DEVELOPMENT OF MUCILAGINOUS SURFACES IN EUGLENOIDS. I. STALK MORPHOLOGY OF COLACIUM MUCRONATUM1

The envelope and stalk of Colacium mucronatum Bourr. & Chad, were examined in living cells with light microscopy and in fixed preparations with scanning electron microscopy using critically point dried (CPD) and freeze dried (FD) preparations. The envelope of palmelloid cells is formed over the entire cell surface by many individual strands attached at right angles to areas of articulation of the pellicular strips. Strands were observed to anastomose on the posterior tip of otherwise naked cells. Stalks of living cells in India ink preparations had an optically dark inner core with a lighter outer sheath. In FD stalks a definite inner core was not evident, whereas CPD stalks had an outer surface composed of thick strands which may be the collapsed and aggregated strands of the FD stalks. In both there was also an amorphous matrix. The stalk forms from the aggregation of many strands from the anterior cell tip back to a point encompassing the cell surface anterior to a cross section of the tip 9 μm diam. The outer surface of the stalk comes from the pellicular surface joining that area and the core from the cell tip in the area of the canal opening. Any possible participation of the inner canal surface in stalk formation could not be determined because of the great density of the mucilage at the cell-tip/stalk junction.     

THE MORPHOLOGY AND DEVELOPMENT OF SOME PROMINENTLY STALKED SOUTHERN AUSTRALIAN HALYMENIACEAE (CRYPTONEMIALES,RHODOPHYTA). I. CRYPTONEMIA KALLYMENIOIDES (HARVEY) KRAFT COMB. NOV. AND C. UNDULATA SONDER1

Several southern Australian red algae of the family Halymeniaceae (Cryptonemiales) are differentiated into hard, massive stalks and considerably softer laminar blades or phyllodes. The taxonomy, morphology and pit-connection ultrastructure of one such species, Cryptonemia kallymenioides (Harvey) Kraft comb. nov., are compared to C. undulata Sonder, which lacks massive stalks. In both species there is extensive periodic secondary cortication of the stalks, resulting in the formation of distinct “growth rings.” The blades of C. kallymenioides appear to be seasonal and its stalks perennial, while plants of C. undulata are apparently perennial but shorter lived than C. kallymenioides. As a result, stalks in the latter can reach 2–3 cm in diameter with up to 18 growth rings, compared to the 1–2 mm diameters and up to 6 rings within the stalks of C. undulata. Heavy secondary thickening of cortical cell walls occurs in both species and confers a “woody” texture to the stalks of C. kallymeniodes. Regardless of the large differences in average stalk diameters between the two species, the pit-connection ultrastructure from cortex to medulla shows much the same sequence of morphological modification. Pit-connections are standard red algal structures in the outer cortex, but become increasingly convoluted on the membrane-bound surfaces abutting cytoplasm and develop wider apertures and less dense cores with increasing distance from the stalk surface. In occasional medullary cells of C. kallymenioides, the cytoplasm disintegrates, leaving cell walls and pit-connections to play an apparently structural role which has not been reported in other red algae. It is suggested that the increase in aperture size and surface areas of pit-connections is compatible with their playing a role in the intercellular transport of solutes towards the inner cell layers which may, in C. kallymenioides, lie many millimeters distant.     

Studies onArtemisia afra Jacq.: The phloem in stem and leaf

Summary The structure of the phloem was studied in stem and leaf ofArtemisia afra Jacq., with particular attention being given to the sieve element walls. Both primary and secondary sieve elements of stem and midvein have nacreous walls, which persist in mature cells. Histochemical tests indicated that the sieve element wall layers contained some pectin. Sieve element wall layers lack lignin. Sieve elements of the minor veins (secondary and tertiary veins) lack nacreous thickening, although their walls may be relatively thick. These walls and those of contiguous transfer cells are rich in pectic substances. Transfer cell wall ingrowths are more highly developed in tertiary than in secondary veins.     

