Quantitative assessment to the structural basis of water repellency in natural and technical surfaces

Many plant surfaces are water-repellent because of a complex 3-dimensional microstructure of the epidermal cells (papillae) and a superimposed layer of hydrophobic wax crystals. Due to its surface tension, water does not spread on such surfaces but forms spherical droplets that lie only on the tips of the microstructures. Studying six species with heavily microstructured surfaces by a new type of confocal light microscopy, the number, height, and average distance of papillae per unit area were measured. These measurements were combined with those of an atomic force microscope which was used to measure the exposed area of the fine-structure on individual papillae. According to calculations based upon these measurements, roughening results in a reduction of the contact area of more than 95% compared with the projected area of a water droplet. By applying water/methanol solutions of decreasing surface tension to a selection of 33 water-repellent species showing different types of surface structures, the critical value at which wetting occurs was determined. The results impressively demonstrated the importance of roughening on different length scales for water-repellency, since extremely papillose surfaces, having an additional wax layer, are able to resist up to 70% methanol. Surfaces that lack papillae or similar structures on the same length scale are much more easily wetted.     

Seasonal changes of leaf surface contamination in beech, oak, and ginkgo in relation to leaf micromorphology and wettability

The leaf surfaces of beech, oak and ginkgo have been investigated with respect to contamination with particles during one growing season. Based on the observation that particles are removed from water-repellent leaves by rain (Lotus effect) the three species were selected because they differ in leaf surface micromorphology and wettability. Leaves of beech are smooth, lacked wax crystals and were ±wettable. Those of ginkgo were rough because their cells were convex and were densely covered by wax crystals, resulting in permanent water repellency. Leaves of oak were covered by waxes and were water repellent when young, but, a few weeks after leaf expansion had ceased the waxes were rapidly eroded. These differences in wettability resulted in different amounts of contamination. Ginkgo collected a very small number of particles during the whole vegetation period. In beech the contamination was significantly higher, but fairly constant, whereas oak leaves accumulated particles with age.     

西安市常见绿化植物叶片润湿性能及其影响因素

利用接触角测定仪测定了西安市21种常见绿化植物叶片表面的接触角,探讨了叶片表面特性如蜡质、绒毛、气孔对接触角的影响。结果表明,植物叶片正背面、物种间的接触角差异均显著,叶片正面和背面接触角大小在40°～140°。接触角大小与变异系数呈负相关,可能由于接触角小的润湿叶片在不同的生境和位置下,受到环境条件的影响较大而出现大的变异;接触角较大的非润湿性叶片,环境物质持留时间较短,对叶片形态和组成影响较小,因而出现小的变异。植物叶片表面的接触角随蜡质含量的升高而增大。表皮蜡质去除后大部分叶片接触角明显降低,尤其是疏水性较强的银杏(Ginkgo biloba)、月季(Ro-sa chinensis)和紫叶小檗(Berberis thunbergii)。女贞(Ligustrum lucidum)正背面、加杨(Popu-lus canadensis)背面等亲水型的叶片蜡质去除后接触角反而增大。叶片绒毛的多少及其形态、分布方式对接触角具有重要的影响,不同的作用方式表现出润湿和不润湿的特征,人为将其去除可以增加叶片的润湿性。背面气孔密度与气孔长度、保卫细胞长度呈负相关;接触角则与气孔密度呈负相关,与气孔长度呈正相关。     

Attachment ability of the beetle Chrysolina fastuosa on various plant surfaces

To study the role of different structures of a plant surface preventing insect attachment, a variety of plant surfaces were screened. Attachment ability of the beetle Chrysolina fastuosa Scop. (Coleoptera, Chrysomelidae) was measured on 99 surfaces among them smooth, hairy, felt-like, waxy, and glandular ones of three plant organs (stems, leaves, fruits) of 83 plant species belonging to 45 families. Insects attached successfully to smooth, hairy, and felt-like substrata. These surface types did not effect the further attachment of C. fastuosa, indicating the adhesive system remained intact after contacting these substrata. However, the beetles could not attach properly to surfaces covered with wax crystalloids or glandular hairs. In most experiments on pruinose plant substrata, no influence of the surfaces on the subsequent attachment ability of insects was observed. Only in one case (the stem of Acer negundo), was such an impairment recorded, but recovery of attachment ability was fast. Crystalloids of this plant species probably temporarily disable function of tenent setae of C. fastuosa. Four hypotheses, explaining anti-adhesive properties of plant surfaces, covered with wax crystalloids are proposed. A plant surface with glandular trichomes disabled the attachment system of the beetle for a long time. Secretions of trichomes probably glue tenent setae together making further attachment impossible.     

