Apple Fruit Cuticles and the Occurrence of Pores and Transcuticular Canals

Developmental studies were made on the dewaxed thin-sectionedapple fruit cuticles of 10 Malus sylvestris Mill, cultivarsfrom 4 weeks before anthesis through fruit maturation and harvest.Cuticular development appears to correlate well with the generalgrowth of the fruit. However, no correlation exists betweenfruit size and cuticle thickness. Cuticular pores were evidentas early as 1 week before anthesis and transcuticular canalsbecame evident by 1 week following anthesis. Dewaxed thin-sectioned,as well as isolated, mature fruit cuticles of 16 cultivars andfour crab-apples consistently revealed the distinct presenceof ubiquitous pores and canals. Evidence is provided by lightmicroscopy and scanning electron microscopy (SEM). Measurementswere taken of cuticle thicknesses during development and ofcuticular pore dimensions, and calculations were made of poreand canal numbers. Fruit size alone is not directly indicativeof total pore numbers per surface area. Canal lengths are directlyrelated to the developmental thickness of the cuticle. No correlationwas found between the thickness of the mature cuticle and eitherthe number of pores present or the pore diameters. Malus sylvestris Mill, apple fruit, morphology, cuticle, cuticular flanges, cuticular pores, transcuticular canals, ultrastructure     

Fine structure of the aesthetasc hairs ofPagurus hirsutiusculus dana

Summary The thin-walled aesthetasc pegs on the antennules of a small hermit crab,Pagurus hirsutiusculus, were studied by light and electron microscopy. The lumen of each aesthetasc was found to be filled with the dendrites of 300–500 bipolar neurons whose cell bodies lie beneath the base of the aesthetasc. These dendrites are ciliary in nature, having well developed basal bodies and rootlets.Each basal body gives rise to a cilium which divides to form a cluster of slender branches, each of which contains a microtubule running lengthwise. These structures occupy most of the length of the hair.The cuticle of the aesthetasc wall is thin and tenuous. Except for the pore canals in the basal region, we have found no pores at either light or electron microscope level, but as the hair is extremely permeable, we conclude that the cuticle itself may permit the passage of solutions. This permeability of the cuticle and the large numbers of dendrites within support the hypothesis that the aesthetascs are chemoreceptors.     

Comparative ultrastructural study on tris 1-aziridinyl phosphine oxide prefixed cuticles of some oligochaetes

The cuticle of five species of Oligochaeta, chosen to represent differences in size and a variety of biotopes, was studied electron microscopically after fixation with the acrolein-TAPO-osmium tetroxide method. Five distinct layers in the cuticle of all studied species were found. Staining with lead and uranyl ions or with silver proteinate visualized basically the same structural components of the cuticle, but the degree of electron opacity and the distribution of the electron-opaque stain in these components differed according to the staining method used. Since the acrolein-TAPO-osmium tetroxide method visualized the cuticular zones preferentially stained by Thiéry's silver proteinate method, it was concluded that the TAPO method may be considered suitable for the visualization of polysaccharides. Staining with phosphotungstic acid provided some information on the composition of the cuticle of Oligochaeta not obtained by staining ultrathin sections with lead and uranyl ions nor with silver proteinate. The conclusion is that phosphotungstic acid binds to polysaccharides which do not contain vicglycol groups nor active sites responsible for the positive reaction with lead and uranyl salts. Structural components in the cuticle of the oligochaetes studied were characteristic for each species. The taxonomic value of such components, however, must be confirmed by examination of a larger number of species of oligochaetes.     

Elektronenmikroskopische Untersuchungen anAnaitides mucosa (Annelida,Polychaeta). Cuticula und Cilien,Schleimzellen und Schleimextrusion

Two components, a basal cuticle and an epicuticle, make up the cuticle ofA. mucosa. The basal cuticle consists of collagen fibrils, which are arranged in about 20 layers. The orientation of the fibrils changes rectangularly from one layer to the next. Fine filaments interweave the basal cuticle. The epicuticle, which is covered by a layer of electron dense material, is composed of irregularly arranged thin filaments. Branched microvilli of the epidermal cells penetrate the cuticle. Bacteria are found in the basal cuticle. Dorsally each segment has a band of densely packed smooth cilia. Laterally and partly ventrally aggregates of cilia are observed. These cilia exhibit apically artificial swellings. At least six different mucous cells are observed in the epidermis, morphologically distinguishable by the structure of the secretion products. Mucus is secreted via exocytosis through cuticular pores. During this process the mucus might expand. The secreted mucus consists of filamentous subunits.     

