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.     

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.     

Colleters in Turnera and Piriqueta (Turneraceae)

The anatomy of colleters was examined by light and scanning electron microscopy in 25 species of Turnera and nine species of Piriqueta. Based on morphology, four categories of colleters were recognized: standard, sessile, lachrymiform and trochleariform, all of which differ in shape and length/width ratio. They all have a similar anatomy: they consist of an axis of parenchymatous cells, sheathed by a palisade epidermis. The standard type is the most widespread in the studied taxa; the lachrymiform example was found in those species of Piriqueta with setiform glandular hairs; only one trochleariform example appeared in T: diffusa. The sessile type is considered to be a morphological transitional form between extrafloral nectaries and colleters. This is the first record of sessile, lachrymiform and trochleariform colleters. The anatomy of colleters is compared with other secretory structures such as glandular trichomes and extrafloral nectaries.     

The structure of colleters in several species of Simira (Rubiaceae)

BACKGROUND AND AIMS: Colleters are secretory structures consisting of a parenchymatic middle axis surrounded by a layer of palisade-like epidermal cells. Colleters occur in a large number of rubiaceous species. Their function is to protect the developing shoot apex. They are also taxonomically useful in the Rubiaceae. This study characterized the structure of the colleters of Simira glaziovii, S. pikia and S. rubra and the biochemistry of secretions in S. glaziovii. METHODS: Stipules of the shoot apices of the three species studied were collected at Barragem de Saracuruna, in Rio de Janeiro state, Brazil. The samples were fixed according to the usual methods for light and electron microscopy. Secretion stipules of S. glaziovii were washed with 0.1 m Tris-HCl plus 0.1 %Triton X-100 to extract proteins and carbohydrates. KEY RESULTS: Colleters in these species are located at the base of the stipule. Each species shows a different pattern of distribution. They form as emergentia from the stipules. Simira glaziovii was different from the other two species because it exhibited vascular traces. The epidermal cells of colleters have dense cytoplasm, nuclei, small vacuoles, endoplasmic reticulum, Golgi apparatus, mitochondria and extraplasmic spaces if they are secretory. The outer cell wall of the mature colleters differs from the outer cell wall of stipule cells and immature colleters. Both carbohydrates and proteins were found in secretions from the stipules of S. glaziovii. CONCLUSIONS: Few ultrastructural differences were noted among the three species. These secretory structures not only protect the shoot apex, but also have taxonomic importance below the genus level.     

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     

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     

Foliar secretory trichomes of Ocimum obovatum (Lamiaceae): micromorphological structure and histochemistry

This study characterises the micromorphology, ultrastructure and main chemical constituents of the foliar glandular trichomes of Ocimum obovatum using light and electron microscopy and a variety of histochemical tests. Two types of glandular trichomes occur on the leaves: large peltate and small capitate. The head of each peltate trichome is made up of four broad head cells in one layer. The head of each capitate trichome is composed of two broad head cells in one layer (type I) or a single oval head cell (type II, rare). In peltate heads, secretory materials are gradually transported to the subcuticular space via fracture in the four sutures at the connecting walls of the head cells. Release to the head periphery occurs through opposite fracture in the four sutures in the head cuticle. In type I capitate trichomes, release of the secretions to the subcuticular space occurs via a pore between the two head cells, and release to the head periphery occurs through the opposite pore in the head cuticle. In type II capitate trichomes, the secreted material is released from the head cell through a ruptured particular squared area at the central part of the head cuticle. These secretion modes are reported for the first time in the family Lamiaceae. Histochemical tests showed that the secretory materials in the glandular trichomes are mainly essential oils, lipophilic substances and polysaccharides. Large peltate trichomes contain a large quantity of these substances than the small capitate trichomes. Ultrastructural evidence suggests that the plastids produce numerous lipid droplets, and the numerous polysaccharide small vesicles are derived from Golgi bodies.     

