Peltate trichomes in the dormant shoot apex of Metrodorea nigra,a Rutaceae species with rhythmic growth

• In Metrodorea nigra, a Rutaceae species with rhythmic growth, the shoot apex in the dormant stage is enclosed by modified stipules. The young organs are fully covered with peltate secretory trichomes, and these structures remain immersed in a hyaline exudate within a hood-shaped structure. Our study focused on the morpho-functional characterization of the peltate trichomes and cytological events associated with secretion.
• Shoot apices were collected during both dormant and active stages and processed for anatomical, cytochemical and ultrastructural studies.
• Trichomes initiate secretion early on, remain active throughout leaf development, but collapse as the leaves expand; at which time secretory cavities start differentiation in the mesophyll and secretion increases as the leaf reaches full expansion. The subcellular apparatus of the trichome head cells is consistent with hydrophilic and lipophilic secretion. Secretion involves two vesicle types: the smaller vesicles are PATAg-positive (periodic acid/thiocarbohydrazide/silver proteinate) for carbohydrates and the larger ones are PATAg-negative. In the first phase of secretory activity, the vesicles containing polysaccharides discharge their contents through exocytosis with the secretion accumulating beneath the cuticle, which detaches from the cell wall. Later, a massive discharge of lipophilic substances (lipids and terpenes/phenols) results in their accumulation between the wall and cuticle. Release of the secretions occurs throughout the cuticular microchannels.
• Continued protection of the leaves throughout shoot development is ensured by replacement of the collapsed secretory trichomes by oil-secreting cavities. Our findings provide new perspectives for understanding secretion regulation in shoot apices of woody species with rhythmic growth.

     

Localization and secretory pathways of a 58K-like protein in multi-vesicular bodies in callus of Arabidopsis thaliana

Multi-vesicular bodies in endocytosis and protoplasts are special cellular structures that are consid-ered to be originated from invagination of plasma membranes. However, the genesis and function of multi-vesicular bodies, the relationship with Golgi bodies and cell walls, and their secretory pathways remain controversial and ambiguous. Using a monoclonal antibody against an animal 58K protein, we have detected, by Western blotting and confocal microscopy, that a 58K-like protein is present in the calli of Arabidopsis thaliana and Hypericum perforatum. The results of immuno-electron microscopy showed that the 58K-like protein was located in the cisternae of Golgi bodies, secretory vesicles, multi-vesicular bodies, cell walls and vacuoles in callus of Arabidopsis thaliana, suggesting that the multi-vesicular bodies may be originated from Golgi bodies and function as a transporter carrying substances synthesized in Golgi bodies to cell walls and vacuoles. It seems that multi-vesicular bodies have a close relationship with the development of the cell wall and vacuole. The possible secretory pathways of multi-vesicular bodies might be in exocytosis, in which multi-vesicular bodies carry sub-stances to the cell wall for its construction, and in endocytosis, in which multi-vesicular bodies carry substances to the vacuole for its development, depending on what they carry and where the materials are transported. We hence propose that there is more than one pathway for the secretion of multi-vesicular bodies. In addition, our results provided a paradigm that a plant molecule, such as the 58k-like protein in callus of Arabidopsis thaliana, can be detected using a cross-reactive monoclonal antibody induced by an animal protein, and illustrate the existence of analog molecules in both animal and plant kingdoms.     

Ovary peltate trichomes of Zeyheria montana (Bignoniaceae): developmental ultrastructure and secretion in relation to function

BACKGROUND AND AIMS: Nectar production in the Bignoniaceae species lacking a nectariferous functional disc is ascribed to trichomatic glands around the ovary base and/or on the inner corolla wall. Nevertheless, knowledge about the secretion and function of these glands is very incomplete. The purpose of this paper is to study, from a developmental viewpoint, the ultrastructure, histochemistry and secretory process of the peltate trichomes on the ovary of Zeyheria montana, a species in the Bignoniaceae which has a rudimentary disc. METHODS: Samples of the gynoecium at various developmental stages were fixed and processed for light and electron microscopy. Histochemistry and cytochemistry tests were performed to examine the chemical composition of exudates. Thin layer chromatography was used to determine the presence of alkaloids and terpenes in gynoecium and fruit extracts, and in fresh nectar stored in the nectar chamber. KEY RESULTS: Peltate trichomes at different developmental stages appear side by side from floral budding up to pre-dispersal fruit. Large plastids with an extensive internal membrane system consisting of tubules filled with lipophilic material, abundant smooth endoplasmic reticulum, few Golgi bodies, lipophilic deposits in the smooth endoplasmic reticulum and mitochondria, and scattered cytoplasmic oil droplets are the main characteristics of mature head cells. The secretion which accumulates in the subcuticular space stains positively for hydrophilic and lipophilic substances, with lipids prevailing for fully peltate trichomes. Histochemistry and thin layer chromatography detected terpenes and alkaloids. Fehling's test to detect of sugars in the secretion was negative. CONCLUSIONS: The continuous presence and activity of peltate trichomes on the ovary of Z. montana from early budding through to flowering and fruiting set, and its main chemical components, alkaloids and terpenes, suggest that they serve a protective function and are not related to the floral nectar source or to improving nectar quality.     