Leaf Epidermal Cells： A Trap for Lipophilic Xenobiotics

Plant surfaces are covered by a layer of cuticle, which functions as a natural barrier to protect plants from mechanical damage, desiccation, and microbial invasion. Results presented in this report show that the epicuticular wax and the cuticle of plant leaves also play an important role in resisting xenobiotic invasion. Although the epicuticular wax is impermeable to hydrophilic xenobiotics, the cuticle not only restricts the penetration of hydrophilic compounds into leaf cells, but also traps lipophilic ones. The role of the epidermal cells of plant leaves in resisting xenobiotic invasion has been neglected until now. The present study shows, for the first time, that the epidermal cells may reduce or retard the transport of lipophilic xenobiotics into the internal tissues through vacuolar sequestration. Although the guard cells appear to be an easy point of entry for xenobiotics, only a very small proportion of xenobiotics present on the leaf surface actually moves into leaf tissues via the guard cells .     

The cuticle micromorphology of in situ Erica arborea L. exposed to long-term volcanic gases

Epidermis micromorphology of in situ Erica arborea L. exposed for generations to long-term effect of volcanic gases in Pisciarelli and Solfatara di Pozzuoli areas have been studied by X-ray analyses, SEM and TEM observations. In particular, the aim of this study is to investigate the effects of volcanic gases on extant and possibly fossil plants. Plants of the same species living in a nearby control site were also studied for comparison. SEM coupled with EDX analysis was used to localize different elements within the leaves (mesophyll, cell wall and cuticle). After conventional and cryo preparation, SEM of mature leaves ascertained that the abaxial side is more serrate in fumigated leaves and hairs, epicuticular wax alterations have also been noted. Leaves experiencing chronic fumigation display stomata more sunken with respect to the epidermal surface. TEM of transverse and longitudinal sections of cuticle showed an outer A2 granular amorphous layer and external to a B1 fibrillous layer. Significant statistical variations of ultrastructural components of the cuticle revealed a response of E. arborea to this extreme environment. At the ultrastructural level, significant variations in thickness of the cell wall plus cuticle, cell wall and A2 layer among fumigated and non-fumigated leaves have been found. In the studied localities a positive correlation between atmospheric CO₂ concentration and the thickness of A2 layer also exists. The results are of interest being applicable in the understanding of plant cuticle responses during periods of normal vs. volcanic activity.     

Phylloepiphytic interaction between bacteria and different plant species in a tropical agricultural system

Plant surfaces are a favourable niche for bacterial establishment, and hypothetically, plant species differ in their capacity to harbour epiphytic bacterial communities. This study was conducted to evaluate and describe the structural relationship of a bacterial community at the phyllosphere level with different plant species in a tropical ecosystem. Leaf blades of 47 plant species distributed in 27 botanical families were collected on a typical small Brazilian farm and prepared for observation under light and scanning electron microscopy. Naturally occurring bacteria were the most abundant settlers of the phylloplane, followed by fungal spore or hyphae. All plant species studied were colonized by phylloepiphytic bacteria, which were observed as solitary cells, microcolonies, and biofilms. However, independent of the family, the plant species differed in the pattern of phyllosphere colonization, as reflected in bacteria frequency and presence or absence of anatomical features that would favour the association. The phylloepiphytic bacteria were preferentially established on the following sites: epidermal cell wall junctions, glandular and nonglandular trichomes, veins, stomata, and epidermal cell wall surface. Profuse bacteria and fungi colonization was observed, at a level that was at least comparable with temperate regions. Interestingly, fungi seemed to alter the bacteria colonization pattern, most probably by microenvironmental modifications. The trichome type and density as well as the presence of epicuticular wax on the leaf blade surface seemed to be the most determinant anatomical features for the pattern of phyllosphere colonization. The presence of trichomes has a favourable, and epicuticular wax an unfavourable influence on the plant-bacteria interaction.     

