCA) and Pearson correlation analyses, we found significant relations between SLA and SM, and HE and WD. The SLA was the main predictor of plant strategy in the Caatinga. According to this characteristic, we identified two functional groups: species with a low SLA and species with a high SLA. We also recognized differences between deciduous and evergreen species based on the same trait. The traits measured, which represent the axes of the plant ecological strategy scheme LHS and wood density, are efficient in the discrimination of functional groups in the Brazilian semiarid. These groups relate to leaf phenology of woody species in this vegetation.

Exploring the response of leaf anatomical structure to climate warming is helpful for understanding the adaptive mechanisms of trees to climate change. We conducted a warming experiment by transplanting seedlings of Larix gmelinii from 11 provenances to two common gardens, and examined the response of leaf anatomical structure to climate warming. The results showed that warming significantly increased leaf thickness (T L ), upper epidermal mesophyll thickness (T UEM ), lower epidermal mesophyll thickness (T LEM ), endodermal thickness (T E ), vascular bundle diameter (D VB ), transfer tissue thickness (T TT ), and the percentage of mesophyll thickness to T L (P MT ), and significantly decreased the upper epidermal thickness (T UE ) and the percentage of epidermal thickness to T L (P E ). The mesophyll thickness was positively associated with chlorophyll concentration and maximum net photosynthetic rate. The responses of T L , T UEM , T LEM , T E , D VB , T TT , T UE , P MT and P E to warming differed among all the provenances.As the aridity index of the original site increased, the magnitude of the warming treatment's effect decreased for T L , T UEM , T LEM , T TT and P MT , and increased for T UE and P E . Warming increased the thickness and proportion of profit tissue (e.g., mesophyll) and decreased the thickness and proportion of defensive tissue (e.g., epidermis), and those changes varied among provenances. L. gmelinii could adapt to climate warming by adjusting leaf anatomical structure, and this ability was weak for trees from provenance with high aridity index.

There is almost no scientific literature available on the phytochemistry and pharmacognosy of this plant so the basic aim of the study is to analyze pharmacognostic features of dried leaves of Anemone rupicola Camb. Morphology was studied by using the features like margins, shape, length, width, surface and so forth. For anatomical examination leaves were preserved in formalin acetic alcohol (FAA) and cutting the thin section of leaves and petiole with razor blade. Macroscopic characters of powdered drug were evaluated through organoleptic method. Qualitative phytochemical analysis was done by adopting the standard procedures. The antimicrobial activity of methanolic extract of Anemone rupicola Camb. was performed by Agar well diffusion method. The powder microscopic study showed epidermal cells, trichomes, various type of tracheids, pitted vessels, and sclerenchymatous fibers. Transverse cutting of leaves and petiole revealed the presence of different cells such as epidermis, palisade parenchyma, spongy parenchyma cells, aerenchyma, collenchyma, trichomes, and vascular bundles. The investigation of phytochemicals in distilled water, methanol and in ethanol solvent indicated the presence of various secondary compounds such as phenols, saponins, glycosides, alkaloids, tannins, cardiac glycosides, terpenoids, coumarins, and flavonoids. The methanolic extract of Anemone rupicola Camb. has potential as an antimicrobial agent. However, further studies are needed to confirm its efficacy and safety. This study can be used as powerful tool for the identification and authentication of this plant. For the purity and quality control these results will be helpful tool. This will also shed light into new areas where researchers can intervene in developing new drugs for future use. RESEARCH HIGHLIGHTS: Morphological Features of the Anemone rupicola Anatomical examination of leaves of Anemeone rupicola Microscopic features of powdered drug of leaves of Anemone rupicola Qualitative Phytochemical analysis of the leaves of Anemone rupicola., (© 2024 Wiley Periodicals LLC.)

The homology of the single cotyledon of grasses and the ontogeny of the scutellum and coleoptile as the initial, highly modified structures of the grass embryo are investigated using leaf developmental genetics and targeted transcript analyses in the model grass Zea mays subsp. mays. Transcripts of leaf developmental genes are identified in both the initiating scutellum and the coleoptile, while mutations disrupting mediolateral leaf development also disrupt scutellum and coleoptile morphology, suggesting that these grass-specific organs are modified leaves. Higher-order mutations in WUSCHEL-LIKE HOMEOBOX3 (WOX3) genes, involved in mediolateral patterning of plant lateral organs, inform a model for the fusion of coleoptilar margins during maize embryo development. Genetic, RNA-targeting, and morphological evidence supports models for cotyledon evolution where the scutellum and coleoptile, respectively, comprise the distal and proximal domains of the highly modified, single grass cotyledon., (© 2024 The Authors. New Phytologist © 2024 New Phytologist Foundation.)