Propagation of the neuroid action potential of the carnivorous plantDrosera

Summary The neuroid action potentials ofDrosera rotundifolia recorded from single cells resemble those recorded from the surface of tentacle stalks. They have similar amplitudes and durations and they show the same variation of duration with interval that characterizes the extracellularly recorded action potentials. All living cells of the stalk appear to be excitable since cells from both layers were observed to produce action potentials when intracellular recording techniques were used.Propagation of electrically induced action potentials down the stalk occurred at a rate of 4.3 mm/s±0.6 S.E.M. while propagation up the stalk occurred at a rate of 9.9 mm/s±2.0 S.E.M. The fact that attenuated signals from electrical processes and stimulus artifacts in distant parts of the stalk were detected in recordings indicates that the cells of the stalk were closely coupled and that propagation from cell to cell is probably by an electrotonic mechanism. This hypothesis gains additional support from the observation of numerous cytoplasmic connections (plasmodesmata) through the cell walls separating the cells which are most likely to conduct the action potential.We wish to thank Dr. M.V. Parthasarathy for guidance in the use of the electron microscope. This work was supported in part by NSF Grant Number GB28124.     

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.     

The vacuolar-tubular continuum in living trichomes of chickpea (Cicer arietinum) provides a rapid means of solute delivery from base to tip

Summary A vacuolar continuum exists from base to tip in the secretory trichomes of chickpea (Cicer arietinum). This continuum is seen in living trichomes which have been labeled with Lucifer yellow CH and examined with confocal microscopy. It encompasses the large vacuole of the lower stalk cell, the vacuoles and tubules of the central stalk cell, the thin tubules of the upper stalk cell, and the tubules and vacuoles of the secretory head cells. The vacuolar-tubular system is structurally distinct within each cell, forming a gradient of large vacuoles in the lower stalk cell, thick tubules in the central stalk cell, and thin anastamozing tubules in the upper stalk cell. This membrane system appears to be continuous between trichome cells, as thin tubules emanate from plasmodesmata between stalk cells and between the upper stalk and lower head cell. In the upper stalk cell, the thin tubules of this continuum are streaming up and down the long axis of the cell at 0.67 m/s. The larger vacuolar-tubular system in the central and lower stalk cells is also slowly moving, with apparent peristalsis occurring in the central cell. The vacuolar-tubular system of the secretory head cells is completely labeled with Lucifer yellow when the dye has only partly diffused up the long walls of the trichome, indicating that the streaming tubular system delivers solute through the stalk cells to the secretory head cells faster than diffusion through the trichome walls. In the lower head cells, tubules emanate from the plasmodesmata connecting to the upper stalk cell, and these tubules are continuous with the head cell vacuoles. In addition, another layer of thin tubules forms along the edges of the secretory head cells, at the site of exocytotic secretion. We propose that the continuous vacuolar-tubular system in these trichomes functions to rapidly deliver solute from the base of the trichome to the secretory head cells. This system provides a pathway for the transport of secretory material.Dedicated to Prof. Dr. Dr. h.c. Eberhard Schnepf on the occasion of his retirement     

Differential modes of crosslinking establish spatially distinct regions of peptidoglycan in Caulobacter crescentus

The diversity of cell shapes across the bacterial kingdom reflects evolutionary pressures that have produced physiologically important morphologies. While efforts have been made to understand the regulation of some prototypical cell morphologies such as that of rod‐shaped Escherichia coli, little is known about most cell shapes. For Caulobacter crescentus, polar stalk synthesis is tied to its dimorphic life cycle, and stalk elongation is regulated by phosphate availability. Based on the previous observation that C. crescentus stalks are lysozyme‐resistant, we compared the composition of the peptidoglycan cell wall of stalks and cell bodies and identified key differences in peptidoglycan crosslinking. Cell body peptidoglycan contained primarily DD‐crosslinks between meso‐diaminopimelic acid and D‐alanine residues, whereas stalk peptidoglycan had more LD‐transpeptidation (meso‐diaminopimelic acid‐meso‐diaminopimelic acid), mediated by LdtD. We determined that ldtD is dispensable for stalk elongation; rather, stalk LD‐transpeptidation reflects an aging process associated with low peptidoglycan turnover in the stalk. We also found that lysozyme resistance is a structural consequence of LD‐crosslinking. Despite no obvious selection pressure for LD‐crosslinking or lysozyme resistance in C. crescentus, the correlation between these two properties was maintained in other organisms, suggesting that DAP‐DAP crosslinking may be a general mechanism for regulating bacterial sensitivity to lysozyme.     

