贯叶连翘分泌结构的发育及其内含物积累的研究

应用半薄切片和组织化学观察结果表明,贯叶连翘植物内具有分泌囊和分泌细胞团,叶片,萼片,花瓣中都存在2种分泌结构,分泌囊尚存在于果壁内,而分泌细胞团还在花药和茎棱中 ,2种分泌结构均起源于幼叶的基本分生组织内,其原始细胞形态相似,都垂周分裂为2子细胞,接着分别以不同分裂方式形成原始细胞群,并分化出鞘细胞,以后分 囊原始细胞群以裂生方式发育成由1层鞘细胞,1层上皮细胞包围着分泌腔组成的分泌囊,而分泌细胞团原始细胞群则继续增加细胞数目和体积发育成2－4层鞘细胞包围紧密排列的分泌细胞组成的分泌细胞团,其中给终未出现分泌腔,组织[化学试验表明,前者产生和贮存油类物质,而后者产生和贮存金丝桃素类物质。     

元宝草的分泌结构与其次生产物的研究

利用整体透明,石蜡掉片和组织化学方法对元宝草不同器官中分泌结构的类型和分布进行研究。结果表明：其分泌结构根据形态结构特点可分为分泌细胞团,分泌囊及分泌道3类,并指出了3类分泌结构在各器官的分布位置和密度,组织化学试验表明,前者含金丝桃素等,后二者含挥发油,为此种药用植物的开发利用提供了依据。同时为该属植物的系统分类研究提供一些解剖学依据。     

Toxic Effects of Pb on Anatomy and Hypericin Content in Hypericum perforatum L.

Lead (Pb) is one of the most common heavy metal contaminants in the environment. The present study was therefore undertaken to determine the effects of Pb on structural characteristics and hypericin production in Hypericum perforatum. Mature plants were treated with contaminated soil in seven treatments (75, 150, 300, 600, 800, 1000, and 1500 mg/kg Pb in soil) with three repeats per treatment every 14 days. Maximum observed Pb content in shoot parts was observed in the treatments with 600 and 1500 mg/kg Pb. The Pb concentration in roots was higher than in shoot parts, enhanced with increasing Pb concentration in the soil. In this study, Pb treatment significantly influenced the morphology, anatomy, and hypericin content in the plant. Anatomical characteristics of leaf, stem, and root affected by Pb contamination, as well as scanning electron microscopy (SEM) studies, revealed structural changes in stomata and epicuticular waxes. Under Pb toxicity, anatomical symptoms occurred in leaves, including increase in sizes of epidermal cells, mesophyll tissue, and diameter of stems and roots, as well as amplified vascular bundles and pith area. This, therefore, indicated that metal contamination can change the chemical composition of this plant. Maximum hypericin content was observed in the treatment containing 600 mg/kg Pb in soil, which then decreased.     

金丝桃属植物叶中分泌结构的比较解剖学研究

利用整体透明法、石蜡制片法和半薄切片法,对金丝桃属9组43种l亚种1变种的植物叶分泌 结构的类型、形态、结构和分布进行了比较研究。结果表明,分泌结构是金丝桃属植物叶片普遍的结构特征,根据其分泌结构的特征,可划分为分泌细胞团、分泌囊(道)和韧皮部中分泌小管道等3种分泌结构。其中,分泌细胞团按其在叶片的分布可分为叶缘型和散生型;分泌囊按其在叶横切面中的位置可分为栅栏组织型、海绵组织型、居中型(位于栅栏组织型与海绵组织型之间)和横跨叶肉组织型。根据分泌囊和分泌细胞团在该属植物叶中的分布可划分为3种类型：(1)只有分泌囊的类型;(2)只有分泌细胞团的类型;(3)具分泌囊和分泌细胞团的类型。两种主要分泌结构的类型、分布密度、分布位置及其形态等方面在组间和种间均存在—定的差异,对金丝桃属属以下等级的区分具有一定意义。在此基础上,对该属分泌结构的形态演化以及金丝桃属各组间的亲缘关系进行了探讨。     

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.     

