Leaf Epidermal Cells Research Articles

A network-based framework for shape analysis enables accurate characterization of leaf epidermal cells

Cell shape is crucial for the function and development of organisms. Yet, versatile frameworks for cell shape quantification, comparison, and classification remain underdeveloped. Here, we introduce a visibility graph representation of shapes that facilitates network-driven characterization and analyses across shapes encountered in different domains. Using the example of complex shape of leaf pavement cells, we show that our framework accurately quantifies cell protrusions and invaginations and provides additional functionality in comparison to the contending approaches. We further show that structural properties of the visibility graphs can be used to quantify pavement cell shape complexity and allow for classification of plants into their respective phylogenetic clades. Therefore, the visibility graphs provide a robust and unique framework to accurately quantify and classify the shape of different objects.

Expression of the SPL-like gene LaSPL9 in Japanese larch (Larix leptolepis) is regulated by miR156 during somatic embryogenesis

The Larix leptolepis SPL-like gene LaSPL9 was identified. Additionally, premiR156b and LaSPL9 expression patterns during somatic embryogenesis and the effects of gibberellin and salicylic acid on their transcription revealed the post-transcriptional regulatory functions of the miR156–LaSPL9 module. A full-length cDNA sequence of a SQUAMOSA PROMOTER-BINDING PROTEIN-LIKE (SPL) gene was isolated from Japanese larch (Larix leptolepis) and designated as LaSPL9. The cDNA comprised 2735 bp, including a 1779-bp open reading frame encoding a protein (592 amino acids) with conserved SBP domains. The LaSPL9 promoter region included several gibberellin (GA)- and salicylic acid (SA)-inducible elements. Further analyses revealed that LaSPL9 mRNA is targeted by miR156. An examination of the LaSPL9-GFP fusion protein in tobacco leaf epidermal cells indicated that LaSPL9 is a nuclear protein. Additionally, LaSPL9 expression was induced by GA and the miR156b precursor (premiR156b) transcript accumulation was inhibited by SA. Moreover, abscisic acid (ABA) was identified as the main factor down-regulating LaSPL9 expression during the early stage of somatic embryogenesis (SE), which can induce premiR156b expression. Furthermore, LaSPL9 transcription peaked at 10 days, implying it might encode an important regulator of early embryonic pattern formation. The LaSPL9 transcript level subsequently decreased gradually and was lowest at 42 days, which is consistent with the gradual increase in miR156 expression during SE. The miR156–LaSPL9 module might regulate SE in L. leptolepis by responding to ABA. Overall, our results provide evidence that miR156–LaSPL9 is important for the crosstalk between GA and SA pathways during SE.

Heat-Induced Oxidation of the Nuclei and Cytosol.

The concept that heat stress (HS) causes a large accumulation of reactive oxygen species (ROS) is widely accepted. However, the intracellular compartmentation of ROS accumulation has been poorly characterized. We therefore used redox-sensitive green fluorescent protein (roGFP2) to provide compartment-specific information on heat-induced redox changes of the nuclei and cytosol of Arabidopsis leaf epidermal and stomatal guard cells. We show that HS causes a large increase in the degree of oxidation of both compartments, causing large shifts in the glutathione redox potentials of the cells. Heat-induced increases in the levels of the marker transcripts, heat shock protein (HSP)101, and ascorbate peroxidase (APX)2 were maximal after 15 min of the onset of the heat treatment. RNAseq analysis of the transcript profiles of the control and heat-treated seedlings revealed large changes in transcripts encoding HSPs, mitochondrial proteins, transcription factors, and other nuclear localized components. We conclude that HS causes extensive oxidation of the nucleus as well as the cytosol. We propose that the heat-induced changes in the nuclear redox state are central to both genetic and epigenetic control of plant responses to HS.

Photoperiod-responsive changes in chromatin accessibility in phloem companion and epidermis cells of Arabidopsis leaves.

