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.     

Lipid polymers accumulate in the epidermis and mestome sheath cell walls during low temperature development of winter rye leaves

Summary Winter rye (Secale cereale L cv. Puma) was grown at 20 °C and at 5 °C and the development of epidermal and mestome sheath cells of leaves from plants grown at both temperatures was compared by electron microscopy. At 5 °C, the cells became densely packed with cytoplasm and small vacuoles after 41 days of growth. By day 56 at 5 °C, epidermal and mestome sheath cells were small in diameter and multivacuolate with asymmetrically thickened walls. By day 76 at 5 °C, a new developmental stage had been reached in epidermal and mestome sheath cells. The cells were larger in diameter although the thickened cell walls and multivacuolate cytoplasm were still present. As epidermal and mestome sheath cell walls thickened during low temperature growth of winter rye, an increase in cuticle thickness and the deposition of a lamellar layer could be observed in epidermal and mestome sheath cells, respectively. The lipid-derived polymers from the leaves of rye plants grown at 20 °C were shown by reductive depolymerization and GC-MS analysis to be comprised of 18-hydroxy-9, 10-epoxyoctadecanoic acid (47%) and dihydroxyhexa-decanoic acid (29%). The leaves of plants grown at 5 °C had two to four times as much lipid-derived polymeric material as those grown at 20 °C and the proportion of the major monomer, 18-hydroxy-9,10-epoxyoctadecanoic acid, increased to 73% of the polymeric material. Physical isolation of both epidermal tissue and vascular bundles followed by GC-MS analysis of the monomeric components released by reduction of the respective lipid polymers showed that 18-hydroxy-9,10 epoxyoctadecanoic acid was the major monomer in the polymer of both the epidermis and the mestome sheaths. The presence of this epoxide monomer in both the cuticles and mestome sheath cell walls of rye leaves was confirmed and visualized by using an epoxide-specific staining reaction.     

Development of mestome sheath cells in leaves ofAegilops comosa var.thessalica

Summary The development of mestome sheath cells ofAegilops comosa var.thessalica was studied by electron microscopy. Anatomical and cytological observations show that this grass belongs to the C₃ or non-Kranz plants. In the asymmetrically thickened walls of mestome sheath cells a suberized lamella is present. This lamella is deposited asynchronously. In the midrib and the large lateral bundles it appears first in the outer and inner walls and usually later in the radial walls. In the small lateral bundles its appearance is delayed in the inner walls of those cells situated on the xylem side. At maturity the suberized lamella is observed in all cell walls; however, in the small lateral bundles it is partly or totally absent from the walls of some cells situated on the xylem side. Tertiary wall formation is asynchronous as well, for it generally follows the deposition pattern of the suberized lamella.During the development of the mestome sheath cells microtubules show marked changes in their number and orientation, being fewer and longitudinal during suberin deposition. Dictyosomes are very active and may be involved in primary and tertiary wall formation. Endoplasmic reticulum cisternae are abundant and partly smooth, while plasmalemmasomes may function to reduce the plasmalemma extension. However, cytoplasmic structures that are clearly involved in suberin synthesis could not be identified.Suberized lamellae react strongly with silver hexamine. This is probably due to post-fixation with osmium tetroxide.On the basis of structural characteristics the mestome sheath may be regarded as an endodermis (cf., alsoFahn 1974). The significance of this view for water and assimilate exchange between the mesophyll and the bundle is discussed.This report represents a portion of a doctoral dissertation.     

Symplastic Transfer of Fluorescent Dyes from Mesophyll to Sieve Tube in Stripped Leaf Tissue and Partly Isolated Minor Veins of Commelina benghalensis

We have stripped small (3 × 3 mm) fields of the upper and the opposite lower epidermis of Commelina benghalensis leaves. Pectinase treatment of the resulting chlorenchyma windows produced free-lying viable minor veins with small lumps of mesophyll cells attached. These veins were still connected with the intact remainder of the leaf. Fluorescent dyes were injected into mesophyll cells or mestome sheath cells. Continuous following of the dye from the moment of injection and use of the simple vein system allowed an unhindered and precise assessment of the cell-to-cell route of dye transfer. Disodium fluorescein and Lucifer Yellow CH injected into mesophyll or mestome sheath cells readily moved to the sieve tube. This symplastic dye transfer from mesophyll to sieve tube was also observed after injection into unmacerated stripped leaf tissue. The displacement of fluorescent dyes substantiates a symplastic continuity between mesophyll and sieve tube and therefore supports the possibility of symplastic phloem loading.     