Non-Smooth Morphologies of Typical Plant Leaf Surfaces and Their Anti-Adhesion Effects

The micromorphologies of surfaces of several typical plant leaves were investigated by scanning electron microscopy（SEM）. Different non-smooth surface characteristics were described and classified. The hydrophobicity and anti-adhesion of non-smooth leaf surfaces were quantitatively measured. Results show that the morphology of epidermal cells and the morphology and distribution density of epicuticular wax directly affect the hydrophobicity and anti-adhesion. The surface with uniformly distributed convex units shows the best anti-adhesion, and the surface with regularly arranged trellis units displays better anti-adhesion. In contrast, the surface with randomly distributed hair units performs relatively bad anti-adheslon. The hydrophobic models of papilla-ciliary and fold-setal non-smooth surfaces were set up to determine the impacts of geometric parameters on the hydrophobicity. This study may provide an insight into surface machine molding and apparent morphology design for biomimetics engineering.     

The outer epidermal wall structure of African Stapelieae (Asclepiadaceae)

The outer epidermal walls of 65 species incorporated in 14 genera of the tribe Stapelieae were investigated by light and electron microscopy (TEM, SEM). The species, all of them stem succulents preferring the semi-arid regions of the Palae-otropics, possess an outer epidermal wall, which is relatively simple in structure. The whole plant body is covered by a massive wax layer, in addition some species bear wax crystals. A cutin layer follows beneath, reticulate over its whole expanse and not divided into a cuticle proper and cutinized layers. A cellulose layer, which often exhibits enclosed cutinbodies, delimits the wall against the plasmalemma.     

Anatomical studies of Carex cuchumatanensis,C. decidua,and C. hermannh (Cyperaceae) and comparisons with north American taxa of the C. acuta complex

Foliar anatomy and micromorphology of perigynia and achenes were investigated for three taxa of theCarex acuta complex of sect.Phacocystis Dumort. in Middle and South America. Anatomical characters appear to be conservative, and show little variation amongC. cuchumatanensis Standl. & Steyerm.,C. decidua F. Boott, andC. hermannii Cochrane. Leaves of all species are amphistomatous, with stomates and papillae on both epidermal surfaces, and have a single layer of bulliform cells but differ in the number of vascular bundles. Perigynia have epidermal cells with thin, convex, outer walls that are collapsed in dired specimens, and epidermal cells of achenes each have a single, rounded, nodulose silica-body with a basal platform. Anatomical data are useful in resolving the affinities among these species, and show that they are anatomically indistinguishable fromC. lenticularis Michx. These four New World endemics differe substantially fromC. nigra (L.) Reichard.     

植物角质层蜡质的化学组成研究综述

角质层是植物与外界的第一接触面,而角质层蜡质则是由位于角质层外的外层蜡质和深嵌在角质层中的内层蜡质两部分构成。植物角质层蜡质成分极其复杂,具有重要的生理功能。综述了有关植物角质层蜡质的化学组成信息,探讨了目前植物角质层蜡质化学成分研究中存在的一些问题,展望了角质层蜡质成分的研究前景。     

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.     

Wax as a Mechanism for Protection against Photoinhibition — A Study of Cotyledon orbiculata

The leaves of the Crassulacean acid metabolism plant Cotyledon orbiculata have a waxy coating which is highly reflective but can be easily removed by brushing. This provided an ideal system in which to investigate the role of epidermal wax as a possible photoprotectant. Removal of the wax, prior to exposure to natural sunlight, resulted in substantial decreases in F_v/F_m and in severe cases evidence of photoinhibitory damage, as indicated by a rise in F_o. Leaves from which wax had been removed also showed higher conversion of violaxanthin to zeaxanthin than waxed leaves. Recovery of brushed leaves over a 12 day period was correlated with an increase in the total pool of xanthophyll cycle components. This study suggests that the presence of highly reflective wax on the epidermis may confer significant photoprotection to plants exposed to high solar radiation environments.     