薄荷头状腺毛分泌过程的超微结构研究

电镜观察表明,刚形成的薄荷头状腺毛的头部细胞,细胞核较大细胞质浓,有一些小液泡,质体和线粒体最显著,分泌前期,内质网及高尔基体数量明显     

Studies in the secretory glands of Hiptage sericea (Malpighiaceae)

Hiptage sericea is shown to possess both lipophilic glands and extrafloral nectaries. Both types of glands develop from a group of initials and show similarities in organisation of tissue systems and secretion. The nature of the secretory substances is however different. The occurrence and function of the glands are discussed.     

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.     

Observations on the development of the foliar secretory cavities of Porophyllum lanceolatum (Asteraceae)

The lipophilic secretory cavities observed in the leaf of Porophyllum lanceolatum (Asteraceae) are scattered throughout the lamina and around its crenate margins. In the young leaf the cavities are initiated, and their development completed, while the surrounding tissues are still at early stages of differentiation. The cavity lumen has a lysigenous origin. Cell lysis, expansion of the developing leaf and, probably, the pressure exerted by the accumulation of secretory products, are believed to account for the gradual enlargement of the lumen. Concomitantly with ctll disintegration, which occurs throughout development, divisions take place in all cells of the gland. A mature cavity has a multilayered epithelium. Histochemical tests for RNA, proteins, phenolics and pectic polysaccharides revealed intense staining of the content of the epithelial cells in the early stages of cavity development, and a decrease in staining towards its maturity. Staining for lipids is intense in all developmental stages. Tests on the material observed in the lumen of mature cavities, show positive results for lipids, pectic polysaccharides and phenolics.     

The secretory apparatus of Ginkgo biloba: structure,differentiation and analysis of the secretory product

Summary The differentiation of the secretory cavities of Ginkgo stem and the structural organization of the epithelial cells were followed by light and electron microscopy. The mode of formation of the cavities is schizo-lysigeneous. Functional complexes of leucoplasts and associated endoplasmic reticulum (ER) membranes are assumed to be the site of synthesis and translocation of the lipophilic secretory product. Most of the endoplasmic reticulum membranes are paired. The content of the cavities was directly collected and analysed by low- and high-resolution mass spectrometry. The cavities contain anacardic acids and cardanols, which are long-chain phenol lipids not characteristic of Ginkgo. The relationship between the plastid/ER complexes and the production of these secondary metabolites is discussed.     

椭圆背角无齿蚌外套膜组织培养与未培养细胞分泌活动的扫描电镜观察

作者用扫描电镜及相差显微镜,对椭圆背角无齿蚌外套膜组织培养与未培养细胞的分泌活动进行了研究,观察到两者的分泌活动都是十分旺盛的。培养细胞有局部分泌和顶浆分泌。细胞分泌形态观察到三种:(1)分泌端形成由膜包裹的突起,突起逐渐伸长,基部变成细颈,最后脱离细胞成为分泌泡(局部分泌);(2)细胞端部伸出长足,将分泌物排到较远处分泌后,长足缩回恢复原状;(3)分泌端伸出很多细枝,分泌物随后如液流式涌出细胞(顶浆分泌)。取外套膜色线边组织为材料,培养后在组织块和细胞上有角质素(与贝壳最外层相似)类的茶褐色结晶和无定形分泌物形成;用去掉色线边的外表皮组织块培养,则有珍珠(与贝壳最内层相似)状的白色和淡黄色结晶生成。表明了细胞在适宜的条件下培养,所形成的分泌物的性质可能与活体相同。因此大批量培养细胞可能得到人们希望获得的细胞产物。       

Development and Histochemical Observations of Tapetum and Peritapetal Membrane in Anther of Pulsatilla chinensis

