Embryonic cuticle establishment: The great (apoplastic) divide

The plant cuticle, a dynamic interface between plants and their environment, is formed by the secretion of hydrophobic lipids and waxes into the outer wall of aerial epidermal cells. Cuticle formation is such a ubiquitous feature of epidermal cells, and is of such fundamental importance for plant survival, that identifying and understanding specific developmental roles for this structure has been a major challenge for plant scientists. In recent work, we have tried to understand the functional relationships between a signaling feedback loop required for epidermal cell specification in developing plant embryos, and a seed specific signaling cascade, involving components localized both in the embryo and in the embryo surrounding endosperm, and necessary for embryo cuticle function. Analysis of the strongly synergistic genetic relationships between these 2 independent pathways, combined with mathematical simulations of the behavior of the signaling feedback loop, have allowed us to propose an important, and hitherto unsuspected, role for the embryonic cuticle as an apoplastic diffusion barrier, necessary for preventing the excessive diffusion of developmentally important signaling molecules away from developing embryo into surrounding tissues.     

The biophysical design of plant cuticles: an overview

The outer surfaces of epidermal cell walls are impregnated with an extracellular matrix called the cuticle. This composite matrix provides several functions at the interface level that enable plants to thrive in different habitats and withstand adverse environmental conditions. The lipid polymer cutin, which is the main constituent of the plant cuticle, has some unique biophysical properties resulting from its composition and structure. This review summarizes the progress made towards understanding the biophysical significance of this biopolymer with special focus on its structural, thermal, biomechanical, and hydric properties and relationships. The physiological relevance of such biophysical properties is discussed in light of existing knowledge on the plant cuticle.     

Cuticle micromorphology of leaves of Pinus (Pinaceae) in east and south-east Asia

Cuticle micromorphology of 21 species of Pinus in east and south-east Asia was studied with scanning electron microscopy and leaf morphology was described. Cuticle characters that can be distinguished by their size, shape, and surface texture, as well as by the number of subsidiary cells, the rows of epidermal cells within a stomatal band, the number of cells between stomata in stomatal rows, and the patterns of cuticular flanges are described in detail. Most of these characters have not been considered in sufficient detail previously. Some of the inner cuticular features, such as the texture of periclinal walls, the shape of the top of the anticlinal walls, the comparative size of the lateral and polar subsidiary cells, and the development of the groove near bristles, are diagnostic characters for subgenera. The agreement between cuticle characters and taxa is discussed. Cuticular characters were used to try to resolve the position of taxa that currently appear in different places in different classifications.     

The epidermal-growth-control theory of stem elongation: an old and a new perspective

The botanist G. Kraus postulated in 1867 that the peripheral cell layers determine the rate of organ elongation based on the observation that the separated outer and inner tissues of growing stems spontaneously change their lengths upon isolation from each other. Here, we summarize the modern version of this classical concept, the "epidermal-growth-control" or "tensile skin" theory of stem elongation. First, we present newly acquired data from sunflower hypocotyls, which demonstrate that the expansion of the isolated inner tissues is not an experimental artefact, as recently claimed, but rather the result of metabolism-independent cell elongation caused by the removal of the growth-controlling peripheral walls. Second, we present data showing that auxin-induced elongation of excised stem segments is attributable to the loosening of the thick epidermal walls, which provides additional evidence for the "epidermal-growth-control concept". Third, we show that the cuticle of aerial organs can be thin and mechanically weak in seedlings raised at high humidity, but thick and mechanically important for organs growing under relatively dry air conditions. Finally, we present a modified model of the "tensile skin-theory" that draws attention to the mechanical and physiological roles of (a) the thickened, helicoidal outer cell walls, (b) the mechanical constraint of a cuticle, and (c) the interactions among outer and inner cell layers as growth is coordinated by hormonal signals.     

Tissue abscission and wound healing in the operculum of Pomatoceros lamarckii Quatrefages (Polychaeta: Serpulidae)

Following opercular amputation in Pomatoceros lamarckii Quatrefages, wound healing is initiated from a predetermined point on the peduncle. The events of abscission, cell migration and cuticle deposition during wound healing have been studied by light and electron microscopy. Abscission occurs at a predetermined point on the peduncle indicated by specialized epidermal cells, the easy break-point cells (EBP). Following detachment of tissues distal to the EBP cells, the resultant wound is plugged by a knot of coelomocytes which provide a substratum over which epidermal cells migrate to seal and restore the epidermis. During their migration, the epidermal cells undergo differentiation and deposit a new cuticle. Cuticle formation is initiated by the deposition of a finely filamentous matrix. The fine filaments subsequently coalesce to form thicker fibrils which become aggregated into layers of orthogonally-arranged fibril bundles. The mechanisms involved in abscission, cell migration and cuticle deposition during wound healing of the opercular filament are discussed.     

