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.     

Ultrastructure of transfer cells in spontaneous nodules of alfalfa (Medicago sativa)

Summary Spontaneous nodules were formed on the primary roots of alfalfa plants in the absence ofRhizobium. Histologically, these white single-to-multilobed structures showed nodule meristems, cortex, endodermis, central zone, and vascular strands. Nodules were devoid of bacteria and infection threads. Instead, the larger cells were completely filled with many starch grains while smaller cells had very few or none. Xylem parenchyma and phloem companion cells exhibited long, filiform and branched wall ingrowths. The characteristic features of both types of transfer cells were polarity of wall ingrowths, high cytoplasmic density, numerous mitochondria, abundant ribosomes, well-developed nucleus and nucleolus, and vesicles originated from rough endoplasmic reticulum. These results were compared with normal nodules induced byRhizobium. Our results suggest that xylem parenchyma and phloem companion transfer cells are active and probably involved in the short distance transport of solutes in and out of spontaneous nodules. Since younger nodules showed short, papillate, and unbranched wall ingrowths, and older tissue showed elongated, filiform and branched wall ingrowths, the development of wall ingrowths seemed to be gradual rather then abrupt. The occurrence of both type-A and -B wall ingrowths suggests that phloem companion transfer cells may be active in loading and unloading of sieve elements. Since there were no symbiotic bacteria and thus no fixed nitrogen, it is tempting to speculate that xylem parenchyma transfer cells may be re-transporting accumulated carbon from starch grains to the rest of the plant body by loading xylem vessels. Fusion of ER-originated vesicles with wall ingrowth membrane indicated the involvement of ER in the membrane formation for elongating wall ingrowths. Since transfer cells were a characteristic feature of both spontaneous andRhizobium-induced nodules, their occurrence and development is controlled by the genetic make-up of alfalfa plant and not by a physiological source or sink emanating from symbiotic bacteria.Abbreviations ATP adenosine triphosphate - ATPase adenosine triphosphatase - EH emergent root hair - EM electron microscope - Nar nodulation in the absence of Rhizobium - RT root tip - RER rough endoplasmic reticulum - YEMG yeast extract mannitol-gluconate     

Minor vein structure and sugar transport in Arabidopsis thaliana

Leaf and minor vein structure were studied in Arabidopsis thaliana (L.) Heynh. to gain insight into the mechanism(s) of phloem loading. Vein density (length of veins per unit leaf area) is extremely low. Almost all veins are intimately associated with the mesophyll and are probably involved in loading. In transverse sections of veins there are, on average, two companion cells for each sieve element. Phloem parenchyma cells appear to be specialized for delivery of photoassimilate from the bundle sheath to sieve element-companion cell complexes: they make numerous contacts with the bundle sheath and with companion cells and they have transfer cell wall ingrowths where they are in contact with sieve elements. Plasmodesmatal frequencies are high at interfaces involving phloem parenchyma cells. The plasmodesmata between phloem parenchyma cells and companion cells are structurally distinct in that there are several branches on the phloem parenchyma cell side of the wall and only one branch on the companion cell side. Most of the translocated sugar in A. thaliana is sucrose, but raffinose is also transported. Based on structural evidence, the most likely route of sucrose transport is from bundle sheath to phloem parenchyma cells through plasmodesmata, followed by efflux into the apoplasm across wall ingrowths and carrier-mediated uptake into the sieve element-companion cell complex. Received: 5 October 1999 / Accepted: 20 November 1999     

Zostera capensis setchell: Root structure in relation to function

Root structure of the seagrass Zostera capensis Setchell was investigated by light and electron microscopy. Roots possess conspicuous root hairs which greatly increase the surface area available for absorption. Exodermal cells abutting root-hair bases possess transfer cell characteristics. The strategic location of these cells suggests that they participate in absorptive and/or transfer processes between the epidermis and cortex. Vascular parenchyma cells within the stele also possess transfer cell features. Wall ingrowths of these cells about xylem elements, sieve tubes, companion cells and other vascular parenchyma cells, suggesting that they play a role in absorptive and/or transfer processes between the stele and cortex. Apoplastic barriers in the form of suberin lamellae and Casparian bands occur in walls of both the exodermis and endodermis. However, plasmodesmata perforate the suberin lamellae in these walls, and a symplastic pathway can be traced from the root hairs to vascular parenchyma transfer cells contiguous with conducting elements of the stele. The occurrence of wall ingrowths adjacent to xylem elements implies that transfer processes occur between vascular transfer cells and xylem. Although reduced, xylem could therefore play a role in transport. Structural evidence obtained in this study supports the role of the roots in absorptive processes and shows pathways available for transport from the water column to the conducting tissues of the root.     

Comparative Histocytology of the Petiole and the Main Pulvinus in Mimosa pudica L.

