THE FINE STRUCTURE OF THE GUARD CELLS OF HELIANTHUS ANNUUS

The guard cells of Helianthus annuus contain elements of endoplasmic reticulum and large numbers of mitochondria and dictyosomes. Each guard cell possesses a complex system of small to large vacuoles which contain small, membrane-bound vesicles; the vacuole may actually be one highly invaginated and dissected vacuole extending throughout the cell. A highly developed grana fretwork within the plastids implies full photosynthetic capability and the capability of producing the osmoticulum required for turgor change. No plasmodesmata occur between the sister guard cells or between the guard and epidermal cells. It is postulated that there is a close relationship between plastid development and the presence or absence of plasmodesmata. No microbodies were positively identified in any of the guard cells. Microtubules appear to lie in two planes, thereby giving support to the “two system” observation for microtubules in the guard cells of Pisum sativum.     

PHYSIOLOGICAL IMPLICATIONS OF VICIA FABA AND NICOTIANA TABACUM GUARD-CELL ULTRASTRUCTURE

The guard cells of Vicia faba and Nicotiana tabacum contain numerous mitochondria, elements of endoplasmic reticulum, spherosomes, and peroxisome-like microbodies. A full ribosomal complement appears in young but not in fully mature guard cells. Numerous small lipid droplets external to the plasmalemma were noted in mature Vicia guard cells. Chloroplasts were found in both epidermal and guard cells of both species. Full photosynthetic capacity was indicated by the grana fretwork of guard-cell chloroplasts. A specialized peripheral reticulum was observed in the guard-cell chloroplasts of Vicia. Plasmodesmata were observed in both walls between sister guard cells and between guard and epidermal cells. In the latter case plasmodesmata were found primarily in pit fields of transverse walls. It is postulated that the small volume of guard cells allows them an osmotic advantage over larger neighboring cells in generating turgor.     

ULTRASTRUCTURE AND DEVELOPMENT OF THE INACTIVE STOMATA OF ANABASIS ARTICULATA (FORSK.) MOQ.

The guard cells of Anabasis articulata mature and senesce a short distance from the intercalary meristem in which they form. When the guard cells reach final size, their ultrastructure is similar to that of stomata of other plants. At this stage, they contain clearly definable, numerous mitochondrial profiles, chloroplasts with starch grains and plastoglobuli, active Golgi bodies, a large nucleus that stains deeply for chromatin and large vacuoles. During later stages of development the whole protoplasmic content becomes very dense, with myelin-like figures and crystals appearing in the vacuoles. The cell walls thicken considerably. This is especially true of the tangential walls, where the microfibrils of different lamellae vary in their orientation. It is suggested that as a result of these ultrastructural changes the guard cells lose the ability to move.     

Cellular changes associated with rest and quiescence in winter-dormant vascular cambium of<Emphasis Type="Italic"> Pinus contorta</Emphasis>

In winter, dormant cambial cells contain many small vacuoles interspersed throughout the cytoplasm. This differs dramatically from actively growing cambial cells whose structure is dominated by large central vacuoles. Structure reported in studies using conventional chemical fixation and transmission electron microscopy (TEM) conflicts with that described earlier for live cambial cells using light microscopy. In this study, cryofixation (high-pressure freezing/freeze substitution) was used to preserve dormant Pinus contorta fusiform cambial cells, revealing structure more consistent with that in early micrographs of live cambial cells. At the ultrastructural level, the plasmalemma was consistently smooth and tightly associated with the cell wall, contrary to the highly in-folded plasmalemma seen in chemically fixed cambial cells. In addition, both TEM and live-cell confocal microscopy demonstrated that, in some places, dormant cells were partitioned into more numerous, smaller vacuoles than were observed after chemical fixation. Populations of different vacuoles were apparent based on size, shape and membrane staining. Larger vacuoles had prominent tonoplasts and were often present as axially elongated, interconnecting networks with associated microfilament bundles. Endoplasmic reticulum fragmented during rest into numerous vesicular structures similar to small vacuoles, then with the transition to quiescence reformed into the smooth cisternal form.     

