Leaf Epidermal Cells Research Articles

Association of haloacid dehydrogenase and alcohol dehydrogenase with vegetative growth, virulence and stress tolerance during tea plant infection by Didymella segeticola

Tea leaf spot, caused by the fungus Didymella segeticola, occurs in the high-mountain tea plantations of Southwest China. Due to a limited understanding of the disease's epidemiology and the lack of comprehensive control measures, it has a significant negative impact on tea yield and quality. In this study, we revealed that D. segeticola infection begins when conidia germinate to form a germ tube on the leaf surface. The fungus then grows in the intercellular spaces of the leaf epidermal cells, invading tea tissue and causing necrotic lesions. This infection leads to significant alterations in the cell walls of spongy and palisade mesophyll cells, severely damaging chloroplasts. We employed transcriptomic and metabolomic analyses based on an in vitro infection model using matcha powder to uncover two key genes of D. segeticola: DsHAD (encoding holoacid dehydrogenase) and DsADH (encoding alcohol dehydrogenase). These genes are the first to be associated with conidiation, virulence, and sensitivity to oxidative stress. DsHAD regulates the virulence of D. segeticola by modulating glutamate homeostasis. Our results elucidate the infection strategy of D. segeticola on tea leaves and provide valuable data for future research on control measures for tea leaf spot.

The antagonistic effects of red and blue light radiation on leaf and stem development of pepper (Capsicum annuum L.) seedlings

Light spectrum plays an essential role in influencing the growth and development of vegetable seedlings in industrial seedling raising. Currently, blue light, red light, and their combination are utilized in industrial seedling raising. However, the theoretical basis behind the screening of red and blue light combinations remains unclear. Therefore, we utilized pepper seedlings to investigate the effects mechanism of B (blue light, 450 nm) and R (red light, 660 nm) light-emitting diodes (pc-LED) on stem and leaf development，the full spectrum light was used as a control (W). The growth of pepper seedlings was evaluated by measuring indicators such as leaf, stem growth, and cellular ultrastructure, and try to reveal the regulatory pathways of auxin in leaves at the level of gene expression and transcriptome analysis. The results indicated that compared to W, the blue light led to shorter internodes and reduced plant height, while the red light resulted in larger leaves and elongated internodes of pepper seedlings. Ultrastructural analysis revealed that the antagonism was associated with the longitudinal elongation and transverse expansion of the stem and the expansion efficiency of leaf epidermal cells. Further analysis indicated that cell proliferation and leaf growth were regulated by the phytohormone pathway through light signaling. The blue light upregulated the expression of CaRAX in the phytohormone pathway, while red light increased the expression of CaGRF and CaARF, thereby influencing leaf size. These findings offer new insights into spectral screening for industrial seedling cultivation.

Allelopathic effects of sorghum/sorghum-sudangrass hybrids against Rotylenchulus reniformis.

Reniform nematode (Rotylenchulus reniformis) is a damaging and difficult-to-manage pest on many agricultural crops. Sorghum and sorghum-sudangrass hybrids (SSgH) have shown potential in managing plant-parasitic nematodes by releasing toxic hydrogen cyanide gas through the hydrolysis of dhurrin, a cyanogenic glucoside found in leaf epidermal cells. The objectives of this study were to evaluate the effect of different SSgH varieties and their age on the suppression of R. reniformis and to quantify their dhurrin contents. Shoot biomass of seven SSgH varieties was harvested at 1, 2, and 3 months of growth and used as a soil amendment in two greenhouse cowpea bioassay trials. Dhurrin concentration was analyzed using high-performance liquid chromatography (HPLC) from 2-month-old tissues. The results indicated that energy sorghum 'NX-D-61' and SSgH 'Latte' exhibited the highest dhurrin concentrations (P ≤ 0.05) and suppressed R. reniformis development in cowpea roots (P ≤ 0.05). In Trial I, 2-month-old amendments showed the greatest suppression, while in Trial II, 1-month-old amendments were more suppressive (P ≤ 0.05). Potential effect of environmental stress on dhurrin concentration in SSgH tissue was discussed. Nonetheless, dhurrin concentrations were negatively related to the number of R. reniformis infecting cowpea roots (r2 =0.69; P = 0.02). These findings suggest that high dhurrin SSgH varieties can be integrated into reniform nematode management plans as a cover crop and terminated no more than 2 months after planting.