Study on pathway and characteristics of ion secretion of salt glands of Limonium bicolor

Recretohalophytes with specialized salt-secreting structures, including salt glands and salt bladders, can secrete excess salts from plant tissues and enhance salinity tolerance of plants. However, the pathway and property of salt secretion by the salt gland has not been elucidated. In the article, Limonium bicolor Kuntze was used to investigate the pathway and characteristics of salt secretion of salt gland. Scanning electron microscope micrographs showed that each of the secretory cells had a pore in the center of the cuticle, and the rice grain-like secretions were observed above the pore. The chemical composition of secretions from secretory pores was mainly NaCl using environmental scanning electron microscope technique. Non-invasive micro-test technology was used to directly measure ion secretion rate of salt gland, and secretion rates of Na⁺ and Cl^? were greatly enhanced by a 200-mmol/L NaCl treatment. However, epidermal cells and stoma showed little secretion of ions. In conclusion, our results provide evidence that the salt glands of L. bicolor have four secretory pores and that NaCl is secreted through these pores of salt gland.     

四种植物花蜜腺的解剖学研究

通过对石竹、庐山小檗、活血丹和紫云英四种植物花蜜腺的外部形态、显微切片和扫描电镜观察,了解到石竹花蜜腺是雄蕊类型,蜜汁通过角质层间小孔分泌;庐山小檗花蜜腺是花被类型,其每个分泌表皮细胞的外切向壁中央向内凹陷与角质层之间形成角质层下空隙,蜜汁由表皮细胞分泌后贮存于角质层下空隙中,在蜜汁积累过程中不断增加对角质层的压力,最后冲破角质层分泌到体外;活血丹和紫云英花蜜腺属于花托类型,前者蜜汁从分泌表皮细胞通过角质层直接渗出,后者蜜汁通过变态气孔分泌。本试验的四种植物花蜜腺中维管组织有三种不同类型。     

Polar paths of diffusion across plant cuticles: new evidence for an old hypothesis

BACKGROUND: The plant cuticle is an extracellular lipophilic biopolymer covering leaf and fruit surfaces. Its main function is the protection of land-living plants from uncontrolled water loss. In the past, the permeability of the cuticle to water and to non-ionic lipophilic molecules (pesticides, herbicides and other xenobiotics) was studied intensively, whereas cuticular penetration of polar ionic compounds was rarely investigated. RECENT PROGRESS: Recent work measuring cuticular penetration of inorganic and organic ions is presented; the effects of molecular size of ions, temperature, wax extraction, humidity and plasticizers strongly support the conclusion that ions penetrate cuticles via water-filled pores. The cuticle covering stomata and trichomes forms the preferential site of ion penetration. This indicates that cuticles possess a pronounced lateral heterogeneity: the largest fraction of the cuticle surface is covered by the lipophilic domains of cutin and wax, but to a certain extent polar domains are also present in the cuticle, which form preferential sites of penetration for polar compounds. THE FUTURE: The chemical nature of these polar domains awaits detailed characterization, which will be of major importance in agriculture and green biotechnology, since polar paths of diffusion represent the most important transport routes for foliar-applied nutrients. Furthermore, many compounds acting as inducers of gene expression in transgenic plants are ionic and need to penetrate the cuticle via polar paths in order to be active.     