ULTRASTRUCTURE OF THE SECONDARY PHLOEM OF TILIA AMERICANA

Summer and winter (July and January) samples of secondary phloem of Tilia americana were studied with the electron microscope. Parenchyma cells contain: nuclei, endoplasmic reticulum, ribosomes, plastids, mitochondria and occasional dictyosomes. Well-defined tonoplasts separate vacuoles from cytoplasmic ground substance. Vacuoles often contain tannins. Lipid droplets are common in cytoplasm. Endoplasmic reticulum–connected plasmodesmata are aggregated in primary pit fields. Companion cells differ from parenchyma cells in having numerous sieve-element connections, possibly slime, and in lacking plastids. Mature, enucleate sieve elements possess 1–4 extruded nucleoli. Numerous vesicles occupy a mostly parietal position in association with plasmalemma. The mature sieve element lacks endoplasmic reticulum, organelles (except for few mitochondria) and tonoplast. In OsO_4– and glutaraldehyde-fixed elements, slime has a fine, fibrillar appearance. Normally, these fine fibrils are organized into coarser ones which form strands that traverse the cell and the plasmalemma-lined pores of sieve plates and lateral sieve areas.     

ULTRASTRUCTURAL CHANGES ASSOCIATED WITH UTILIZATION OF METABOLITE RESERVES AND TRICHOME DIFFERENTIATION IN THE PROTOCORM OF VANDA

Certain aspects of protocorm development in Vanda were examined ultrastructurally. The parenchymal cells of the protocorm accumulate substantial quantities of lipid, protein, and carbohydrate reserves which disappear gradually with the senescence of the parenchymatous region. The proteinaceous reserves appear initially as discrete bodies which become intimately associated with clusters of small tubules. The tubules eventually disperse throughout the cytoplasm and disappear following depletion of the protein bodies. The lipid reserves also appear as discrete bodies and are associated with an electron dense, laminated inclusion which appears to increase in size with the disappearance of the lipid bodies. While plastids in the meristematic cells differentiate a well-developed thylakoid system and contain little starch, those of the parenchymal cells contain large starch grains and numerous osmiophilic droplets and develop meager thylakoid systems. Membrane-bound crystalline structures of hexagonal and rhomboid cross section occur frequently in the cytoplasm of senescent parenchyma cells. Trichome initials, which differentiate from the epidermis, contain few conventional organelles and exhibit numerous membrane-bound structures containing many small crystalline inclusions. Numerous vesicles accumulate at the tips of the trichomes in spaces between the cell wall and the plasmalemma.     

THE LATERAL MERISTEM AND ITS DERIVATIVES IN THE CORM OF ISOETES MURICATA

Corm tissue of Isoetes muricata Dur. was fixed in glutaraldehyde and postfixed in osmium tetroxide for electron microscopy. Very young secondary sieve elements can be distinguished from contiguous cambial cells by their distinctive plastids and by the presence of crystalline and/or fibrillar proteinaceous material in dilated cisternae of rough endoplasmic reticulum (ER). At maturity, the sieve elements are lined by the plasmalemma and a parietal, anastomosing network of smooth ER. Degenerate nuclei persist in all mature sieve elements. In addition, mature sieve elments contain plastids and mitochondria. Sieve-area pores are present in all walls. The lateral meristem of I. muricata consists of 2–3 layers of cells year-round. Judging from numerous collections made between October 1972 and July 1975, new sieve-element differentiation precedes cambial activity by about a month. Early in May, 1–2 cells immediately adjacent to already mature sieve elements differentiate directly into sieve elements without prior division. In early June, at about the time sieve-element differentiation is completed, cambial division begins. Division is sporadic, not uniform throughout the meristem. Dormancy callose accumulates in the secondary sieve elements in late October, and is removed in early May, at about the same time new sieve-element differentiation begins. Cells of the dormant cambium are characterized by the presence of numerous small vacuoles and large quantities of storage materials, including lipid droplets, starch grains, and tannin. By contrast, active cambial cells contain few large vacuoles with little or no tannin, and they have little storage material.     