The SBP-box transcription factor PlSPL2 negatively regulates stem development in herbaceous peony.

The SBP-box transcription factor PlSPL2 silencing in herbaceous peony enhanced stem strength by regulating xylem development, whereas its overexpression in tobacco resulted in weaker stem strength and undeveloped xylem. The strength of plant stems is a critical determinant of lodging resistance of plants, which significantly affects crop yield and cut-flower quality. Squamosa promoter binding (SBP) protein-like (SPL) transcription factors (TFs), participate in multiple regulatory processes, particularly in stem development. In this study, PlSPL2, an orthologous gene of Arabidopsis AtSPL2 in herbaceous peony, was isolated and found to contain a conserved SBP domain featuring two typical Zn-binding sites, as well as a nuclear localization sequence (NLS). Subsequently, transient infection of tobacco leaf epidermal cells using Agrobacterium confirmed the nuclear localization of PISPL2 protein. Additionally, gene expression analyses revealed that PlSPL2 was preferentially expressed in stems, and demonstrated a download trend in expression levels within vascular bundles during stem cell wall development. Furthermore, silencing of PlSPL2 in herbaceous peony enhanced stem strength. The silenced plants exhibited more developed xylems with wider radii and higher numbers of cell layers. Overexpression of PlSPL2 in tobacco, however, resulted in weaker stem strength, accompanied by a narrower radius of the xylem. These findings suggested that PlSPL2 was a negative regulator of herbaceous peony stem development, and its discovery and research could significantly contribute to a deeper understanding of stem growth and development mechanisms.

Pollen and Leaf Micromorphological Characteristics of Spiny Almonds (Prunus subgenus Amygdalus) in Iran.

This study investigates the micromorphological characteristics of pollen grains and leaf epidermal cells from 20 accessions across four species of spiny almonds using scanning electron microscopy. Thirteen quantitative traits of pollen grains, including exine sculpturing, were analyzed alongside qualitative features such as shape, exine sculpture type, and aperture type. Additionally, four quantitative and five qualitative features of the leaf epidermis were examined, focusing on cuticular ornamentation patterns, types of epicuticular wax, and stomatal measurements. The pollen grains were found to be isopolar monads, radially symmetric, medium-sized, varying from prolate spheroidal to prolate, and exhibiting 3-colporate to 3-colpate structures. The exine sculpturing was generally striate with short and long ridges, with or without perforations, and could be classified into three types. Notably, in the Isfahan population of Prunus lycioides and the North Khorasan population of Prunus spinosissima, the exine sculpture types were distinctly different, being rugulate and reticulate, respectively. Multivariate statistical analysis identified equatorial diameter, colpus length and width, and ridge width as key diagnostic markers for species identification within spiny almonds. Principal component analysis and hierarchical clustering further highlighted the significance of stomatal length, cuticular ornamentation patterns, and epicuticular wax types in differentiating among taxa. We conclude that groupings recognized in recent classifications of the subgenus Amygdalus (spiny almonds) remain challenging to delineate solely based on palynological data, as diverse pollen types are present across different clades and subclades. Furthermore, micromorphological leaf traits proved valuable in distinguishing certain spiny almond taxa, and the traits of epidermal cells may reflect the ecological adaptations of spiny almond species.

Image-Based Quantitative Analysis of Epidermal Morphology in Wild Potato Leaves.