Bacterial leaf nodule symbiosis in angiosperms with emphasis on Rubiaceae and Myrsinaceae

Over 400 species of three genera of Rubiaceae and one genus of Myrsinaceae reportedly have bacterial leaf nodules. Light and/or electron microscope studies of a few species have shown that bacteria exist in spaces within buds filled with mucilage secreted by glands. These bacteria enter substomatal chambers (Rubiaceae) or marginal hydathodes (Myrsinaceae) and establish short-lived colonies, in intercellular spaces, that die out almost before full leaf expansion. Bacteria occur in seeds between endosperm and embryo, but only two studies have followed bacteria into flowers and ovules. Previous work on the physical relations of bacteria and host plants is discussed critically. Reviewing work done on isolation and identification of presumed endophytes leads to the conclusion that there is no agreement whether one or several bacterial taxa are the endophyte, and no unambiguous identifications, although four genera are suggested as possibilities. Nitrogen fixation was considered as the bacterial contribution until quite recently, but a review of such studies reveals that fixation has been detected almost exclusively in isolated presumed endophytes, whereas almost all studies involving the bacterium in intact leaves have failed to detect nitrogen fixation. Studies of particular substances (besides combined nitrogen) contributed by the endophyte have been inconclusive, although the most recent works suggest that cytokinins are involved. Host plants lacking the endophyte have been reportedly produced many times, either spontaneously or by seed treatment. Such “cripples”, used for several aspects of symbiosis study, frequently revert to a nodulated condition, and a more reliable method of producing them is needed. Tissue culture may offer the best potential, but this approach has not yet produced whole bacteria-free plants. A proposed scheme for the evolution of the symbiosis suggests that a variety of bacteria entered buds first, and only in rare instances were compatible with the host bud mucilage. In a few of these cases, specific bacteria, compatible with the microenvironment, contributed a useful substance to the host, and bud mucilage and those bacteria co-evolved until large numbers of bacteria thrived in the buds. Nodules may have resulted from accidental entry of bacteria into leaves, with the possibility that some host plant nodules are merely pathogenic responses, whereas in others the bacteria are beneficial and further selection has resulted in numerous, regularly produced nodules. This review deals with taxonomy of host plants and endophytes, morphology of the symbiosis, its physiology, and speculation on the evolution of the symbiosis.     

木香薷腺毛形态结构发生发育规律的研究

采用常规石蜡切片法及扫描电镜技术对木香薷(Elsholtzia stauntoni Benth)腺毛发生发育及其规律进行了研究。结果表明：木香薷表皮上主要有两种表皮毛：无分泌细胞的表皮毛与有分泌细胞的腺毛。前者包括单细胞乳头状毛、2~3细胞管状毛、分枝状毛及多细胞管状毛;后者包括头状腺毛与盾状腺毛。成熟头状腺毛头部由1、2或4个分泌细胞构成,头部呈圆球形或半圆球形;成熟盾状腺毛头部由8~12个分泌细胞构成,分泌细胞横向扩展形成盾状头部。木香薷腺毛主要在茎端幼叶处大量发生,从茎端第一对幼叶处开始产生;从幼叶期到成熟期均有腺毛发生,大部分腺毛在幼叶期发生发育,只有极少部分在叶的成熟期进行发生发育。     

Structural investigation of the glandular trichomes of <Emphasis Type="Italic">Salvia argentea</Emphasis>

The morphology, anatomy and distribution of glandular trichomes on the aerial organs of Salvia argentea L. has been investigated. Two morphologically distinct types of glandular trichomes were determined. Capitate glandular trichomes forming a base 1–7 celled, a stalk 1–5 celled or no stalk and a head uni- or bicellular had various types. In capitate trichomes, the neck cell that has an important role especially for xeroformic plants, acting to prevent the backflow of secreted substance through the apoplast has been distinctively observed in the investigated species. The capitate trichomes were present abundantly on all aerial organs of S. argentea. Peltate glandular trichomes had a large secretory head forming 1–5, 8 central and 8–10, 12, 14 peripheral cells. Peltate trichomes are present on all aerial organs, except petiole, being the most abundant on calyx and corolla. Results were shown by tables and photographs.     

Development and Essential Oil Content of Secretory Glands of Sage (Salvia officinalis)

Scanning electron microscopy of sage (Salvia officinalis L.) leaves confirmed the presence of two basic types of glandular trichomes consisting of a capitate stalked form containing a multicellular stalk and surmounted by a unicellular secretory head, and a capitate sessile form containing a unicellular stalk and unicellular, or multicellular, secretory head. In the latter type, secretory activity and filling of the subcuticular cavity may begin at virtually any stage of the division cycle affording fully developed glands containing from one to twelve cells in the secretory head. Gas liquid chromatographic analysis of the oil content of the most numerous gland species (capitate stalked, capitate sessile with one and with eight secretory cells) indicated only minor quantitative differences in essential oil composition. Thus, each gland type is capable of producing the four major monoterpene families (p-menthanes, pinanes, bornanes and thujanes) characteristic of sage.     

