Participation of the mitochondrial permeability transition pore in nitric oxide-induced plant cell death

In the present study, we investigated the involvement of the mitochondrial permeability transition pore (PTP) in nitric oxide (NO)-induced plant cell death. NO donors such as sodium nitroprusside (SNP) and S-nitroso-N-acetylpenicillamine inhibited growth and caused death in suspension-cultured cells of Citrus sinensis. Cells treated with SNP showed chromatin condensation and fragmentation, characteristic of apoptosis. SNP caused loss of the mitochondrial membrane electrical potential, which was prevented by cyclosporin A (CsA), a specific inhibitor of PTP formation. CsA also prevented the nuclear apoptosis and subsequent Citrus cell death induced by NO. These findings indicate that mitochondrial PTP formation is involved in the signaling pathway by which NO induces apoptosis in cultured Citrus cells.     

Ultrastructural studies on the secretory cavities ofCitrus deliciosa ten. II. Development of the essential oil-accumulating central space of the gland and process of active secretion

Summary Oil glands ofCitrus deliciosa are multicellular secretory structures, globular to oval in shape, in the centre of which an essential oil-accumulating space is formed. Opening of this space begins from a single cell. It undergoes lysis which later extends to the neighbouring gland cells.Secretory material in form of droplets is produced in plastids, from where it is transported to the parietal cytoplasm of the secretory cells via numerous ER-elements. After fusion of the ER-membranes with the plasmalemma, the exudate reaches the apoplast, through which it is driven to the central cavity of the gland.Peripheral cells of the secretory complex are modified into a protective sheath with thick walls and large vacuoles, while their plastids are differentiated from leucoplasts into typical amyloplasts.     

Ultrastructural studies on the secretory cavities ofCitrus deliciosa ten. I. Early stages of the gland cells differentiation

Summary Secretory cavities ofCitrus deliciosa seem to originate from a pair of meristematic cells (an epidermal cell and a second one placed under it). These cells undergo successive divisions resulting in the formation of a globular/oval gland situated in the parenchyma, and a conical stalk, which joins the gland with the epidermis. The young gland consists of a central group of polyhedral cells ensheathed by layers of radially flattened cells.During the early differentiation stages of the gland cells a close association of cytoplasmic microtubules with various organelles is observed. Plastids increase progressively in number and size and their matrix locally contains tubular networks accompanied by small oil droplets. In peripheral cytoplasm numerous myelin-like lomasomes have been observed. Peripheral cells of the gland are gradually modified from the inner cells following a different developmental pattern. Their walls become thicker and plastids do not contain tubular complexes, but only a few thylakoids partly surrounding the newly formed starch grains.     

A phytochemical investigation of Citrus halimii

Essential leaf oils, isoenzymes, protein and polyphenol oxidase-catalyzed browning patterns of Citrus halimii (Rutaceae) are investigated and compared with those of other species of Citrus in order to support the species standing of the recently described C. halimii.     

Effects of Citrus sinensis (L.) Osbeck epicarp essential oil on growth and morphogenesis of Aspergillus niger (L.) Van Tieghem

Essential oils from different plant parts are known for their antimicrobial activity but the antifungal effects of essential oil from Citrus sinensis (L.) Osbeck epicarp on growth and morphogenesis of Aspergillus niger has not been observed so far. The mycelial growth was inhibited at 2.5 and 3.0 μg/ml of oil in Potato Dextrose Broth and Agar medium, respectively. These concentrations were fungicidal under the test conditions. The fungitoxicity of oil did not change even at exposure to 100 °C and autoclaving. The main changes observed under light and scanning electron microscopy after oil treatment were loss of cytoplasm in fungal hyphae, and budding of hyphal tip. The hyphal wall and its diameter became markedly thinner, distorted and resulted in cell wall disruption. The flattened and empty hyphal tips bifurcated into bud like structures. GC-MS studies of the oil revealed the presence of 10 chemical constituents. Limonene has been found to be major component (84.2%).     

Some physiological and morphological characteristics of citrus plants for drought resistance

Tolerance and avoidance mechanisms to drought stress were studied in 6-month-old plants of Newhall orange (Citrus sinensis (L.) Osbeck) and Ellendale tangor (orange × mandarin hybrid) (Citrus sinensis (L) Osbeck × Citrus reticulata Blanco) during a drought/rewatering cycle under controlled conditions. Drought stress did not promote osmotic adjustment, while elastic adjustment (tissue elasticity increase) was noted in stressed orange and tangor plants. Both citrus plants showed a parallel decrease in leaf conductance (g₁) and leaf water potential (Ψ₁) under water stress. Tangor plants had a more efficient water conservative strategy than orange, based on the characteristics of canopy architecture (lower canopy area and a more closed canopy with leaves nearly vertically oriented) together with a significant decrease in cuticular transpiration rates (TRc) under stress.     

