Secretory Epidermal Cells Research Articles

Cell-type specific localization and biological activity of the volatiles from the endemic species Chaerophyllum coloratum L.

Main conclusionNew findings are presented for Chaerophyllum coloratum L. on the volatile composition of the essential oil, based on data of hydrosol and fresh plant material, light and electron microscopy of leaves, and cytotoxic and antiviral activity.The widespread Apiaceae family includes many well-known and economically important plants that are cultivated as food or spices. Many produce essential oils and are generally a source of secondary metabolites and compounds that have numerous applications in daily life. In this study, the chemical composition of volatile organic compounds (VOCs), ultrastructure and biological activity of the Mediterranean endemic species Cheaerophyllum coloratum L. are investigated, as literature data for this plant species are generally very scarce. The essential oil and hydrosol were extracted from the air-dried leaves by hydrodistillation and the chemical composition of both extracts was analysed by GC–MS in conjunction with headspace solid-phase microextraction (HS-SPME) of VOCs from the hydrosol and the fresh plant material. In the composition of the essential oil, the oxygenated sesquiterpenes spathulenol and caryophyllene oxide were the most abundant components. In the fresh plant material, non-oxygenated sesquiterpenes dominated, with β-caryophyllene and germacrene D being the main components. The hydrosol was dominated by monoterpenes, with the oxygenated monoterpene p-cymen-8-ol being the most abundant. Light and electron micrographs of the leaf of C. coloratum show secretory structures, and we hypothesize that glandular leaf trichomes, secretory epidermal cells and secretory canals are involved in the production of volatiles and their secretion on the leaf surface. Since the biological potential of C. coloratum is poorly investigated, we tested its cytotoxic activity on cancer and healthy cell lines and its antiviral activity on plants infected with tobacco mosiac virus (TMV). Our results dealing with the composition, ultrastructure and biological activity show that C. coloratum represent a hidden valuable plant species with a potential for future research.

Transcription Factor RrANT1 of Rosa rugosa Positively Regulates Flower Organ Size in Petunia hybrida.

The flower is the main organ that produces essential oils in many plants. The yield of raw flowers and the number of secretory epidermal cells are the main factors for essential oil production. The cultivated rose species “Pingyin 1” in China was used to study the effect of RrANT1 on floral organ development. Eighteen AP2 transcription factors with dual AP2 domains were identified from Rosa rugosa genome. RrANT1 belonged to euANT. The subcellular localization results showed that RrANT1 protein is localized in the nucleus. The relative expression level of RrANT1 in the receptacle is higher than that in petals in the developmental stages, and both decreased from the initial phase to senescence. Compared with the RrANT1 expression level in petals in the blooming stage, RrANT1 expression level was significant in petals (~48.8) and highest in the receptacle (~102.5) in the large bud stage. It was only highly expressed in the receptacle (~39.4) in the blooming period. RrANT1 overexpression significantly increased petunia flower and leaf sizes (~1.2), as well as flower fresh weight (~30%). The total number of epidermis cells in the petals of overexpressing plants significantly increased (>40%). This study concluded that RrANT1 overexpression can increase the size and weight of flowers by promoting cell proliferation, providing a basis for creating new rose germplasm to increase rose and essential oil yield.

Floral nectaries and pseudonectaries in Eranthis (Ranunculaceae): petal development, micromorphology, structure and ultrastructure.

Flowers are an innovative characteristic of angiosperms, and elaborate petals usually have highly specialized structures to adapt to different living environments and pollinators. Petals of Eranthis have complex bilabiate structures with nectaries and pseudonectaries; however, the diversity of the petal micromorphology and structure is unknown. Petal development, micromorphology, structure and ultrastructure in four Eranthis species were investigated under SEM, TEM and LM. The results show that petals undergo 5 developmental stages, and accessory structure formation (stage 4) mainly determines the diversity of final mature petal morphology and pseudonectaries; the central depression formed in stage 2 will develop into nectary tissues. Petals are bilabiate and have hidden nectaries in nectary grooves; they consist of one layer of rounded and raised secretory epidermal cells and 3-14 layers of secretory cells with abundant plasmodesmata between cells. A large number of sieve tubes are distributed between the cells and extend to the epidermis; in addition, the vessel elements are located below the secretory area. Nectar is stored in the intercellular space between secretory parenchyma cells and escapes through microchannels or cell rupture. Pseudonectaries in all species of Eranthis except for E. hyemalis consist of smooth, ornamented epidermal cells and 9-12 layers of parenchyma cells with sparse cytoplasm, which may have the function of attracting pollinators.