Epicuticular wax structures on stems and comparison between stems and leaves – A survey

The cuticle, forming the outermost layer of plant tissues and being in direct contact with the environment, consists of waxes and cutin. Waxes are hydrophobic substances that are divided in two groups: intra- and epicuticular, depending on their localisation. Epicuticular waxes appear as smooth coverings, however, many plants also produce superimposed wax structures of a crystalline nature. While studies of waxes have almost exclusively focused on leaves, here a survey of epicuticular wax structures on stems is presented. The stem surface of 343 higher plant taxa, representing 80 families, was examined using scanning electron microscopy. The adaxial and abaxial surfaces of leaves of 319 taxa were also examined to determine the relationship between wax structures on stems and leaves. Wax structures are classified, described and discussed. The results of the study indicate that stems exhibit the same main wax crystal types that have been described for leaves. Seventy percent of the examined taxa produced wax crystals on their stems. In ∼24% of the taxa, wax crystals were absent on leaves and found only on stems. In plant taxa that produce wax crystals, 40% exhibit the same type on either side of their leaves and on their stem. However, a much stronger morphological similarity exists between crystal shapes present on the adaxial and abaxial surfaces of leaves than between those present on the stem and those on leaves. In general, these observations suggest that stems are quite different than leaves in terms of their epicuticular wax structures.     

植物角质层内外蜡质的差异及其与抗逆性的关系

植物角质层是覆盖在植物地上部分的叶、花和非木质茎等器官表面的保护层,包括角质和蜡质。其中蜡质根据分布位置不同又分为表皮蜡质和内部蜡质。大量研究表明,表皮蜡质含量和结构在植物生长发育和抗逆性申发挥着重要作用。近年来有研究发现构成蜡质的成分在内外蜡质层中的分布存在差异,角质层蜡质成分影响植物抗逆性。本文针对角质层结构和内外蜡质差异性以及角质层结构和组成与植物抗逆性之间的关系进行了综述。     

Comparative Epidermal Ultrastructure of Cotton (Gossypium hirsutum L.) Leaf, Bract and Capsule Wall

Cotton leaves are more physiologically active than the bractand the capsule wall of the fruiting structures. To elucidatethe disparities in their physiological behaviour, epidermalcell density, stomatal index, stomatal size, trichome densityand type, and epicuticular wax ultrastructure of cotton leaf,bract and capsule wall were delineated using scanning electronmicroscopy (SEM). The epidermal cells of the outer periclinalwalls on both surfaces of the leaf and bract were raised andconvex. Conversely, the capsule wall epidermal cells were polygonalwith flat outer periclinal walls. The stomatal complex in theleaf and bract was paracytic, whereas in the capsule wall thestomatal complex was anomocytic. The adaxial and abaxial stomataof the leaf were coplanar to the epidermal surface, as opposedto the raised adaxial stomata on the bract. On the contrary,the stomata on the capsule wall surface appeared to be slightlysunken. Furthermore, the capsule wall stomata were larger thanthe stomata on either surface of both the leaf and the bract.The stomatal index was greater on the abaxial surfaces of theleaf and the bract (18.4 and 9.4, respectively) than their correspondingadaxial surfaces (14.4 and 4.7, respectively). Leaves had thehighest stomatal index followed by the bract and the capsulewall. The indumentum consisted of glandular and nonglandulartrichomes, the density of which was greater on the abaxial surfacesthan on the adaxial surfaces of the leaf and bract. The capsulewall indumentum lacked nonglandular trichomes. Epicuticularwax occurred in the form of striations. However, the striationpattern varied among the organs. This study clearly illustratesmorphological disparities in the epidermal features of leaf,bract and capsule wall, helping to explain their physiologicaldivergence. Copyright 2000 Annals of Botany Company Gossypium hirsutum, epicuticular wax, raised stomata, scanning electron microscopy, stomatal index, trichomes     