Embolism resistance of xylem tissue varies among species and is an important trait related to drought resistance, with anatomical attributes like pit membrane thickness playing an important role in avoiding embolism spread. Grafted Citrus trees are commonly grown in orchards, with the rootstock being able to affect the drought resistance of the whole plant. Here, we evaluated how rootstocks affect the vulnerability to embolism resistance of the scion using several rootstock/scion combinations. Scions of 'Tahiti' acid lime, 'Hamlin', 'Pera' and 'Valencia' oranges grafted on a 'Rangpur' lime rootstock exhibit similar vulnerability to embolism. In field-grown trees, measurements of leaf water potential did not suggest significant embolism formation during the dry season, while stomata of Citrus trees presented an isohydric response to declining water availability. When 'Valencia' orange scions were grafted on 'Rangpur' lime, 'IAC 1710' citrandarin, 'Sunki Tropical' mandarin or 'Swingle' citrumelo rootstocks, variation in intervessel pit membrane thickness of the scion was found. The 'Rangpur' lime rootstock, which is known for its drought resistance, induced thicker pit membranes in the scion, resulting in higher embolism resistance than the other rootstocks. Similarly, the rootstock 'IAC 1710' citrandarin generated increased embolism resistance of the scion, which is highly relevant for citriculture., (© 2024 The Authors. Plant, Cell & Environment published by John Wiley & Sons Ltd.)

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., (© 2024 Wiley Periodicals LLC.)

Drought-induced xylem embolism is a primary cause of plant mortality. Although c. 70% of cycads are threatened by extinction and extant cycads diversified during a period of increasing aridification, the vulnerability of cycads to embolism spread has been overlooked. We quantified the vulnerability to drought-induced embolism, pressure-volume curves, in situ water potentials, and a suite of xylem anatomical traits of leaf pinnae and rachises for 20 cycad species. We tested whether anatomical traits were linked to hydraulic safety in cycads. Compared with other major vascular plant clades, cycads exhibited similar embolism resistance to angiosperms and pteridophytes but were more vulnerable to embolism than noncycad gymnosperms. All 20 cycads had both tracheids and vessels, the proportions of which were unrelated to embolism resistance. Only vessel pit membrane fraction was positively correlated to embolism resistance, contrary to angiosperms. Water potential at turgor loss was significantly correlated to embolism resistance among cycads. Our results show that cycads exhibit low resistance to xylem embolism and that xylem anatomical traits - particularly vessels - may influence embolism resistance together with tracheids. This study highlights the importance of understanding the mechanisms of drought resistance in evolutionarily unique and threatened lineages like the cycads., (© 2024 The Authors. New Phytologist © 2024 New Phytologist Foundation.)

Premise: Increases in genome size in plants-often associated with larger, low-density stomata and greater water-use efficiency (WUE)-could affect plant ecophysiological and hydraulic function. Variation in plant genome size is often due to polyploidy, having occurred repeatedly in the austral sedge genus Schoenus in the Cape Floristic Region (CFR), while species in the other major schoenoid genus in the region, Tetraria, have smaller genomes. Comparing these genera is useful as they co-occur at the landscape level, under broadly similar bioclimatic conditions. We hypothesized that CFR Schoenus have greater WUE, with lower maximum stomatal conductance (g wmax ) imposed by larger, less-dense stomata., Methods: We investigated relationships between genome size and stomatal parameters in a phylogenetic context, reconstructing a phylogeny of CFR-occurring Schoeneae (Cyperaceae). Species' stomatal and functional traits were measured from field-collected and herbarium specimens. Carbon stable isotopes were used as an index of WUE. Genome size was derived from flow-cytometric measurements of leafy shoots., Results: Evolutionary regressions demonstrated that stomatal size and density covary with genome size, positively and negatively, respectively, with genome size explaining 72-75% of the variation in stomatal size. Larger-genomed species had lower g wmax and C:N ratios, particularly in culms., Conclusions: We interpret differences in vegetative physiology between the genera as evidence of more-conservative strategies in CFR Schoenus compared to the more-acquisitive Tetraria. Because Schoenus have smaller, reduced leaves, they likely rely more on culm photosynthesis than Tetraria. Across the CFR Schoeneae, ecophysiology correlates with genome size, but confounding sources of trait variation limit inferences about causal relationships between traits., (© 2024 The Authors. American Journal of Botany published by Wiley Periodicals LLC on behalf of Botanical Society of America.)