花椒果实分泌囊发育过程的超微结构研究

电镜观察结果表明,花椒(Zanthoxylum bungeanum Maxim.)果实分泌囊是由裂生方式形成,由鞘细胞、上皮细胞和油腔构成。对分泌囊的原始细胞、油腔发生和扩大以及发育成熟3个时期的超微结构研究表明,其精油是在分泌囊油腔发生时开始积累,以油滴形态存在于上皮细胞的质体内及其周围的细胞质中。根据各细胞器的变化规律分析,质体是精油合成的主要场所,内质网参与精油的合成和转运,线粒体为上述活动提供能量。上皮细胞内积累的精油可能通过两种途径排出细胞,分泌至油腔内贮存。鞘细胞内也积累精油,其主要合成场所也与质体有关,以后转运至上皮细胞内。成熟分泌囊的质体由于功能改变,其内出现蛋白质结晶和淀粉粒。     

Glandular scale development and essential oil secretion in Origanum dictamnus L.

Glandular scales of Origanum dictamnus L. originate from a single protodermal cell. They are composed of a 12-celled head and an unicellular stalk and foot. During the early stages of gland differentiation, the head cells possess a small number of plastids which contain globular inclusions. Similar inclusions are also observed in the plastids of the stalk and the foot cell. The lateral walls of the stalk cell progressively undergo cutinization which does not extend to the upper and lower periclinal walls. At the onset of secretion the electron density of the plasmalemma region lining the apical walls of the head cells remarkably increases. These walls are impregnated with an osmiophilic substance identical in appearance to the content of the subcuticular space. In a following stage of the secretory process osmiophilic droplets of various size arise in the cytoplasm of the secretory cells which undergoes simultaneously a reduction of its initial density. After secretion has been concluded the protoplast of the head cells becomes gradually degenerated. The chlorenchyma cells of the mesophyll possess numerous microbodies closely associated with various organelles. In the cytoplasm of these cells crystalloids occasionally occur.     

Gland cells associated with the alimentary tract of a monogenean, Diclidophora merlangi

Three distinct groups of unicellular glands open into the buccal cavity, prepharynx, and oesophagus, respectively, of Diclidophora merlangi. Buccal glands produce a dense, acidophilic secretion of protein-rich droplets that are discharged from duct-like extensions of the glands by eccrine secretion. Prepharyngeal glands are extensive and contain basiphilic secretory droplets of varying density and characterized by the presence of one or two dense core-like inclusions. The droplets are PAS-positive and reactive for protein, and accumulate in the gland cell apices which form part of the lining to the prepharynx lumen. They are released in large numbers by apocrine secretion or, individually, by an eccrine secretory mechanism. Oesophageal glands are acidophilic and contain electronlucid, PAS-positive droplets that develop a crystalline appearance prior to release via either eccrine or apocrine secretion. Differences in ultrastructure and histochemistry indicate the glands are also functionally separate, and their probable role in feeding and extracellular digestion of blood in the worm is discussed.     

Tegument and apical end organ fine structure in the metacestode and adult Proteocephalus ambloplitis

The tegument of plerocercoid and adult P. ambloplitis was examined. Differences in tegument structure existed between these two stages. Plerocercoids of P. ambloplitis lacked extensive vacuolization and unicellular gland cells characteristic of adult tegument. Plerocercoid microtriches were short and conoid; adult microtriches were lenticular with an extended, whip-like shaft. An inclusion, not previously reported from proteocephalid cestodes, is described. Adult tegument had ducts, originating from underlying unicellular glands, extending through the distal cytoplasm and opening to the exterior between microtriches.

The apical end organ cavity of P. ambloplitis contained numerous labyrinth-like spherical bodies. These structures appeared to be synthesized and secreted into the end organ by a thin cellular lining of the end organ. This lining was composed of discrete, filamentous cells believed to be modified subtegumental cell bodies. Spherical structures identical to those observed within the end organ cavity occurred within this cellular lining. The spherical bodies may be associated with enzymes necessary for tissue migration by the metacestode.     