Surface wax,structure and function in leaves of Polytrichaceae

Abstract

The form and distribution of epicuticular wax has been examined by SEM in 35 species of Polytrichaceae. Australian and New Zealand species of Dawsonia have a dense covering of prominent angular platelets (or more irregular wax) on the lamella-margins; Polytrichadelphus magellanicus is very similar. In Polytrichum the epicuticular wax forms an angular to rounded granularity which varies in quantity from species to species; abundant and prominent on the lamella-margins of, e.g., P. formosum and P. commune, hardly detectable in, e.g., P. sexangulare. Pogonatum spp. show sparse to dense angular to rounded granularity on both surfaces, generally more abundant on the upper surface but (with the exceptions of P. grandifolium and (especially) P. urnigerum) less differentiated on the lamella-margins than in Dawsonia or Polytrichum. The parallel between Polytrichum alpinum, Pogonatum grandifolium, P. urnigerum, Psilopilum australe and P. crispulum is noteworthy. Oligotrichum, Batramiopsis, Atrichum and the other species of Psilopilum show either no apparent surface wax or only a fine surface granularity, although their leaves are generally water-repellent, sometimes strongly so. Dendroligotrichum dendroides has almost smooth lamella-margins; D. squamosum has abundant and prominent surface wax.

A major function of epicuticular wax in Polytrichaceae is probably to exclude water from the ‘pseudo-mesophyll’ formed by the lamellae, a function supplemented by the inflexed leaf margins of Polytrichum section Juniperifolia. It is best developed on the lamella-margins of the species of Dawsonia, Polytrichadelphus, Polytrichum and Pogonaturn which probably most nearly approach mesophytic vascular plants in their pattern of adaptation.     

Composition differences between epicuticular and intracuticular wax substructures: how do plants seal their epidermal surfaces?

The protective wax coating on plant surfaces has long been considered to be non-uniform in composition at a subcellular scale. In recent years, direct evidence has started to accumulate showing quantitative compositional differences between the epicuticular wax (i.e. wax exterior to cutin that can be mechanically peeled off) and intracuticular wax (i.e. wax residing within the mechanically resistant layer of cutin) layers in particular. This review provides a first synthesis of the results acquired for all the species investigated to date in order to assign chemical information directly to cuticle substructures, together with an overview of the methods used and a discussion of possible mechanisms and biological functions. The development of methods to probe the wax for z-direction heterogeneity began with differential solvent extractions. Further research employing mechanical wax removal by adhesives permitted the separation and analysis of the epicuticular and intracuticular wax. In wild-type plants, the intracuticular (1-30 μg cm(-2)) plus the epicuticular wax (5-30 μg cm(-2)) combined to a total of 8-40 μg cm(-2). Cyclic wax constituents, such as triterpenoids and alkylresorcinols, preferentially or entirely accumulate within the intracuticular layer. Within the very-long-chain aliphatic wax components, primary alcohols tend to accumulate to higher percentages in the intracuticular wax layer, while free fatty acids and alkanes in many cases accumulate in the epicuticular layer. Compounds with different chain lengths are typically distributed evenly between the layers. The mechanism causing the fractionation remains to be elucidated but it seems plausible that it involves, at least in part, spontaneous partitioning due to the physico-chemical properties of the wax compounds and interactions with the intracuticular polymers. The arrangement of compounds probably directly influences cuticular functions.     

Interfacial Effects of Superhydrophobic Plant Surfaces： A Review

Nature is a huge gallery of art involving nearly perfect structures and properties over the millions of years of development. Many plants and animals show water-repellent properties with fine micro-structures, such as lotus leaf, water skipper and wings of butterfly. Inspired by these special surfaces, the artificial superhydrophobic surfaces have attracted wide attention in both basic research and industrial applications. The wetting properties of superhydrophobic surfaces in nature are affected by the chemical compositions and the surface topographies. So it is possible to realize the biomimetic superhydrophobic surfaces by tuning their surface roughness and surface free energy correspondingly. This review briefly introduces the physical-chemical basis of superhydrophobic plant surfaces in nature to explain how the superhydrophobicity of plant surfaces can be applied to different biomimetic functional materials with relevance to technological applications. Then, three classical effects of natural surfaces are classified： lotus effect, salvinia effect, and petal effect, and the promising strategies to fabricate biomimetic su- perhydrophobic materials are highlighted. Finally, the prospects and challenges of this area in the future are proposed.