Anatomical inferences on aerial bud protection of three Eugenia shrub species from the Cerrado

• Location and degree of protection of aerial buds are important functional traits in disturbance- or stress-prone environments since aerial buds ensure the development of new organs under favourable growing conditions. This study was carried out in a Brazilian Cerrado area under regeneration after long-term Pinus cultivation, where the trees were clear-cut in 2012 and the remaining material was burned in 2014.
• After the fire treatment, several species resprouted from belowground organs and their aboveground organs were directly exposed to full sunlight. We collected 15 terminal branches with fully expanded leaves from three individuals of each of three Eugenia species to investigate if those with well-developed belowground organs invest in bark for aboveground bud protection. The samples were analysed using light and electron microscopy.
• In addition to terminal and axillary buds, all species presented accessory buds, and the number varied according to the node analysed. None of the aerial buds were protected by bark, but all were well protected by cataphylls and densely pubescent leaf primordia. There were also inter- and intra-petiolar colleters that released a mucilaginous protein exudate. The distance between the shoot apical meristem and the outer surface was longer in the terminal bud than in axillary buds. The bud leaf primordia covering the shoot apical meristem had a thick cuticle, unicellular non-glandular trichomes that accumulate phenolic and lipophilic compounds, and secretory cavities.
• Our study shows that all three Eugenia species studied here had highly protected aerial buds allocated from belowground organs. These morphological traits may improve the chances of the species' persistence in areas subjected to frost events, low relative humidity, high irradiance and harmful UV levels.

     

A flower with several secretions: anatomy,secretion composition,and functional aspects of the floral secretory structures of Chelonanthus viridiflorus (Helieae—Gentianaceae)

Floral secretory structures have been reported for Gentianaceae; however, morphoanatomical studies of these glands are rare. We described the development and secretory activity of the colleters and nectaries throughout the floral development of Chelonanthus viridiflorus. We collected flower buds, flowers at anthesis, and fruits to be investigated using light and scanning electron microscopy. We performed histochemical tests on the secretion of colleters and used glycophyte to confirm the presence of glucose in nectar. Colleters are located on the ventral surface of sepals and nectaries occur in four regions: (i) the dorsal and (ii) ventral surfaces of sepals; (iii) apex of petals; and (iv) base of ovary. The colleters have a short peduncle and a secretory portion with homogeneous cells. They are active in flower buds and secrete polysaccharides and proteins. In flowers at anthesis, they begin to senescence presenting protoplast retraction, cell collapse, and lignification; these characteristics are intensified in fruit. The nectaries of sepals and petals have two to five cells surrounding a central cell through which the secretion is released. Nectaries are numerous, forming a nectariferous area on the dorsal surface of sepals, like that observed on petals, and can form isolated units on the ventral surface of sepals. They are active from flower buds to fruits. A region with secretory activity was identified at the base of the ovary. The secretion of colleters acts in the protection of developing organs, while nectaries are related to defenses against herbivores and the supply of nectar to potential robbers or pollinators.

     

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.     

羽叶薰衣草表皮毛的发育解剖学研究

对羽叶薰衣草(LavandulapinnataL.)茎和叶上两种表皮毛(腺毛和非腺毛)发育的解剖学观察表明,两者的发生都源于茎或叶的原表皮细胞,但外部形态、发育过程及功能明显不同。腺毛有头状腺毛和盾状腺毛两种类型,均由1个基细胞、1个柄细胞和头部细胞构成。头状腺毛的头部只有1个或2个分泌细胞,盾状腺毛由8个分泌细胞构成头部。非腺毛由3-20个细胞组成,可分为三种类型:单列不分枝、二叉分枝和三叉及三叉以上多分枝的树状分枝。非腺毛的顶部细胞由基部到顶部逐渐变细,先端成尖形。腺毛发育由原表皮细胞经两次平周分裂形成,由于柄细胞和头部细胞所处的分化状态不同而发育成两类腺毛。非腺毛由非腺毛原始细胞经二次或多次平周分裂和不均等分裂,再发育成数个至二十多个子细胞。     