The epidermal leaf patterns of plants exhibit remarkable diversity in cell shapes, sizes, and arrangements, driven by environmental interactions that lead to significant adaptive changes even among closely related species. The Solanaceae family, known for its high diversity of adaptive epidermal structures, has traditionally been studied using qualitative phenotypic descriptions. To advance this, we developed a workflow combining multi-scale computer vision, image processing, and data analysis to extract digital descriptors for leaf epidermal cell morphology. Applied to nine wild potato species, this workflow quantified key morphological parameters, identifying descriptors for trichomes, stomata, and pavement cells, and revealing interdependencies among these traits. Principal component analysis (PCA) highlighted two main axes, accounting for 45% and 21% of variance, corresponding to features such as guard cell shape, trichome length, stomatal density, and trichome density. These axes aligned well with the historical and geographical origins of the species, separating southern from Central American species, and forming distinct clusters for monophyletic groups. This workflow thus establishes a quantitative foundation for investigating leaf epidermal cell morphology within phylogenetic and geographic contexts.

Cadmium-induced protein AS8: A protein to improve Cd accumulation and transport via Cd uptake in poplar

The pollution of soil with heavy metals (HMs) has become an environmental problem of global concern. Phytoremediation, whereby plants extract HMs from soil, can efficiently and substantially reduce HM pollution in soil in an environmentally friendly manner. Cadmium-induced protein AS8 (CIPAS8) is present in many plants and its expression is induced by HMs. In this study, PeCIPAS8 and SlCIPAS8 were transformed into 84K poplar to study their effects on tolerance to, and translocation of, cadmium (Cd) in woody plants. Localization analyses showed that two CIPAS8 proteins were localized at the plasma membrane when transiently expressed in tobacco leaf epidermal cells. Compared with wild-type 84K poplar seedlings, transgenic poplar lines overexpressing PeCIPAS8 or SlCIPAS8 showed increased Cd contents and decreased Cd tolerance. Transgenic poplar lines overexpressing PeCIPAS8 or SlCIPAS8 accumulated more Cd in the roots, stems, and leaves, but the plant height did not differ significantly, compared with wild-type 84K poplar under Cd stress during the vegetative stage. CIPAS8 increased the Cd influx rate of transgenic poplar roots compared with that of the wild type, and affected the transcription levels of other metal transporters. These findings show that CIPAS8 increases Cd flux into plant tissues and demonstrate moderate Cd sensitivity of the plant. Therefore, CIPAS8 is an influx transporter with the potential to increase the uptake of toxic HMs by woody plants growing in HM-contaminated soils.

Genome-wide identification of metal tolerance protein genes in Quercus dentata and their roles in response to various heavy metal stresses

Metal tolerance protein (MTP) is a cation transporter that plays an important role in tolerance to heavy metal stress. However, thus far, there has been no genome-wide investigation of the MTP gene family in Quercus plants. Quercus dentata is one of the main constructive species of forest in northern China. It has strong tolerance to a variety of heavy metal stresses. In this study, 25 MTPs were identified from the Q. dentata genome and classified into three subfamilies and seven groups according to their sequence characteristics and phylogenetic relationships. Both tandem and segmental duplication events contributed to the expansion of the QdMTP gene family. Interestingly, all 10 tandem duplication events contributed to the expansion of the Mn-CDF subfamily. The expression of Mn-CDF subfamily members in different organs and tissues of Q. dentata was different, and they responded differently to manganese, iron, zinc and cadmium stress treatments. QdMTP10.7, a member of the Mn-CDF subfamily, enhanced yeast growth under manganese, zinc and iron stresses. The subcellular localization in tobacco leaf epidermis cells showed that QdMTP10.7 was located in vacuoles. These data generated from this study provide an important foundation to elucidate the biological roles of QdMTP genes related to heavy metal tolerance in Q. dentata.

Localization of the MTP4 transporter to trans-Golgi network in pollen tubes of Arabidopsis thaliana.