Development and structure of four different stamens in Clematis macropetala (Ranunculaceae): particular emphasis on staminodes and staminal nectary.

The stamens of angiosperms are diverse in number, colour and structure. The morphological and structural changes of stamens show important evolutionary significance for improving pollination efficiency. In Clematis macropetala, the androecium consists of fertile stamens and tepaloid staminodes. However, studies on the developmental features, structures and possible functions of stamens are few. In this study, the stamen ontogeny, micromorphology and nectary structure of C. macropetala were studied by scanning electron microscopy, light microscopy and transmission electron microscopy. The results indicate that the stamens can be divided into four forms according to shape and anther size: tepaloid staminode (St1), spatulate staminode (St2), linear-spatulate fertile stamen (St3) and linear fertile stamen (St4). The characteristics of stamen development are similar in the early stage but gradually differentiate in the later stage. St1 has delayed development and no anther differentiation. St2 develops abnormally at the early stage of anther differentiation. St3 and St4 are fertile, but their anther sizes are different. Nine epidermal cell types were observed in stamens, with only 4 types in St1 and 6-7 types in St2, St3 and St4. Nectary tissue appears on the adaxial side of the filament base. The nectary is composed of only one layer of secretory epidermal cells, which have a large nucleus, dense cytoplasm and well-developed wall ingrowth. Nectar is released through micro-channels in the cuticle of the outer wall. In Ranunculaceae, the staminal nectary is often located on fertile or sterile stamens, and the position, structure and micromorphology of secretory tissues of the stamen within Ranunculales are discussed.

Wax glands of the horned gall aphid, Schlechtendalia chinensis, at different stages

The horned gall aphid, Schlechtendalia chinensis, inhabits the productive species of Chinese gallnuts, which have economic value. Aphid wax glands are crucial for the survival of the insects, since the secreted waterproofing wax is important to protect the aphids from predators, pathogens and honeydew contamination. In this study, we investigated the structure of wax glands and their role in different aphid stages using light and electron microscopy. Our results show that aphids of all stages except the newly hatched fundatrix possess six parallel dorsal lines and have a total of 56 wax gland plates. Although no wax glands occur on the dorsum of the newly hatched fundatrix (first instar), the glands do appear once a fundatrix enters the second instar. The wax gland plate is composed of 2–22 polygonal depressions, each of which corresponds to a secretory cell covered by cuticle. The wax glands of this aphid belong to the class 1 glands, which are formed by epidermal secretory cells. The structure of the wax glands varies in the different stages and these changes may be adaptive to the changeable microenvironments in which the aphids live.

Cytological Evidence for the Pathway of Synthesis, Accumulation, and Secretion of Rose Essential Oil

Rosa rugosa Thunb. is an important ornamental and economic plant. Essential oil extracted from its flowers is known as “liquid gold” and has a high economic and health value. In this study, the developmental process of secretory epidermal cells of petals, the location of essential oil synthesis and accumulation, and the associated secretory pathway were studied by using Chinese R. rugosa as a test material. Results showed that rose essential oil might be synthesized in two pathways: direct synthesis in plastids and conversion from starch grains. Although starch grains acted as a precursor for essential oil synthesis and provided energy during the synthesis process, the endoplasmic reticulum initially processed a portion of osmiophilic oil for transport and reprocessing through the vesicles of the golgi apparatus. The energy for this process originated from the mitochondria, and the oils were stored in central large vacuoles. Essential oils passed through the cytoplasmic membrane in two ways: through breaks of the cytoplasmic membrane and through exocytosis. This study laid a cytological foundation for regulating the synthesis and metabolism of rose essential oil and provided an important reference for cytologically studying volatile oils in other plants.

