Leaf blade structure of Verbesina macrophylla (Cass.) F. S. Blake (Asteraceae): ontogeny,duct secretion mechanism and essential oil composition

• Secretory structures are common in Asteraceae, where they exhibit a high degree of morphological diversity. The species Verbesina macrophylla, popularly known as assa‐peixe, is native to Brazil where it is widely used for medicinal purposes. Despite its potential medical importance, there have been no studies of the anatomy of this species, especially its secretory structures and secreted compounds. This study examined leaves of V. macrophylla with emphasis on secretory structures and secreted secondary metabolites.
• Development of secretory ducts and the mechanism of secretion production are described for V. macrophylla using ultrastructure, yield and chemical composition of its essential oils.
• Verbesina macrophylla has a hypostomatic leaf blade with dorsiventral mesophyll and secretory ducts associated with vascular bundles of schizogenous origin. Histochemistry identified the presence of lipids, terpenes, alkaloids and mucopolysaccharides. Ultrastructure suggests that the secretion released into the duct lumen is produced in plastids of transfer cells, parenchymal sheath cells and stored in vacuoles in these cells and duct epithelial cells. The essential oil content was 0.8%, and its major components were germacrene D, germacrene D‐4‐ol, β‐caryophyllene, bicyclogermacrene and α‐cadinol.
• Secretory ducts of V. macrophylla are squizogenous. Substances identified in tissues suggest that both secretions stored in the ducts and in adjacent parenchyma cells are involved in chemical defence. The essential oil is rich in sesquiterpenes, with germacrene D and its derivatives being notable components.

     

Allelopathic potential of Tagetes minuta terpenes by a chemical, anatomical and phytotoxic approach

The dominant terpenes in the essential oils of the leaves and reproductive structures of Tagetes minuta L. (Asteraceae) were studied throughout its life cycle in a natural population. The anatomy of the secretory cavities was described in order to correlate the changes in terpene content with structural changes. Finally, the phytotoxic effect of ocimenones on germination was also evaluated. Monoterpenes increased in both green leaves and reproductive structures throughout the plant's life cycle, whereas the opposite occurred in senescent leaves. Spathulenol was the main component in senescing leaves. The highest content of ocimenones, the bioactive components of the essential oil, was found in the reproductive structures. Bioassays showed that both pure ocimenones and fruit material from T. minuta delayed and inhibited the germination of cohabitant species. A relationship between allelopathy, biosynthesis, catabolism and terpene release is proposed for the chemical ecological effect of T. minuta.     

Oleoresin glands in copaíba (<Emphasis Type="Italic">Copaifera trapezifolia</Emphasis> Hayne: Leguminosae), a Brazilian rainforest tree

Although studies have addressed the chemical analysis and the biological activity of oleoresin in species of Copaifera, the cellular mechanisms of oleoresin production, storage, and release have rarely been investigated. This study detailed the distribution, ontogeny, and ultrastructure of secretory cavities and canals distributed in leaf and stem, respectively, of Copaifera trapezifolia, a Brazilian species included in a plant group of great economic interest. Axillary vegetative buds, leaflets, and portions of stem in primary and secondary growth were collected and processed in order to study the anatomy, histolocalization of substances, and ultrastructure. Secretory cavities are observed in the foliar blade and secretory canals in the petiolule and stem. They are made up of a uniseriate epithelium delimiting an isodiametric or elongated lumen. Biseriate epithelium is rarely observed and is a novelty for Leguminosae. Cavities and canals originate from ground meristem cells and the lumen is formed by schizogenesis. The content of the cavities and canals of both stem and leaf is oily and resinous, which suggests that the oleoresin could be extracted from the leaf instead of the stem. Phenolic compounds are also detected in the epithelial cell cytoplasm. Cavities and canals in the beginning of developmental stages have polarized epithelial cells. The cytoplasm is rich in smooth and rough endoplasmic reticula connected to vesicles or plastids. Smooth and rough endoplasmic reticulum and plastids were found to be predominant in the epithelial cells of the secretory cavities and canals of C. trapezifolia. Such features testify the quantities of oleoresin found in the lumen and phenolic compounds in the epithelial cell cytoplasm of these glands. Other studies employing techniques such as correlative light electron microscopy could show the vesicle traffic and the compartmentalization of the produced substances in such glands.     