Zinc (Zn) is an essential element for plants. Numerous proteins in different cellular compartments require Zn for their structure and function. Zn can be toxic when it accumulates in high levels in the cytoplasm. Therefore, Zn homeostasis at tissue, cell, and organelle levels is vital for plant growth. A part of the metal tolerance protein (MTP) / Cation Diffusion Facilitator (CDF) transporters functions as Zn transporters, exporting Zn from the cytosol to various membrane compartments. In Arabidopsis thaliana, MTP1, MTP2, MTP3, MTP4, MTP5, and MTP12 are classified as Zn transporters (Zn-CDF). In this study, we systematically analyzed the localization of GFP-fused Zn-CDFs in the leaf epidermal cells of Nicotiana benthamiana. As previously reported, MTP1 and MTP3 were localized to tonoplast, MTP2 to endoplasmic reticulum, and MTP5 to Golgi. In addition, we identified the localization of MTP4 to trans-Golgi Network (TGN). Since MTP4 is specifically expressed in pollen, we analyzed the localization of MTP4-GFP in the Arabidopsis pollen tubes and confirmed that it is in the TGN. We also showed the Zn transport capability of MTP4 in yeast cells. We then analyzed the phenotype of an mtp4 T-DNA insertion mutant under both limited and excess Zn conditions. We found that their growth and fertility were not largely different from the wild-type. Our study has paved the way for investigating the possible roles of MTP4 in metallating proteins in the secretory pathway or in exporting excess Zn through exocytosis. In addition, our system of GFP-fused MTPs will help study the mechanisms for targeting transporters to specific membrane compartments.

Cell type specific regulation of phenolic acid and flavonoid metabolism in Taxus mairei leaves

The Taxus tree is an important industrial resource for the extraction of Taxol, a natural anticancer drug. Recently, numerous active compounds, such as phenolic acids and flavonoids, were identified in the leaves of Taxus mairei. However, the distribution and biosynthesis of phenolic acids and flavonoids in Taxus leaves are largely unknown. Integrated MALDI-IMS, ScRNA-seq and ScATAC-seq analysis was used to reveal the cell type-specific regulation of phenolic acid and flavonoid biosynthesis. Based on the cluster-specifically expressed genes, ScRNA-seq grouped the cells into 12 clusters, while ScATAC-seq grouped the cells into 9 clusters, indicating a high degree of leaf cell heterogeneity. Several marker genes were selected for cell type annotation, some of which were effectively verified by in situ hybridization. ScRNA-seq identified four phenolic acid and flavonoid biosynthesis-related genes (each one > 5 % cell coverage), which significantly expressed in the leaf epidermal cells. The cell-specific open chromatin regions surrounding the four phenolic acid and flavonoid biosynthesis-related genes were enriched in the epidermal cells. We further identified several cell type-specifically expressed transcription factors that might be involved in the biosynthesis of phenolic acids and flavonoids. Our study provided valuable information for studying the cell type specific regulation of phenolic acid and flavonoid biosynthesis in Taxus leaves.

Hierarchical global and local auxin signals coordinate cellular interdigitation in Arabidopsis.

The development of multicellular tissues requires both local and global coordination of cell polarization, however, the mechanisms underlying their interplay are poorly understood. In Arabidopsis, leaf epidermal pavement cells (PC) develop a puzzle-piece shape locally coordinated through apoplastic auxin signaling. Here we show auxin also globally coordinates interdigitation by activating the TIR1/AFB-dependent nuclear signaling pathway. This pathway promotes a transient maximum of auxin at the cotyledon tip, which then moves across the leaf activating local PC polarization, as demonstrated by locally uncaged auxin globally rescuing defects in tir1;afb1;afb2;afb4;afb5 mutant but not in tmk1;tmk2;tmk3;tmk4 mutants. Our findings show that hierarchically integrated global and local auxin signaling systems, which respectively depend on TIR1/AFB-dependent gene transcription in the nucleus and TMK-mediated rapid activation of ROP GTPases at the cell surface, control PC interdigitation patterns in Arabidopsis cotyledons, revealing a mechanism for coordinating a local cellular process with the development of whole tissues.

Morphological studies on leaf epidermis in six species of Nekemias Raf. (Vitaceae) from China

Nekemias Raf. (Vitaceae) is a re-defined genus isolated from the traditional Ampelopsis in the grape family of Vitaceae, including eight species mainly distributed in eastern Asia, and one in North America. In this study, the leaf epidermis of 28 samples representing six species of Nekemias and one species from the related genus of Ampelopsis are investigated using light microscopy. The results show that the shapes of leaf epidermal cells are polygonal or irregular. The variations of anticlinal walls are found commonly among species of Nekemia, especially within N. cantoniensis and N. grossedentata, which is probably associated with their habitats. Irregular stomata are found on the abaxial surface. The epidermal hair styles are multicellular, single-row on both sides and common in N. cantoniensis, N. grossedentata, and N. rubifolia, not in N. chaffanjonii and N. hypoglauca. Our results highlight that the variation of leaf epidermis in Nekemias are possibly related to the environment and habitats.

