Development, Structure and Cytochemistry of Resin-secreting Colleters of Gardenia gummifera (Rubiaceae)

The structure, ontogeny and cytochemistry of stipular colletersof Gardenia gummifera are described. The repeated division ofa few adaxial protodermal and subprotodermal cells of the youngstipule and the subsequent elongation by the latter resultsin the formation of a standard-type colleter. A developed colleterhas outer palisade-like glandular cells covered by a continuouscuticle and inner layers of elongated columnar cells. Enzymelevels in the palisade-like epidermal cells in the colletersindicate their active role in secretion. The secretion is resinous,occurs in the young colleters, and accumulates between the cellsand the cuticle. The cuticle later ruptures and releases thesecretory product, which drenches the young shoot apex. Gardenia gummifera L, stipule, colleters, resin, ontogeny, cytochemistry     

Anatomical structure and secretion compounds of colleters in nine Ilex species (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. © 2009 The Linnean Society of London, Botanical Journal of the Linnean Society, 2009, 160 , 197–210.     

Comparative systematic study of colleters and stipules of Rhizophoraceae with implications for adaptation to challenging environments

Colleters are multicellular secretory structures found on various organs in flowering plants. Colleters on the adaxial sides of stipules have been hypothesized to play a role in protecting the developing shoot. Rhizophoraceae is a stipulate family with a broad distribution from mangrove to montane environments, which makes the family well suited for the examination of this hypothesis, but the colleters of Rhizophoraceae are not well known. We compared species from all three tribes of Rhizophoraceae, including five inland genera and all four mangrove genera. In all species, several to hundreds of colleters, sessile or stalked, arranged in rows aggregated in genus‐specific shapes, are found at the adaxial bases of open and closed stipules. Pellacalyx uniquely has additional colleters at the stipule margins. Colleters are all of the standard type, comprising a central axis of core parenchyma with large vacuoles and tannins, and an outer palisade‐like epidermis with organelles involved in secretory activity. An exception is Pellacalyx axillaris, in which colleters appear as extremely small epidermal protrusions. Kandelia obovata has a tracheary element in some colleters. Pellacalyx uniquely has an unusual fleshy outgrowth on the adaxial stipule base. We propose an evolutionary sequence in which Macarisia has plesiomorphic stipule and colleter traits and the mangrove Kandelia obovata with colleter vascular traces is most derived. Colleter and stipule structures are largely concordant with habitat and phylogeny, and show taxonomic value. The strong alignment of colleter and stipule patterns with habitat is suggestive that colleters have a protective function, although some components of these patterns may be phylogenetically determined. © 2013 The Linnean Society of London, Botanical Journal of the Linnean Society, 2013, 172 , 449–464.     

Anatomy and histochemistry of colleters in Roupelia grata (Apocynaceae)

The colleters of Roupelia grata are present on the petiole of the leaf, bract, bracteole and calyx. A mature colleter has a distinct head and a stalk. The parenchymatous central core is surrounded by epithelial cells. The senescence of the colleter occurs at its apex first and then proceeds towards the base.     

Colleter types in Rubiaceae, especially in relation to the bacterial leaf nodule symbiosis

Vegetative buds were removed from herbarium specimens, and a small number of living plants, of 116 species of 111 genera of Rubiaceae and processed for light-microscopic examination of colleter form and anatomy. The results were combined with those of earlier studies. Most colleters in this family are of the 'standard type' described by others. Colleters are, however, lacking in eight genera. dendroid colleters occur in six genera. Two other genera have an intermediate type of colleter. Three genera have bacterial leaf nodules; dendroid colleters are not restricted to these genera, but their pattern of occurrence suggests a causal relationship between bacterial infection and evolution of dendroid colleters. Coupled with the geographic distribution of nodulared species within these genera, this pattern indicates that Pavetta became infected very early in its evolutionary history, Psychotria relatively later, and Neorosea most recently.     

