The suberin lamella,a possible barrier to water movement from the veins to the mesophyll ofThemeda triandra forsk

Summary Precipitation of ferrous ions by ferricyanide in transpiring leaves ofThemeda triandra Forsk. produced crystalline deposits, which were visible with the light and electron microscope. Prussian blue crystals were formed within the lumina of the tracheary elements and the apoplast, or cell wall continuum of the vascular tissues and bundle-sheath cells. Little if any deposition was noted within the lignified secondary thickenings of the tracheary elements. The localization pattern suggests that the ferrous ions moved from the lumina of the tracheary elements via the exposed primary walls. Prussian blue crystals were abundant in the outer tangential and radial walls of the bundle-sheath cells. By contrast, crystals were lacking in the walls of neighbouring mesophyll cells, suggesting that the suberin lamella in the bundle-sheath walls effectively inhibited the apoplastic movement of ferrous ions and possibly may impede, or restrict the movement of water across the bundle-sheath/mesophyll interface.     

PATHWAY OF WATER MOVEMENT IN HYDROIDS OF POLYTRICHUM COMMUNE HEDW. (BRYOPSIDA)

The pathway of water movement in hydroids of Polytrichum was determined by the precipitation of an electron-dense crystal (Prussian blue) in the transpirational stream. Hydrolysed end walls appear highly water permeable since Prussian blue granules were localized within the loose fibrillar network. Electron-dense granules were found free in the lumen but not in the lateral wall or in the middle lamella. These results are compared with data from vascular plant tracheary elements.     

An Application of the Prussian Blue Technique to a Light Microscope Study of Water Movement in Transpiring Leaves of Cotton (Gossypium hirsutum L.)

Prussian blue deposition in transpiring leaves of Gossypiumhirsutum L. indicated that water moved rapidly from the trachearyelements by a path along the bundle sheath parenchyma to thebundle sheath extension parenchyma and then laterally throughthe epidermis to the sub-stomatal cavities or to the tnchomes.A slower pathway of water movement to the substomatal cavitiesoccurred through the mesophyll. The path of Prussian blue depositionindicated that waterleft the leaf through the stomata and toa lesser extent through the trichomes. In the outer periclinalwall of epidermalcells Prussian blue was deposited below butnot within the cuticle.     

Free-space marker studies on the leaf ofZea mays L.

Summary Two free-space marker procedures (Prussian blue and lanthanum nitrate) were employed to determine the pathway(s) followed by water and solutes in the transpiration stream after their introduction into the xylem of small and intermediate bundles, and the effectiveness of the suberin lamellae of the bundle-sheath cells as a barrier to the movement of tracer ions (Fe³⁺ and La³⁺). Judged from the distribution of Prussian-blue crystals (insoluble, crystalline deposits resulting from the precipitation of ferric ions by ferrocyanide anions) and lanthanum deposits, water and the tracer ions moved readily from the lumina of the vessels into the apoplast (cell wall continuum) of the phloem and bundle-sheath cells via portions of vessel primary walls not bearing lignified secondary wall thickenings. Prussian blue and lanthanum deposits were abundant on the bundlesheath cell side of the bundle sheath/mesophyll interface but few occurred on that of the mesophyll, indicating that the suberin lamella is an effective barrier to apoplastic movement of both ferric and lanthanum ions. The presence of Prussian-blue crystals and lanthanum deposits in the compound middle lamella of the radial wall of the bundle-sheath cells indicates that the compound middle lamella provides an apoplastic pathway for transpirational water from the xylem to the evaporating surfaces of the mesophyll and epidermal cells.     

Water pathways in wheat leaves. III. The passage of the mestome sheath and the function of the suberised lamellae

The fluorochrome sulphorhodamine G, when present in the transpiration stream in wheat leaves, passes rapidly out of the veins and produces fluorescence in the mesophyll and epidermal cell walls. The path of movement of the dye out of the tracherary elements and across the mestome sheath to the parenchyma sheath cells was followed by rapid freezing, freeze-subsitution, dry embedding in resin, sectioning and epifluorescence microscopy. The sulphorhodamine solution was visible in tracheary elements, and, where it had passed out of the tracheary elements, strongly fluorescent in some of the cell walls. The patterns of wall fluorescence are used to chart the movements of water from the xylem through some of the radial walls of mestome sheath cells near the xylem to the free space of the mesophyll. The suberised lamellae of the mestome sheath cells must form an incomplete barrier near the xylem to permit passage of the dye. A hypothesis is formulated that the function of the suberised lamellae is to keep separate the oppositely directed fluxes of water and assimilates through the sheath. It is further proposed that the function of pits in living cells is a similar insulation of the symplastic traffic from the wayward waters of the apoplast.     