Photoperiod plays a key role in controlling the phase transition from vegetative to reproductive growth in flowering plants. Leaves are the major organs perceiving day-length signals, but how specific leaf cell types respond to photoperiod remains unknown. We integrated photoperiod-responsive chromatin accessibility and transcriptome data in leaf epidermis and vascular companion cells of Arabidopsis thaliana by combining isolation of nuclei tagged in specific cell/tissue types with assay for transposase-accessible chromatin using sequencing and RNA-sequencing. Despite a large overlap, vasculature and epidermis cells responded differently. Long-day predominantly induced accessible chromatin regions (ACRs); in the vasculature, more ACRs were induced and these were located at more distal gene regions, compared with the epidermis. Vascular ACRs induced by long days were highly enriched in binding sites for flowering-related transcription factors. Among the highly ranked genes (based on chromatin and expression signatures in the vasculature), we identified TREHALOSE-PHOSPHATASE/SYNTHASE 9 (TPS9) as a flowering activator, as shown by the late flowering phenotypes of T-DNA insertion mutants and transgenic lines with phloem-specific knockdown of TPS9. Our cell-type-specific analysis sheds light on how the long-day photoperiod stimulus impacts chromatin accessibility in a tissue-specific manner to regulate plant development.

Leaf morphoanatomical traits of Jacquinia armillaris Jacq. (Theophrastoideae - Primulaceae) in two xeric shrublands from Venezuela

ABSTRACT This study aimed to investigate the leaf anatomy of Jacquinia armillaris plants in two different Venezuelan xeric shrublands to evaluate possible variations caused by the environments, which differ in rainfall and soil salinity. Leaf samples were collected in two sites: La Tortuga Island, a dryer and salty environment than the other collecting site, Turpialito, a coastal location in the mainland. The epidermis and the mesophylls were observed with a light microscope and measured with an ocular micrometer inserted in one of the eyepieces. The results show that J. armillaris has many characteristic anatomical traits of xerophytes, among them: thick cuticles and thick epidermis in both leaf surfaces, stomata only in the abaxial side and epidermal depressions lodging a glandular trichome. Leaves are bifacial and present multistratified palisade parenchyma facing the adaxial epidermis and abaxial spongy parenchyma with abundant intercellular spaces. Furthermore, the hypodermis is composed of one or two cell layers below the upper epidermis. Leaf lamina dimension, petiole, cuticles, epidermis and hypodermis cells, as well as the stomatal occlusive cells and number of trichomes, had differences in size and number when the plants of both sites were compared, being higher in plants from La Tortuga. On the other hand, palisade parenchyma and stomatal density were higher in Turpialito plants. Plants collected in La Tortuga Island showed characteristics that suggest adaptation to the island’s saline and more arid conditions in comparison to plants from Turpialito, which presented more leaf traits related to adaptations to water deficit. These results demonstrate the phenotypic plasticity of J. armillaris plants that grow in two different xeric shrublands.

Chemokine-like MDL proteins modulate flowering time and innate immunity in plants

Human macrophage migration inhibitory factor (MIF) is an atypical chemokine implicated in intercellular signaling and innate immunity. MIF orthologs (MIF/D-DT-like proteins, MDLs) are present throughout the plant kingdom, but remain experimentally unexplored in these organisms. Here, we provide an in planta characterization and functional analysis of the three-member gene/protein MDL family in Arabidopsis thaliana. Subcellular localization experiments indicated a nucleo-cytoplasmic distribution of MDL1 and MDL2, while MDL3 is localized to peroxisomes. Protein–protein interaction assays revealed the in vivo formation of MDL1, MDL2, and MDL3 homo-oligomers, as well as the formation of MDL1-MDL2 hetero-oligomers. Functionally, Arabidopsismdl mutants exhibited a delayed transition from vegetative to reproductive growth (flowering) under long-day conditions, but not in a short-day environment. In addition, mdl mutants were more resistant to colonization by the bacterial pathogen Pseudomonas syringae pv. maculicola. The latter phenotype was compromised by the additional mutation of SALICYLIC ACID INDUCTION DEFICIENT 2 (SID2), a gene implicated in the defense-induced biosynthesis of the key signaling molecule salicylic acid. However, the enhanced antibacterial immunity was not associated with any constitutive or pathogen-induced alterations in the levels of characteristic phytohormones or defense-associated metabolites. Interestingly, bacterial infection triggered relocalization and accumulation of MDL1 and MDL2 at the peripheral lobes of leaf epidermal cells. Collectively, our data indicate redundant functionality and a complex interplay between the three chemokine-like Arabidopsis MDL proteins in the regulation of both developmental and immune-related processes. These insights expand the comparative cross-kingdom analysis of MIF/MDL signaling in human and plant systems.