Colleter morphology in Pavetta, Neorosea and Tricalysia (Rubiaceae) and its relationship to the bacterial leaf nodule symbiosis

Buds removed from herbarium specimens were processed for light microscopic examination of colleter form and anatomy. Most Pavetta species have dendroid colleters and bacterial leaf nodules, a correlation also found in an earlier study of Psychotria. Colleters of Neorosea , another genus with leaf-nodulated species, are more like the standard rubiaceous type that predominates in the family except that they have irregular, bulging epidermal cells. Tricalysia , a nodule-free genus closely related to Neorosea , shows a range of colleter form from standard to dendroid, and some species have the Neorosea type of colleter. Such morphological correlation between the bacterial leaf nodule symbiosis in Pavetta and Psychotria and dendroid colleters may indicate a chemical change in colleter secretion. Apart from their involvement with the bacterial symbiosis, rubiaceous colleters have now been shown to vary sufficiently in certain taxa to be considered as additional useful taxonomic characters.     

Leaf colleters in Tontelea micrantha (Celastraceae,Salacioideae): Ecological,morphological and structural aspects

The colleter secretion can be useful to protect plants of Cerrado (Brazilian savanna) biome during the long and pronounced dry season. This study describes the presence of colleters in Tontelea micrantha and represents the first record of these structures in Celastraceae. To investigate colleter structure and their secretory processes, young leaves were collected, fixed, and processed according to conventional techniques for light, and electron microscopy. Colleters were observed at the marginal teeth on the leaf. They produce mucilaginous secretions that spread over the leaf surface. After secretory phase, colleters abscise. The secretory epithelium is uniseriate and composed of elongated cells whose dense cytoplasm is rich in organelles. The ultrastructure of the secretory cells is compatible with the pectin-rich secretion. Observations of the young leaves surface revealed the presence of superficial hydrophilic secretion films that appeared to have the function of maintaining the water status of those organs.     

Structural, Functional and Phylogenetic Aspects of the Colleter

This article surveys the structural, functional and phylogeneticsignificance of colleters in different dicotyledonous families.Colleters are multicellular secretory structures attached tothe stipule, petiole, lamina, bract, bracteole, calyx and corolla.Colleters are grouped into standard (S), dendroid (D) and brush-like(B) types on the basis of their morphology and structure. Dand B-type colleters occur in certain members of Rubiaceae thatalso have bacterial leaf nodules. Besides the normal structure,epithelial hairs, thin-walled subepidermal cells, laticifersand vasculature are present in many colleters of Apocynaceae.It is probable that the colleter functions to protect the developingmeristem by secreting a viscous fluid. Exudate of D-type colletersare mucilaginous, providing the substrate necessary for thenutrition of endophytic bacteria. Colleter, structure, phylogeny     

MORPHOLOGY AND DISTRIBUTION OF COLLETERS AND CRYSTALS IN RELATION TO THE TAXONOMY AND BACTERIAL LEAF NODULE SYMBIOSIS OF PSYCHOTRIA (RUBIACEAE)

Earlier investigators established that a finger-like colleter with elongate axial cells and a palisade epidermis is the standard type in the Rubiaceae. This type, with some variation, also prevails in Psychotria, based on a worldwide anatomical survey of vegetative buds from herbarium specimens of 296 species (about 50 % of total). It is virtually the only type found outside of continental Africa. Among African species, it is most common in subgenus Psychotria, with mostly nodule-free species. In subgenus Tetramerae, with only leaf-nodulated species, there is a strong tendency toward brushlike and dendroid colleters in which epidermal cells are extremely elongate and separated from each other. It is speculated that this change in colleter morphology associated with presence of nodule bacteria may be correlated with a change in secretion product more suitable for support of bacteria. Three morphological forms of crystal occur: raphides, styloids, clustered crystals. They may occur singly or in combination. Several patterns and trends were noted in crystal distribution that could be of taxonomic significance.     