Design of the labial cuticle in Cenocorixa bifida Hung. (Hemiptera: Corixidae) with reference to ionic transport

The surface topography and ultrastructure of the labial cuticle of Cenocorixa bifida were examined by scanning and transmission electron microscopy. The dorsal wall of the labium consists of seven sclerotized transverse bars each displaying two rows of semicircular grooves and pores. The cuticle is about 20 microm thick and is composed of epicuticle and lamellate exocuticle and endocuticle, the latter separated from the underlying epidermis by subcuticle containing amorphous material. The epicuticle is subdivided into an electron-dense very thin outer epicuticle and a homogenous thick inner epicuticle, which is penetrated by grooves. The exocuticle is filled with electron-dense blocks of material, which may provide mechanical support to the labial wall. The elongate epidermal cells display extensive infoldings of the apical plasma membrane (facing the cuticle) and contain abundant mitochondria in the cytoplasm. The presence of deep epicuticular grooves and pores in the thin labial cuticle and extensive apical membrane infolding and abundant mitochondria in the epidermal cells suggest that the labium in C. bifida is the site of osmoregulatory ionic uptake.     

The Prevalence of Pores and Canals in Leaf Cuticular Membranes

Ubiquitous, visibly discrete, natural cuticular pores and transcuticularcanals were found in the dewaxed leaf (and one herbaceous stem)cuticular membranes of 27 out of 32 taxa among 14 families.Clear evidence for their existence is provided by light photomicrographs.Both adaxial and abaxial leaf surfaces were investigated usingthin transections and chemically isolated cuticular membranes,in conjunction with ordinary staining techniques and light microscopymethods. No correlations were found between cuticle thicknessesand either the frequency of pore or the pore and canal diameters. Leaf cuticles, cuticle morphology, cuticular pores, transcuticular canals, cuticular flanges     

Giant cuticular pores in Eidothea zoexylocarya (Proteaceae) leaves

Ubiquitous, large diameter pores have not previously been adequately demonstrated to occur in leaf cuticles. Here we show conclusively that such structures occur in Eidothea zoexylocarya, a rainforest tree species of Proteaceae restricted to the Australian Wet Tropics. The pores are abundant, large-diameter apertures (～1 μm), that extend perpendicularly most of the way through the cuticle from the inside. They occur on both sides of the leaf, but are absent from the cuticle associated with stomatal complexes on the abaxial side. No such pores were found in any other species, including the only other species of Eidothea, E. hardeniana from New South Wales, and other species that have previously been purported to possess cuticular pores. To determine whether these pores made the cuticles more leaky to water vapor, we measured astomatous cuticular conductances to water vapor for E. zoexylocarya and seven other Proteaceae species of the Wet Tropics. Cuticular conductance for E. zoexylocarya was relatively low, indicating that the prominent pores do not increase conductance. The function of the pores is currently obscure, but the presence of both pores and an adaxial hypodermis in E. zoexylocarya but not E. hardeniana suggests evolution in response to greater environmental stresses in the tropics.     

Cuticular Pores and Transcuticular Canals in Diverse Fruit Varieties

Dewaxed thin-sectioned and dewaxed isolated mature fruit cuticlesrevealed the unequivocal presence in situ of visibly discrete,ubiquitous, cuticular pores or orifices concomitant with anticlinally-orientedtranscuticular canals in 51 varieties of fruit among 20 plantfamilies. More than 66 per cent of the fruit cuticles have poresand/or canals. No correlation exists between either fruit sizeor pore size and cuticle thickness. Dewaxed cuticles rangedfrom 1.25–22.5 µm in thickness. Canal lengths aredirectly related to cuticle thickness. Cuticular occlusionsof the epidermal cells were found in 76 per cent of the fruitsexamined. Evidence is provided by light microscopy photomicrographs. Fruit cuticles, cuticle morphology, cuticular pores, transcuticular canals     

Electron microscopic and preparative methods for the analysis of isopod cuticle

The crustacean cuticle consists of a complex organic matrix and a mineral phase. The physical and chemical properties of the cuticle are corellated to the specific functions of cuticular elements, leading to a large variety in its structure and composition. Investigation of the structure-function relationship in crustacean cuticle requires sophisticated methodological tools for the analysis of different aspects of the cuticular architecture. In the present paper we report improved preparation methods that, in combination with various electron microscopic techniques, have led to new insights of cuticle structure and composition in the tergite cuticle of Porcellio scaber. We used thin sections of non-decalcified tergites and decalcified resin embedded material for transmission electron microscopy and scanning transmission electron microscopy. Etched sagittal planes of bulk tergite samples were analysed with field emission scanning electron microscopy. We have found a distinct distal region within the exocuticle that differs from the subjacent proximal exocuticle in the arrangement of fibres. Within this distal exocuticle chitin-protein fibrils assemble to fibres with diameters between 15 and 50 nm that are embedded in a mineral matrix. In the proximal exocuticle and the endocuticle fibrils do not assemble to fibres and are surrounded by mineral individually. Furthermore, we show that the pore canals are filled with mineral, and demonstrate that mild etching of polished sagittal cuticle surfaces reveals regions containing mineral of diverse solubility.     