慈菇匍匐茎中分泌道的初步研究

慈茹匍蔔茎的分泌道是裂生的胞间道,分布于匍匐茎的基本组织中。单个分泌道原始细胞起始于离茎端约1毫米处的基本分生组织中,原始细胞经分裂形成5—7个上皮细胞包围着中央的裂生腔隙,成为管道系统。上皮细胞无鞘细胞包围。上皮细胞中高尔基体和内质网发达,并溢出小囊泡向着分泌道腔隙面壁的质膜附近迁移,乳汁中亦存在大量完整的小囊泡。上皮细胞和外围薄壁细胞之间的壁层具有大量胞间连丝,小囊泡和内质网的膜结构与胞间连丝末端相接,同时可见上皮细胞的质膜在数处反折内陷,形成袋状结构,在与上皮细胞相对的薄壁细胞内也有同样现象出现,袋状结构内含小形颗粒或囊泡,并在结构上显示出上皮细胞与相邻薄壁细胞间存在着活跃的物质交流。由此认为。代谢物质以整体小囊泡的形式经胞间连丝或内陷的质膜向分泌道迁移是物质运输和分泌的可能方式之一。在电镜下观察,液泡中的积聚物与乳汁十分相似,液泡可能是乳汁的贮存场所之一。     

STRUCTURE OF THE VASCULAR PARENCHYMA IN THE STEM OF LYCOPODIUM LUCIDULUM

At maturity the vascular cylinder of the stem of Lycopodium lucidulum contains two distinct types of parenchyma cells, one which is always associated with sieve cells, the other with tracheids. The remaining parenchyma cells have characteristics intermediate between the two extremes. The most conspicuous feature of the sieve cell-associated parenchyma cell is the very dense appearance of its protoplast, due to a high ribosome population and absence of large vacuoles. The large, ramifying nuclei of these cells have numerous connections with the endoplasmic reticulum (ER). The tracheid-associated parenchyma cells, which are light in appearance, contain many small vacuoles and a relatively small ribosome population. These cells also contain relatively small nuclei and considerable ER cisternae. The parenchymatous elements which have characteristics intermediate between sieve cell- and tracheid-associated parenchyma may or may not be contiguous to the sieve cells or tracheids. An intergradation in wall thickness occurs among parenchyma cells of the vascular cylinder, the thicker-walled cells being adjacent to the sieve cells, the thinner-walled ones next to the tracheids. An intergradation also occurs in the frequency of plasmodesmata between the various parenchyma cells. The closer parenchyma cells are to the sieve cells the greater the number of connections between them. No plasmodesmata were found between the tracheid-associated parenchyma cells.     

THE FINE STRUCTURE OF EPENDYMA IN THE BRAIN OF THE RAT

The ciliated ependyma of the rat brain consists of a sheet of epithelial cells, the luminal surface of which is reflected over ciliary shafts and numerous evaginations of irregular dimensions. The relatively straight lateral portions of the plasmalemma of contiguous cells are fused at discrete sites to form five-layered junctions or zonulae occludentes which obliterate the intercellular space. These fusions occur usually at some distance below the free surface either independently or in continuity with a second intercellular junction, the zonula adhaerens. The luminal junction is usually formed by a zonula adhaerens or, occasionally, by a zonula occludens. The finely granular and filamentous cytoplasm contains supranuclear dense bodies, some of which are probably lysosomes and dense whorls of perinuclear filaments which send fascicles toward the lateral plasmalemma. The apical regions of the cytoplasm contain the basal body complexes of neighboring cilia. These complexes include a striated basal foot and short, non-striated rootlets emanating from the wall of each basal body. The rootlets end in a zone of granules about the proximal region of the basal body, adjacent to which may lie a striated mass of variable shape. All components of the basal body complex of adjacent cilia are independent of each other.     