Autolysis during in vitro tracheary element differentiation: formation and location of the perforation

Tracheary elements differentiated from isolated Zinnia: mesophyll cells were observed at various times of culture under a scanning electron microscope. Perforation occurred on the primary wall at one of the longitudinal ends in single tracheary elements. In double tracheary elements, which both of two cells derived from a single cell differentiated into, the pore opened on the primary walls both at the junction of the two tracheary elements and at a longitudinal end of one of the two tracheary elements. These results suggest not only that a single tracheary element has its own program to form a perforation at one end without being affected by neighboring cells, but also that isolated cells indeed hold some traces of polarity and cell-cell communication.     

Activity of cell-wall degradation associated with differentiation of isolated mesophyll cells of Zinnia elegans into tracheary elements

Cell walls were prepared from cultured mesophyll cells of Zinnia elegans L. that were transdifferentiating into tracheary elements and incubated in a buffer to undergo autolysis. The rate of autolysis of cell walls was determined by measuring the amount of carbohydrate released from the cell walls into the buffer during incubation. During the course of culture of mesophyll cells, the autolysis rate increased markedly at the time when thickenings of secondary cell walls characteristic of tracheary elements became visible (after 48-72 h of culture), and thereafter the rate remained at a high level. Comparative studies on the autolysis rate of cell walls using various control cultures, in which tracheary element differentiation did not take place, revealed a close relationship between the autolysis rate around the 60th hour of culture and differentiation. Sugar analysis by colorimetric assays and gas chromatography of carbohydrates released from the cell walls detected uronic acid, arabinose, galactose, glucose, xylose, rhamnose, fucose, and mannose. Among these sugars, uronic acid was the most abundant, and accounted for approximately half of the total released sugars. The decrease of acidic polysaccharides in the primary cell walls during tracheary element differentiation was visualized by staining cultured cells with alcian blue at pH 2.5. These results suggest that active degradation of components of primary cell walls, including pectin, is integrated into the program of tracheary element differentiation.     

Regulation of secondary cell wall development by cortical microtubules during tracheary element differentiation in Arabidopsis cell suspensions

Cortical microtubules participate in the deposition of patterned secondary walls in tracheary element differentiation. In this study, we established a system to induce the differentiation of tracheary elements using a transgenic Arabidopsis (Arabidopsis thaliana) cell suspension stably expressing a green fluorescent protein-tubulin fusion protein. Approximately 30% of the cells differentiated into tracheary elements 96 h after culture in auxin-free media containing 1 mum brassinolide. With this differentiation system, we have been able to time-sequentially elucidate microtubule arrangement during secondary wall thickening. The development of secondary walls could be followed in living cells by staining with fluorescein-conjugated wheat germ agglutinin, and the three-dimensional structures of the secondary walls could be simultaneously analyzed. A single microtubule bundle first appeared beneath the narrow secondary wall and then developed into two separate bundles locating along both sides of the developing secondary wall. Microtubule inhibitors affected secondary wall thickening, suggesting that the pair of microtubule bundles adjacent to the secondary wall played a crucial role in the regulation of secondary wall development.     

Study on the Convergent Passages of Water Conducting Elements in Secondary Xylem of Fagaceae

Structural variations in the convergent passages of water conducting elements were described in secondary xylem of 70 species of Fagaceae. Two types of water conducting systems were recognized on the ground of water-flow directions. The convergent passages of the longitudinal system mainly exhibited in the forms as perforations between the ends of Vessel elements, lateral wall pitting between the lateral walls of vessel elements, pitting be-tween vessel elements and imperforate tracheary elements, and pitting between imperforate tracheary elements. The convergent passages in the transverse system consisted of pitting between vessel elements and ray cells, pitting between imperforated tracheary elements and ray cells and simple pitting between ray cells. In addition, the roles of vasicentric tracheids and broad rays played in terms of conducting effectiveness and efficiency as proposed by Zimmermann were discussed.     

壳斗科次生木质部水分输导分子间连接通道结构的研究

壳斗科次生木质部水分运输聚合体及其连接通道结构,根据水分运输方向可分纵向和横向两大系统。水分纵向运输通道有导管分子间的穿孔,导管分子侧壁间的管间纹孔,导管分子与无穿孔管状分子间的侧壁纹孔,无穿孔管状分子与无穿孔管状分子间的侧壁纹孔。     

Further observations on hydrolysis of the cell wall in the xylem

Summary Hydrolyzed walls (birefringent, Periodic acid/Schiff negative, remnants of primary walls that also lack polyuronides with free carboxyl groups) are demonstrated in the primary xylem of wheat and bean leaves. Walls with similar properties have been found in the primary xylem of a variety of tissues from different species, and are believed to be ubiquitous. It is shown that the pit membrane of intervessel pits between tracheary elements of willow is also a hydrolyzed wall. Combined with the observation byLiese (1965) it seems likely that the removal of non-cellulosic polysaccharides from primary walls unprotected by lignin is a general phenomenon that occurs late in the autolysis of all tracheary elements. Parenchyma cells that abut autolyzing tracheary elements appear to react to hydrolytic attack in a number of ways that are illustrated and discussed.     