Ultrastructural aspects of leaves inFestuca ovina andPoa sphondylodes (C-3 Poaceae)

Structural aspects of the leaves of two common festucoids,Festuca ovina andPoa sphondylodes, have been examined employing the electron microscopy. The nature of vascular bundles and of sheaths that surround vascular tissues was discussed in the study. The festucoids exhibited a non-Kranz C-3 anatomy with more than four mesophyll cells separating the bundle sheaths of a leaf blade. Vascular tissues in theseFestuca andPoa leaves were surrounded by a double sheath: an inner distinct mestome sheath (MST) and an outer indistinctive layer of parenchymatous bundle sheath (PBS) cells. The PBS cells were much larger than the MST and had thin walls. The MST cells were relatively small and rectangular inP. sphondylodes and more or less hexangular in transverse sections ofF. ovina. InP. sphondylodes, MST had conspicuously thickened inner tangential walls with asymmetrically uninterrupted suberized lamellae in radial and tangential walls. In most differentiated MST cells, all walls were highly suberized. During suberin deposition, MST cells were quite vacuolated and most of the cytoplasm was present as a thin peripheral layer. However, MST walls inF. ovina revealed very thin suberized lamellae with translucent striations. No chloroplasts were detected inP. sphondylodes, whereas the MST inF. ovina contained small chloroplasts. Plasmodesmata were well developed in the primary pit fields of walls between MST and vascular cells, and between adjacent MST cells. Plasmodesmata were less frequent in the walls between the inner and outer sheath cells. Suberized lamellae were totally absent from the PBS cell walls in all veins. External to the PBS, the mesophyll comprised thin walled cells with abundant intercellular spaces. Peripherally arranged chloroplasts in the mesophyll were numerous and often larger than those of PBS and MST cells. Characteristics associated with C-3 and other ultrastructural features were also discussed in the study.     

Antifreeze proteins in winter rye

Six antifreeze proteins, which have the unique ability to adsorb onto the surface of ice and inhibit its growth, have been isolated from the apoplast of winter rye leaves where ice forms at subzero temperatures. The rye antifreeze proteins accumulate during cold acclimation and are similar to plant pathogenesis-related proteins, including two endoglucanase-like, two chitinase-like and two thaumatin-like proteins. Immunolocalization of the glucanase-like antifreeze proteins showed that they accumulate in mesophyll cell walls facing intercellular spaces, in pectinaceous regions between adjoining mestome sheath cells, in the secondary cell walls of xylem vessels and in epidermal cell walls. Because the rye antifreeze proteins are located in areas where they could be in contact with ice, they may function as a barrier to the propagation of ice or to inhibit the recrystallization of ice. Antifreeze proteins similar to pathogenesis-related proteins were also found to accumulate in closely-related plants within the Triticum group but not in freezing-tolerant dicotyledonous plants. In winter wheat, the accumulation of antifreeze proteins and the development of freezing tolerance are regulated by chromosome 5. Rye antifreeze proteins may have evolved from pathogenesis-related proteins, but they retain their catalytic activities and may play a dual role in increasing both freezing and disease resistance in overwintering plants.     

Leaf ultrastructure of C4 species possessing different Kranz anatomical types in the cyperaceae

The leaf ultrastructure of NADP-malic enzyme type C₄ species possessing different anatomical features in the Cyperaceae was examined: types were the Rhynchosporoid type, a normal Kranz type in which mesophyll cells are adjacent to Kranz cells, and Fimbristyloid and Chlorocyperoid types, unusual Kranz types in which nonchlorophyllous mestome sheath intervenes between the two types of green cells. They show structural characteristics basically similar to the NADP-malic enzyme group of C₄ grasses, that is, centrifugally located chloroplasts with reduced grana and no increase of mitochondrial frequency in the Kranz cells. However, the Kranz cell chloroplasts of the Fimbristyloid and Chlorocyperoid types exhibit convoluted thylakoid systems and a trend of extensive development of peripheral reticulum, although those of the Rhynchosporoid type do not possess such particular membrane systems. The suberized lamella, probably a barrier for CO₂ diffusion, is present in the Kranz cell walls of the Rhynchosporoid type and in the mestome sheath cell walls of the other two types, and tightly surrounds the Kranz cells (sheaths) that are the sites of the decarboxylation of C₄ acids. These ultrastructural features are discussed in relation to C₄ photosynthetic function.     