The Development, Structure and Function of Dendroid Colleters in Psychotria kirkii Hiern (Rubiaceae)

The dendroid colleters of the leaf nodulated Rubiaceous shrubPsychotria kirkii Hiern. have been studied with respect to theirdevelopment, structure and function. The colleters, which arisefrom the adaxial surface of stipules of apical and lateral shoots,secrete a protein/carbohydrate mucilaginous substance in whichis maintained a colony of leaf nodule bacteria. The colletersare multicellular and multiseriate, consisting of a two to fourcell thick stalk from which radiate up to 70 elongate secretorybranch cells. Cuticle envelops both stalk and branch cells inearly developmental stages and as secretory activity increasesthis cuticle is largely lost in two different ways. The majorpart is forced off the branch cell surface through the passageof a largely carbohydrate component of the mucilage which surroundsthe colleters: a second method of cuticle loss involves thepassage from the cell of small electron-translucent bodies whichbecome coated with cuticle as they exit the cell. The mucilagein which the bacterial cells are found provides a vehicle wherebythe bacteria are able to enter the leaves, thus leading to theinitiation of leaf nodules. Psychotria kirkii Hiern., secretory dendroid colleters, symbiosis, ultrastructure, trichome development     

Ontogenesis, structure and ultrastructure of Hymenaea stigonocarpa (Fabaceae: Caesalpinioideae) colleters

The genus Hymenaea is characterized by a great diversity of secretory structures, but there are no reports of colleters yet. The objectives of this study are to report the occurrence and describe the origin and structure of colleters in Hymenaea stigonocarpa Mart. ex Hayne. Shoot apex samples were collected, fixed, and processed for light microscopy, scanning electron microscopy, and transmission electron microscopy as per usual methods. Colleters occur predominantly on the stipule's adaxial side. These structures are found at the base on a narrow strip, corresponding to the median vein up to half the length of the stipule. When present on the abaxial side, they are concentrated at the base and restricted to the margins. Colleters develop from the protoderm; they are elongate and club-shaped. Their body has no stratification; their surface cells differ from the inner cells only in position and presence of cuticle. Colleter cells have thin walls, dense cytoplasm, large nuclei, many mitochondria, rough endoplasmic reticulum, and abundant dictyosomes. Histochemical tests with Ruthenium red showed pectic compounds in the cytosol. In H. stigonocarpa, colleter arrangement is compatible with the hypothesis that they protect shoot apex. In this species, protection is reinforced by the sheath formed by the stipule pairs.     

An Electron Microscopic Study of Double Fertilization in Allohexaploid Wheat Triticum aestivum L.

Double fertilization has been examined by electron microscopyin allohexaploid wheat, Triticum aestivum L. cv. Mardler. Serialsections through fertilized ovules revealed that following dischargeof the pollen tube contents into the degenerate synergid thelatter extended to form a continuum between the egg and centralcells. The two naked sperm nuclei were seen at the far end ofthis extended synergid. These observations suggest that thedegenerate synergid may play a role in transporting the spermnuclei to the site where they can be accepted by the egg andcentral cell. In comparison with double fertilization in Plumbagozeylanica L., we also suggest that the degenerate synergid preventsmale cytoplasms from being transmitted to the egg and centralcell. The present study also confirms that in the fertilizedcentral cell the maternal and paternal genomes remain physicallyseparate at least until the first nuclear division of the coenocyticphase of endosperm development. Double fertilization, degenerate synergid, Triticum aestivum     

Dendroid colleters on vegetative and reproductive apices in Alibertia sessilis (Rubiaceae) differ in ultrastructure and secretion

In this work we compare the structure and secretion of dendroid colleters on stipules, bracts and sepals of Alibertia sessilis, a non-nodulate Rubiaceae species from Brazilian cerrado, with notes on the plant phenology. Samples were processed according to usual methods for anatomy, histochemistry and ultrastructure. Colleters are conical and constituted by a central axis of elongated parenchyma cells from which radiate numerous epidermal cells. Epidermal cells are cylindrical on the vegetative apex and digitiform or bulbous on reproductive apex. Both colleters produce hydrophilic and lipophilic compounds. On the vegetative apex, epidermal cells present numerous well-developed Golgi bodies associated with a network of smooth endoplasmatic reticulum (SER), scarce oil bodies and profiles of rough endoplasmatic reticulum (RER), indicating the involvement of these glands in the production of mainly polysaccharides in addition to protein and lipids. Differently, epidermal cells on bracts and sepals present abundant and prominent oil drops, fewer Golgi bodies and a well developed network of SER with locally dilated cisterns indicating predominance of lipids. Ecrine and granulocrine mechanisms are common to colleters of both apices. We hypothesize that exudates protect vegetative meristems and developing organs against desiccation in the dry season and against insects and pathogens during the wet season. Predominantly lipidic secretion protects the floral organs against dehydration in the dry season and can attract floral visitants. These aspects are relevant if one considers that A. sessilis inhabits the cerrado, an environment characterized by a well-delimited dry season, high irradiance and elevated vapor pressure deficits.     