Salt excretion through the cuticle without disintegration of fine structures in the salt glands of Rhodes grass (Chloris gayana Kunth)

Some salt-tolerant plants belonging to the Poaceae possess bicellular salt glands that excrete salt-containing water. To clarify the excretion process from the outer cell of the bicellular salt gland (cap cell), unwashed and washed fresh leaves of Rhodes grass (Chloris gayana Kunth), Poaceae, were cryo-fixed rapidly, and the surface fine structures of the leaves were observed by cryo-scanning electron microscopy with high resolution. The cuticle on the cap cell did not have any pores or signs of rupturing. The cuticle of the cap cells lacks the epicuticular waxes that cover most surfaces of leaf epidermis. After excreted droplets on the salt glands were completely removed by washing with water, re-excreted droplets were observed 3 h later. These findings indicate that the bicellular salt glands of Rhodes grass excrete salt-containing water continuously through the wax-free cuticle of the cap cell without disintegration of the cuticular structure.     

Characterization of aqueous pores in plant cuticles and permeation of ionic solutes

Plant cuticles are lipid membranes with separate diffusion paths for lipophilic non-electrolytes and hydrated ionic compounds. Ions are lipid insoluble and require an aqueous pathway across cuticles. Based on experimental data, the aqueous pathway in cuticles has been characterized. Aqueous pores arise by hydration of permanent dipoles and ionic functional groups. They can be localized using ionic fluorescent dyes, silver nitrate, and mercuric chloride. Aqueous pores preferentially occur in cuticular ledges, at the base of trichomes, and in cuticles over anticlinal walls. Average pore radii ranged from 0.45 to 1.18 nm. Penetration of ions was a first order process as the fraction of the salt remaining on the cuticle surface decreased exponentially with time. Permeability of cuticles to ions depended on humidity and was highest at 100% humidity. Wetting agents increased rate constants by factors of up to 12, which indicates that the pore openings are surrounded by waxes. The pores in cuticular ledges of Helxine soleirolii allowed passage of berberine sulphate, which has a molecular weight of 769 g mol(-1). Increasing the molecular weight of solutes from 100 to 500 g mol(-1) decreased the rate constants of penetration by factors of 7 (Vicia faba) and 13 (Populus canescens), respectively. Half-times of penetration of inorganic salts and organic ions across Populus cuticles and Vicia leaf surfaces varied between 1 and 12 h. This shows that penetration of ionic compounds can be fairly rapid, and ions with molecular weights of up to 800 g mol(-1) can penetrate cuticles that possess aqueous pores.     

BINDING SITES FOR INORGANIC IONS AND UREA ON ISOLATED CUTICULAR MEMBRANE SURFACES

Binding sites for calcium and chloride ions, and urea on cuticular membrane surfaces were identified by microautoradiography. There was no localization of binding on either the smooth outer or irregular inner surfaces of the astomatous tomato fruit cuticle, and urea was bound to a lesser degree than inorganic ions. Calcium and chloride binding sites on surfaces of green onion leaf cuticles were closely associated with stomatal pores and above the periclinal cell walls. Binding of calcium was more pronounced than of chloride. These results are discussed in light of possible sites of entry for foliar applied nutrients.     

Cluster-forming property correlated with hemolytic activity by staphylococcal γ-hemolysin transmembrane pores

Staphylococcal γ-hemolysin (Hlg) is a pore-forming toxin consisting of two separate components, LukF (34kDa) and Hlg2 (32kDa). Here we show that Hlg pores aggregate and form clusters on human erythrocyte membranes in association with increasing hemolytic activity. Quantitative analysis using transmission electron microscopy and image processing revealed that the formation of single pores and clusters is related to the release of potassium ions and of hemoglobin from erythrocytes, respectively. This is the first study to suggest a novel and unique property which can facilitate hemolysis by the cluster formation of Hlg pores.     