Ultrastructure of leaves in C4 Cyperus iria and C3 Carex siderosticta

The ultrastructural aspects ofCyperus iria leaves showing the C₄ syndrome and the typical C₃ species,Carex siderosticta, in the Cyperaceae family were examined.C. iria exhibited the chlorocyperoid type, showing an unusual Kranz structure with vascular bundles completely surrounded by two bundle sheaths. The cellular components of the inner Kranz bundle sheath cells were similar to those found in the NADP-ME C₄ subtype, having centrifugally arranged chloroplasts with greatly reduced grana and numerous starch grains. Their chloroplasts contained convoluted thyla-koids and a weakly-developed peripheral reticulum, although it was extensive mostly in mesophyll cell chloroplasts. The outer mestome bundle sheath layer was sclerenchymatous and generally devoid of organelles, but had unevenly thickened walls. Suberized lamellae were present on its cell walls, and they became polylamellate when traversed by plasmodesmata. Mesophyll cell chloroplasts showed well-stacked grana with small starch grains. InC. siderosticta, vascular bundles were surrounded by the inner mestome sheath and the outer parenchymatous bundle sheath with intercellular spaces. The mestome sheath cells degraded in their early development and remained in a collapsed state, although the suberized lamellae retained polylamellate features. Plastids with a crystalline structure, sometimes membrane-bounded, were found in the epidermal cells. The close interveinal distance was 35–50 μm inC. iria, whereas it was 157–218 μm inC. siderosticta. These ultrastructural characteristics were discussed in relation to their photosynthetic functions.     

Suberized bundle sheaths in grasses (Poaceae) of different photosynthetic types I. Anatomy,ultrastructure and histochemistry

Summary We investigated the histochemistry and ultrastructure of the cell walls of mestome sheaths and parenchymatous bundle sheaths of ten species of grasses. The species surveyed included representatives from all the major photosynthetic types: C₃-Bromus tectorum, Phalaris arundinacea; C₄/NAD-ME-Eragrostis cilianensis, Panicum capillare; C₄/NAD-ME/PCK-Bouteloua curtipendula; C₄/PCK-Chloris gayana, Sporobolus elongatus; C₄/NADP-ME-Echinochloa crus-galli, Setaria glauca, Themeda triandra. All vein orders (designated here as major, minor and transverse) from mature leaves of each species were tested histochemically for lipids and phenols, and the majority of species were also examined with the electron microscope. A suberized lamella was detected ultrastructurally in at least some walls of major vein bundle sheath cells of all species examined. These lamellae were also present in some cells associated with the minor veins of the C₃ species and in the minor and transverse veins of the C₄/NADP-ME species. Histochemical tests for lipids and phenols consistently failed to differentiate this layer. Based on these tests, none of the vein orders in any species showed evidence of a Casparian band. In all suberized bundle sheaths, the compound middle lamella between cells with suberin lamellae is modified by the presence of phenols. These did not, however, confer resistance to acid digestion to the cell layer, in contrast to cell layers with Casparian bands. Therefore, although the mestome sheath has some features in common with the root endodermis (i.e. cells with a suberized lamella and thick, cellulosic walls which may be further modified), we could find no substantive anatomical or ultrastructural evidence for the presence of a Casparian band in any of the grass leaves investigated. The significance of these observations is discussed in the context of apoplastic permeability of these walls.     

Movement of Lucifer Yellow CH in potato tuber storage tissues: A comparison of symplastic and apoplastic transport

The fluorescent dye Lucifer Yellow CH (LYCH) was introduced directly into the symplast of potato (Solanum tuberosum L.) tuber storage parenchyma by microinjection and also into the apoplast through cuts made in the stolon cortex. Microinjected LYCH moved away rapidly from a single storage cell and spread radially via the symplast. When the microinjected tissue was subsequently fixed in glutaraldehyde and sectioned the dye was seen clearly to be localised in the cytoplasm but not in the vacuole. In comparison, when LYCH was introduced into cuts made in the stolon cortex the dye entered the tuber by the xylem and subsequently spread apoplastically. No movement of dye was observed in the phloem. In glutaraldehyde-fixed tissues, in which LYCH was introduced to the apoplast, the dye was found within xylem vessels, in the cell walls and in intercellular spaces. Wall regions, possibly associated with plasmodesmata, became stained by the dye as it moved through the apoplast. Three hours after introduction of the dye to the stolon, intense deposits of LYCH were found in the vacuoles of all cells in the tuber, many aligned along the tonoplast. Differentiating vascular parenchyma elements contained large amounts of dye within enlarging vacuoles. However, with the exception of plasmolysed and-or damaged cells, LYCH was absent from the cytoplasm following its introduction to the plasmalemma it is suggested that the most likely pathway from the cell wall to the vacuole was by endocytosis, the dye being transported across the cytoplasm in membrane-bound vesicles. Clathrin-coated vesicles were abundant in the storage cells, providing a possible endocytotic pathway for dye movement. The significance of these observations is discussed in relation to the movement of LYCH in plant tissues and to the movement of solutes within and between storage cells of the tuber.Abbreviation LYCH Lucifer Yellow CH     