Effects of light intensity on the anatomical structure,secretory structures,histochemistry and essential oil composition of Aeollanthus suaveolens Mart. ex Spreng. (Lamiaceae)

Aeollanthus suaveolens Mart. ex Spreng belongs to Lamiaceae family, and in the Amazon this species is cultivated by natives’ people, this medicinal plant is popularly known as Catinga-de-mulata, being used by the population for general pain treatment. The present study analyzed the effects of light intensity on the anatomy, secretory structures, histochemistry and composition of essential oil of leaf and stem of A. suaveolens. The anatomical structure were observed in response to two light intensities, namely 50% (half shade, HS) and 100% (full sun, FS) light. Histochemical analyses were performed to detect lipids, essential oils, alkaloids, phenolic compounds, sesquiterpene lactones, mucilage, and tannins. Secretory structures were observed under a scanning electron microscope. The results obtained in the present work indicate that the light intensity can affect the histochemistry and structures of A. suaveolens. Cross sections of the leaves and stem revealed glandular trichomes on both leaf surfaces as well as the stem surface. Essential oil was detected by histochemical analyses in all types of secretory trichomes. These anatomical and histochemical responses suggest modifications to protect the photosynthetic apparatus from excess light, in addition we note that in the chemical composition of the essential oil the class of hydrocarbons sesquiterpene prevailed.     

Chemical Constituents of the Fssential Oils from the Leaves of Laurus nobilis and Tendency in Changes of the Constituents Month by Month

The chemical constituents of the essential oils from the leaves of Laurus nobilis L., have been identified both by capillary GC-MS and fused silica capillary GC Kovats retention in,lex of components. From laurel leaf oil, 45 compounds have been identified, among which, 19 compounds such as 1,4-cineole, sabinene hydrate etc. have not been found in the leaf oil previously. This paper studies systematically the constituents of essential oil from dry, fresh leaves, and those of annual, perennial, different district, and reported the changes of the chemical constituents month by month. Results showed that the yield of essential oil and 1,8-cineole content is the highest in July.     

Comparative anatomy of secretory structures of leaves in Hypericum L.

The structure, type, morphology and location of secretory structures in leaves of 43 species, 1 subspecies and 1 variety of 9 sections in Hypericum L. were comparatively studied using tissue clearing, paraffin sectioning and thin sectioning. The results have shown that the presence of secretory structures is a common feature of leaves in this genus. According to their anatomical characteristics, the secretory structures can be divided into nodules, secretory cavities (canals) and tiny secretory tubes. In their distribution in leaves the nodules fall into two types: the leaf edge type and the scattered type. According to the location of cavities in the cross sections of leaves, the cavities can be divided into 4 types: the median type which is situated between the palisade tissue and spongy tissue, the palisade tissue type, the spongy tissue type and the across-mesophyll type. Based on the location of cavities and nodules in leaves, the species in Hypericum can be divided into 3 groups: group Ⅰ , in which only cavities are present; group Ⅱ, in which only nodules are present; group Ⅲ, in which both cavities and nodules are present. The type, location, distribution density and morphology of secretory structures are of some taxonomic value at the level of species and of section in Hypericum L. From these observations, the evolutionary trends concerning the morphology and anatomy of secretory structures and the affinity among sections in the genus Hypericum ate dis-cussed.     