FINE STRUCTURAL STUDIES ON THE INTERPHASE AND DIVIDING APICAL CELLS OF SPHACELARIA TRIBULOIDES (PHAEOPHYTA)1

The apical cells of Sphacelaria tribuloides Menegh. are larger than other thallus cells, contain more organelles and appear polarized. Their tip portion, where they grow, contains a well developed Golgi apparatus, abundant endoplasmic reticulum (ER) membranes, mitochondria, chloroplasts and a large number of small vacuoles. It seems likely that a continuous flow of membranous material from the ER membranes to the dictyosomes and from the latter to the plasmalemma of the extending tip portion takes place. In contrast, the basal pole possesses fewer organelles and is occupied mainly by large-sized, sometimes central vacuoles. The apical cells undergo two distinct types of highly asymmetrical differential divisions giving rise to cells of the thallus and hair initials. During the early stages of mitosis the nuclear envelope remains intact, except for fenestrated poles. Microtubules pass through the fenestrae into the nucleoplasm. During meta-phase, a typical chromosome plate is organized. The sites of attachment of spindle microtubules to the chromosomes are structurally different from the rest of the chromosomes. At late anaphase, the nuclear envelope breaks down completely. During telophase, a new membrane encloses the chromosomes which are decondensed and the nucleoli are reorganized. Cytokinesis proceeds long after mitosis at a stage in which the nuclei have increased in size and have moved farther apart. A membranous furrow develops centripetally, without the participation of microtubules. However, microtubules traverse the thin cytoplasmic strands which, in both interphase and cytokinetic cells, meander among the vacuoles of the basal pole of the cell and the internuclear space. Dictyosomes appear to be involved in the subsequent wall deposition.     

THE ULTRASTRUCTURE OF DEVELOPING LATEX DUCTS IN MAMMILLARIA HEYDERI (CACTACEAE)

Electron microscopy was used to investigate early development of latex ducts in Mammillaria heyderi (Cactaceae). Numerous vesicles (secondary vacuoles) form from invaginations of the plasmalemma near sites of wall thinning, from endoplasmic reticulum (ER), and from vesiculate grana of degenerate plastids. Dictyosomes, though they occur in young duct cells, do not seem to be responsible for the formation of vesicles. Cytoplasmic vesicles may contain fibrillar, globular, or crystalline materials, or may be devoid of any type of particulate matter. They may be responsible for storage of numerous laticiferous components. Lysosomal materials could be stored in some vesicles and contribute to the degradation of the protoplast. Some nuclei contain condensed chromatin and are subject to deformation and collapse. Mitochondria and lipid bodies are common in young duct cells but ER is rare. When ducts form in young tissues, plastids in the lumen do not produce starch grains or extensive membranous networks. The plastids eventually degenerate to become a part of latex. If ducts form in older, established tissues having mature plastids, the plastids undergo extreme modification.     

The fine structure and histochemistry of the caecal epithelium of Calicotyle kröyeri (Monogenea: Monopisthocotylea)

The caecal epithelium of Calicotyle kröyeri consists of a single cell type which functions in the uptake and intracellular digestion of host epidermis and associated mucus. Each cell is columnar with a small basal nucleus and prominent nucleolus. Perinuclear cytoplasm contains narrow profiles of GER and mitochondria with numerous cristae. Golgi complexes are small and indistinct. Most of the cell is filled with vacuoles of heterogeneous content, the largest occupying the cell apex. There is in each cell an apical endocytotic complex comprising cell surface lamellae, apical vesicles and numerous tubular invaginations of the plasmalemma. The limiting membrane of all these components is structurally modified and bears a highly organized array of peg-like structures on its luminal surface. The complex is capable of ingesting particulate food material from the gut lumen for transfer, via vesicles, to the vacuoles for digestion. Most of the vacuoles represent the digestive elements of the cell and, histochemically, are reactive for protein, mucus and carboxylic esterases. Indigestible residues and lipid droplets accumulate in the large apical vacuole and are periodically released to the lumen by exocytosis. Small, undifferentiated caecal cells were occasionally observed in the epithelium, but their development has not been recorded.     