Anatomical traits of Artemisia umbelliformis subsp. eriantha (asteraceae) alpine glacial relict from Mt. Durmitor (Montenegro)

In this study, the vegetative anatomy of Artemisia umbelliformis subsp. eriantha, a Central European alpine glacial relict, wild-growing on Mt. Durmitor (Montenegro) was examined for the first time. The aim was to investigate the general anatomy and particular anatomical traits which might have possible taxonomic value. Microscopic slides were prepared according to the standard histological procedures. The adventitious young root showed a primary structure, while the older root showed a secondary structure with a well-developed periderm on its surface. The rhizome showed a primary structure with elements of a secondary structure (periderm). The stem cross section is characterized by a round shape with a well-developed periderm at certain stages, and collateral vascular bundles arranged in a circle. The petiole is concave in shape with a single-layered epidermis and parenchyma tissue with one large and two small vascular bundles. Druses and rhombohedral crystals are observed inside some petiole parenchyma cells. The leaf lobe cross section has an oblong-linear shape and is isolateral and amphistomatous in structure. The anticlinal walls of the leaf epidermal cells are sinuate. Secretory canals are present in the root cortex parenchyma (endodermal secretory canals) and the rhizome cortical parenchyma. The stem, petiole and leaf lack secretory canals. The stem and leaf are covered with T-shaped non-glandular and glandular trichomes. The taxonomic value of the analyzed characteristics is briefly discussed.

Time-Resolved Observation of the Destination of Microinjected Potato Spindle Tuber Viroid (PSTVd) in the Abaxial Leaf Epidermal Cells of Nicotiana benthamiana.

Viroids are single-stranded noncoding RNA molecules of 250–400 nucleotides that cause plant diseases. One of the two families of viroids is Pospiviroidae, the members of which replicate in the nuclei of host cells. To replicate in plants, viroids of Pospiviroidae must enter the nucleus. However, the nuclear import of viroids remains understudied. In this work, we documented the time-dependent characteristics of the changes in microinjected fluorescently labeled potato spindle tuber viroid (PSTVd). The cytoplasmic fluorescence disappeared gradually, with only nuclear fluorescence remaining as the PSTVd injected in the cytoplasm was imported into the nucleus. Through this work, we determined that the time for half-maximal nuclear accumulation of the viroid was about 23 min. Interestingly, we found some cells where the nuclear import did not occur, despite the high level of cytosolic viroid injected. In some cells, the injected viroids disappeared within 10–20 min. The nuclear import of PSTVd is not a simple concentration-dependent process but was probably under the regulation of diverse factors that may be missing from some cells used for our observation.

Atomic Force Microscopy to Study Cell Wall Mechanics in Plants.

Atomic force microscopy (AFM) is an indentation technique used to reconstruct the topography of various materials and organisms. AFM can also measure the mechanical properties of the sample. In plants, AFM is applied to image cell wall structural details and measure the elastic properties in the outer cell walls. Here, I describe the use of high-resolution AFM to measure the elasticity of resin-embedded ultrathin sections of leaf epidermal cell walls. This approach allows to access the fine details within the wall matrix and eliminate the influence of the topography or the turgor on mechanical measurements. In this chapter, the sample preparation, AFM image acquisition, and processing of force curves are described. Altogether, these methods allow to measure the wall stiffness and compare different cell wall regions.