两种类型栓皮栎软木细胞微观排列与形态特征

采用扫描电镜对秦岭北坡楼观台地区的厚皮、薄皮两种类型栓皮栎软木进行细胞微观构造观察分析,并与欧洲栓皮槠进行比较,以阐明厚皮、薄皮栓皮栎软木的相关特性,为中国栓皮栎软木的合理利用提供依据。结果表明:(1)两种类型栓皮栎软木细胞的排列结构较一致,均由内部中空的封闭型薄壁细胞紧密排列组成;在弦切面上呈蜂窝状排列,径切面和横切面上呈砖墙状排列;在径切面上,软木细胞侧高整齐地排列成行,且与树干轴向垂直;在横切面上,软木细胞侧高整齐地处于以树干轴为中心散发出来的射线上。(2)栓皮栎软木细胞大小、细胞壁和侧壁褶皱等受生长季节的影响;从软木细胞形态特征上看,厚皮类型软木细胞壁薄、细胞体积大,其软木质量优于薄皮类型。(3)与欧洲栓皮槠比较,发现厚皮类型栓皮栎早软木细胞棱柱高较小(20.6μm vs.(对比)30~40μm),软木细胞壁略厚(1.7μm vs.1~1.5μm),细胞实体积(细胞壁体积占细胞总体积比例)略大(18.75%vs.10%),厚皮类型栓皮栎软木比欧洲栓皮槠的软木质量差一些。(4)受树皮生长应力的影响,两种类型栓皮栎软木细胞侧高壁上多发生褶皱,早软木细胞褶皱严重,晚软木细胞没有褶皱,但在早晚软木交界或含有杂质处褶皱特别严重,表明厚皮类型软木细胞的侧壁褶皱程度高于薄皮类型。(5)对细胞形态特征及软木特性等的分析表明,薄皮类型栓皮栎软木质量比厚皮类型差,未来对软木资源的开发利用应更注重厚皮类型。     

Development and ultrastructure of glandular trichomes in<Emphasis Type="Italic">pelargonium x fragrans</Emphasis> ‘mabel grey’ (geraniaceae)

The glandular trichomes of leaves fromPelargonium xfragrans ‘Mabel Grey’ (Geraniaceae) were examined by light, scanning, and transmission electron microscopy. These trichomes had unicellular globular heads and stalks of different lengths and features. Two types were classified: Type I, with an elongated, large head and a short (100 μm), cylindrical stalk that was more apparent on the adaxial surface; and Type II, with a spherical, small head and a long (300μm), conical stalk that was more pronounced on the abaxial surface. The ultrastructure of secretory cells from both types was distinguished by a well-developed endoplasmic reticulum, mitochondria, plastids, dictyosomes, and numerous vacuoles that likely were involved in the storage and transport of lipophilic substances. Plasmodesmata were frequent on the walls of the secretory and stalked cells. Here, we discuss the implication of structural differentiation in these trichomes.     

THE DEVELOPMENT OF CELLULAR STALKS IN BACTERIA

Extensive stalk elongation in Caulobacter and Asticcacaulis can be obtained in a defined medium by limiting the concentration of phosphate. Caulobacter cells which were initiating stalk formation were labeled with tritiated glucose. After removal of exogenous tritiated material, the cells were subjected to phosphate limitation while stalk elongation occurred. The location of tritiated material in the elongated stalks as detected by radioautographic techniques allowed identification of the site of stalk development. The labeling pattern obtained was consistent with the hypothesis that the materials of the stalk are synthesized at the juncture of the stalk with the cell. Complementary labeling experiments with Caulobacter and Asticcacaulis confirmed this result. In spheroplasts of C. crescentus prepared by treatment with lysozyme, the stalks lost their normal rigid outline after several minutes of exposure to the enzyme, indicating that the rigid layer of the cell wall attacked by lysozyme is present in the stalk. In spheroplasts of growing cells induced with penicillin, the stalks did not appear to be affected, indicating that the stalk wall is a relatively inert, nongrowing structure. The morphogenetic implications of these findings are discussed.     