Water Pathways in Higher Plants: III. THE TRANSPIRATION STREAM WITHIN LEAVES

Water in the transpiration stream is distributed throughoutthe leaves in the vascular bundles. In wheat, water appearsto be confined to the main veins by the mestome sheath and toenter the mesophyll through the walls of the smaller veins.Within the mesophyll the water in the transpiration stream movesin the free space of the cell walls to the evaporating surfacesof the leaf. The lead chelate, which is used to trace the transpirationstream, accumulates at the final points of evaporation at themargin of the leaf. Lead chelate accumulates beneath and onthe surface of the cuticle, being partly associated with theanticlinal walls of the epidermal cells, the walls of the stomatalguard cells and specialized epidermal cells. Chelate does notaccumulate at the base of substomatal cavities, indicating thatthe cuticle of the epidermis is the main evaporating surfaceof the leaf. The behaviour in broad bean, laurel, and plantainis essentially the same. The rate of peristomatal and cuticulartranspiration is closely related to the size of the stomatalaperture. Conditions which control stomatal aperture also causechanges in the dimensions of the epidermal cells.     

Distribution and frequency of plasmodesmata in relation to photoassimilate pathways and phloem loading in the barley leaf

Large, intermediate, and small bundles and contiguous tissues of the leaf blade of Hordeum tvulgare L. ‘Morex’ were examined with the transmission electron microscope to determine their cellular composition and the distribution and frequency of the plasmodesmata between the various cell combinations. Plasmodesmata are abundant at the mesophyll/parenchymatous bundle sheath, parenchymatous bundle sheath/mestome sheath, and mestome sheath/vascular parenchyma cell interfaces. Within the bundles, plasmodesmata are also abundant between vascular parenchyma cells, which occupy most of the interface between the sieve tube-companion cell complexes and the mestome sheath. Other vascular parenchyma cells commonly separate the thick-walled sieve tubes from the sieve tube-companion cell complexes. Plasmodesmatal frequencies between all remaining cell combinations of the vascular tissues are very low, even between the thin-walled sieve tubes and their associated companion cells. Both the sieve tube-companion cell complexes and the thick-walled sieve tubes, which lack companion cells, are virtually isolated symplastically from the rest of the leaf. Data on plamodesmatal frequency between protophloem sieve tubes and other cell types in intermediate and large bundles indicate that they (and their associated companion cells, when present) are also isolated symplastically from the rest of the leaf. Collectively, these data indicate that both phloem loading and unloading in the barley leaf involve apoplastic mechanisms.     

FITC-dextran for measuring apoplast pH and apoplastic pH gradients between various cell types in sunflower leaves

The liquid in the free space of leaf cell walls, the apoplast, is in direct contact with the plasma membrane and its nutrient uptake systems. Therefore, the pH of the apoplast is of utmost interest. We have elaborated a non-destructive method by which excised sunflower leaves ( Helianthus annuus cv. Erika) were perfused with fluorescein isothiocyanate-dextran (FITC-dextran) (4 000 Da) via the transpiration stream. We showed that leaf apoplast pH can be measured by using the fluorescence ratio technique together in conjunction with this dye. Evidence is provided that FITC-dextran does not penetrate the plasma membrane over a period of ca 17 h from the beginning of dye perfusion. Dye enrichment in the leaf apoplast did not cause an 'inner filter effect' and thus the fluorescence ratio was only dependent on pH. In vivo calibration yielded a pKa of 5.92, which was virtually identical to the pKa of 5.93 calculated for dye solutions. Hence, FITC-dextran can be detected in complex environments and covers a pH range prevailing in the leaf apoplast.
Based on this method we developed a microscope image technique visualizing pH gradients between various cell types. The pH in the lumen of the xylem vessel was ca 0.3–0.5 units lower than that of the apoplast of surrounding cells. Nitrate present in the leaf apoplast caused an increase in pH, especially in the dark. Under these conditions, in the intercostal area, the apoplast pH around the stomata was ca 0.5–1.0 units higher than that of the surrounding epidermal cells.     