Armadillo-repeat kinesin1 interacts with Arabidopsis atlastin RHD3 to move ER with plus-end of microtubules

In living cells, dynamics of the endoplasmic reticulum (ER) are driven by the cytoskeleton motor machinery as well as the action of ER-shaping proteins such as atlastin GTPases including RHD3 in Arabidopsis. It is not known if the two systems interplay, and, if so, how they do. Here we report the identification of ARK1 (Armadillo-Repeat Kinesin1) via a genetic screen for enhancers of the rhd3 mutant phenotype. In addition to defects in microtubule dynamics, ER organization is also defective in mutants lacking a functional ARK1. In growing root hair cells, ARK1 comets predominantly localize on the growing-end of microtubules and partially overlap with RHD3 in the cortex of the subapical region. ARK1 co-moves with RHD3 during tip growth of root hair cells. We show that there is a functional interdependence between ARK1 and RHD3. ARK1 physically interacts with RHD3 via its armadillo domain (ARM). In leaf epidermal cells where a polygonal ER network can be resolved, ARK1, but not ARK1ΔARM, moves together with RHD3 to pull an ER tubule toward another and stays with the newly formed 3-way junction of the ER for a while. We conclude that ARK1 acts together with RHD3 to move the ER on microtubules to generate a fine ER network.

Comparative Anatomical Features Study of the Some Medicinal Rumex Species Distributed in Turkey

Distributed in Turkey Rumex crispus L., Rumex patientia L., anatomical features of Rumex obtusifolius species were comparatively investigated in this study. For anatomical studies, cross-sections were taken from root, stem and leaf. In addition, superficial sections were taken from the leaf and stomata index and stomata features were determined. Similarities and differences of taxa are revealed. There were druse crystals at the root of the R. obtusifolius plant. The radial-shaped vascular bundles in the root of R. crispus had 4-5 arms. Glandular hair was found only in the body of R. obtusifolius. It is also characteristic that vascular bundles were scattered in the stem of this species. There was cuticle in the stem of R. crispus. Stem corner numbers of taxa can also be seen as a distinguishing feature in terms of species. R. crispus was 10, R. obtusifolius was 8, R. patientia was 14 cornered. The number and type of collenchyma layer in the stem of the species were also different Outward bulge was detected in the middle-vascular region of leaves of the taxa. This bulge was less pronounced in R. crispus and the most prominent in R. obtusifolius. At least the number of stomata was observed on the R. obtusfolius leaf. Leaf epidermis cell walls were wavy in R. patientia.

A phosphoinositide 5-phosphatase from Solanum tuberosum is activated by PAMP-treatment and may antagonize phosphatidylinositol 4,5-bisphosphate at Phytophthora infestans infection sites.

Potato (Solanum tuberosum) plants susceptible to late blight disease caused by the oomycete Phytophthora infestans display enhanced resistance upon infiltration with the pathogen-associated molecular pattern (PAMP), Pep-13. Here, we characterize a potato gene similar to Arabidopsis 5-phosphatases which was identified in transcript arrays performed to identify Pep-13 regulated genes, and termed StIPP. Recombinant StIPP protein specifically dephosphorylated the D5-position of phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P2 ) in vitro. Other phosphoinositides or soluble inositolpolyphosphates were not converted. When transiently expressed in tobacco (Nicotiana tabacum) pollen tubes, a StIPP-YFP fusion localized to the subapical plasma membrane and antagonized PtdIns(4,5)P2 -dependent effects on cell morphology, indicating in vivo functionality. Phytophthora infestans-infection of N.benthamiana leaf epidermis cells resulted in relocalization of StIPP-GFP from the plasma membrane to the extra-haustorial membrane (EHM). Colocalizion with the effector protein RFP-AvrBlb2 at infection sites is consistent with a role of StIPP in the plant-oomycete interaction. Correlation analysis of fluorescence distributions of StIPP-GFP and biosensors for PtdIns(4,5)P2 or phosphatidylinositol 4-phosphate (PtdIns4P) indicate StIPP activity predominantly at the EHM. In Arabidopsis protoplasts, expression of StIPP resulted in the stabilization of the PAMP receptor, FLAGELLIN-SENSITIVE 2, indicating that StIPP may act as a PAMP-induced and localized antagonist of PtdIns(4,5)P2 -dependent processes during plant immunity.