Osmophores and floral fragrance in Anacardium humile and Mangifera indica (Anacardiaceae): an overlooked secretory structure in Sapindales.

Flowers of Anacardiaceae and other Sapindales typically produce nectar, but scent, often associated with a reward for pollinators, has surprisingly been mentioned only rarely for members of the family and order. However, flowers of Anacardium humile and Mangifera indica produce a strong sweet scent. The origin and composition of these floral scents is the subject of this study. Screening of potential osmophores on the petals and investigations of their anatomy were carried out by light, scanning and transmission electron microscopy. The composition of the floral fragrance was characterized by gas chromatography–mass spectrometry. In both species, the base of the adaxial side of each petal revealed specialized secretory epidermal cells which are essentially similar in structure and distinct from all other neighbouring cells. These cells also showed evidence of granulocrine secretory mechanisms and slight specific variations in their subcellular apparatus coinciding with the respective composition of the floral fragrance, predominantly composed of sesquiterpenes in A. humile and monoterpenes in M. indica. This study reports the presence of osmophores for the first time in flowers of Anacardiaceae and confirms the link between the ultrastructural features of their secretory cells and the volatiles produced by the flowers. The flowers of most Sapindales, including Anacardiaceae, are nectariferous. However, the presence of osmophores has only been described for very few genera of Rutaceae and Sapindaceae. Both the occurrence of osmophores and fragrance may have largely been overlooked in Anacardiaceae and Sapindales until now. Further studies are needed to better understand the nature and diversity of the interactions of their nectariferous flowers with their pollinators.

Pharmacognostical study of Cynanchum stauntonii and Cynanchum glaucescens, botanical sources of TCM Baiqian

Objective: Cynanchum stauntonii and Cynanchum glaucescens are botanical species of Baiqian (Cynanchi Stauntonii Rhizoma et Radix) in Chinese Pharmacopoeia, in which, however, there are no microscopic identification. Therefore, we provided the morphological and microscopic identification of the crude drug for updating Chinese Pharmacopoeia.Methods: Twelve batches of C. stauntonii and three batches of C. glaucescens and their crude drugs were taxonomically, morphologically, and microscopically examined.Results: Taxonomically, C. stauntonii had narrowly lanceolate leaves with acuminate apex and 5mm long petiole; Whereas C. glaucescens was oblong-lanceolate or oblong with rounded or acute apex in leaves, and had very short or no petiole. Morphologically, rhizomes of C. stauntonii and C. glaucescens both had hollow pith, but the hollow pith occupied about a half of the rhizome's diameter in C. stauntonii, whereas only a very small proportion of the overall diameter in C. glaucescens. Moreover, microscopic observation showed the difference in the proportion of xylem and in rhizome transverse-sections of the two species along with the difference in the size of the pith. Finally, laticifers and rhizome epidermal secretory cells were present in the powders of C. stauntonii, but absent from C. glaucescens.Conclusion: Based on observation of morphological and microscopic characteristics, the two species can be distinguished by the size of the pith, proportion of xylem of rhizomes, and crude drug powder characters such as laticifers and secretory cells.

Comparative studies on petals structure, micromorphology and ultrastructure in two species of Stephania (Menispermaceae)