Differences in demographic and environmental niches facilitate plant species coexistence in tropical forests. However, the adaptations that enable species to achieve higher demographic rates (e.g. growth or survival) or occupy unique environmental niches (e.g. waterlogged conditions) remain poorly understood. Anatomical traits may better predict plant environmental and demographic strategies because they are direct measurements of structures involved in these adaptations. We collected 18 leaf and twig traits from 29 tree species in a tropical freshwater swamp forest in Singapore. We estimated demographic parameters of the 29 species from growth and survival models, and degree of association toward swamp habitats. We examined pairwise trait-trait, trait-demography and trait-environment links while controlling for phylogeny. Leaf and twig anatomical traits were better predictors of all demographic parameters than other commonly measured leaf and wood traits. Plants with wider vessels had faster growth rates but lower survival rates. Leaf and spongy mesophyll thickness predicted swamp association. These findings demonstrate the utility of anatomical traits as indicators of plant hydraulic strategies and their links to growth-mortality trade-offs and waterlogging stress tolerance that underlie species coexistence mechanisms in tropical forest trees., (© 2024 The Authors. New Phytologist © 2024 New Phytologist Foundation.)

The order Sapindales is comprised of nine families and in Brazil it is represented by six, including Rutaceae Juss., which constitutes the largest group of this order. A variety of species of Zanthoxylum L. are distributed throughout the country, and among them is the species Zanthoxylum kleinii (R.S. Cowan) P.G. Waterman, which is found in the states of Brazil. This study aimed to characterize the morphoanatomy of the leaf, petiole, rachis, and stem of the species Z. kleinii. Histochemical tests were performed, and the sections were visualized under optical and scanning electron microscopy. The analysis showed that the morphoanatomical characteristics of the species are: hypoestomatic leaflets; stomata classified as anomocytic, tetracytic, and anisocytic; dorsiventral mesophyll; cavities that produce a secretion of lipid nature, present in the leaflet, rachis, and petiole; colleters distributed in the leaf; presence of simple non-glandular trichomes in all structures; and prismatic crystals in the petiole. Histochemical tests indicated the presence of phenolic and lipophilic compounds, mucilage, and lignin. With the result of this research, it was possible to identify the nature of the compounds secreted by the secretory structures of the leaves; in addition, the morphoanatomical characterization of Z. kleinii can provide relevant data for future studies for other organs of the species not yet described. Furthermore, contributing concomitantly with data for the genus, in this way, supporting to differentiate them. RESEARCH HIGHLIGHTS: Ultrastructural features observed by microscopic techniques. Calcium oxalate crystals present in the rachis. Microchemical tests confirmed the presence of colleters in the leaflet., (© 2024 Wiley Periodicals LLC.)

Mesophyll resistance for CO 2 diffusion (r m ) is one of the main limitations for photosynthesis and plant growth. Breeding new varieties with lower r m requires knowledge of its distinct components. We tested new method for estimating the relative drawdowns of CO 2 concentration (c) across hypostomatous leaves of Fagus sylvatica. This technique yields values of the ratio of the internal CO 2 concentrations at the adaxial and abaxial leaf side, c d /c b , the drawdown in the intercellular air space (IAS), and intracellular drawdown between IAS and chloroplast stroma, c c /c bd . The method is based on carbon isotope composition of leaf dry matter and epicuticular wax isolated from upper and lower leaf sides. We investigated leaves from tree-canopy profile to analyse the effects of light and leaf anatomy on the drawdowns and partitioning of r m into its inter- (r IAS ) and intracellular (r liq ) components. Validity of the new method was tested by independent measurements of r m using conventional isotopic and gas exchange techniques. 73% of investigated leaves had adaxial epicuticular wax enriched in 13 C compared to abaxial wax (by 0.50‰ on average), yielding 0.98 and 0.70 for average of c d /c b and c c /c bd , respectively. The r IAS to r liq proportion were 5.5:94.5% in sun-exposed and 14.8:85.2% in shaded leaves. c c dropped to less than half of the atmospheric value in the sunlit and to about two-thirds of it in shaded leaves. This method shows that r IAS is minor but not negligible part of r m and reflects leaf anatomy traits, i.e. leaf mass per area and thickness., (© 2024 The Authors. Plant Biology published by John Wiley & Sons Ltd on behalf of German Society for Plant Sciences, Royal Botanical Society of the Netherlands.)