Structure and cuticle formation of the reticulated carapace of the ostracode Bicornucythere bisanensis

Electron microscopic examination revealed fine structures of the adult carapace of Bicornucythere bisanensis , which consists mainly of epicutiele, procuticle, membranous layer, epidermal cells and subdermal cells. Piled membrane structure is recognized as an organic framework of the procuticle. Newly formed cuticle examined with specimens at various stages of the molt cycle clarified the process of cuticle formation. Analysis of the complicated process of cuticular ridge formation permits morphogenetic discussion of surface features of the ostracode carapace of many species. Cuticle formation in close relation to cell boundaries clearly indicates that each polygon of the carapace reticulation is formed by one epidermal cell. The cell-polygon correspondence suggests that ostracode reticulation is important for elucidation of the phylogenic and ontogenic development of multicellular organisms at cell level.
□ Ostracoda, carapace structure, biomineralization, cuticle formation.     

Rumen bacterial interrelationships with plant tissue during degradation revealed by transmission electron microscopy

The mode of rumen bacterial degradation of cell walls in coastal bermudagrass [Cynodon dactylon (L) Pers.] differed with the plant tissue type. Bacteria degraded thin, primary cell walls of mesophyll and phloem apparently by extracellular enzymes and without prior attachment; thick-walled bundle sheath and epidermal cells apparently were degraded after bacterial attachment, in some types by an extracellular substance, to the plant cell walls. Rumen bacteria split the nondegraded cuticle from the epidermis by preferentially attacking the cell just underneath the cuticle. The propensity for bacterial attachment to lignified cells of the vascular tissue was low, and bacterial degradation of these cells did not occur after 72 h of incubation.     

Transgenic Arabidopsis plants expressing a fungal cutinase show alterations in the structure and properties of the cuticle and postgenital organ fusions

A major structural component of the cuticle of plants is cutin. Analysis of the function of cutin in vivo has been limited because no mutants with specific defects in cutin have been characterized. Therefore, transgenic Arabidopsis plants were generated that express and secrete a cutinase from Fusarium solani f sp pisi. Arabidopsis plants expressing the cutinase in the extracellular space showed an altered ultrastructure of the cuticle and an enhanced permeability of the cuticle to solutes. In addition, pollen could germinate on fully differentiated leaves of cutinase-expressing plants but not on control leaves. These differences coincided with strong postgenital organ fusions. The junctions of the fusions contained pectic polysaccharides. As fused organs grew apart from each other, organ deformations and protrusions of epidermal cells developed at positions with high mechanical stress. These results demonstrate that an intact cutin layer not only is important for plant-environment interactions but also prevents fusions between different plant organs and is therefore necessary for normal epidermal differentiation and organ formation.     

The presence of cutan limits the interpretation of cuticular chemistry and structure: Ficus elastica leaf as an example

Plant cuticles have been traditionally classified on the basis of their ultrastructure, with certain chemical composition assumptions. However, the nature of the plant cuticle may be misinterpreted in the prevailing model, which was established more than 150 years ago. Using the adaxial leaf cuticle of Ficus elastica, a study was conducted with the aim of analyzing cuticular ultrastructure, chemical composition and the potential relationship between structure and chemistry. Gradual chemical extractions and diverse analytical and microscopic techniques were performed on isolated leaf cuticles of two different stages of development (i.e. young and mature leaves). Evidence for the presence of cutan in F. elastica leaf cuticles has been gained after chemical treatments and tissue analysis by infrared spectroscopy and electron microscopy. Significant calcium, boron and silicon concentrations were also measured in the cuticle of this species. Such mineral elements which are often found in plant cell walls may play a structural role and their presence in isolated cuticles further supports the interpretation of the cuticle as the most external region of the epidermal cell wall. The complex and heterogeneous nature of the cuticle, and constraints associated with current analytical procedures may limit the chance for establishing a relationship between cuticle chemical composition and structure also in relation to organ ontogeny.     