Anatomy of the vegetative organs and secretory structures of Rhaponticum carthamoides (Asteraceae)

Roots, stems, rhizomes and leaves of Rhaponticum carthamoides (Willd.) Iljin (a Siberian adaptogenic plant, originating from the Altai and Saian Mountains) of different ages were investigated by means of light and electron microscopy. Schizogenous secretory reservoirs occurred in every organ, and were located within the secondary xylem (adventitious roots and rhizome of young plants), at the interface of endodermis/cortical parenchyma (roots and hypocotyl), along phloem and primary xylem (older rhizome), around the vascular bundles (inflorescence stem, petiole and leaf midrib veins) and along phloem (cotyledonary and leaf veins). At the interface of endodermis/inner parenchyma, secretion accumulated in the intercellular spaces prior to the formation of proper epithelial cells. The secretion as observed by transmission electron microscopy comprised three components: soluble (i.e. absent from sections; probably phenolic), insoluble and strongly osmiophilic (probably phenolic) and insoluble, moderately osmiophilic (probably lipidic). Numerous osmiophilic oil droplets, similar to the lipidic secretion inside the reservoirs, local proliferation of rough endoplasmic reticulum and numerous multivesicular bodies characterized epithelial cells in all organs. Leucoplasts (in subterranean organs) with osmiophilic inclusions and peroxisomes with crystalloid inclusions were specific for parenchyma cells. Peltate glandular hairs were formed on leaf blades.  © 2004 The Linnean Society of London, Botanical Journal of the Linnean Society , 2004, 144 , 207–233.     

贯叶连翘的水培及其代谢产物检测

水培可诱导贯叶连翘组培苗生根能力强,根活力也增加;生根苗在1/6MS培养液中培养6周后的金丝桃素(HP)、假金丝桃素(PHP)和贯叶金丝桃素(HF)含量分别比基质[腐质土 蛭石(1:1)]中培养的提高10.13%、16.00%和61.36%。     

突脉金丝桃的花器官发生及其系统学意义

利用扫描电镜（SEM）观察了突脉金丝桃（Hypericum przewalskii）（金丝桃科）的花部器官发生发育过程。结果表明,突脉金丝桃2枚苞片原基首先发生,花原基在苞片原基的包裹中完成发育。在苞片原基发生后,5枚萼片原基沿2/5圆周依次发生。萼片原基发生近完成时,5枚雄蕊—花瓣共同原基在萼片原基之间的角隅处近同时发生,此后,雄蕊—花瓣共同原基下部向外伸展形成花瓣原基,上部向上凸起形成与花瓣原基相对的雄蕊原基,之后雄蕊原基由内向外依次分化发育产生次生雄蕊原基,随着次生雄蕊原基的发育和数目的增多,形成了5束雄蕊。次生雄蕊原基发生的同时,5枚心皮原基近同时发生。突脉金丝桃雄蕊束的发生方式表明,金丝桃属的雄蕊束可能起源于5基数的单轮雄蕊。金丝桃科与藤黄科植物花瓣及雄蕊原基发生方式的显著不同,支持了APG Ⅲ系统将金丝桃亚科从藤黄科中独立为金丝桃科的观点。     

Leaf anatomy inCaesalpinia andHoffmannseggia (Leguminosae,Caesalpinioideae) with emphasis on secretory structures

Leaflets of 65 species ofCaesalpinia s.l. and seven species ofHoffmannseggia were studied in clearings supplemented by resin sections and scanning electron microscopy. Three types of secretory structure occurred among 46 species; in 43 species they were distributed mutually exclusively (external glands: 8 species; internal cavities: 5 species; idioblastic cells: 30 species); three other species each had two types. Species with secretory structures conform mostly to proposed subgenera and informal groups. Other unusual features were external glands with internal spaces, thickened walls or conspicuous localized wall thickenings in epidermal cells or mesophyll cells of certain species, and differentially stained epidermal cells surrounding stomata. Prismatic crystals predominate but druse crystals also occur.     