THE GYNOECIAL DEVELOPMENT AND SYSTEMATIC POSITION OF ALLAMANDA (APOCYNACEAE)

Allamanda exhibits an unusual type of gynoecial development in which the two carpels are free at initiation, but fuse completely during development, resulting in a unilocular ovary with parietal placentation at maturity. Whereas the majority of the Apocynaceae are characterized by an advanced type of gynoecium that is secondarily apocarpous, in Allamanda gynoecial evolution has proceeded one step further to secondary syncarpy. This condition is not known to occur in any other genus in the Apocynaceae and provides further evidence of the isolated position of Allamanda within the family.     

Leaf Nodule Development in Psychotria kirkii Hiern. (Rubiaceae)

The initiation, development and structure of the leaf nodulesof the Rubiaceous shrub Psychotria kirkii Hiern. has been studiedin detail at the ultrastructural level. Bacteria, maintainedin the shoot tip in the secretions from dendroid colleters,invade the substomatal chamber of stomatal pores which formprecociously on the abaxial leaf surface. Proliferation of theepidermis around the pore pushes the bacterial cavity deep intothe lamina, thus forming a small internal nodule. Endophyte-mediatedschizogeny of the cells surroundng the nodule causes it to expandwhile at the same time giving rise to an interconnected reticulumof invasive host cells which are involved in metabolite exchangebetween microoganisms and host plant. Bacterial morphology changesafter entry of the microsymbiont into the host plant and, bythe time the nodule is mature, the bacteria exhibit distinctpleomorphism. Senescent nodules are shown to accumulate lipidand starch. The developmental process is discussed in the lightof existing information on this symbiosis. Psychotria kirkii, leaf nodule development, symbiosis, ultrastructure     

Structure and Histochemistry of Stomata and Epidermal Cells in Five Species of Polypodiaceae

The epidermal structure of the five species of ferns, Arthromeriswallichiana (Spr.) Ching., Drymoglossum piloselloides (Prest.),Drynaria quercifolia (L.) J. Smith, Lepisorus nudus (Hook.)Ching. and Pyrrosia nuda (Gies.) Ching., has been investigated.Fifteen types of stomatal structures have been identified ofwhich copolo-desmocytic and coperi-desmocytic are new types.Four more possible stomatal structures: ccpolo-peri-, codesmo-polo-,codesmo-peri- and duplodesmocytic, are suggested. Localizationof starch, insoluble polysaccharides, protein and lipids hasbeen examined histochemically in the guard cells, subsidiarycells and epidermal cells. In Drynaria starch plastids and plastidscontaining both starch and protein are present in guard cells.Starch plastids are present in the subsidiary cells of all speciesexcept in Arthromeris, whereas, they are present in epidermalcells of only Drymoglossum and Lepisorus. Granular or amorphousinsoluble polysaccharides (other than starch) are present inguard cells of all the species, in the subsidiary cells of Arthromeris,Drynaria and Pyrrosia, and in the epidermal cells of Pyrrosia.Except in Pyrrosia lipids are present in the guard cells. Subsidiarycells of Drynaria and the epidermal cells of Arthromeris andDrynaria show lipid bodies. The presence of plasmodesmata andectodesmata is demonstrated in the epidermal cells of Drymoglossum.     

Colleters in Bathysa cuspidata (Rubiaceae): Development,ultrastructure and chemical composition of the secretion