Living jewels: iterative evolution of iridescent blue leaves from helicoidal cell walls.

Structural colour is responsible for the remarkable metallic blue colour seen in the leaves of several plants. Species belonging to only ten genera have been investigated to date, revealing four photonic structures responsible for blue leaves. One of these is the helicoidal cell wall, known to create structural colour in the leaf cells of five taxa. Here we investigate a broad selection of land plants to understand the phylogenetic distribution of this photonic structure in leaves. We identified helicoidal structures in the leaf epidermal cells of 19 species using transmission electron microscopy. Pitch measurements of the helicoids were compared to the reflectance spectra of circularly polarised light from the cells to confirm the structure-colour relationship. By incorporating species examined with a polarising filter, our results increase the number of taxa with photonic helicoidal cell walls to species belonging to at least 35 genera. These include 23 monocot genera, from the orders Asparagales (Orchidaceae) and Poales (Cyperaceae, Eriocaulaceae, Rapateaceae) and 17 fern genera, from the orders Marattiales (Marattiaceae), Schizaeales (Anemiaceae) and Polypodiales (Blechnaceae, Dryopteridaceae, Lomariopsidaceae, Polypodiaceae, Pteridaceae, Tectariaceae). Our investigation adds considerably to the recorded diversity of plants with structurally coloured leaves. The iterative evolution of photonic helicoidal walls has resulted in a broad phylogenetic distribution, centred on ferns and monocots. We speculate that the primary function of the helicoidal wall is to provide strength and support, so structural colour could have evolved as a potentially beneficial chance function of this structure.

Single-Cell Transcriptome Atlas of Leaves at Different Developmental Stages in Populus alba × Populus glandulosa Clone 84K

Leaves are crucial photosynthetic plant organs. The development of poplar leaves has spatio-temporal specificity and it is of great significance to study the single-cell transcription atlas of leaves to reveal the temporal regulation of gene expression in different cell types. Here, single-cell RNA sequencing was performed on 17,768 tender leaf and 5846 functional leaf cells of Poplar 84K to construct a transcriptome atlas and developmental trajectory. The results showed that there were five and six cell types in tender and functional leaves, respectively. According to a pseudo-time trajectory analysis and the clustering of expressed genes into different cell types, the development of tender and functional leaves was divided into two temporal stages. Tender leaf epidermal cells developed earliest and were enriched with genes related to cell division and growth, indicating that tender leaves were in the stage of cell expansion and functional differentiation. Functional leaf palisade mesophyll cells were enriched with genes related to photosynthesis and carbon metabolism and cell types performing different functions tended to mature, indicating that functional leaves were in the stage of leaf development and the initial formation of photosynthesis. Our in-depth analysis of the transcriptional regulation at the single-cell level during leaf development provides an important basis for studying the mechanisms involved in cell differentiation and leaf development in poplar as well as other plants.

ScRNA-seq reveals dark- and light-induced differentially expressed gene atlases of seedling leaves in Arachis hypogaea L.

Although the regulatory mechanisms of dark and light-induced plant morphogenesis have been broadly investigated, the biological process in peanuts has not been systematically explored on single-cell resolution. Herein, 10 cell clusters were characterized using scRNA-seq-identified marker genes, based on 13 409 and 11 296 single cells from 1-week-old peanut seedling leaves grown under dark and light conditions. 6104 genes and 50 transcription factors (TFs) displayed significant expression patterns in distinct cell clusters, which provided gene resources for profiling dark/light-induced candidate genes. Further pseudo-time trajectory and cell cycle evidence supported that dark repressed the cell division and perturbed normal cell cycle, especially the PORA abundances correlated with 11 TFs highly enriched in mesophyll to restrict the chlorophyllide synthesis. Additionally, light repressed the epidermis cell developmental trajectory extending by inhibiting the growth hormone pathway, and 21 TFs probably contributed to the different genes transcriptional dynamic. Eventually, peanut AHL17 was identified from the profile of differentially expressed TFs, which encoded proteinlocated in the nucleus promoted leaf epidermal cell enlargement when ectopically overexpressed in Arabidopsis through the regulatory phytohormone pathway. Overall, our study presents the different gene atlases in peanut etiolated and green seedlings, providing novel biological insights to elucidate light-induced leaf cell development at the single-cell level.