DEVELOPMENT OF THE CUTICULAR LAYERS IN ANGIOSPERM LEAVES

Schieferstein , R. H., and W. E. Loomis . (Iowa State U., Ames.) Development of the cuticular layer in angiosperm leaves. Amer. Jour. Bot. 46(9): 625–635. Illus. 1959.—The cuticularized layers of leaves and other plant surfaces consist of a primary cuticle, formed by the oxidation of oils on exposed cell walls, plus various surface and subsurface wax deposits. The primary cuticle appears to form rapidly on the walls of any living cell which is exposed to air. Surface wax is present on the mature leaves of about half of the 50 or 60 species studied. In general, wax is extruded at random through the newly formed cuticle of young leaves and accumulated in various reticulate to semicrystalline patterns. No wax pores through the cuticle or primary wall can be observed in electron-micrographs of dewaxed mature leaves. Wax accumulations on older leaves are generally subcuticular and may involve the entire epidermal wall. These deposits appear to be of considerably greater ecological significance than those on the surface. Isolated cuticular membranes from Hedera helix increased slightly in permeability to water with age of the leaf, but permeability to 2,4-D decreased 50 times. Evidence based on the patterns of cellulose in primary walls, of surface wax on growing leaves, of the appearance of the cuticle at the margins of growing epidermal cells, of the forms of the cuticle plates digested from growing and older leaves, and of the marginal location of new wax deposits on growing maize leaves is presented to support the thesis that the enlargement of the outer surface of the epidermal cells of leaves occurs at the margins of the surface. Earlier formed cuticle and wax are thus undisturbed during growth. These observations, coupled with evidence for apical growth in fibers, root hairs, etc. suggest that the primary walls of angiosperm cells are formed in specific, localized growth regions, rather than by plastic extension and apposition.     

Epidermal and hypodermal characteristics in North American Cactoideae (Cactaceae)

Dermal and hypodermal anatomical features of 70 species representing 21 genera of North American Cactoideae were studied. Results show that all species examined have parallelocytic stomata and anticlinal wall surface varies from straight to undulate. Cuticle thickness is mostly narrow (1–10 μm) contrary to the general opinion that cuticle is thick in most cacti; however, few species such as Ariocarpus fissuratus and several species of Pachycereus show a distinctive thick cuticle. More than 80% of the species studied have a single-layered epidermis. Papillae occur in eight species belonging to four genera. Notable papillae are a feature shared by all members of Peniocereus subg. Peniocereus. Other species show a bullate surface produced by irregular patches of secondary epidermal cell divisions. Commonly, the hypodermis is composed of more than two cell layers with distinctive collenchymatous walls as reported in many South American species. Silica bodies, prismatic crystals, druses, sphaerocrystals, and tannins are the most common cellular inclusions that distinguish several genera and appear to have taxonomic value. However, a more thorough search in species of Cephalocereus, Coryphantha, Echinocereus, Mammillaria, Neobuxbaumia, Pilosocereus, and Turbinicarpus is needed to support the previous assertion. Electronic Publication     

Epidermal structure and development of stomata in vegetative and floral organs ofHybanthus enneaspermus (Linn.)F. Muell

The present paper deals with the epidermal structure and ontogeny of stomata in vegetative and floral organs ofHybanthus enneaspermus. The epidermal cells are either polygonal or elongated with straight, sinuous or arched thick anticlinal walls. The surface of the cuticle shows parallel striations radiating from the guard cells or hair bases. Unicellular and uniseriate bicellular trichomes with verrucose margins have been observed on all organs. The mature stomata are anisocytic, paracytic, anomocytic and transitional between anisocytic and paracytic. The ontogeny of anisocytic and paracytic stomata is syndetocheilic or mesogenous, anomocytic is haplocheilic or perigenous, while that of the transitional type is mesoperigenous. Four types of stomata have been observed on all the vegetative and floral organs and their ontogeny from organ to organ of this plant is constant. Stoma with a single guard cell is the result of disintegration of one of the guard cells before or after pore formation. Contiguous stomata are also occasionally noticed.     

Partial purification of tomato fruit peroxidase and its effect on the mechanical properties of tomato fruit skin

Peroxidase (EC 1.11.1.7)-mediated stiffening of cell walls within the fruit skin of tomato is hypothesized to regulate fruit growth. However, to date, there is no experimental evidence demonstrating that peroxidase affects the mechanical properties of skin tissue. Here, the mechanical properties of skin strips excised from a range of fruits at different ages were determined using an 'Instron' universal material testing instrument. The stiffness of tomato fruit skin strips increases 3-fold with increasing fruit age. Application of partially-purified peroxidase from the cell walls of mature tomato fruit skin significantly increased the stiffness of fruit skin irrespective of the age of fruit. Furthermore, the application of hydrogen peroxide significantly increased the stiffness of skin strips excised from fruit of an age when endogenous peroxidase isozymes associated with the termination of growth are first detected. The results support the hypothesis that the tomato fruit skin plays an integral role in the regulation of tomato fruit growth, and that changes in its mechanical properties may be mediated by peroxidase. As far as is known, this is the first demonstration that peroxidases alter the mechanical properties of the plant cell wall.     