This paper describes the development of colleters of Bathysa cuspidata, Rubiaceae, considering anatomical, histochemical and ultrastructural aspects and going from first differentiation stages until senescence. Further, the chemical composition of the secretion is investigated. The samples were prepared according to the usual techniques for light microscopy and scanning and transmission electron microscopy. Electrophoresis and thin layer chromatography (TLC) were used to confirm the results obtained in the histochemical tests. The colleters occur at the ventral surface of the stipules which protect the leaf primordia as well as the shoot meristem. The origin of the colleters is mixed, involving protoderm and ground meristem. The Bathysa colleters are of the standard type or are bifurcated; this latter type is documented here for the first time for Rubiaceae. Colleter secretion is a mucilage rich in protein, as determined by histochemical tests and confirmed by chemical analysis. Phenolic compounds and terpenes were detected only in the colleters themselves, but not in the secretion. The epithelial cells present conspicuous nuclei and nucleoli and the cytoplasm is rich in dictyosomes, endoplasmic reticulum, mitochondria, vesicles and small vacuoles with a fibrillar content. The accumulation of phenolic compounds and terpenes, the formation of a large central vacuole, the increase of the intercellular and subcuticular spaces occupied by the secretion and, finally, the darkening and the wilting of the colleters characterize the senescence of these structures. The secretion process of the colleters of B. cuspidata suggests a process of programmed cell death.     

Ontogenesis secretion and senescence of Tocoyena bullata (Vell.) Mart. (Rubiacaeae) colleters

Colleters are secretory structure present on many families including Rubiaceae. Particular characteristics have been described about colleters secretory cells, however senescence process are still under debate. Tocoyena bullata (Vell.) Mart. (Rubiaceae) shoot apex were collected at Jardim Botânico do Rio de Janeiro, RJ/Brazil. Stipules were separated and fragments were fixed in 2.5% glutaraldehyde and 4.0% formaldehyde in 0.05 m sodium cacodylate buffer, pH 7.2, post fixed in 1.0% osmium tetroxide in the same buffer, dehydrated in acetone, critical‐point‐drying, sputtered coated and observed. For light microscopy fragments were fixed and dehydrated, infiltrated with historesin and stained with 1% toluidine blue. For transmission electron microscopy, the samples were infiltrated with Epoxi resin. Colleters are present on stipule adaxial surface. On the beginning of development, these structures are recognized as small projections. Later on, colleters differentiated and secrete by cuticle rupture. The colleters senescence occurs in a concomitant and indissoluble way of programmed cell death. Ultrastructural analyses during the process strongly suggest the senescence is based on a non‐autolitic programmed cell death. T. bullata colleters, present at stipule abaxial surface are cylindrical secretory structures. Colleters secretory cells originated as stipule projections; differentiate; secrete and senesce by programmed cell death. The secretion and the cell dead occurs in a concomitantly and indissoluble way.     

Localization of Peroxidized Lipids in Non-Sedimentable Microvesicles of Senescing Bean Cotyledons

Fluorescent peroxidized lipids are present in lipid extractsof microsomal membranes and cytosol from young and senescingbean (Phaseolus vulgaris) cotyledon tissue. In young tissue,the peroxidized membrane lipids are mainly phospholipids, whereasthose in the cytosol are primarily free fatty acids. With advancingsenescence, microsomal peroxidized lipids increase by 200% relativeto membrane protein and by 50% on a per cotyledon basis, andthe increase is mainly attributable to enhanced levels of peroxidizedfree fatty acids. Cytosolic peroxidized lipids expressed ona per cotyledon basis decline by 55% over the same period. Fractionationof the cytosol revealed that, for both young and senescing tissue,about 50% of the cytosolic fluorescent peroxidized lipids areassociated with non-sedimentable microvesicles, which are formedfrom membranes and enriched in phospholipid catabolites. Moreover,the decline in cytosolic peroxidized lipids with advancing senescencecorrelates with progressive impairment of the formation of thesenon-sedimentable microvesicles. Key words: Phaseolus vulgaris, senescence, lipid peroxidation, fluorescence     

Storage Lipid Accumulation by Zygotic and Somatic Embryos in Culture

In vitro accumulation of storage lipids occurs in zygotic andsomatic embryos of Brassicu napus L. The concentration of sucrosein the medium modified the pattern of storage lipid accumulationin zygotic and somatic embryos. The sucrose concentration atwhich the maximum amount of lipid is accumulated by the twotypes of embryos is different Analysis of fatty acid compositionshowed that the same fatty acids are present in embryos in vivoand those cultured in vitro although there are quantitativedifferences. The possibility of using this type of system forin vitro production of valuable plant metabolites is discussed Embryo cloning, somatic embryogenesis, in cilro culture, storage lipids, Brussica napus, oilseed rape