ULTRASTRUCTURE OF OSMOPHORES IN RESTREPIA (ORCHIDACEAE)

Osmophores of the myophilous genus Restrepia (Orchidaceae) were studied developmentally at the ultrastructural level. They are located at petal apices and on the adaxial surface of the dorsal sepal apex. Up to and through anthesis a dense, osmiophilic exudate is synthesized probably in endoplasmic reticulum or in plastids of papillose epidermal cells, and then appears to be transported through the plasmalemma by granulocrine elimination. As the exudate is amassed, the cuticle ruptures to form numerous pores that extend from the cell wall. Mitochondria and amyloplasts are especially abundant at anthesis. From anthesis to post-anthesis, lipid droplets appear in the cytoplasm and vacuoles of epidermal cells, and frequency of cell organelles drops markedly with increasing vacuolation.     

STRUCTURE AND DEVELOPMENT OF THE SIEVE ELEMENTS IN PSILOTUM NUDUM

Shoot tissue of Psilotum nudum (L.) Griseb. was fixed in glutaraldehyde and postfixed in osmium tetroxide for electron microscopy. Young sieve elements can be distinguished from contiguous parenchyma cells by their distinctive plastids, the presence of refractive spherules, and the overall dense appearance of their protoplast. The refractive spherules apparently originate in the intracisternal spaces of the endoplasmic reticulum (ER). With increasing age the sieve-element wall undergoes a marked increase in thickness. Concomitantly, a marked increase occurs in the production of dictyosome vesicles, many of which can be seen in varying degrees of fusion with the plasmalemma. Other fibril- and vesicle-containing vacuoles also are found in the cytoplasm. In many instances the delimiting membrane of these vacuoles was continuous with the plasmalemma. Vesicles and fibrillar materials similar to those of the vacuoles were found in the younger portions of the wall. At maturity the plasmalemma-lined sieve element contains a parietal network of ER, plastids, mitochondria, and remnants of nuclei. The protoplasts of contiguous sieve elements are connected by solitary pores on lateral walls and pores aggregated into sieve areas on end walls. All pores are lined by the plasmalemma and filled with numerous ER membranes which arise selectively at developing pore sites, independently of the ER elsewhere in the cell. P-protein and callose are lacking at all stages of development.     

ULTRASTRUCTURE OF THE SHOOT APEX OF TOMATO (LYCOPERSICON ESCULENTUM)

The vegetative shoot apical meristem of tomato (Lycopersicon esculentum Mill.) was examined at the ultrastructural level. The meristem consisted of a surface layer that was different from the rest of the meristem and was unique among the dicotyledonous species. The cells of the surface layer contained large distal vacuoles with relatively large electron-dense inclusions, proplastids with membrane-bound inclusions (MB), and differentiating chloroplasts. In addition, periclinal and oblique divisions were observed in the surface layer cells along with anticlinal divisions. The cells of the subsurface layers contained small vacuoles with fewer inclusions as well as proplastids of various shapes but without MB. Differentiating chloroplasts were not observed in these cells, but autophagic vacuoles at various stages of development were present. The normal complement of cell inclusions, e.g., the mitochondria, golgi bodies, endoplasmic reticulum (ER), ribosomes, and microtubules were observed in subsurface layers, and in many cells the ER was observed to be continuous with the outer membrane of the nuclear envelope and with the plasmalemma. Further below in the meristem, cells contained both the proplastids and differentiating chloroplasts with MB. In the latter, the outer membrane of the MB was found to be continuous with the developing lamellae, suggesting that MB probably serve as the storage centers for lamellae membranes. Near the base of the meristem, in the pith-rib meristem, enlarged cells containing large vacuoles and differentiated chloroplasts were present.     