Leaf biomechanics, morphology, and anatomy of the deciduous mesophyte Prunus serrulata (Rosaceae) and the evergreen sclerophyllous shrub Heteromeles arbutifolia (Rosaceae)

Leaf tensile properties were compared between the mesic deciduous tree Prunus serrulata (var. "Kwanzan") and the xeric and sclerophyllous chaparral evergreen shrub Heteromeles arbutifolia (M. Roem). All values for biomechanical parameters for H. arbutifolia were significantly greater than those of P. serrulata. The fracture planes also differed between the two species with P. serrulata fracturing along the secondary veins, while H. arbutifolia most often fractured across the leaf irrespective of the vein or mesophyll position, thus yielding qualitative differences in the stress-strain curves of the two species. Anatomically, P. serrulata exhibits features typical for a deciduous mesophytic leaf such as a thin cuticle, a single layer of palisade mesophyll, isodiametric spongy mesophyll, and extensive reticulation of the laminar veins. Heteromeles arbutifolia leaves, however, are typically two- to three-fold thicker with a 35% higher dry mass/fresh mass ratio. The vascular tissue is restricted to the interface of the palisade and spongy mesophyll near the center of the leaf. Both epidermal layers have a thick cuticle. The palisade mesophyll is tightly packed and two to three layers thick. The spongy mesophyll cells are ameboid in shape and tightly interlinked both to other spongy cells as well as to the overlying palisade layer. We conclude that the qualitative and quantitative biomechanical differences between the leaves of these two species are likely due to a complex interaction of internal architectural arrangement and the physical/chemical differences in the properties of their respective cell walls. These studies illustrate the importance that morphological and anatomical correlates play with mechanical behavior in plant material and ultimately reflect adaptations present in the leaves of chaparral shrubs that are conducive to surviving in arid environments.     

植物角质层基因研究进展

角质层是形成于陆生植物表皮细胞壁外表面的脂质保水层。角质层的基本功能是保水,同时也在响应逆境胁迫、自我清洁及器官发育等方面发挥作用。角质层通常由角质和蜡质组成。角质是角质层的主要结构成分,其主要组分是聚酯。蜡质成分主要为极长链饱和脂肪酸及其衍生物。这些组分在内质网上合成后被转运到细胞表面,进一步形成完整的角质层结构。近年来通过对角质层相关突变体及相应基因的研究,人们对角质层在合成、转运、形成及调控等各个阶段都有了较为深入的认识。蜡质和角质的合成途径已在角质层相关基因功能的解释下逐渐浮出水面。有关角质层前体转运方面的研究,主要的突破在于ABCG全转运蛋白的发现和功能解析。在角质层形成的机理方面,角质层基因中的酯酶和脂酶类基因的研究有助于进一步认识这个复杂的过程。在基因调控方面,新的转录因子基因和角质层与环境之间的相互关系研究,也为已知的调控网络增加了新内容。该文综述了目前关于角质层相关基因的最新研究进展。     

The stabilization of locust cuticle

Cuticle from the metathoracic femur of adult locusts (Locusta migratoria) is characterized with respect to changes in water content and in protein extractability during maturation. The swelling behaviour and extractability of fully-sclerotized cuticle are compared to those of chemically-modified, unsclerotized cuticle.It is concluded that although dehydration may contribute to the stabilization of cuticle, it cannot account for the observed differences. The properties of mature cuticle can best be explained by the assumption that covalent cross-links are present between protein molecules.     

The importance of leaf cuticle for carbon economy and mechanical strength

Cuticle thickness of leaves varies >?100 times across species, yet its dry mass cost and ecological benefits are poorly understood. It has been repeatedly demonstrated that thicker cuticle is not superior as a water barrier, implying that other functions must be important. Here, we measured the mechanical properties, dry mass and density of isolated cuticle from 13 evergreen woody species of Australian forests. Summed adaxial and abaxial cuticle membrane mass per unit leaf area (CMA) varied from 2.95 to 27.4?g m(-2) across species, and accounted for 6.7-24% of lamina dry mass. Density of cuticle varied only from 1.04 to 1.24?g?cm(-3) ; thus variation in CMA was mostly due to variation in cuticle thickness. Thicker cuticle was more resistant to tearing. Tensile strength and modulus of elasticity of cuticle were much higher than those of leaf laminas, with significant differences between adaxial and abaxial cuticles. While cuticle membranes were thin, they could account for a significant fraction of leaf dry mass due to their high density. The substantial cost of thicker cuticle is probably offset by increased mechanical resistance which might confer longer leaf lifespans among evergreen species.