Petals in Ranunculales are nectariferous organs referred to as “nectary leaves” and show diversity in shape, color and structure due to various positions and structure of nectary tissue. Menispermaceae are in the core Ranunculales and have green and short nectary leaves, but the knowledge on structure and functions of the nectary leaves is limited. We use scanning electron microscopy, light microscopy and transmission electron microscopy to investigate nectary leaves structure, micro-morphology and ultrastructure in two species of Stephania in Menispermaceae. Our results show that secretory tissues present in the upper part of abaxial side of the nectary leaves and the specialized secretory epidermal cells are distinguished from other cells. In Stephania cepharantha, clusters of secreting epidermal cells are raised slightly above other cells to form some “bulges” (8–25 cells arranged in a cluster) and connected to distinct huge sieve tube elements. In contrast, in Stephania japonica, secretory epidermal cells are lower than non-secreting cells and result in many “well-like” structures (comprising 6–20 cells per “well”), and have no sieve tube element connection. Secretory epidermal cells have dense cytoplasm, large nucleus and abundant organelles. Nectar secretions are exuded via micro-channels or pores of cuticle on outer walls. The type of secretory tissue in Stephania is the variant of nectarioles.

A structural review of foliar glands in Passiflora L. (Passifloraceae).

Extrafloral glands in Passifloraceae species have aroused the interest of many researchers because of their wide morphological diversity. The present work analyzed the foliar glands on 34 species of Passiflora from samples containing glands in the petiole and foliar blade fixed in 50% solution of formaldehyde-ethanol-acetic acid and stored in a 70% ethanol solution. For anatomical analyses, part of the material was embedded in Paraplast, longitudinally sectioned and double stained with safranin and astra blue. Scanning electron microscopy analysis was also carried out. To analyze the presence of sugars in the secretion of foliar glands, a glucose strip test was used. Based on the results of morphological, anatomical and glucose strip tests, the foliar secretory glands in Passiflora can be grouped into two categories: Type I glands, defined as nectaries, can be elevated or flattened, and can have a sugar content high enough to be detected by the glucose strip test analysis. Type II glands are elevated and did not show a positive reaction to the glucose strip test. From an anatomical viewpoint, glands characterized as extrafloral nectaries show a multistratified secretory epidermis, typically followed by two flat layers of nectariferous parenchyma with dense content. Internal to these layers, vascular bundles are immersed in the subsecretory parenchyma and terminate in phloem cells. On the other hand, type II glands show a single layer of elongated secretory epidermal cells. Internal to this single layer, parenchyma and vascular tissue with both phloem and xylem elements can be observed. The analyzed species show a wide diversity of gland shape and distribution, and the combined analysis of morphology, anatomy and preliminary tests for the presence of glucose in the exudate in different Passiflora subgenera suggests the occurrence of two categories of glands: nectaries and resin glands.

Colleters inCasearia(Salicaceae): a new interpretation for the theoid teeth

Salicaceae possess a range of leaf teeth types, many characterized as nectariferous salicoid or violoid teeth. One large genus, Casearia, deviates from this generalization in having theoid teeth. Although taxonomic descriptions of Casearia have included reports of glands at the apex of teeth, there have been no descriptions of their anatomy or functional role. Here, we aim to describe the anatomy of the theoid teeth of 43 Casearia spp. Leaf samples from herbarium specimens were processed for light and scanning electron microscopy. Leaves of C. sylvestris were collected and fixed in the field for histochemical tests, revealing only polysaccharides in secretory epidermal cells and outside of these cells. The glands have precocious development and are composed of a non-vascular central axis covered with a palisade-like secretory epidermis. During senescence, the cells of the palisade-like secretory epidermis and central axis appear to lose their typical form, with some cell walls disintegrating. In the mature leaf, an abscission zone at the base of the gland causes detachment of the gland. The placement, anatomical structure, precocious activity and polysaccharide secretion (in C. sylvestris) allow the recognition of the glands associated with the marginal teeth in Casearia as colleters. Similar studies are needed for other genera in order to clarify how colleters have evolved in Salicaceae and how they are functionally and anatomically related to other teeth types in the family.