The leaf internal morphology and ultrastructure of Zostera muelleri irmisch ex aschers. (Zosteraceae): a comparative study of the intertidal and subtidal forms

The leaf anatomy, histochemistry and ultrastructure of the intertidal and subtidal seagrass Zostera muelleri Irmish ex Aschers. from Westernport Bay, Victoria were studied. Unusual anatomical and ultrastructural features are compared with other seagrasses and their functional significance is assessed. Subcuticular cavities are present in the young blade, but not observed in the older blade nor young and old leaf sheath. Wall ingrowths occur in the blade epidermal cells particularly on the inner tangential walls and the lower portions of the radial walls. Plasmodesmata are present between adjacent epidermal cells and between the epidermal and mesophyll cells, suggesting that solutes could transfer between these tissues both symplastically and apoplastically. Each leaf has three longitudinally aligned vascular bundles, each of which comprises a single xylem element isolated from the phloem tissue. The phloem consists of nacreous-walled sieve elements accompanied by phloem parenchyma cells which also process wall ingrowths. The xylem walls are completely hydrolysed and the middle lamella borders directly on the xylem lumen. Leaves have prominent air lacunae bisected transversely by septa at regular intervals along their length. Each septum consists of a file of small parenchyma cells with wall protuberances projecting into intercellular space. There are no major structural differences between the subtidal and intertidal plants, but the former have larger leaves and more leaves per shoot than the latter. In addition, a network of unusual reticulated fungal hyphae is present in the leaf intercellular spaces of the subtidal form and this network may facilitate solute transfer in these plants.     

Suberized bundle sheaths in grasses (Poaceae) of different photosynthetic types. II. Apoplastic permeability

Summary The relative hydraulic conductivities of major and minor longitudinal veins, and the apoplastic permeability of the bundle sheaths surrounding all longitudinal and transverse veins were investigated in representatives of the C₃, C₄/NAD-ME, C₄/NAD-ME/PCK intermediate, C₄/PCK and C₄/NADP-ME photosynthetic types. Using the Hagen-Poiseuille equation and measurements of tracheary element diameters, the number of elements in each vein type and the numbers of each vein type, we calculated that 87–99% of the water flow in a longitudinal direction would be expected to occur in the major veins. The permeability of the mestome sheaths and parenchymatous bundle sheaths surrounding the veins was tested using the negatively-charged, fluorescent dye, trisodium 3-hydroxy-5,8,10-pyrenetrisulfonate (PTS). This dye proved nontoxic to plant tissue at a concentration of 0.5%, according to a deplasmolysis test with onion epidermal strips. The PTS concentration achieved in the tested grass leaves was about 0.035%, well below the toxic limit. When a solution of PTS was fed to the leaves by means of a basal cut, the dye moved into the veins of all orders. From there, it moved outward into the surrounding tissues, indicating that the sheaths surrounding the veins of all orders in all species tested were permeable. Therefore, contrary to previous predictions based on structural observations and some tracer studies, bundle sheaths with suberized cell walls do not function as endodermal layers.     

Ultrastructure of and plasmodesmatal frequency in mature leaves of sugarcane

Vascular bundles and contiguous tissues of leaf blades of sugarcane (Saccharum interspecific hybrid L62–96) were examined with light and transmission electron microscopes to determine their cellular composition and the frequency of plasmodesmata between the various cell combinations. The large vascular bundles typically are surrounded by two bundle sheaths, an outer chlorenchymatous bundle sheath and an inner mestome sheath. In addition to a chlorenchymatous bundle sheath, a partial mestome sheath borders the phloem of the intermediate vascular bundles, and at least some mestome-sheath cells border the phloem of the small vascular bundles. Both the walls of the chlorenchymatous bundlesheath cells and of the mestome-sheath cells possess suberin lamellae. The phloem of all small and intermediate vascular bundles contains both thick- and thin-walled sieve tubes. Only the thin-walled sieve tubes have companion cells, with which they are united symplastically by pore-plasmodesmata connections. Plasmodesmata are abundant at the Kranz mesophyll-cell-bundlesheath-cell interface associated with all sized bundles. Plasmodesmata are also abundant at the bundle-sheathcell-vascular-parenchyma-cell, vascular-parenchyma-cellvascular-parenchyma-cell, and mestome-sheath-cell-vascular-parenchyma-cell interfaces in small and intermediate bundles. The thin-walled sieve tubes and companion cells of the large vascular bundles are symplastically isolated from all other cell types of the leaf. The same condition is essentially present in the sieve-tube-companion-cell complexes of the small and intermediate vascular bundles. Although few plasmodesmata connect either the thin-walled sieve tubes or their companion cells to the mestome sheath of small and intermediate bundles, plasmodesmata are somewhat more numerous between the companion cells and vascular-parenchyma cells. The thick-walled sieve tubes are united with vascular-parenchyma cells by pore-plasmodesmata connections. The vascular-parenchyma cells, in turn, have numerous plasmodesmatal connections with the bundle-sheath cells.This study was supported by National Science Foundation grants DCB 87-01116 and DCB 90-01759 to R.F.E. and a University of Wisconsin-Madison Dean's Fellowship to K. R.-B. We also thank Claudia Lipke and Kandis Elliot for photographic and artistic assistance, respectively.     