Calycine colleters of seven species of Apocynaceae (Apocynoideae) from Brazil

The structure and organization of calycine colleters belonging to four genera of Apocynaceae are described. Calycine colleters were found in all species. All colleters had the same histological composition, but morphological differences were observed in four species. Most colleters were of the standard type, and possible deviations resulted in three morphological types: bifurcated, laminar, and sessile. Mechanisms of cell separation, proliferation, and elongation may explain the origin of the new types. As a consequence of morphological changes, the secretory surface was expanded in the bifurcated, laminar, and sessile glands. Most colleters were in a post-secretory phase, but a hydrophilic secretion was observed in Mandevilla pycnantha and Mesechites mansoana . A larger secretory surface, together with the extension of the secretory phase in adult flowers, were major shifts in the calycine colleters of Apocynaceae, and are possibly correlated with the functional and ecological aspects of these organs. Three patterns of distribution of calycine colleters were observed: alternate, opposite, or indefinitely distributed. The alternate pattern was not homogeneous, and three subtypes were recognized: four-, five-, and ten-grouped colleters. The number of calycine colleters was variable between taxa and even within the same species.  © 2006 The Linnean Society of London, Botanical Journal of the Linnean Society , 2006, 152 , 387–398.     

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.     

Structure and function of trichornes in the shoot tip of Ardisia crispa (Thunb.) A.DC. (Myrsinaceae)

MILLER, I. M., GARDNER, I. C. & SCOTT, A., 1984. Structure and function of trichomes in the shoot tip of Ardisia crispa (Thunb.) A.DC. (Myrsinaceae). The trichomes in the shoot tip of the myrsinaceous leaf-nodulated species Ardisia crispa (Thunb.) A.DC. have been studied at the ultrastructural level. Two distinct kinds of trichomes are found. Small peltate scales arise from the abaxial protoderrn of the developing leaves. Multicellular stellate trichomes on the adaxial surface of young leaves are uniseriate, rotate and consist of up to eight multicellular arms radiating from a central stalk. The multicellular arms terminate distally in swollen club-shaped tips. The stellate trichomes secrete large quantities of carbohydrate-containing mucilage. Colonies of leaf nodule bacteria are found resident in this mucilage. The role of the peltate scales and the stellate trichomes n the maintenance of the cyclic leaf nodule symbiosis is discussed.     

Trichome micromorphology of the East Asiatic genus Chelonopsis (Lamiaceae) and its systematic implications

The micromorphology of trichomes of the leaves of 17 taxa (including two varieties) of the genus Chelonopsis Miq. and of six species representing four additional genera (Bostrychanthera deflexa Benth., Colquhounia coccinea Wall. var. coccinea, Co. seguinii Vaniot. var. seguinii, Gomphostemma chinense Oliv. var. chinense, G. crinitum Wall. ex Benth. and Physostegia virginiana (L.) Benth.) was surveyed by light and scanning electron microscopy. Two basic types of trichomes can be identified: non-glandular and glandular trichomes. The non-glandular trichomes can be subdivided into two subtypes: simple unbranched and branched trichomes. Based on the cell number, simple unbranched trichomes are further divided into four shapes (unicellular, two-celled, three-celled, and more than three cells), whilst branched trichomes are separated into three shapes (biramous, stellate, and dendroid trichomes). The glandular trichomes can in turn be subdivided into four subtypes: subsessile, capitate, clavate, and sunken. Non-glandular trichomes with two cells (NGTW) and subsessile glandular trichomes (GSU) are most widespread in all taxa examined. The indumentum shows considerable variation among different sections or species. Consequently, trichome micromorphology and distribution have high taxonomic value for Chelonopsis at both infrageneric and interspecific levels. The presence of capitate glandular trichomes (GCA) provides an additional morphological character to clarify the boundaries between subgenus Chelonopsis and Aequidens Wu and Li. Within subgenus Aequidens, non-glandular trichomes with more than three cells (NGMT) and clavate glandular trichomes (GCL) are important characters for sectional division between sect. Aequidens Wu and Li and sect. Microphyllum Wu and Li. Again, three forms of three-celled trichomes can be used as a distinctive taxonomic character at specific level between C. albiflora Pax et K. Hoffm. ex Limpr., C. forrestii J. Anthony, and C. souliei (Bonati) Merr. in sect. Aequidens. This study supports Wu's delimitation of subgenus and sections and the subsequent review work by Xiang et al. Additionally, distribution of trichome types is correlated with the altitudinal distribution and habitats of some species in Chelonopsis.