Structure-function analysis of the maize bulliform cell cuticle and its potential role in dehydration and leaf rolling.

The hydrophobic cuticle of plant shoots serves as an important interaction interface with the environment. It consists of the lipid polymer cutin, embedded with and covered by waxes, and provides protection against stresses including desiccation, UV radiation, and pathogen attack. Bulliform cells form in longitudinal strips on the adaxial leaf surface, and have been implicated in the leaf rolling response observed in drought‐stressed grass leaves. In this study, we show that bulliform cells of the adult maize leaf epidermis have a specialized cuticle, and we investigate its function along with that of bulliform cells themselves. Bulliform cells displayed increased shrinkage compared to other epidermal cell types during dehydration of the leaf, providing a potential mechanism to facilitate leaf rolling. Analysis of natural variation was used to relate bulliform strip patterning to leaf rolling rate, providing further evidence of a role for bulliform cells in leaf rolling. Bulliform cell cuticles showed a distinct ultrastructure with increased cuticle thickness compared to other leaf epidermal cells. Comparisons of cuticular conductance between adaxial and abaxial leaf surfaces, and between bulliform‐enriched mutants versus wild‐type siblings, showed a correlation between elevated water loss rates and presence or increased density of bulliform cells, suggesting that bulliform cuticles are more water‐permeable. Biochemical analysis revealed altered cutin composition and increased cutin monomer content in bulliform‐enriched tissues. In particular, our findings suggest that an increase in 9,10‐epoxy‐18‐hydroxyoctadecanoic acid content, and a lower proportion of ferulate, are characteristics of bulliform cuticles. We hypothesize that elevated water permeability of the bulliform cell cuticle contributes to the differential shrinkage of these cells during leaf dehydration, thereby facilitating the function of bulliform cells in stress‐induced leaf rolling observed in grasses.

Astilbe uljinensis (Saxifragaceae), a new species from South Korea.

A new species Astilbe uljinensis B.U.Oh & H.J.Choi is described from Gangwon-do and Kyeongsangbuk-do in South Korea based on its morphological characteristics and distributional pattern. A. uljinensis is easily distinguished not only from three other Astilbe species in South Korea, but all other species in the genus by possessing a green young rachis, dense long whitish glandular hairs on the young rachis, dense long brownish glandular hairs on the mature inflorescence, and a slightly undulated margin of leaf epidermal cells. Specific comparisons of morphological features such as the type of the trichome, the shape of the leaf epidermis cell, and the color of the young rachis that differentiate A. uljinensis from Astilbe chinensis, another South Korean Astilbe species, are provided.

Developing a tool to shoot genes by a man-made air pressure

BackgroundBiolistic systems are used to shoot exogenous DNA, RNA, protein, and other macromolecules to transfer them into cells for genetic transformation, genome editing, and drug delivery. Such systems are especially useful for plants and other organisms that are incompatible with other macromolecule delivery methods. Commercially available, conventional biolistic systems consist of a shooting device (or “gun”) and a cylinder bottle for high-pressure helium gas. These cost a lot for installation and have low portability. ResultsWe assembled an inexpensive air duster gun and a hand pump into a portable tool to shoot genes by a man-made air pressure (TSGAMAP). TSGAMAP allows to shoot DNA-coated gold particles with the 3-MPa maximum air pressure. When DNA with a fluorescent protein gene, GFP, was shot by TSGAMAP into leaf epidermal cells of onion, leaf lettuce, and Chinese cabbage, for all of these species, some cells in all became to exhibit GFP signals. When GFP was shot with another fluorescent protein gene, mCherry, into Chinese cabbage cells, both GFP and mCherry signals were detected in some cells. When a transcription factor gene AoAMS was fused with GFP and shot into Chinese cabbage cells, nuclear-localized GFP signals were detected in some cells. These results suggest that TSGAMAP can be used for protein coexpression and protein subcellular localization analyses. ConclusionsTSGAMAP is a cost-saving and portable tool to shoot DNA and other microparticles into cells. This can expand the use of biolistics in research and education.