Survey of leaf anatomy,especially secretory structures,of tribeCaesalpinieae (Leguminosae,Caesalpinioideae)

We studied leaflet anatomy, emphasizing secretory structures, from herbarium specimens of 128 species of 44 genera of tribeCaesalpinieae, using clearings, resin sections, and scanning electron microscopy. These observations, combined with those from our three earlier papers, provide a survey of 210 species representing all genera. Seventy-three species had secretory structures: 21 had glands or gland-like trichomes, 40 had living mesophyll idioblasts, and nine had cavities (three species each had two different types). Five additional species, all inCercidium (Caesalpinia group), had paired or clustered large spheroidal, thick-walled, empty cells (veinlet idioblasts) interconnected by perforation plate-like gaps. Secretory structures have systematic significance at various taxonomic levels.     

Signal interaction between nitric oxide and hydrogen peroxide in heat shock-induced hypericin production of Hypericum perforatum suspension cells

Heat shock(HS, 40℃, 10 min) induces hypericin production, nitric oxide(NO) generation, and hydrogen peroxide(H2O2) accumulation of Hypericum perforatum suspension cells.Catalase(CAT) and NO spe-cific scavenger 2-4-carboxyphenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide(cPTIO) suppress not only the HS-induced H2O2 generation and NO burst, but also the HS-triggered hypericin produc-tion.Hypericin contents of the cells treated with both NO and H2O2 are significantly higher than those of the cells treated with NO alone, although H2O2 per se has no effects on hypericin production of the cells, which suggests the synergistic action between H2O2 and NO on hypericin production.NO treatment enhances H2O2 levels of H.perforatum cells, while external application of H2O2 induces NO generation of cells.Thus, the results reveal a mutually amplifying action between H2O2 and NO in H.perforatum cells.CAT treatment inhibits both HS-induced H2O2 accumulation and NO generation, while cPTIO can also suppress H2O2 levels of the heat shocked cells.The results imply that H2O2 and NO may enhance each other's levels by their mutually amplifying action in the heat shocked cells.Membrane NAD(P)H oxidase inhibitor diphenylene iodonium(DPI) and nitric oxide synthase(NOS) inhibitor S,S′-1,3-phenylene-bis(1,2-ethanediyl)-bis-isothiourea(PBITU) not only inhibit the mutually amplifying action between H2O2 and NO but also abolish the synergistic effects of H2O2 and NO on hypericin production, showing that the synergism of H2O2 and NO on secondary metabolite biosynthesis might be dependent on their mutual amplification.Taken together, data of the present work demonstrate that both H2O2 and NO are essential for HS-induced hypericin production of H.perforatum suspension cells.Furthermore, the results reveal a special interaction between the two signal molecules in mediating HS-triggered secondary metabolite biosynthesis of the cells.     

Matrix‐free UV‐laser desorption/ionization (LDI) mass spectrometric imaging at the single‐cell level: distribution of secondary metabolites of Arabidopsis thaliana and Hypericum species

The present paper describes matrix‐free laser desorption/ionisation mass spectrometric imaging (LDI‐MSI) of highly localized UV‐absorbing secondary metabolites in plant tissues at single‐cell resolution. The scope and limitations of the method are discussed with regard to plants of the genus Hypericum. Naphthodianthrones such as hypericin and pseudohypericin are traceable in dark glands on Hypericum leaves, placenta, stamens and styli; biflavonoids are also traceable in the pollen of this important phytomedical plant. The highest spatial resolution achieved, 10 μm, was much higher than that achieved by commonly used matrix‐assisted laser desorption/ionization (MALDI) imaging protocols. The data from imaging experiments were supported by independent LDI‐TOF/MS analysis of cryo‐sectioned, laser‐microdissected and freshly cut plant material. The results confirmed the suitability of combining laser microdissection (LMD) and LDI‐TOF/MS or LDI‐MSI to analyse localized plant secondary metabolites. Furthermore, Arabidopsis thaliana was analysed to demonstrate the feasibility of LDI‐MSI for other commonly occurring compounds such as flavonoids. The organ‐specific distribution of kaempferol, quercetin and isorhamnetin, and their glycosides, was imaged at the cellular level.     