Ultrastructural Studies on the Secretory Cavity Development and Essential Oil Accumulation in the Fruits of Evodia rutaecarpa

The secretory cavity in fruits of Evodia rutaecarpa (Juss.) Benth. developed schizogenously through a separation of the walls of the central initial cells. Electron micrographs revealed that in the early stages of cavity development there was an apparent increase in the number and volume of the plastids in which esmiophilic droplets and tubular dements were observed. This suggested that the essential oils might be synthesized in the plastids.. The essential oils were then transport ed through the plastid membrane to the surrounding endoplasmic reticulum or into the vacuoles, becoming vesicles approaching the plasmalemma, and finally releasing their contents into the oil chamber by plasmalemma invaginations.     

鹅掌楸油细胞发育过程中超微结构的变化与挥发油产生的关系

鹅掌楸[Liriodendron chinense(Hemsl.)Sargent.]油细胞的发育过程可依据细胞壁的结构变化依次划分为3个阶段,即仅具初生纤维素壁层阶段、木栓质化壁层形成阶段和内纤维素壁层形成阶段。在发育早期,仅具初生纤维素壁层时,油细胞因其体积大,核仁显著,含极少淀粉粒和质体几乎无类囊体而与周围的组织细胞不同。对其3个发育阶段中内部结构变化分析表明,挥发油合成于细胞质和质体中。细胞质中,挥发油就以小滴形式产生,然后逐渐与油囊融合直接贮入油囊,与此同时,在各种细胞器中,质体的变化最为明显,质体中合成的锇物质,随质体解体进入细胞质中,再经转化通过杯形构造积累入油囊。油囊中积累的油经OsO4染色后呈灰色,且分为2层,外层较内层深,推测与油的2种来源有关。     

Ontogenesis of trichome-like cavities in Dictamnus dasycarpus

The ontogeny of a special type of glandular hairs, namely the trichome-like cavities in Dictamnus dasycarpus, characterized by many morphological similarities to non-glandular hairs, capitate glandular hairs, and secretory cavities, was studied using light and electron microscopy. These trichome-like cavities originate from a single, initial epidermal cell that undergoes a periclinal division, with one cell developing into the internal cells and the other into the outer, epidermal cells. A beak-shaped apex is formed on the head of the trichome-like cavity. The histochemical test shows that the trichome-like cavities are important sites for lipid production. By ultrastructural analysis it becomes evident that formation of these trichome-like cavities starts with a disorganization of the cytoplasm that is accompanied by formation of odd shaped nuclei with condensed chromatin. The process continues bringing about plasmolytic processes, and disintegration of the plasma membrane system follows leading finally to autolysis, when mitochondria and degenerated plastids with disorganized membrane systems are engulfed by vacuoles, multivesicular bodies, and double-membranous autophagosomes within the vacuoles. A strong structural twist and swelling of the internal cell walls ultimately leads to complete breakdown of the structures. Nuclei of the inner internal cells within the trichome-like cavity become TUNEL-positive and DAPI-negative first; later this is detected also in the outer internal cells, indicating a centrifugal nuclear degradation process. On the basis of this work, it can be assumed that the lysigenous formation of the trichome-like cavity is a typical programmed cell death process.     

Ultrastructure of Resin Ducts in Pinus halepensis Development, Possible Sites of Resin Synthesis, and Mode of its Elimination from the Protoplast

Development of the resin duct cavity, sites of resin synthesisin the epithelial cells and elimination of resin from the protoplastwere studied in roots of young Pinus halepensis seedlings. Itis suggested that the Golgi bodies are involved in dissolutionof the middle lamella in the region of the future duct cavityby secretion of lytic enzymes into the cell walls. In earlystages of duct development osmiophilic droplets were observedin plastids, periplastidal and cytoplasmic ER, Golgi vesicles,mitochondria, nuclear envelope and in the cytoplasm. In thelatter they were often observed to be surrounded by a membrane.Electron micrographs suggested that elimination of resin dropletsfrom the protoplast occurs by their becoming surrounded by plasmalemmainvaginations.     