SlTLFP8 reduces water loss to improve water-use efficiency by modulating cell size and stomatal density via endoreduplication.

Improving plant water-use efficiency (WUE) is important to plant survival and crop yield in the context of water limitation. In this study, SlTLFP8 (Tubby-like F-box protein 8) was identified as an osmotic-induced gene in tomato. Transgenic tomato with up-regulated expression of SlTLFP8 showed enhanced water-deficient resistance, whereas knockout mutants generated by CRISPR/Cas9 were more sensitive to water deficit. SlTLFP8 overexpression significantly enhanced WUE by suppressing transpiration under both water-sufficient and water-deficient conditions. Further study showed that overexpressing SlTLFP8 significantly increased leaf epidermal cell size and thereby decreased stomatal density 10-20%, conversely SlTLFP8 knockout resulted in decreased cell size and thereby increased stomatal density 20-50%. SlTLFP8 overexpression and knockout modulated ploidy levels in leaf cells. Changes in expression of cell cycle related genes also indicated that SlTLFP8 affected cell size and stomatal density through endocycle transition. Despite changes in stomata density and transpiration, altering the expression of SlTLFP8 did not change photosynthesis. Additionally, biomass was not altered and there was little difference in fruit yield for transgenic and wild type lines under water-sufficient and water-deficient conditions. Our results demonstrate the effect of SlTLFP8 on endoreduplication and the potential of SlTLFP8 for improvement of WUE. BRIEF SUMMERY: This work found a new mechanism of TLP (Tubby like protein) response to water-deficient stress. SlTLFP8, a member of TLP family, regulates water-deficient resistance by modulating water loss via affecting stomatal density. Expression of SlTLFP8 was induced by osmotic stress. Transgenic tomato lines with SlTLFP8 overexpression or SlTLFP8 knockout showed significantly differences in water-use efficiency (WUE) and water-deficient resistance. The difference of leaf water loss caused by transpiration is the main explanation of the difference in WUE and water-deficient resistance. Additionally, overexpressing SlTLFP8 significantly decreased stomatal density, while SlTLFP8 knockout resulted in increased stomatal density, and SlTLFP8 affected stomatal density through endoreduplication and altered epidermal cell size. Despite changes in stomata density, altering the expression of SlTLFP8 did not result in distinct changes in photosynthesis, biomass and yield of tomato.

Growing straight and strong

Plant Science Wood is made of fibrils spun out of cellulose, which are major load-bearing polymers. The cellulose synthase complexes, embedded in the cell membrane, are aligned with intracellular microtubules that lie just inside the plant cell membrane. Those connections are enabled by a protein called CSI1. Bunder et al. found that aspen trees with less than the normal complement of CSI1 had twisted leaves, leaf epidermal (pavement) cells that lacked their normally refined puzzle shapes, and wood that was brittle and mechanically weak. The cause seemed to be less cellulose polymerization than normal, which undermined the sturdy, fined-grained structure of aspen wood. Plant J. 10.1111/tpj.14873 (2020).

Fine-mapping of the BjPur gene for purple leaf color in Brassica juncea.