Bacterial leaf glands in Styrax camporum (Styracaceae): first report for the family

In this study, we recorded, for the first time, the occurrence of leaf glands in a member of Styracaceae and their association with bacteria. Samples of Styrax camporum Pohl shoot apices and leaves at different developmental stages were prepared according to the conventional techniques for light and electron microscopy. Glands are emergences constituted by epidermal and parenchyma cells and are differentiated into a secretory body on a short nonsecretory stalk supplied with phloem. Actively secreting glands occur from leaf primordia to mature leaves and produce mucilage that accumulates inside schizogenous intercellular spaces. The epidermal secretory cells have abundant cytoplasm rich in hyperactive dictyosomes, an extensive endoplasmic reticulum, and modified plastids. Bacteria enter the gland via the intact surface and proliferate in the intercellular spaces of the glands. Once inside the intercellular spaces of the glands, bacteria enter the cells owing to the weakening of the anticlinal and inner periclinal cell walls and by phagocytosis. Strands of actin filaments occur near the endocytical vesicles containing degenerating bacteria. Accumulations of phenolic compounds and callose could explain the absence of bacteria in the stalk cells. The presence of bacteria inside the leaf glands of S. camporum is a regular and cyclic trait. The significance of the bacteria (not yet identified) and the type of interaction between these two organisms remain unknown.

Comparative anatomy of the floral elaiophore in representatives of the newly re-circumscribed Gomesa and Oncidium clades (Orchidaceae: Oncidiinae)

Recently, molecular approaches have been used to investigate the phylogeny of Oncidiinae. This has resulted in the transfer of taxa previously considered to be species of Oncidium Sw. into Gomesa R. Br. and the re-circumscription of both genera. In this study, the structure of the floral elaiophore (oil gland) is described and compared for Gomesa echinata (Barb. Rodr.) M.W. Chase & N.H. Williams, G. ranifera (Lindl.) M.W. Chase & N.H. Williams, Oncidium amazonicum (Schltr.) M.W. Chase & N.H. Williams and O. oxyceras (Königer & J.G. Weinm.) M.W. Chase & N.H. Williams in order to determine whether phylogenetic revision is supported by differences in its anatomy. The floral elaiophore structure was examined and compared at three developmental stages (closed bud, first day of anthesis and final stage of anthesis) for all four species using light microscopy, fluorescence microscopy, scanning electron microscopy, transmission electron microscopy and histochemistry. In all species investigated, the floral elaiophore occurs on the labellar callus and is of the epithelial type, comprising cuboidal to palisade-like, secretory epidermal cells and a layer of sub-epidermal cells, both tissues enclosing ground parenchyma supplied with collateral vascular bundles and containing idioblasts, often with raphides or phenolic contents. A bi-layered cuticle comprising an outer, lamellate and an inner, reticulate layer is present, and sub-cuticular accumulation of secreted material results in distension of the cuticle. Secretion-filled cavities are present at anthesis in the elaiophore cell walls and, in most species, the outer, tangential walls of the elaiophore have small, peg-like projections that protrude into the cytoplasm. In all taxa examined, the elaiophore organelle complement, especially the smooth endoplasmic reticulum (SER), is typical of lipid-secreting cells. In terms of location, morphology, anatomy and ultrastructure, the floral elaiophores of both Gomesa and Oncidium species examined are very similar, and distinction between these genera is not possible based on elaiophore features alone. Furthermore, many of these elaiophore characters are shared with representatives of other clades of Oncidiinae, including the Ornithocephalus clade. Consequently, elaiophores are considered homoplasious and of limited value in investigating the phylogeny of this subtribe.

Post-embryonic Development of Intramandibular Glands in Pachycondyla verenae (Forel) (Hymenoptera: Formicidae) workers

The current knowledge of intramandibular glands in Hymenoptera is focused on occurrence and morphology in adult insects. This is the first report regarding the post-embryonic development of intramandibular glands in a “primitive” ant, Pachycondyla verenae. In this study, we analyzed mandibles of prepupae, white-eyed, pink-eyed and black-eyed pupae, pupa of pigmented body pupae, and adults. Adult workers of P. verenae have intramandibular glands with epidermal secretory cells of class I and isolated glands of class III, and both glands have onset differentiation in pink-eyed pupae. Some histological sections were submitted to histochemical test for total proteins and neutral polysaccharides. Histochemical tests showed occurrence of polysaccharides and proteins in epidermal secretory cells of class I from the white-eyed pupae, polysaccharides and proteins in pink-eyed pupae to black-eyed pupae in both glands classes I and III and presence of polysaccharides in adult ants also in both gland classes I and III. Intramandibular glands of classes I and III in P. verenae workers differentiate during pupation, with onset occurring in pink-eyed pupae, and completion occurring in black-eyed pupae.