Vessel origins in Nymphaeaceae: Euryale and Victoria

Metaxylem tracheary elements of roots have differentiation between end walls and lateral walls in both Euryale and Victoria End walls have narrower, more closely spaced bars and scalariform plates. primary walls of end walls (and, to a lesser extent, lateral walls) have striations that are thickened primary wall portions orientated in an axial direction. These striations are less common in Victoria than in Euryale. Although secondary wall strands between perforations occur in some dicotyledons, the report of primary wall striations is new; these can be seen with scanning electron microscopy (SEM) but not with light microscopy. Perforations occur irregularly and sometimes sparsely on end walls of tracheary elements of Victoria , but perforations were not observed in Euryale. Thus, Euryale satisfies one criterion for the presence of vessel (end wall different from lateral wall), whereas Barclaya satisfies another (perforations in end walls) and Victoria satisfies both. Vessel origins in Nymphaeaceae are important in illustrating that there may be multiple vessel origins in dicotyledons.     

Fine Structure of Hydrolysed Primary Walls in Tracheary Elements of Petiolar Xylem in Eucalyptus delegatensis

The hydrolysed lateral primary walls of tracheary elements ofthe petiolar xylem of Eucalyptus delegatensis were examinedby electron microscopy. Vessel-vessel and vessel—tracheidhydrolysed walls were strikingly different in appearance fromtracheid—tracheid walls. The difference seemed to be inthe degree to which the primary walls were hydrolysed. The observationssuggest the wall hydrolysis to be an ordered and controlledprocess. Eucalyptus delegatensis, hydrolysed wall, petiolar xylem, tracheary elements     

Permeability of the suberized mestome sheath in winter rye

Mestome sheath cells of winter rye (Secale cereale L. cv Puma) deposit suberized lamellae in their secondary cell walls. Histochemical tests including acid digestion and staining with Sudan IV and Chelidonium majus root extract were used to detect the presence of suberin in the primary cell wall. There was no evidence of a Casparian band between adjacent mestome sheath cells. Fluorescent dye techniques were used to trace solute movement through the rye leaf apoplast. Calcofluor white M2R, a fluorescent dye which binds to cell walls as it moves apoplastically, proved to be too limited in its mobility in leaves to test mestome sheath permeability. Trisodium 3-hydroxy-5,8,10 pyrene trisulfonate, a fluorescent dye which is mobile in the apoplast, moved easily up the vascular bundles in the transpiration stream, and diffused outward from the veins to the epidermal cell walls within minutes of reaching a particular level in the leaf. We conclude that the suberized mestome sheath of rye leaves is freely permeable to solutes moving apoplastically through radial primary cell walls.     

Hydrolysed walls in the water-conducting cells of Dendroligotrichum (bryophyta): Histochemistry and ultrastructure

Summary Histochemical techniques and electron microscopy have been used to investigate the nature of the oblique primary end-walls of the water-conducting cells (hydroids) of Dendroligotrichum dendroides. (Hedw.) Broth. The observed properties (weakly birefringent; IKI-H₂SO₄-positive; periodic acid/Schiff negative; toluidine blue O-negative) support the conclusion that these end-walls are the cellulose residue of a primary wall that has been hydrolysed during autolysis of the hydroids. The walls are now referred to as hydrolysed end-walls. The unhydrolysed lateral-walls appear to be protected from hydrolytic attack by lignin or a lignin-like compound within those walls. The similarities between the hydrolysed end-walls of the hydroids and the hydrolysed walls of vascular plant tracheary elements are discussed.Contribution No. 109 from the Department of Biology, The Pennsylvania State University.     

LEAF VASCULATURE IN BARLEY,HORDEUM VULGARE (POACEAE)

The vascular system of the Hordeum vulgare L. leaf consists of multiple longitudinal strands interconnected by transverse bundles. In any transverse section, the longitudinal strands can be categorized into three bundle types: small, intermediate, and large. Individual longitudinal strands intergrade structurally from one bundle type into another as they descend the leaf. At their distal ends, they have the anatomy of a small bundle. As they descend the leaf, most intergrade into intermediate bundle and then into large bundle types. All strands with large bundle anatomy extend basipetally into the stem. Typically, the other longitudinal strands, which do not intergrade structurally into large bundles, do not enter the sheath, but fuse with other longitudinal strands above the junction of the blade with the sheath. Despite the decrease in number of longitudinal bundles entering the sheath, an increase takes place in the total crosssectional area of sieve tubes and tracheary elements. A linear relationship exists between leaf width and total bundle number in the blade but not in the sheath. Moreover, a linear relationship exists between cross-sectional area of vascular bundles and both total and mean cross-sectional area of tracheary elements and thin-walled sieve tubes.     