Effects of 15N application frequency on nitrogen uptake efficiency in citrus trees

Two irrigation systems were used to compare nitrogen uptake efficiency in citrus trees and to evaluate the NO₃⁻ runoff in «Navelina» orange trees [Citrus sinensis (L.) Osbeck] on Carrizo citrange rootstock (Citrus sinensis × Poncirus trifoliata Raf.). These were fertilized with 125 g N as labelled K¹⁵NO₃ and grown outdoors in containers filled with a sand-loamy soil. Two groups of 3 trees received this N dose either in five equally split applications by a flooding irrigation system or in 66 applications by drip. Trees were harvested at the end of the vegetative cycle (December) and the isotopic ratios of ¹⁵N/¹⁴N were measured in the soil-plant system. The N uptake efficiency of the whole tree was higher with drip irrigation (75 percnt;) than with flooding system (64 percnt;). In the 0-90 cm soil profile, the N immobilized in the organic fraction was similar for both irrigation methods (around 13 percnt;), whereas the N retained as NO₃⁻ was 1 percnt; of the N applied under drip and 10 percnt; under flooding. In the last case, most of NO₃⁻ remained under root system and it could be lost to leaching either by heavy rainfalls or excessive water applications. These results showed that a drip irrigation system was more efficient for improving water use and N uptake from fertilizer, in addition to potentially reduced leaching losses.     

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.     

The forebody glands and surface features of the metacercariae and adults of Microphallus similis

Numerous unicellular glands are present in the forebody of the metacercariae and adults of Microphallus similis. The gland cell bodies lie beneath the tegumental perinuclear cytoplasm and ducts pass upwards penetrating the tegumental distal cytoplasm. Electron dense granules are secreted onto the forebody surface and into the oesophagus via small pits in the tegument. Histochemical tests revealed that the glands contain diastase-resistant neutral mucosubstances, RNA, protein, esterase and small amounts of acid phosphatase; inhibitor studies indicated that the esterase was acetylcholinesterase (EC 3.1.1.7). Ultracytochemistry showed that the mucosubstances were located in the general cytoplasm of the gland cells and that cholinesterase was present in the granules. The forebody of M. similis is covered by large toothed spines whereas the hindbody only bears short peg-like spines. It is suggested that the forebody spines may have an irritating, abrasive effect on the host's intestinal mucosa and that the secretion of acetylcholinesterase in the region of these spines may produce a ‘local anaesthetic’ effect on the host's gut by reducing movement in the immediate vicinity of the fluke, so decreasing the likelihood of expulsion.     

Subcellular localization of beta-glucosidase in rye, maize and wheat seedlings

The subcellular compartmentation of β -glucosidase was studied in rye, maize and wheat seedlings by immunocytochemical methods. For detection, we used a 10 nm gold-labeled secondary antibody, and results were observed using transmission electron microscopy. In all three species, β -glucosidase was found in plastids, cytoplasm and cell walls. In rye, gold particles were seen on cell walls and cytoplasm in epidermal cells of the root tip and shoot, in bundle sheath cells of the shoot and in all cells, except the vascular bundle cells of the coleoptile. Gold labeling was also observed in plastids of the bundle sheath cells of rye shoot tips and in cortical cells of root tips. In wheat, gold labeling was observed on cell walls and cytoplasm of epidermal cells in the shoot base and coleoptile, and on cell walls and plastids in epidermal cells of the root tip. In maize, gold labeling was mainly found in plastids or proplastids in vascular bundle cells and bundle sheath cells of the shoot, in bundle sheath cells of the coleoptile and in epidermal cells of the root. Some gold particles were also found in cell walls and cytoplasm of stomatal guard cells of the shoot base and vascular bundle cells of the shoot tip and in the cell walls of bundle sheath cells of the shoot tip and root tip epidermal cells. Results are discussed in relation to the role of β -glucosidase in hydroxamic acid release and overall defense mechanism of monocotyledons.     

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.     

Structure and ultrastructure of leaf and calyx glands in Galphimia brasiliensis (Malpighiaceae)

The present study describes the anatomical structure of calyx and leaf glands in Galphimia brasiliensis and analyzes the mechanism of secretion. The glands are marginal and suprabasal, cup-shaped, sessile, and scarcely visible with the naked eye. Light microscopy reveals the following features: a thin, smooth cuticle; unistratified secretory cells; subglandular parenchyma; and vascular bundle supply composed of phloem and xylem with abundant druses of calcium oxalate. Transmission electron microscopy reveals the presence of secretory cells with conspicuous nuclei, dense cytoplasm, lipid droplets, numerous vesicles, mitochondria, Golgi, rough endoplasmic reticulum (RER), and elongated plastids with osmiophilic contents. The secretion reaches the apoplastic space and accumulates beneath the cuticle. Finally, the viscous, translucent exudate is eliminated by mechanical rupture of the cuticle. Histochemical analysis confirms that lipids are the main constituent. Small amounts of polysaccharides were also identified.