Tapetum of Pulsatilla chinensis is of secretory type. Its development proceeds rapidly in following sequence: (1) The stage of initiation-differentiation. At this stage cytological and histochemical features have been described in detail in this paper. (2) The stage of growth- synthesis: This stage appears to be the most important anabolic phase during the development of the tapetum. The salient features are that the tapetal cells become relatively enlarged and form two polyploid nuclei or aberrent polyploid nuclei resulting in synthetizing maximum proteins, fluorescing substances and maximum fluorescent Pro-Ubisch bodies in the tapetal cytoplasm. (3) The stage of secretion-disorganization: After the disintegration of the tapetal wall the enlarged naked cells appear at once. This is an important secretion period in which Pro-Ubisch bodies as well as all other fluorescing substances, carbohydrate or some enzymes are released into anther loculus. The naked cell layer becomes disorgnized until the beginning divition of the pollen grains into two ceils. As to peritapetal membrane of P. chinensis, mainly based on the membrane being on the outer side of the tapetum enclosing both the pollen, tapetal cytoplasm and Ubisch bodies, and the cellular configurations facing the pollen, Authors postulate that peritapetal membrane might be survival of the cytoplasmic membrane of tapetal cells. However, the peritapetal membrane of P. chinensis is similar to that of plasmodial, tapetum reported in certain Compositae and that of secretory tapetum reported in Pinus banksiana. Heslop-Harrison and Gupta et al. had conceded that the tapetal and peritapetal membrane belong to the general class of sporopollenin. On the contrary in P. chinensis the sporopollenin property of peritapetal membrane is only confined to its inner surface. But the thin mem- brane itself with the reticulate sporopollenin attched on its inner side appears negative staining reactions for sporopollenin though it has an ability to resist the acetolysis as well. In P. chinensis the Ubisch body is short necked flask shaped and their size is very similar. Ubisch body is either single or 2–5 in a group, resulting in compound bodies. When the Pro-Ubisch body is still within the tapetal cell it shows positive fluorescent reaction, while it eomletely unstains with Teluidine blue O. So Authors infer that the sporopollenin precur- sors may have permeated through Pro-Ubisch bodies. Finally, How sporopollenin precursor is synthesized in the tapetal cells, transported to pollen locula and polymerized into the sporopollenin on pollen, Ubisch body and peritapetal membrane? Future works along these problems may yield fruitful results.     

Feinstrukturelle Untersuchungen zur Entwicklung der Ölzellen bei dem LebermoosRiella

Summary Liverworts are characterized by the possession of typical cell elements, the oil bodies. In the submerse, thalloid liverwortRiella helicophylla (Sphaerocarpales) oil bodies are existing in idioblastic cells only, called oil body cells. Each oil body cell contains only one oil body. The oil body originates from small vacuoles. Their membranes are extremely high in contrast and asymmetric. The lipophilic substances are probably produced inside the oil bodies. At the end of the development of an oil body cell lipophilic and hydrophilic material will be separated from each other inside the oil body. The result is an oil body, consisting of one large spherical oil globule surrounded by a thin layer of hydrophilic matrix.

     

Androecia in two Clusia species: development,structure and resin secretion

Clusia fluminensis and C. lanceolata are dioecious shrubs having resiniferous flowers with strongly distinct androecia. The aim of this study was to investigate the development and anatomy of their androecia and the ultrastructure, histochemistry and secretory process of their androecium resin glands, examining whether the cellular aspects of resin secretion differed between these two morphologically distinct androecia. Stamens differ, being free in C. fluminensis and clustered in a synandrium in C. lanceolata. Staminode sterility is due to the undifferentiated nature of the anthers in C. lanceolata and degeneration of meiocytes and anther indehiscence in C. fluminensis. Resin is produced in subepidermal cavities and canals with wide lumens. In the secretory stage, epithelial cells present sinuous walls, voluminous nuclei, polymorphic plastids associated with periplastidial reticulum, mitochondria, oil bodies, multivesicular bodies, endoplasmic reticulum and dictyosomes. The resin is released through rupture points on the distal surface of stamens and staminodes, associated with disrupted cavities and canals. Our results show morphological diversity associated with functional similarity. Also, a secretion pattern shared by the two species includes initiation of the secretory process in young floral buds, compartmentalisation of the secretion in pre‐anthesis buds and release of secretions at anthesis. Cellular aspects of resin secretion in these species are quite similar, as are the chemical identities of the main components of the floral resins of the genus.     