Subcellular localization of silicon and germanium in grass root and leaf tissues by SIMS: evidence for differential and active transport

Silicon transport and incorporation into plant tissue is important to both plant physiological function and to the influence plants have on ecosystem silica cycling. However, the mechanisms controlling this transport have only begun to be explored. In this study, we used secondary ion mass spectrometry (SIMS) to image concentrations of Si in root and shoot tissues of annual blue grass (Poa annua L.) and orchard grass (Dactylis glomerata L.) with the goal of identifying control points in the plant silica uptake pathway. In addition, we used SIMS to describe the distributions of germanium (Ge); the element used to trace Si in biogeochemical studies. Within root tissue, Si and Ge were localized in the suberized thick-walled region of endodermal cells, i.e. the proximal side of endodermal cells which is in close association to the casparian strip. In leaves, Si was present in the cell walls, but Ge was barely detectable. The selective localization of Si and Ge in the proximal side of endodermal cell walls of roots suggests transport control is exerted upon Si and Ge by the plant. The absence of Si in most root cell walls and its presence in the cell walls of leaves (in areas outside of the transpiration terminus) suggests modifications in the chemical form of Si to a form that favors Si complexation in the cell walls of leaf tissue. The low abundance of Ge in leaf tissue is consistent with previous studies that suggest preferential transport of Si relative to Ge.     

Distribution and structure of the plasmodesmata in mesophyll and bundle-sheath cells of Zea mays L.

In leaf blades of Zea mays L. plasmodesmata between mesophyll cells are aggregated in numerous thickened portions of the walls. The plasmodesmata are unbranched and all are characterized by the presence of electron-dense structures, called sphincters by us, near both ends of the plasmodesmatal canal. The sphincters surround the desmotubule and occlude the cytoplasmic annulus where they occur. Plasmodesmata between mesophyll and bundle-sheath cells are aggregated in primary pit-fields and are constricted by a wide suberin lamella on the sheath-cell side of the wall. Each plasmodesma contains a sphincter on the mesophyll-cell side of the wall. The outer tangential and radial walls of the sheath cells exhibit a continuous suberin lamella. However, on the inner tangential wall only the sites of plasmodesmatal aggregates are consistently suberized. Apparently the movement of photosynthetic intermediates between mesophyll and sheath cells is restricted largely or entirely to the plasmodesmata (symplastic pathway) and transpirational water movement to the cell walls (apoplastic pathway).Abbreviation ER endoplasmic reticulum     

VARIATIONS IN ANATOMY,ASSOCIATIONS, AND ORIGINS OF KRANZ TISSUE

Although the unique tissue required for C₄ photosynthesis in nonsucculent plants is often described as being modified leaf parenchyma sheath, which is positioned meaningfully between the mesophyll externally and the vascular tissues internally, the actual range of locations and known associations make that concept untenable. In origin the Kranz tissue develops from procambium as well as ground parenchyma. It is found in stems as well as leaves. In position Kranz tissue can lie in the parenchyma sheath, in the mestome sheath, isolated in the mesophyll, peripherally in some thick leaves, or within the veins. It can be associated with mesophyll only, mesophyll and colorless parenchyma, mesophyll and sclerenchyma, other Kranz tissue and vascular tissues, mesophyll and mestome sheath, mesophyll and phloem, mesophyll and xylem, epidermis, and, finally, mestome sheath and xylem and phloem. The use of the term Kranz is expounded.