Comparative floral micromorphology in four sympatric species of Serapias (Orchidaceae)

A great variety of colours, shapes, scents, energy-rich rewards and other floral traits form an integrated system that increases pollination efficiency or reduces pollen loss. Flowers lacking energetic rewards possess a more elaborate morphology to ensure co-specific pollination. Despite the prevalence of deceptive species among Euro-Mediterranean orchids, the study of their floral micromorphology is still in its infancy. In this work, we examine the floral micromorphology of four sympatric species of the widespread Mediterranean genus Serapias. Electron microscopy (scanning, transmission) was performed on flowers of four sympatric species of the deceptive genus Serapias, the widespread S. cordigera, S. lingua, S. parviflora and S. vomeracea. Each species has a characteristic combination of specialized, secretory epidermal cells and trichomes on the labellum and, to a lesser extent, the petals. Serapias lingua is highly specialized in that it produces olfactory signals and has two types of trichome with secretory apical cells, one of which is also present in S. parviflora. In addition to striated, conical secretory cells, S. cordigera and S. vomeracea also have characteristic, atypical secretory trichomes with bilobulate or dome-like protuberances. The labellar micromorphology of S. lingua and S. parviflora is adapted for scent production, whereas S. vomeracea and S. cordigera possess trichomes that may also provide visual and/or tactile cues. Owing to the combined effect of the specific floral fragrance and labellar micromorphology in pollinator selection, the pollination ecology of Serapias spp. is even more specialized than previously thought. © 2012 The Linnean Society of London, Botanical Journal of the Linnean Society, 2012, 169, 714–724.

Comparative structure of the osmophores in the flowers of Stanhopea graveolens Lindley and Cycnoches chlorochilon Klotzsch (Orchidaceae)

The structure of the osmophores in <i>Stanhopea graveolens</i> and <i>Cycnoches chlorochilon</i> was studied by means of light microscopy (LM), scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The scent glands are located in the basal part of the labellum. The surface of the osmophores is wrinkled or rugose, which increases the area of fragrance emission. On the surface of the epidermis, remnants of secretion are noticeable in <i>S. graveolens</i>, but these are absent in <i>C. chlorochilon</i>. The osmophore tissue is composed of secretory epidermal cells and several layers of subepidermal parenchyma, and it is supplied by vascular bundles that run in ground parenchyma. The secretory cells have large nuclei, a dense cytoplasm with numerous ER profiles, lipid droplets, and plastids with a substantial amount of starch, which are probably involved in the synthesis of volatile substances. In the cell walls of the osmophore cells, numerous pits with plasmodesmata occur that are likely to take part in symplastic transport of the scent compounds. The structure of the osmophores is similar in both investigated species. Both <i>S. graveolens</i> and <i>C. chlorochilon</i> are pollinated by euglossine bees, and such similarity results from adaptation to effective scent emission and attraction of pollinators.

Biotic versus abiotic pollination in Oeceoclades maculata (Lindl.) Lindl. (Orchidaceae)

AbstractGeographical variation in the reproductive biology of widespread species often occurs at their distributional boundaries. We sought to determine whether such variation has occurred in an invasive orchid, Oeceoclades maculata, across its naturalized range. We compared its reproductive biology in a Brazilian population with that published for a population on the Caribbean island of Puerto Rico. In the state of São Paulo, O. maculata flowers between December and February, at the height of the rainy season. Similar fruit sets were observed in manual self (76%) and cross (70.4%) pollination treatments. The fruit set of plants protected from both pollinators and rainfall was 6.1%, whereas plants exposed only to rainfall had a fruit set of 41.4%, slightly less than the controls (48.3%). Like the Puerto Rico population, reproduction is primarily through rain‐assisted autogamy, but unlike observations made on the island, outcrossing can eventually occur. We observed two butterfly species (Heliconius ethilla narcaea and Heliconius erato phyllis) pollinating O. maculata. Secretory epidermal cells and trichomes of the spur lumen produced 0.7 µL of 25% (sucrose equivalents) nectar per flower each morning, which was stored in a dilated basal portion of the spur and reabsorbed by the afternoon. Thus, geographical variation in reproductive biology exists across the broad invasive range of O. maculata.