Factors affecting growth of cell suspension cultures of hypericum perforatum L. (St. John's wort) and production of hypericin

Summary Use of Hypericum perforatum L., commonly known as St. John's wort, has increased recently due to the pharmaceutical potential of hypericin, found in its leaves. Hypericin has been reported to effect a natural treatment for mild and moderate depression by increasing the concentration of neurotransmitters in the central nervous system. We have developed a novel cell culture system for in vitro growth and production of this species, suggesting a possible technology for large-scale production of hypericin. Leaf explants grown in Murashige and Skoog salts supplemented with 2,4-dichlorophenoxyacetic acid (0.90 μM) and kinetin (0.11 μM) gave maximum percentage callus formation compared to other medium treatments evaluated. Hypericin localization in cell phase and leaves was found to vary, with cell phase accumulating hypericin in special organelles and leaves accumulating it in vacuoles. Light and dark conditions, with cell aggregate size, played important roles in growth and hypericin production in cell suspension cultures.     

Leaf and twig anatomy of the Pterostemonaceae (Engl.) Small: ecological and systematic features

An anatomical investigation of the leaves and twigs of Pterostemonaceae (Engl.), a monogeneric family of two species, has been made. The following anatomical characters in the leaf are of particular interest: glandular hairs, hydathodes on the marginal dentations and secretory substances in the glands and palisade cells. Characters of interest in the twig xylem include: vessel lumina of very small tangential diameter and with simple perforation plates; fibriform vessels with scalariform plates having one to six bars and also plates with perforations in irregular patterns.     

Comparative leaf anatomy of Alpinia Roxb. species (Zingiberaceae) from China

The leaf anatomy of 20 Alpinia species from China was investigated. Results show that there is interspecific variation in the structure of the leaf midrib and petiole which can be used for species identification. Adaxial hypodermis is present in the lamina in all species of subgenus Catimbium and absent from all species of subgenera Dieramalpinia and Aobolocalyx and Alpinia , excepting A. conchigera, A. galanga and A. aquatica , which appear to be closely allied in having subepidermal fibres in midribs and petioles, which are absent from the rest of the species.     

All in an afternoon: mixed breeding system in one-day lasting flowers of Hypericum elodes L. (Hypericaceae)

Knowledge about mixed mating systems can improve our understanding of the evolutionary dynamics of reproductive systems. Here we report a study of the floral and reproductive biology of Hypericum elodes, an Atlantic-European soft-water pools specialist which shows a floral architecture consistent with both self- and cross-pollination. Controlled pollination experiments were performed in a natural population during three consecutive years. Marked flowers were monitored until fruit production, and laboratory germination experiments were conducted with the seeds produced. Plants were self-compatible (SCI>0.75), however, compared with selfing, cross-pollination enhanced fruit-set, seed-set and seedling growth, but not seed germination. Inbreeding depression (δ) was mild in the pre-dispersal stages (δ = 0.22 for fruit set, 0.18 for seed set and 0.13 for seed mass), low for germination percentage (δ = 0.003) and mild for seedling growth (δ = 0.23). The breeding system of H. elodes promotes outcrossing and assures reproductive success by means of competitive autogamy. Our results suggest a mixed mating strategy for the studied population, characterized by mild inbreeding depression (cumulative δ = 0.57), highlighting the benefit of this reproductive mode in unpredictable habitat, as the typical shallow-water meadows where H. elodes grows.     

Foliar anatomy ofPolygonum (Polygonaceae): Survey of epidermal and selected internal structures

Several anatomical characters in leaves were described, and their distribution determined, for 153 species ofPolygonum, mostly from herbarium specimens. Structures surveyed were epidermal (glandular and nonglandular trichomes, nodules, specialized parenchyma, stomatal apparatus) and internal (cavities, crystals, laticifer-like cells, nodules, subepidermal fibers). Cleared leaves were supplemented by resin-embedded sections and SEM preparations of selected species. No feature defines any taxonomic section, but some features occur only within one section. Laticifer-like cells, epidermal and internal nodules, resin cups, and unique epidermal and subepidermal cavities seem to be unknown elsewhere; other features (invaginated epidermal cells; enlarged crystal cells confined to paraveinal layer) are rare among angiosperms.