THE ASSOCIATION OF DRUSE CRYSTALS WITH THE DEVELOPING STOMIUM OF CAPSICUM ANNUUM (SOLANACEAE) ANTHERS

Each of the two stomiums in the anther of Capsicum annuum (sweet pepper) consists of a single layer of cells immediately below the epidermis between two adjacent locules. Each stomium extends the entire length of the anther and splits open at pollen maturity. Many calcium oxalate druse crystals form within the vacuoles of the stomium cells in association with membrane complexes and paracrystalline bodies. These latter structures are reported here for the first time and each is considered to be a nucleation site for druse crystal formation. Prior to the appearance of membrane complexes and crystals within the vacuoles, plasmalemmasomes are visible next to the stomium cell walls and contain vesicles and fibrous material. We propose that these bodies carry wall materials, including calcium ions and possibly oxalate ions, into the vacuoles. Their presence coincides with crystal formation. Two other types of crystals occur in the connective tissue between stomiums and the single vascular strand. These crystals, along with those in the two stomiums, form at precise times during anther development. Contrary to the more numerous suggestions that crystals protect against predators or are metabolic waste products, we believe their formation aids in degradation and weakening of the cell walls between the locules and, thus, contributes to the release mechanism for the pollen.     

Microtubules and coated vesicles in guard-cell protoplasts ofAllium cepa L.

Protoplasts were prepared from the guard cells ofA. cepa. Epidermal peels taken from expanding green leaves and largely free of mesophyll were treated with Cellulysin, and protoplasts were harvested after 18 h of digestion. That the protoplasts were derived from guard cells was ascertained from their characteristic vacuolar autofluorescence and from observations showing that all other epidermal cells are killed in the peeling procedure. The protoplasts proved to be a good system with which to view the cell cortex and inner surface of the plasmalemma. The lysis of cells adhering to polylysine-treated, Formvar-coated grids, followed by negative staining in uranyl acetate, showed that many microtubules normally present in ordered arrays in situ remain closely applied to the inner surface of the plasmalemma in protoplasts. In addition, numerous vesiculate elements including coated vesicles and/or pits are present amongst the microtubules. Similar vesicles are evident in thin sections of fixed, embedded guard cells and protoplasts. The significance of these structures in the cell cortex is discussed.     

在分化条件下甜菊愈伤组织分生区细胞超微结构研究

对甜菊（Ｓｔｅｖｉａｒｅｂａｕｄｉａｎａ）愈伤组织中尚未发生器官分化的分生细胞团进行了超微结构研究．结果表明,在器官分化条件下,愈伤组织中形成的分生区域的细胞体积小,细胞核大,核仁明显,且具核仁泡,部分细胞核中含有核内含物．大量小液泡分布在细胞的边周或散布于整个细胞中．液泡中通常含有陷入的细胞质成分和膜状物．部分液泡的形成与内质网膨大有密切关系．同时也观察到由内质网形成的多圈膜和双层膜包围细胞质成分的同心环结构．高尔基体及其小泡丰富,有时聚集分布在细胞某一区域．核糖体密集,有的聚集成多聚核糖体．因此,愈伤组织中分生区的细胞与分生组织中的液泡化和分裂的细胞类似．分生区细胞的另一明显特征是出现质膜内陷．推测这些超微结构特征可能反映了甜菊愈伤组织器官分化前的某些形态变化。     

天麻大型细胞消化蜜环菌过程中溶酶体小泡的作用

蜜环菌(Armillaria mellea Fr.)菌丝由天麻(Gastrodia elata Bl.)皮层细胞经纹孔侵入大型细胞。初期大型细胞的原生质膜凹陷,同时细胞壁产生乳突状加厚阻止菌丝侵入。当菌丝侵入大型细胞以后,凹陷的质膜将菌丝紧密包围,大量由单位膜围成的小泡聚集在其周围。随后这些小泡的膜与质膜融合并将其内含物释放到菌丝周围的空间中,凹陷质膜逐渐膨大成为一个包围菌丝的消化泡。小泡和消化泡中均具酸性磷酸酶活性反应产物,证实其分别相当于植物溶酶体系统中的初级和次级溶酶体。菌丝在消化泡中被彻底消化。