Laticiferous canal formation in fruits of Decaisnea fargesii: a programmed cell death process?

Programmed cell death (PCD), a topic of abiding interest, remodels plants at the cell, tissue, and organ levels involving various developmental processes of plants. The aim of this study is to provide a morphological characterization of evidence of PCD involvement in the laticiferous canal formation in fruit of Decaisnea fargesii. Several ultrastructural features of PCD have been observed including disintegration of vacuole and plasma membranes, cell wall degeneration, degenerated cytoplasm, abundant membrane structures and flocculent material, mitochondria and misshapen nuclei coupled with degraded plastids in vacuoles, and nuclei enveloped by rubber granule. In D. fargesii, the nuclei of the secretory epidermal cells become TUNEL-positive from the sunken stage to the late expanding stage, then DAPI-negative during the mature stage, indicating an early event of deoxyribonucleic acid (DNA) cleavage and a late event of complete DNA degeneration. Gel electrophoresis indicates that DNA cleavage is random and does not result in the laddering pattern indicating multiples of internucleosomal units. During the PCD of secretory epidermal cells, the rubber granules continue to be synthesized and accumulated in the secretory epidermal cells despite nuclear degradation. The PCD's role in laticiferous canal formation suggests that PCD may play important roles in gland development of plants.

Anatomical structure and secretion compounds of colleters in nineIlexspecies (Aquifoliaceae) from southern South America

In order to clarify whether the structures observed at the base of the petiole of the genus Ilex are colleters resulting from stipules, the anatomy, vascularization and secretions of these supposed glandular structures were analysed in nine species. This is the first report of colleters in Ilex. Stipular colleters replace the stipules in all species studied and are characterized by the presence of vascular traces. In addition to the stipular colleters, three other types of colleter were distinguished: standard and lachrymiform colleters found on the leaf teeth or crenations, and sessile colleters found on the margins of the floral bracts. Their basic structure consists of a central core of parenchymatous cells surrounded by one layer of palisade secretory epidermal cells. Histochemical tests were also performed on secretions; proteins were found in the secretions studied, but glucose was not. The glandular origin of the stipular colleters is confirmed on the basis of their position, secretions and anatomy. Analyses of the colleter-secreted proteins distinguished two different groups of Ilex species.

Morphological characterization of the venom secretory epidermal cells in the stinger of marine and freshwater stingrays

Marine and freshwater stingrays are characterized by the presence of one to three mineralized serrated stingers on the tail, which are covered by epidermal cells secreting venom. When these animals are dorsally touched, the stinger can be introduced into the aggressor by a whip reflex mechanism of the tail, causing severe mechanical injuries and inoculating the venom. Accidents in humans are frequent causing intense local pain, oedema and erythema. Bacterial secondary infection is also common. In addition, injuries involving freshwater stingrays frequently cause a persistent cutaneous necrosis. The exact localization of the venom secretory epidermal cells in the stinger is controversial, but it is known that it is preferentially located in the ventrolateral grooves. A comparative morphological analysis of the stinger epidermal tissue of different marine and freshwater Brazilian stingray species was carried out. The results indicate that in freshwater species there is a larger number of protein secretory cells, of two different types, spread over the whole stinger epidermis, while in marine species the protein secretory cells are located only around or inside the stinger ventrolateral grooves. These differences between the stingers of the two groups can justify the more severe envenomation accidents with the freshwater species when compared with the marine species.