Ultrastructure of the Resin Ducts of Mangifera indica L. (Anacardiaceae). 1. Differentiation and Senescence of the Shoot Ducts

A detailed electron microscope study has led to the conclusionthat the resin ducts of the shoot of Mangifera indica L. developlysigenously. This study has also established several characteristicswhich can serve as criteria for a clear distinction betweenschizogenous and lysigenous cavities. The main characteristicsof lysigenous cavities are: (1) The presence of disorganized cytoplasm in the duct cavity. (2) The presence of wall remains attached to the wall of livingepithelial cells facing the cavity. (3) The presence of specific intercellular spaces at the cellcorners facing the duct lumen. Duct development starts with the disintegration of a file ofcells forming an initial cavity. Later the cells lining thiscavity differentiate into cells secreting lipophilic compounds.As a result of growth and differentiation of the tissues aroundthe duct, its lumen becomes compressed and comes to resemblea branched narrow slit. Such a slit may wrongly be regardedas an initial stage of a schizogenous duct. Disintegration ofepithelial cells occurs throughout all stages of development.Neighbouring cells partly fill the space which is released bydisintegrating cells. At the end of the stage of secretion thecytoplasm of all epithelial cells darkens, preceding their disintegration.This darkening is a gradual process which begins in the vicinityof ribosomes. When all dark epithelial cells disintegrate thecavity widens and the neighbouring parenchyma cells substitutefor the secretory epithelium without undergoing any significantchange in their cytoplasm. Mangifera indica L., mango, resin ducts, ultrastructure     

ULTRASTRUCTURAL DEVELOPMENT OF CAPITATE GLANDULAR HAIRS OF CANNABIS SATIVA L. (CANNABACEAE)

The capitate-sessile and capitate-stalked glands of the glandular secretory system in Cannabis, which are interpreted as lipophilic type glandular hairs, were studied from floral bracts of pistillate plants. These glands develop a flattened multicellular disc of secretory cells, which with the extruded secretory product forms the gland head and the auxiliary cells which support the gland head. The secretory product accumulates beneath a sheath derived from separation of the outer wall surface of the cellular disc. The ultrastructure of secretory cells in pre-secretory stages is characterized by a dense ground plasm, transitory lipid bodies and fibrillar material, and well developed endoplasmic reticulum. Dictyosomes and dictyosome-derived secretory vesicles are present, but never abundant. Secretory stages of gland development are characterized by abundant mitochondria and leucoplasts and by a large vacuolar system. Production of the secretory product is associated with plastids which increase in number and structural complexity. The plastids develop a paracrystalline body which nearly fills the mature plastid. Material interpreted as a secretion appears at the surface of plastids, migrates, and accumulates along the cell surface adjoining the secretory cavity. Extrusion of the material into the secretory cavity occurs directly through the plasma membrane-cell wall barrier.     

Phytochrome Control of Nyetinasty in Samanea as Modified by Oxgen, Submergence, and Chemicals

Samanea saman has rapid phytochrome-regulated nyctinasty: red light preceding darkness causes pinnules to close while far-rod light allows opening- Not only the initial angle of the pinnules, but the degree of control by phytochrome depends on the “subjective time of day” at which the tissue is exposed. Excised pairs of pinnules close rapidly when submerged in water; such closure is prevented by bubbling oxygen through the water. However, if submergence closure were due solely to low oxygen levels, then nonsubmerged pinnules in a pure nitrogen atmosphere should also close. Instead, they neither close nor respond to light, but they do respond when air is readmitted, indicating that oxygen is necessary for movement rather than for photoreception. The closure of submerged pinnules remains unexplained. Attempts to detect effects of red or far-red on oxygen uptake by pulvinus tissue were unsuccessful. The following method has been used to test the effects of various inhibitors and other substances: pinnules are excised at the first hour of the day, trimmed, submerged in a sealed chamber, given far-red light, and left in darkness while oxygen is bubbled through the medium. Concentrated test solutions are injected either initially or when the red and far-red exposures are given. Thus far, the substances tested have had little or no effect on the phytochrome response.     

ULTRASTRUCTURE OF THE SECRETORY DUCTS OF RHUS GLABRA L.

The infrastructure and development of the secretory ducts were studied in the secondary phloem of Rhus glabra L. The ducts were found to develop schizogenously. The electron microscope observations may explain the view of several previous authors of schizo-lysigenous development of the duct lumen in the Anacardiaceae. The secreted material consists of lipophilic and polysaccharide substances. The electron micrographs suggest that the lipophilic substances arise in plastids, ER cisternae, Golgi vesicles, and mitochondria. The polysaccharide constituents apparently originate from the outer wall layers of the epithelial cells. The wall layers facing the lumen of the duct disintegrate and form, together with the secreted osmiophilic droplets, the gum-resin. Numerous microtubules were found along walls of the epithelial cells.     