Identification of bZIP Transcription Factors That Regulate the Development of Leaf Epidermal Cells in Arabidopsis thaliana by Single-Cell RNA Sequencing.

Epidermal cells are the main avenue for signal and material exchange between plants and the environment. Leaf epidermal cells primarily include pavement cells, guard cells, and trichome cells. The development and distribution of different epidermal cells are tightly regulated by a complex transcriptional regulatory network mediated by phytohormones, including jasmonic acid, and transcription factors. How the fate of leaf epidermal cells is determined, however, is still largely unknown due to the diversity of cell types and the complexity of their regulation. Here, we characterized the transcriptional profiles of epidermal cells in 3-day-old true leaves of Arabidopsis thaliana using single-cell RNA sequencing. We identified two genes encoding BASIC LEUCINE-ZIPPER (bZIP) transcription factors, namely bZIP25 and bZIP53, which are highly expressed in pavement cells and early-stage meristemoid cells. Densities of pavement cells and trichome cells were found to increase and decrease, respectively, in bzip25 and bzip53 mutants, compared with wild-type plants. This trend was more pronounced in the presence of jasmonic acid, suggesting that these transcription factors regulate the development of trichome cells and pavement cells in response to jasmonic acid.

Do Cuticular Gaps Make It Possible to Study the Composition of the Cell Walls in the Glands of Drosophyllum lusitanicum?

Carnivorous plants can survive in poor habitats because they have the ability to attract, capture, and digest prey and absorb animal nutrients using modified organs that are equipped with glands. These glands have terminal cells with permeable cuticles. Cuticular discontinuities allow both secretion and endocytosis. In Drosophyllum lusitanicum, these emergences have glandular cells with cuticular discontinuities in the form of cuticular gaps. In this study, we determined whether these specific cuticular discontinuities were permeable enough to antibodies to show the occurrence of the cell wall polymers in the glands. Scanning transmission electron microscopy was used to show the structure of the cuticle. Fluorescence microscopy revealed the localization of the carbohydrate epitopes that are associated with the major cell wall polysaccharides and glycoproteins. We showed that Drosophyllum leaf epidermal cells have a continuous and well-developed cuticle, which helps the plant inhibit water loss and live in a dry environment. The cuticular gaps only partially allow us to study the composition of cell walls in the glands of Drosophyllum. We recoded arabinogalactan proteins, some homogalacturonans, and hemicelluloses. However, antibody penetration was only limited to the cell wall surface. The localization of the wall components in the cell wall ingrowths was missing. The use of enzymatic digestion improves the labeling of hemicelluloses in Drosophyllum glands.

External jasmonic acid isoleucine mediates amplification of plant elicitor peptide receptor (PEPR) and jasmonate-based immune signalling.