Anatomical analysis of Saccharomyces cerevisiae stalk-like structures reveals spatial organization and cell specialization

Recently we reported an unusual multicellular organization in yeast that we termed stalk-like structures. These structures are tall (0.5 to 3 cm long) and narrow (1 to 3 mm in diameter). They are formed in response to UV radiation of cultures spread on high agar concentrations. Here we present an anatomical analysis of the stalks. Microscopic inspection of cross sections taken from stalks revealed that stalks are composed of an inner core in which cells are dense and vital and a layer of cells (four to six rows) that surrounds the core. This outer layer is physically separated from the core and contains many dead cells. The outer layer may form a protective shell for the core cells. Through electron microscopy analysis we observed three types of cells within the stalk population: (i) cells containing many unusual vesicles, which might be undergoing some kind of cell death; (ii) cells containing spores (usually one or two spores only); and (iii) familiar rounded cells. We suggest that stalk cells are not only spatially organized but may undergo processes that induce a certain degree of cell specialization. We also show that high agar concentration alone, although not sufficient to induce stalk formation, induces dramatic changes in a colony's morphology. Most striking among the agar effects is the induction of growth into the agar, forming peg-like structures. Colonies grown on 4% agar or higher are reminiscent of stalks in some aspects. The agar concentration effects are mediated in part by the Ras pathway and are related to the invasive-growth phenomenon.     

Fine Structure of Fruiting Bodies of Stigmatella aurantiaca (Myxobacterales)

The fruiting body of Stigmatella aurantiaca consists of a thick stalk supporting a number of individual cysts. The stalk is made of discontinuous tubules, of dimensions known for vegetative cells, which are oriented parallel to the longitudinal axis of the stalk. The red-brown cysts contain numerous, randomly oriented myxospores which are surrounded by thick, fibrous capsules. Their cell walls are wavy or ruffled and exhibit fewer budlike infoldings than reported for myxospores induced in liquid. We suggest that the extended time period available for metabolic and regulatory adjustments by the cell during morphogenesis within cysts accounts for the presence of considerably fewer deep cell wall infoldings than in glycerol-induced myxospores.     

Importance of organic phosphate hydrolyzed in stalks of the lotic diatom <Emphasis Type="Italic">Didymosphenia geminata</Emphasis> and the possible impact of atmospheric and climatic changes

The aquatic colonial stalked diatom, Didymosphenia geminata, has acquired notoriety in recent years because of huge increases in many rivers of temperate regions. However, in some streams in northern England it has probably or, in the case of the R. Coquet (Northumberland), certainly been abundant for many decades. The paper describes the nutrient environment and phosphatase activities of Didymosphenia in Stony Gill (N. Yorkshire), a fast-flowing stream draining an upland catchment with peaty soils overlying limestone. Organic phosphate formed 85% of the filtrable phosphate in the water during the study (January–August 2000), with a maximum in April. Colonies were most abundant in June, but had disappeared by August. Surface phosphomonoesterase (PMEase) and phosphodiesterase activities assayed from March to July showed low PMEase activity in early March, but otherwise both were high throughout the period and especially so in June and July. Use of BCIP-NBT staining procedure showed that PMEase activity occurred in the stalks. A more detailed study of colony structure and staining with material from the R. Coquet in June 2006 also showed marked PMEase activity, with staining localized in the upper part of the stalks and the cells remaining unstained. It is suggested that organic phosphate is hydrolyzed in the stalk and the inorganic phosphate passes to the cell via a central tube in the stalk. It seems likely that organic phosphate as a major P source is a key factor favouring the success of Didymosphenia. The possible impact of environmental changes in the catchment such as climatic warming, C loss from peat and atmospheric N deposition is discussed. Handling editor: L. Naselli-Flores     