Ontogenesis of internal secretory cells in leaves of Lantana camara (Verbenaceae)

Internal secretory cells may be morphologically indistinguishable from their neighbours except for the presence of secreted material, or they may differ to such an extent that could be classified as secretory idioblasts. Several authors have reported the presence of glandular trichomes secreting essential oil in Verbenaceae, including Lantana . However, none have reported internal secretory cells. Anatomical and histochemical methods applied to Lantana camara leaves revealed the occurrence of internal secretory cells whose ontogenesis and chemical nature are described in this paper. According to leaf developmental analysis, L. camara secretory cells originated from the ground meristem, started to differentiate in the third node leaves, and were actively secreting in the fourth node leaves. The content of the secretory cells was of a lipidic nature, and a terpenoid essence of their secretion was also identified. Based upon differences in size and shape from neighbouring cells and on detection of nonvolatile terpenes, they were confirmed as true internal secretory idioblasts.  © 2005 The Linnean Society of London, Botanical Journal of the Linnean Society , 2005, 148 , 427–431.     

白鲜营养器官解剖结构及其与白鲜碱的积累关系

应用植物解剖学、组织化学及植物化学方法对白鲜营养器官根、茎、叶的结构及其生物碱的积累进行了研究。结果显示:(1)白鲜根的次生结构以及茎和叶的结构类似一般双子叶植物;白鲜多年生根主要由周皮、次生韧皮部、维管形成层以及次生木质部组成,根次生韧皮部中可见大量的淀粉、草酸钙簇晶、韧皮纤维以及油细胞;茎由表皮、皮层、维管组织和髓组成;叶由表皮、栅栏组织、海绵组织和叶脉组成;在茎和叶初生韧皮部的位置均分布有韧皮纤维,在叶表皮上分布有头状腺毛和非腺毛;在茎和叶紧贴表皮处分布有分泌囊。(2)组织化学分析结果显示:在白鲜多年生根中,生物碱类物质主要分布在周皮、次生韧皮部、维管形成层和木薄壁细胞中;在茎中,生物碱主要分布在表皮、皮层、韧皮部、木薄壁细胞及髓周围薄壁细胞中;在叶中,生物碱主要分布在表皮细胞、叶肉组织和维管组织的薄壁细胞;此外在分泌囊和头状腺毛中亦含有生物碱类物质。(3)植物化学结果显示,秦岭产白鲜根皮/白鲜皮、根木质部、茎和叶中白鲜碱含量分别为0.041%、0.012%、0.004%和0.002%,其中木质部中白鲜碱含量和其他部分地区白鲜皮中白鲜碱含量类似。研究表明,在秦岭产白鲜营养器官中,除根皮/白鲜皮外,在根木质部亦含有大量的白鲜碱,且在茎和叶中亦含有一定的白鲜碱,具有潜在的开发利用价值。     

Structure and development of the secretory cavities of Myrtus communis leaves

The structure and development of Myrtus communis L. secretory cavities has been studied in young and expanded leaves, using light and scanning electron microscope. Secretory cavities are continuously formed during leaf development, but in mature leaves the rhythm of their appearance shows steep decrease. Each secretory cavity is developed from a single epidermal cell, which undergoes a periclinal division followed by anticlinal and several oblique cell divisions. The lumen of the secretory cavity is initiated by cell wall separation, i.e., schizogenously. The secretory cells line the cavity, where the secreted material is collected. Secretory cavities are covered by modified epidermal cells, which do not seem to form any special aperture. Essential oils seem to be discharged after mechanical treatment of the leaf.     

月桂叶精油成分及逐月动态变化

采用毛细柱 GC-MS 和弹性石英毛细柱 GC Kovats 指数的鉴定方法,从月桂(Laurusnobilis L.)叶精油中鉴定出45个化合物,其中1,4-桉叶油素、水合桧烯等19个系该种植物中首次报道。系统地研究了月桂干叶与鲜叶、不同产地、当年生与多年生成分,首次报道月桂叶精油成分逐月动态变化,表明其得率和主成分1,8-桉叶油素含量均以7月最高。     

Ultrastructure and cytochemistry of the pearl gland in Piper regnellii (Piperaceae)