Jasmonic acid-isoleucine (JA-Ile) is a plant defence hormone whose cellular levels are elevated upon herbivory and regulate defence signalling. Despite their pivotal role, our understanding of the rapid cellular perception of bioactive JA-Ile is limited. This study identifies cell type-specific JA-Ile-induced Ca2+ signal and its role in self-amplification and plant elicitor peptide receptor (PEPR)-mediated signalling. Using the Ca2+ reporter, R-GECO1 in Arabidopsis, we have characterized a monophasic and sustained JA-Ile-dependent Ca2+ signature in leaf epidermal cells. The rapid Ca2+ signal is independent of positive feedback by the JA-Ile receptor, COI1 and the transporter, JAT1. Microarray analysis identified up-regulation of receptors, PEPR1 and PEPR2 upon JA-Ile treatment. The pepr1 pepr2 double mutant in R-GECO1 background exhibits impaired external JA-Ile induced Ca2+ cyt elevation and impacts the canonical JA-Ile responsive genes. JA responsive transcription factor, MYC2 binds to the G-Box motif of PEPR1 and PEPR2 promoter and activates their expression upon JA-Ile treatment and in myc2 mutant, this is reduced. External JA-Ile amplifies AtPep-PEPR pathway by increasing the AtPep precursor, PROPEP expression. Our work shows a previously unknown non-canonical PEPR-JA-Ile-Ca2+ -MYC2 signalling module through which plants sense JA-Ile rapidly to amplify both AtPep-PEPR and jasmonate signalling in undamaged cells.

HDA6 modulates Arabidopsis pavement cell morphogenesis through epigenetic suppression of ROP6 GTPase expression and signaling.

The transcriptional regulation of Rho-related GTPase from plants (ROPs), which determine cell polarity formation and maintenance during plant development, still remains enigmatic. In this study, we elucidated the epigenetic mechanism of histone deacetylase HDA6 in transcriptional repression of ROP6 and its impact on cell polarity and morphogenesis in Arabidopsis leaf epidermal pavement cells (PCs). We found that the hda6 mutant axe1-4 exhibited impaired jigsaw-shaped PCs and convoluted leaves. This correlated with disruptions in the spatial organizations of cortical microtubules and filamentous actin, which is integral to PC indentation and lobe formation. Further transcriptional analyses and chromatin immunoprecipitation assay revealed that HDA6 specifically represses ROP6 expression through histone H3K9K14 deacetylation. Importantly, overexpression of dominant negative-rop6 in axe1-4 restored interdigitated cell morphology. Our study unveils HDA6 as a key regulator in Arabidopsis PC morphogenesis through epigenetic suppression of ROP6. It reveals the pivotal role of HDA6 in the transcriptional regulation of ROP6 and provides compelling evidence for the functional interplay between histone deacetylation and ROP6-mediated cytoskeletal arrangement in the development of interdigitated PCs.

Stomatal variations and their position relative to leaf epidermal cells in ten Maple species

Abstract In the present study, we investigated the structure of stomata in seven native species of Hyrcanian forests (Acer hyrcanum, A. velutinum Boiss., A. campestre, A. platanoides L., A. cappadocicum, A. monspessulanum, A. amazandaranicum), as well as non-native species that have fully adapted (A. negundo, A. negundo variegatum, and A. palmatum). We used light and electron microscopy to determine the form and position of the stomata in relation to the leaf epidermal cells. The length, width, shape, area, perimeter, and stomatal density were all measured. Our findings revealed that the stomata type of A. negundo varengiayum, A. campestre, A. hyrcanum, A. mazandaranicum and A. monsspesulanum is anomocytic, A. platanoides and A. cappadocicum have anomocytic stomata with wavy subsidiary cells, while A. palmatum has anisocytic stomata and A. velutinum has parasitic stomata. A. negundo has actinocytic stomata. Regarding the location of stomata relative to adjacent epidermal cells, we identified three types. In the first type, the stomata were flush with adjacent epidermal cells (A. cappadocicum, A. negundo, A. platanoides). In the second type, the stomata were higher (A. negundo variegatum), and in the third type, the stomata were lower (A. velutinum, A. monspesulanom, A. campestre, A. mazandaranicum, A. hyrcanum). The principal component analysis was used to determine the essential stomatal traits in differentiating between species. We also investigated the distribution of trees in the coordinate axis space based on two main components and performed cluster analysis based on stomatal characteristics. A. platanoides, A. negundo, A. negundo variegatum were in one cluster, while the other species were in separate clusters. The calculation of dissimilarity among the studied species revealed the lowest similarity between A. negundo and A. hyrcanum and the highest similarity between A. campestre and A. mazandaranicum. The results of the discriminant analysis identified stomatal density as the essential factor in differentiation between the studied species.