An Enzymic Method for Measuring the Molecular Weight Exclusion Limit of Plasmodesmata of Bundle Sheath Cells of C4 Plants

Bundle sheath cell strands have been prepared from four C₄ plantspecies and used to study the molecular weight exclusion limitof plasmodesmata located in the cell wall of bundle sheath cells.By measuring the activity and the inhibition of enzymes locatedwithin the bundle sheath cells of the strands in the absenceor presence of a variety of inhibitors of different molecularweight, the molecular weight exclusion limit of the plasmodesmatalocated within the cell walls of bundle sheath cells has beendetermined. Using a variety of Reactive dyes (of different molecularweight) which inhibit a number of cytosohc enzymes, as wellas a graded series of Reactive Yellow 2 derivatives as probes,it has been shown that compounds with molecular weights greaterthan about 900 daltons do not pass through the plasmodesmataof bundle sheath cells of C₄ plants. Key words: Plasmodesmata, molecular weight exclusion limit, bundle sheath cells     

In vitro induction of secondary xylem-like tracheary elements in calli of hybrid poplar (Populus sieboldii × P. grandidentata)

The formation of tracheary elements was induced in calli derived from petioles of hybrid poplar (Populus sieboldii × P. grandidentata) after 10 days of culture on medium that lacked auxin but contained 1 μM brassinolide. Some differentiated cells formed broad regions of cell walls and bordered pits, which are typical features of tracheary elements of secondary xylem. Other differentiated cells resembled tracheary elements of primary xylem, with spiral or reticulate thickening of cell walls. The tracheary elements that developed in calli were formed within cell clusters. This induction system provides a new model for studies of the mechanism of differentiation of secondary xylem cells in vitro.     

Ultrastructure of graniferous tracheary elements in the haustorium of Exocarpus bidwillii, a root hemi-parasite of the Santalaceae.

The xylem in the body of the haustorium of E. bidwillii has the shape of an inverted conical flask with the expanded portion being known as the vascular core. The tracheary elements of the vascular core are notable for the occurrence of numerous granules within their lumina and the presence of mostly imperforate walls. Elsewhere in the haustorium graniferous tracheary elements are absent and the cells are usually ordinary vessel elements. Thin sections for transmission electron microscopy, post-stained in potassium permanganate, show that the secondary wall thickenings of the graniferous tracheary elements consist of eccentric layers in which the microfibrils of each successive layer run alternately longitudinally and transversely. The granules of the tracheary elements average 2 micrometer in diameter and consist of a homogeneous matrix which shows a fine fibrillar structure on high resolution. The granules are naked and mostly remain as separate structures within the lumen of the cell, but occasionally they fuse into small groups or irregular masses. In some cells the granules become transformed into fibrillar material that disperses throughout the lumen. This dispersed material may accumulate in vessels of the interrupted zone proximal to the vascular core. Occasionally, the granules also change into compacted amorphous masses that adhere to the walls of the cell. Ultrastructural cytochemistry confirms that the granules are protein and not starch as was originally believed for the Santalaceae. The function of the vascular core and its graniferous tracheary elements is discussed and we suggest that it might help regulate the pressure and flow of xylem sap entering the parasite from the host. Graniferous tracheary elements in the Santalaceae and in root parasites of the Serophulariaceae are compared and it is concluded that they represent examples of convergent evolution.     

The Vascular System of Maize Stems Revisited: Implications for Water Transport and Xylem Safety

The plexus of vascular bundles in the nodes of grasses is notoriouslycomplex, where long axial bundles pass through a network oftransverse bundles. The xylem pathways for water in maize stemshave been investigated anatomically and with dye and particulatetracers, revealing some of the details of this complexity. Onlyapprox. 3% of axial vessels pass through nodes without beinginterrupted by end walls. Axial bundles at nodes differ fromthose in internodes in having the metaxylem and protoxylem vesselsconnected by small tracheary elements. So it is only at nodesthat exchange of sap occurs between the large vessels withina bundle. End walls, acting as filters for particles and gasbubbles, always separate axial vessels from vessels in transversebundles. The high redundancy of bundle connections in the nodalplexus is interpreted as providing alternative water pathwaysto bypass embolisms and damaged or diseased sections of thexylem. The pores in the filters at the base of nodes and betweenaxial and transverse vessels within nodes are <20 nm in diameter.Where axial vessels connect to transverse vessels, a varietyof unusual shapes of vessel elements mediate two- and three-wayconnections within the plexus.Copyright 2000 Annals of BotanyCompany Zea mays, cryoSEM, maize, node, pits, pit membranes, vessel ends, vessels, xylem embolism, xylem pathogens