Pearl glands are scattered throughout the lamina of developing leaves and rarely found on adult leaves of Piper regnellii (Piperaceae). The pearl gland is a bicellular secretory trichome composed of a short broad basal cell and a spatula-like, semiglobular apical cell. Four different stages of the pearl gland were determined during its ontogenesis: origin, pre-secretory, secretory and post-secretory. During the pre-secretory stage, mitochondria, ribosomes, dictyosomes, rough endoplasmic reticulum, and plastids with electron dense inclusions were present in the cytoplasm of the apical cell. During the secretory stage, the most remarkable characteristics of the apical cell are the proliferation of dictyosomes and their vesicles, rough endoplasmic reticulum, and modified plastids. At this stage, electron-dense oil drops occur in the plastids as well as scattered within the cytoplasm, proteins and polysaccharides are seen in the plastids, vesicles, and vacuoles. Only polysaccharides are present in the periplasmic space, wall cavities, and on the surface of the apical cell. The polysaccharides are one of the main components of the mucilagenous exudate that covers the developing leaf structures. The apical cell of the senescing trichomes undergoes a progressive degeneration of its cellular components, the plastids being the first organelles to undergo lysis.     

Non-volatile components of the essential oil secretory cavities of Eucalyptus leaves: Discovery of two glucose monoterpene esters, cuniloside B and froggattiside A

The essential oils extracted from the embedded foliar secretory cavities of many Eucalyptus species are of economic value as pharmaceuticals and fragrance additives. Recent studies have indicated that Eucalyptus secretory cavities may not be exclusively involved in the biosynthesis and storage of essential oils. Therefore, we selected three species upon which to perform an examination of the contents of foliar secretory cavities: Eucalyptus froggattii, E. polybractea and E. globulus. This paper describes the isolation and structural characterization of two non-volatile glucose monoterpene esters, which we have named cuniloside B and froggattiside A, from within the secretory cavities of these species, and shows the presence of these compounds in solvent extracts of the leaves from two other species of Eucalyptus. Both compounds were found in high proportions relative to the essential oils extracted from the leaves. We propose that many other carbohydrate monoterpene esters previously isolated from bulk leaf extracts of various Eucalyptus species may also be localized within the non-volatile fraction of foliar secretory cavities.     

Glandular Trichomes in Satureja thymbra Leaves

The leaves of the aromatic plant Satureja thymbra have numerousglandular trichomes of two morphologically distinct types glandularhairs and glandular scales Investigations of the anatomy ofthese glandular trichomes with serial thick sections revealedthat the glandular hairs consist of three cells a foot, stalkand head cell Glandular scales also have a unicellular footand stalk Their heads, however, are composed of 12 cells Fourof these cells are small, occupying the central region of thehead, whereas the remainder are large and peripherally arrangedMorphometric analysis showed that, in leaf surface view, glandularscales are about 17-fold larger than glandular hairs In addition,glandular scales were found to occupy 5 7 % of the entire leafsurface area In each glandular scale the total amount of essentialoil, contained within both the subcuticular space and the interiorof the secretory cells, was calculated to be 2 51 x 10^–4mm³ The volume of the essential oil produced by all glandularscales on a single mature leaf was correspondingly determinedto be 0.059 mm³ Finally, the theoretical essential oil yieldof 100 g dry leaves of S thymbra was estimated to be 3 54 %(secretory activity of glandular scales only) Satureja thymbra, glandular trichomes, morphology, morphometry     

四种裸子植物叶精油化学成分的研究

采用气相色谱—质谱—计算机联用仪对穗花杉、南方红豆杉、三尖杉和罗汉松叶精油化学成分的研究发现,三尖杉和穗花杉叶精油的组成特点极为相似,相同成分１３个,占各自精油组成的４８．０７％和３３．３２％．三尖杉和南方红豆杉叶精油的相同成分４个,占各自精油组成的１６．１４％和４０．５９％．在一定程度上支持三尖杉科和红豆杉科的亲缘关系接近,红豆杉科可能是通过穗花杉属和三尖杉科相联系的观点．罗汉松和穗花杉叶精油的相同成分４个,占各自叶精油组成的比例为２４．０９％和２０．８２％,比罗汉松和南方红豆杉、三尖杉之间组分的相似性要高．反映出罗汉松科和红豆杉科之间有一定联系,穗花杉属是认识红豆杉科、三尖杉科和罗汉松科之间系统关系的关键属     