The secretory apparatus of Xerophyta viscosa (Velloziaceae): Epidermis anatomy and chemical composition of the secretory product

In this study, secretory activity on the adaxial surface of the leaves of the desiccation tolerant plant, Xerophyta viscosa Baker was investigated, using light, scanning and transmission electron microscopy. Glandular activity was associated with sunken cavities which appear to be modifications of infolded epidermal cells. The secretory cavity consisted of a globose lumen surrounded by two layers of cells. The cells of the outer layer were flattened with thickened walls, while those of the inner layer (epithelium), exhibited ultrastructural features of intense metabolic activity. Epithelial cells were larger with a large nucleus, numerous vacuoles, and a dense cytoplasm with abundant mitochondria, endoplasmic reticulum (ER), plastids and a few dictysomes. Lipophilic droplets were abundant in the cytoplasm, mitochondria, ER, plastids and in infolding sites of the plasmalemma outside the protoplast. ER appeared to be involved in the synthesis and transport of lipophilic substances. The mechanism of secretion in X. viscosa appeared to be granulocrine. The chemical composition of hexane and methanol extracts of the leaves, analysed by gas chromatography coupled with mass spectrometry (GC–MS), revealed the presence of diterpenes, phenolic compounds and fatty acids. Compounds in the hexane fraction included velloziolone, manoyl oxide, asperdiol and an unknown substance. Compounds in the methanol fraction included 5-(hydroxymethyl)-2-furancarboxaldehyde, 2,6-dimethoxyphenol, 4-(3-hydroxy-1-propenyl)-2-methoxyphenol and stearic acid.     

杭白芷根的分泌道超微结构与其挥发油分泌的关系

利用光镜及透射电子显微镜技术研究了杭白芷根中分泌道结构及其挥发油的分泌,并重点探讨分泌道中挥发油的分泌过程。结果显示:(1)杭白芷的分泌道是由上皮细胞围绕着的伸长的胞间隙,腔道内贮存着挥发油。(2)分泌道细胞的质体、细胞基质以及线粒体参与挥发油或其前体物质的合成。(3)在分泌道发育的后期,大量小泡与分泌细胞的液泡膜和细胞质膜融合,将其内的物质释放进入空腔。研究认为,杭白芷分泌道中挥发油主要合成部位为质体及细胞基质,之后以扩散渗透或通过膜质小泡与液泡及质膜融合这两种方式分泌到空腔内,丰富的线粒体可能为这一系列过程提供能量。     

LYSOSOME FUNCTION IN THE REGULATION OF THE SECRETORY PROCESS IN CELLS OF THE ANTERIOR PITUITARY GLAND

The nature and content of lytic bodies and the localization of acid phosphatase (AcPase) activity were investigated in mammotrophic hormone-producing cells (MT) from rat anterior pituitary glands. MT were examined from lactating rats in which secretion of MTH¹ was high and from postlactating rats in which MTH secretion was suppressed by removing the suckling young. MT from lactating animals contained abundant stacks of rough-surfaced ER, a large Golgi complex with many forming secretory granules, and a few lytic bodies, primarily multivesicular bodies and dense bodies. MT from postlactating animals, sacrificed at selected intervals up to 96 hr after separation from their suckling young, showed (a) progressive involution of the protein synthetic apparatus with sequestration of ER and ribosomes in autophagic vacuoles, and (b) incorporation of secretory granules into multivesicular and dense bodies. The content of mature granules typically was incorporated into dense bodies whereas that of immature granules found its way preferentially into multivesicular bodies. The secretory granules and cytoplasmic constituents segregated within lytic bodies were progressively degraded over a period of 24 to 72 hr to yield a common residual body, the vacuolated dense body. In MT from lactating animals, AcPase reaction product was found in lytic bodies, and in several other sites not usually considered to be lysosomal in nature, i.e., inner Golgi cisterna and associated vesicles, and around most of the immature, and some of the mature secretory granules. In MT from postlactating animals, AcPase was concentrated in lytic bodies; reaction product and incorporated secretory granules were frequently recognizable within the same multivesicular or dense body which could therefore be identified as "autolysosomes" connected with the digestion of endogenous materials. Several possible explanations for the occurrence of AcPase in nonlysosomal sites are discussed. From the findings it is concluded that, in secretory cells, lysosomes function in the regulation of the secretory process by providing a mechanism which takes care of overproduction of secretory products.