Fluctuation analysis of Caulobacter crescentus adhesion

The aquatic bacterium Caulobacter crescentus divides asymmetrically to a flagellated swarmer cell and a cell with a stalk. At the end of the stalk is an adhesive organelle known as the holdfast, which the stalked cell uses to attach to a solid surface. Often there are two or more cells with their stalks attached to the same holdfast. By analyzing the fluctuations in the stalk angle for a pair of cells attached to a single holdfast, we determine the elastic stiffness of the holdfast. We model the holdfast as three torsional springs in series and find that the effective torsional spring constant for the holdfast is of the order of (10(-17)-10(-18)) Nm, with unequal spring constants. The asymmetry suggests the sequence in which the cells attach to each other, and in some cases suggests that strong crosslinks form between the stalks as they make a shared holdfast.     

A Caulobacter gene involved in polar morphogenesis.

At specific times in the cell cycle, the bacterium Caulobacter crescentus assembles two major polar organelles, the flagellum and the stalk. Previous studies have shown that flbT mutants overproduce flagellins and are unable to form chemotaxis swarm rings. In this paper, we report alterations in both the stalk and the flagellar structure that result from a mutation in the flagellar gene flbT. Mutant strains produce some stalks that have a flagellum, produce some stalks that have an extra lobe protruding from their sides, have filaments lacking the 29-kilodalton flagellin, and produce several unusual cell types, including filamentous cells as well as predivisional cells with two stalks and predivisional cells with no stalk at all. We propose that flagellated stalks arise as a consequence of a failure to eject the flagellum at the correct time in the cell cycle and that the extra stalk lobe is due to a second site for the initiation of stalk biogenesis. Thus, a step in the pathway that establishes the characteristic asymmetry of the C. crescentus cell appears to be disrupted in flbT mutants. We have also identified a new structural feature at the flagellated pole and the tip of the stalk: the 10-nm polar particle. The polar particles appear as a cluster of approximately 1 to 10 stain-excluding rings, visible in electron micrographs of negatively stained wild-type cells. This structure is absent at the flagellar pole but not in the stalks of flbT mutant predivisional cells.     

Caulobacter and Asticcacaulis stalk bands as indicators of stalk age.

The prosthecae (stalks) of dimorphic caulobacters of the genera Caulobacter and Asticcacaulis are distinguished among such appendages by the presence of disk-like components known as stalk bands. Whether bands are added to a cell's stalk(s) as a regular event coordinated with the cell's reproductive cycle has not been settled by previous studies. Analysis of the frequency of stalks with i, i + 1, i + 2, etc. bands 'among more than 7,000 stalks of Caulobacter crescentus revealed that in finite (batch) cultures (in which all offspring accumulate), the proportion of stalks with i + 1 hands was regularly 50% of the proportion of stalks with i bands. This implied that the number of bands correlated with the number of reproductive cycles completed by a stalked cell. In chemostat-maintained perpetual cultures, the proportion was greater than 50% because stalked cells, with their shorter reproductive cycle times, contributed a larger proportion of offspring to the steady-state population than did their swarmer siblings. In Asticcacaulis biprosthecum cells, which bear twin prosthecae, the twins on a typical cell possessed the same number of bands. For both genera, stalk bands provide a unique morphological feature that could be employed in an assessment of age distribution and reproductive dynamics within natural populations of these caulobacters.     

Electron-translucent angular areas in developing tapetum cell walls and pollen grains ofTilia platyphyllos

Summary InTilia platyphyllos, the anther tapetal cell walls undergo significant modifications from the tetrad stage onwards. During the tetrad stage the inner tangential and radial parts of the tapetal walls begin to dissolve, while the distal parts swell. After the tetrad stage, the distal and outer radial tapetal cell walls become covered by a thick, irregular, highly electron-dense, polysaccharide layer. Striking features of the maturing tapetal walls (microspore stage and later) are electron-translucent, structureless, unstainable angular areas of variable dimensions. Similar electron-translucent areas occur in the exine arcades and apertures, but also isolated in the locular fluid ofT. platyphyllos. Electron-translucent areas, that are also found in the exine arcades and tapetal cells of other angiosperms, can be interpreted as the products of poorly understood metabolic processes.     