The Secretory Structure of Hypericum perforatum and Its Relation to Hypericin Accumulation

Three different kinds of internal secretory structures, secretory cell globules (black dots), secretory cavities (translucent dots) and secretory ducts (translucent streaks) were observed in Hypericum perforatum L. The secretory cell globule, which occurred in flower, leaf and stem, consisted of a core of large secretory cells surrounded by two layers of flattened sheath cells. The secretory cavities were present throughout the lamina and the secretory ducts throughout the flower, both consisted of one or two layers of flattened cells walling an oil chamber or duct. Histochemical and fluorescence microscopy revealed that hypericin was accumulated in the secretory cell globules. Microscopic and ultrastmctural observation further demonstrated that hypericin was accumulated in the large central vacuole of the mature secretory cells. Numerous dictyosome, endoplasmic reticulum and small vacuoles were observed in the dense cytoplasm surrounded the large central vacuole. The process of hypericin accumulation in the secretory cells was preliminarily discussed.     

贯叶连翘的分泌结构及其与金丝桃素积累的关系

贯叶连翘（ＨｙｐｅｒｉｃｕｍｐｅｒｆｏｒａｔｕｍＬ．）地上器官分布着分泌细胞球（黑色腺点）、分泌囊（半透明腺点）和分泌道（半透明腺条）３类内部分泌结构。分泌细胞球在茎、叶和花器官中均有分布,由２层鞘细胞包围多个分泌细胞构成实心的分泌细胞团。分泌囊主要分布于叶片中,分泌道则分布于花器官中,它们都是由１～２层切向扁平细胞围绕圆形或长形腔道构成,腔道的贮存物为精油。利用组织化学方法,结合荧光显微镜观察,证实金丝桃素类物质是由分泌细胞球（黑色腺点）所合成和积累的。通过用戊二醛和锇酸固定样品的显微和超微结构观察,发现金丝桃素类物质积累在成熟腺体分泌细胞的中央大液泡中,细胞周围浓厚的细胞质中分布着大量小液泡和高尔基体、内质网等细胞器。在此基础上对金丝桃素类物质的积累过程进行了初步探讨     

Non-invasive localization of thymol accumulation in Carum copticum (Apiaceae) fruits by chemical shift selective magnetic resonance imaging

Magnetic resonance imaging was used to localize the site of essential oil accumulation in fruit of Carum copticum L. (Apiaceae). A chemical shift method is described that utilized the spectral properties of the aromatic monoterpene thymol, the major component of the essential oil, to image thymol selectively. The presence of essential oil secretory structures in the fruit and an essential oil containing a high proportion of thymol were confirmed with optical microscopy and gas chromatography-mass spectrometry, respectively. Selective imaging of whole C. copticum fruits showed that thymol accumulation was localized to the secretory structures (canals) situated in the fruit wall. The technique was considered non-invasive as the seeds used in the imaging experiments remained intact and viable.     

Essential Oil Gland Number and Ultrastructure During Mentha arvensis Leaf Ontogeny

Alterations in essential oil gland number, distribution and fine structure, and the oil content in the leaf of Mentha arvensis L. were examined during its growth and senescence. Accumulation of essential oil occurred predominantly during the rapid leaf expansion phase followed by a similar decline. The oil gland (trichome) number increased upto leaf maturation and declined thereafter. Initially, cuticle remains tightly apposed to the secretory head of oil glands but progressively a sub-cuticular space appears to be created for the oil. Considerable enlargement of vacuole with ageing is witnessed, whereas cytoplasm gradually decreases to a thin peripheral layer. Some secretory cells from senescing leaf were found almost empty, having only a few remnant oil droplets. This revised version was published online in June 2006 with corrections to the Cover Date.