Increasing planting density is a key strategy for enhancing maize yields 1-3 . An ideotype for dense planting requires a 'smart canopy' with leaf angles at different canopy layers differentially optimized to maximize light interception and photosynthesis 4-6 , among other features. Here we identified leaf angle architecture of smart canopy 1 (lac1), a natural mutant with upright upper leaves, less erect middle leaves and relatively flat lower leaves. lac1 has improved photosynthetic capacity and attenuated responses to shade under dense planting. lac1 encodes a brassinosteroid C-22 hydroxylase that predominantly regulates upper leaf angle. Phytochrome A photoreceptors accumulate in shade and interact with the transcription factor RAVL1 to promote its degradation via the 26S proteasome, thereby inhibiting activation of lac1 by RAVL1 and decreasing brassinosteroid levels. This ultimately decreases upper leaf angle in dense fields. Large-scale field trials demonstrate that lac1 boosts maize yields under high planting densities. To quickly introduce lac1 into breeding germplasm, we transformed a haploid inducer and recovered homozygous lac1 edits from 20 diverse inbred lines. The tested doubled haploids uniformly acquired smart-canopy-like plant architecture. We provide an important target and an accelerated strategy for developing high-density-tolerant cultivars, with lac1 serving as a genetic chassis for further engineering of a smart canopy in maize., (© 2024. The Author(s), under exclusive licence to Springer Nature Limited.)

The genus Tilia L. belongs to Malvaceae and it is represented by four species in Turkey. These are T. cordata, T. platyphyllos, T. rubra subsp. caucasica, and T. tomentosa. Tilia species have medicinal importance, especially to treat colds, flu, and coughs, and are also used as sedatives. Inflorescence with bract is recorded in the pharmacopeia as used parts. This study aimed to investigate the differences in leaf and bract characteristics in distinguishing Tilia species. Leaves and bracts were examined anatomically and micromorphologically under light and scanning electron microscopes (LM and SEM). There is no comparative anatomical and micromorphological study of Tilia species from Turkey. Microscopic examination allows for the observation of stomata types, trichomes, and other anatomical structures. These features can vary among species and can aid in differentiation. Features of bract that is medicinally used part were evaluated to find efficient diagnostic characters. The leaves also possess distinct characteristics, and leaf morphology is already used in the identification key in Flora. The emphasis is on identifying the species of medically used drugs. A diagnostic key was created using LM and SEM based on bract features to differentiate taxa for the first time. RESEARCH HIGHLIGHTS: Leaf and bract anatomy of Tilia taxa was studied via LM and SEM. The raw plant material is inflorescence with bract so identification by bract is important for species adulteration. An diagnostic key based on bract features was created for the first time., (© 2024 The Author(s). Microscopy Research and Technique published by Wiley Periodicals LLC.)

Transformation of plants using wild strains of agrobacteria is termed natural transformation and is not covered by GMO legislation in e.g. European Union and Japan. In the current study, offspring lines (A11 and B3) of Rhizobium rhizogenes naturally transformed oilseed rape (Brassica napus) were randomly selected to characterize the morphological traits, and analyze the implications of such morphological changes on plant drought resilience. It was found that the introduction of Ri-genes altered the biomass partitioning to above- and under-ground parts of oilseed rape plants. Compared to the wild type (WT), the A11 and B3 lines exhibited 1.2-4.0 folds lower leaf and stem dry weight, leaf area and plant height, but had 1.3-5.8 folds greater root dry weight, root length and root surface area, resulting in a significantly enhanced root: shoot dry mass ratio and root surface area: leaf area ratio. In addition, the introduction of Ri-genes conferred reduced stomatal pore aperture and increased stomatal density in the B3 line, and increased leaf thickness in A11 line, which could benefit plant drought resilience. Finally, the modulations in morphological traits as a consequence of transformation with Ri-genes are discussed concerning resilience in water-limited conditions. These findings reveal the potential of natural transformation with R. rhizogenes for drought-targeted breeding in crops., Competing Interests: Declaration of Competing Interest No conflict of interest declared, (Copyright © 2024 Elsevier B.V. All rights reserved.)