Studies on the ultrastructure of the integument of the rotifer Habrotrocha rosa Donner (Aschelminthes)

Summary The integument of the rotifer Habrotrocha rosa Donner is provided with pores and formed by an extrasyncytial cuticle and a syncytial hypodermis. The hypodermis peripherally contains 3 layers of dense cytoplasm and borders the cuticle by an asymmetric cell membrane. The wall of the pores is stiffened proximally like an annulus. The pores lead into cytoplasmic invaginations which are surrounded by vesicles. Close to and also beneath the condensed cytoplasmic layers microbodies are found, which are interpreted as microperoxisomes. Subhypodermal layers of muscles are connected with the cytoplasm of the hypodermis by desmosome-like structures.I am indebted to Dr. H. Breucker, Anatomisches Institut Hamburg, and Prof. Dr. W. Becker, Zoologisches Institut Hamburg, under whose direction this work was carried out. Supported by the Deutsche Forschungsgemeinschaft (Be 464/10)     

The surface morphology of locust hindgut cuticle*

The surface architecture of hindgut cuticle in Locusta migratoria is described. Infundibulate pores are present in the midrectal pads, the ileum and the posterior rectum. It is suggested that they lead to canals traversing the cuticle. Larger occluded crater-like pores are present on the posterior rectal pads and may be glandular. The cuticle of the ileum and colon is invested with numerous spikes pointing backwards, and the interpad region of the rectum with upright spikes pointing inwards. These spikes probably serve to direct the faeces backwards from the ileum and colon, and to impale the faecal pellet during dehydration in the rectum.     

苹毛丽金龟触角嗅感器超微结构

利用扫描电镜和透射电镜对苹毛丽金龟Proagopertha lucidula (Faldermann)雌雄成虫触角嗅感器超微结构特征进行研究。结果表明:嗅感器集中于触角鳃片上,分布在触角鳃片表皮内陷形成的凹腔里,有5种感器,以板状感器为主,以外还有锥状感器、腔状感器、腔锥状感器和毛状感器。板状感器根据盘体形状的不同可分为4种类型,锥状感器根据锥体形状的差异可分为2种类型,腔锥状感器根据形状分为2种类型。嗅感器表皮有微孔和孔道微管。嗅感器内神经元的数目并不一致,1～2个不等。雄虫鳃片嗅感器总数显著多于雌性,是雌虫嗅感器总数1.8倍。其中雄性锥状感器的数目是雌性2倍,雌虫腔锥状感器数量是雄虫的4倍,雄虫板状感器数量是雌虫的2倍,雌雄腔状感器的数目无显著性差异。     

The exocytotic fusion pore modeled as a lipidic pore.

Freeze-fracture electron micrographs from degranulating cells show that the lumen of the secretory granule is connected to the extracellular compartment via large (20 to 150 nm diameter) aqueous pores. These exocytotic fusion pores appear to be made up of a highly curved bilayer that spans the plasma and granule membranes. Conductance measurements, using the patch-clamp technique, have been used to study the fusion pore from the instant it conducts ions. These measurements reveal the presence of early fusion pores that are much smaller than those observed in electron micrographs. Early fusion pores open abruptly, fluctuate, and then either expand irreversibly or close. The molecular structure of these early fusion pores is unknown. In the simplest extremes, these early fusion pores could be either ion channel like protein pores or lipidic pores. Here, we explored the latter possibility, namely that of the early exocytotic fusion pore modeled as a lipid-lined pore whose free energy was composed of curvature elastic energy and work done by tension. Like early exocytotic fusion pores, we found that these lipidic pores could open abruptly, fluctuate, and expand irreversibly. Closure of these lipidic pores could be caused by slight changes in lipid composition. Conductance distributions for stable lipidic pores matched those of exocytotic fusion pores. These findings demonstrate that lipidic pores can exhibit the properties of exocytotic fusion pores, thus providing an alternate framework with which to understand and interpret exocytotic fusion pore data.