Purple leaves are rich in health-protecting anthocyanins and food colorants in Brassica juncea. But the causal gene, which is related to leaf color formation, have not been reported in B. juncea. Anthocyanins mainly accumulated throughout the adaxial and abaxial epidermal leaf cells of purple leaves. A genetic analysis indicated that an incompletely dominant gene controls the purple leaf trait in B. juncea. Furthermore, the BjPur gene, which increased anthocyanin accumulation in purple-leafmustard, was cloned. Blast and phylogenetic analyses revealed that BjPur encodes a new R2R3-MYB transcription factor. Sequence analysis of two alleles revealed a DNA sequence insertion in the first intron of BjPur in green leaves parent line (LY) when compared with the BjPur gene in the purple-leaf parent line (ZY). And this insertion greatly reduced the transcription of BjPur in green leaves. In purple-leaf plants, the transcript level of BjPur was significantly higher in leaves than in roots, stems, siliques, and flower buds. Additionally, molecular markers linked to leaf color were developed to distinguish different genotypes of B. juncea. These results will be helpful for the genetic improvement of the purple leaf color in B. juncea.

Taxonomic Implication of Integrated Chemical, Morphological, and Anatomical Attributes of Leaves of Eight Apocynaceae Taxa

Up to now, the taxonomic conflict of the Apocynaceae family has attracted the attention of scientists and researchers worldwide. Recently, this family was divided into five subfamilies. The present study aims to investigate the implication of interlacing macro-, micro-morphological, anatomical, and chemical characteristics of the leaves of eight Apocynaceae plants (Adenium obesum, Dipladenia boliviensis, Carissacarandas, Nerium oleander, Asclepias curassavica, Calotropisprocera, Acokanthera oblongifolia, and Thevetia neriifolia), and to provide valuable taxonomic differentiation of these species. The macro-morphological investigation includes shape, apex, base, and venation of leaves, while the micro-morphological study includes leaf epidermal cells, stomata, and trichomes. The anatomical features of the leaf blade were studied by scanning electron microscope (SEM). Additionally, the chemical composition of the silylated methanolic extract was analyzed by Gas chromatography–mass spectroscopy (GC-MS). Sixty-three compounds were characterized from the silylated extracts of the eight plants, where quinic acid, sucrose, D-pinitol, and D-(−)-fructopyranose were determined as major compounds. The Principal Component Analysis (PCA) based on the chemical composition revealed a significant chemical correlation among all species with the presence of sugars and amino acids, as well as phenolic acids and iridoid glycosides. The cluster analysis, based on all merged characters, showed that the eight species can be categorized into three clusters. The first cluster comprises A.obesum, A. curassavica, and T. neriifolia, while the second cluster contains D. boliviensis, N. oleander, A. oblongifolia, and C. carandas, and the third cluster consists of C. procera alone. This cluster revealed some similarities to the recent classification of Apocynaceae, while it showed inconsistency regarding A.obesum, C. procera, and N. oleander. Due to the obtained inconsistent data and observed variation among the studied species, further study is recommended for more characterization of these species, based on additional parameters, including molecular characteristics, particularly A.obesum, C. procera, and N. oleander.

Inspired by the growth characteristic of leaf epidermis cell walls: A bio-ITCLC core for supporting a flexible skin with combined morphing applications

Inspired by low transverse and high vertical stiffness characteristics of leaf epidermis cell walls, a biomimetic Isosceles Trapezoid Corrugated Lattice Cellular (bio-ITCLC) core is designed and fabricated. The constitutive model of the bio-ITCLC sandwich panel under out-of-plane compression is built to demonstrate great potential of the bio-ITCLC core for decoupling the mutual restriction of small tangential and large normal stiffness of a flexible skin. After that, the variable stiffness, equating to the continually changing stiffness characteristic of epidermis cell walls, is introduced in the above constitutive model to capture the growth behavior of a leaf shape via epidermis cell divisions to study morphing mechanisms of a flexible skin. On this basis, by the supporting effect of self-adaption units in the bio-ITCLC core, with new circumstances, is proposed to be one promising approach for designing a flexible skin with the small tangential and large normal stiffness for multi-morphing applications.