Co‐ordinate synthesis and protein localization in a bacterial organelle by the action of a penicillin‐binding‐protein

Organelles with specialized form and function occur in diverse bacteria. Within the Alphaproteobacteria, several species extrude thin cellular appendages known as stalks, which function in nutrient uptake, buoyancy and reproduction. Consistent with their specialization, stalks maintain a unique molecular composition compared with the cell body, but how this is achieved remains to be fully elucidated. Here we dissect the mechanism of localization of StpX, a stalk‐specific protein in Caulobacter crescentus. Using a forward genetics approach, we identify a penicillin‐binding‐protein, PbpC, which is required for the localization of StpX in the stalk. We show that PbpC acts at the stalked cell pole to anchor StpX to rigid components of the outer membrane of the elongating stalk, concurrent with stalk synthesis. Stalk‐localized StpX in turn functions in cellular responses to copper and zinc, suggesting that the stalk may contribute to metal homeostasis in Caulobacter. Together, these results identify a novel role for a penicillin‐binding‐protein in compartmentalizing a bacterial organelle it itself helps create, raising the possibility that cell wall‐synthetic enzymes may broadly serve not only to synthesize the diverse shapes of bacteria, but also to functionalize them at the molecular level.     

Developmental plasticity and regenerative capacity in the renal ureteric bud/collecting duct system

Branching morphogenesis of epithelia is an important mechanism in animal development, being responsible for the characteristic architectures of glandular organs such as kidney, lung, prostate and salivary gland. In these systems, new branches usually arise at the tips of existing branches. Recent studies, particularly in kidney, have shown that tip cells express a set of genes distinct from those in the stalks. Tip cells also undergo most cell proliferation, daughter cells either remaining in the tip or being left behind as the tips advance, to differentiate and contribute to new stalk. Published time-lapse observations have suggested, though, that new branches may be able to arise from stalks. This happens so rarely, however, that it is not clear whether this reflects true plasticity and reversal of differentiation, or whether it is just an occasional instance of groups of tip cells being "left behind" by error in a mainly stalk zone. To determine whether cells that have differentiated into stalks really do retain the ability to make new tips, we have removed existing tips from stalks, verified that the stalks are free of tip cells, and assessed the ability of tip-free stalks to initiate new branches. We find stalks to be fully capable of regenerating tips that express typical tip markers, with these tips going on to form epithelial trees, at high frequency. The transition from tip to stalk is therefore reversible, at least for early stages of development. This observation has major implications for models of pattern formation in branching trees, and may also be important for tissue engineering and regenerative medicine.     

Origin and function of the stalk-cell vacuole in Dictyostelium

Large vacuoles are characteristic of plant and fungal cells, and their origin has long attracted interest. The cellular slime mould provides a unique opportunity to study the de novo formation of vacuoles because, in its life cycle, a subset of the highly motile animal-like cells (prestalk cells) rapidly develops a single large vacuole and cellulosic cell wall to become plant-like cells (stalk cells). Here we describe the origin and process of vacuole formation using live-imaging of Dictyostelium cells expressing GFP-tagged ammonium transporter A (AmtA-GFP), which was found to reside on the membrane of stalk-cell vacuoles. We show that stalk-cell vacuoles originate from acidic vesicles and autophagosomes, which fuse to form autolysosomes. Their repeated fusion and expansion accompanied by concomitant cell wall formation enable the stalk cells to rapidly develop turgor pressure necessary to make the rigid stalk to hold the spores aloft. Contractile vacuoles, which are rich in H⁺-ATPase as in plant vacuoles, remained separate from these vacuoles. We further argue that AmtA may play an important role in the control of stalk-cell differentiation by modulating the pH of autolysosomes.