In Mimosa pudica, the main pulvinus, which brings about leafmovements, presents unusual structural characteristics in comparisonwith the petiole. Peculiar cellular features which exist inthe cortex, epidermis, parenchyma and endodermal regions includethe shape of the cells, their disposition and the location ofthe organelles. The central cylinder of the petiole is surrounded only by afew parenchyma layers whereas the central cylinder of the pulvinusforms a narrow central core enclosed in numerous cortical parenchymalayers. The phloem of the pulvinus contains collenchymatouscells towards the outside and possesses companion cells withwall ingrowths; these phloem members do not exist in the petiole.Xylem and protoxylem parenchyma cells of the petiole possesswall ingrowths which do not occur in homologous cells of thepulvinus. Moreover the pith of the pulvinus is composed of smallfibriform elements similar to the xylem fibriform elements ofthe organ. The structures observed may facilitate exchanges between cellsin the petiole and in the pulvinus. The predominant functionsof the organs relative to lateral and longitudinal transferof nutrients and conduction of stimuli are discussed. Mimosa pudica L., sensitive plant, pulvinus, ultrastructure, conduction of stimuli, leaf movement     

GIGANTEA is a component of a regulatory pathway determining wall ingrowth deposition in phloem parenchyma transfer cells of Arabidopsis thaliana

Transfer cells are specialised transport cells containing invaginated wall ingrowths that generate an amplified plasma membrane surface area with high densities of transporter proteins. They trans‐differentiate from differentiated cells at sites at which enhanced rates of nutrient transport occur across apo/symplasmic boundaries. Despite their physiological importance, little is known of the molecular mechanisms regulating construction of their intricate wall ingrowths. We investigated the genetic control of wall ingrowth formation in phloem parenchyma transfer cells of leaf minor veins in Arabidopsis thaliana. Wall ingrowth development in these cells is substantially enhanced upon exposing plants to high‐light or cold treatments. A hierarchical bioinformatic analysis of public microarray datasets derived from the leaves of plants subjected to these treatments identified GIGANTEA (GI) as one of 46 genes that are commonly up‐regulated twofold or more under both high‐light and cold conditions. Histological analysis of the GI mutants gi‐2 and gi‐3 showed that the amount of phloem parenchyma containing wall ingrowths was reduced 15‐fold compared with wild‐type. Discrete papillate wall ingrowths were formed in gi‐2 plants but failed to develop into branched networks. Wall ingrowth development in gi‐2 was not rescued by exposing these plants to high‐light or cold conditions. In contrast, over‐expression of GI in the gi‐2 background restored wall ingrowth deposition to wild‐type levels. These results indicate that GI regulates the ongoing development of wall ingrowth networks at a point downstream of inputs from environmental signals.     

PLASMODESMATAL FREQUENCY AND OTHER STRUCTURAL ASPECTS OF ASSIMILATE COLLECTION AND PHLOEM LOADING IN LEAVES OF SONCHUS OLERACEUS (ASTERACEAE), A SPECIES WITH MINOR VEIN TRANSFER CELLS

Leaves of Sonchus oleraceus (Asteraceae) were examined with the electron microscope to determine plasmodesmatal frequencies and other structural features relating to the collection of photoassimilate and its subsequent loading into minor veins. Few plasmodesmata occur between mesophyll cells, which contain chloroplasts that are sometimes connected to both the plasmalemma and the tonoplast by membranous tubules. The minor veins consist of tracheary elements, sieve-tube members, vascular parenchyma cells, and companion cells. The latter two cell types are transfer cells, with some of the fingerlike wall ingrowths in companion cells being traversed lengthwise by plasmodesmata. The frequencies of plasmodesmata at the mesophyllbundle sheath boundary and within are higher at some interfaces than at corresponding interfaces in nine other species, including some that previously had been characterized as loading assimilate via the symplast. It is thus premature to designate all species containing transfer cells in their minor veins as loading assimilate only via the apoplast.     

Transfer cells and nematode induced giant cells inHelianthemum

Summary The morphology of wall ingrowths in xylem and phloem transfer cells inHelianthemum is different. It is possible to use nematode infection to induce the formation of giant cells which abut both xylem and phloem elements to test whether ingrowth morphology is controlled by the solutes presumed to be transported across the plasmalemma of the cells. This experiment has been done and it is found that although wall ingrowths develop against both xylem and phloem, the giant cells exhibit only the ingrowth structure characteristic of xylem transfer cells.     

Internodes of grain legumes — New location for xylem parenchyma transfer cells

Summary Xylem parenchyma transfer cells were observed in the primary and secondary vascular tissue of stem internodes of 21 in 28 species of grain legumes. Their structural features were similar to those of other transfer cells. The relationships of these cells to transfer cells at nodes were investigated. Non-nodulated seedlings ofPhaseolus vulgaris L. formed internode transfer cells if provided mineral nutrients through their roots, but not if grown in distilled water or fed nutrients entirely through their leaves. Wall ingrowths formed in parenchyma of primary xylem ofPhaseolus just before full extension of an internode. The significance of this new location for transfer cells was discussed.     

Transfer cells and flange cells in sinkers of the mistletoePhoradendron macrophyllum (Viscaceae), and their novel combination

Summary The unusual thick-walled cells in contact with host and parasite vessels, first noted by Calvin 1967 in sinkers (structures composed of tracheary elements and parenchyma that originate from parasite bark strands that grow centripetally to the host vascular cambium and become embedded by successive development of xylem) of the mistletoePhoradendron macrophyllum (Englem.) Cockerell, have been investigated by modern methods of microscopy. The wall is thickest in cells abutting large-diameter host vessels, less so against smaller host vessels and those abutting sinker vessels. Transmission electron microscopy reveals the wall to be complex, consisting of a basement primary wall, upon which two developments of secondary-wall material occur. These are represented by lignified thickenings, in the form of flanges, and a labyrinth of wall ingrowths characteristic of a transfer cell. The wall ingrowths occur mostly in the primary-wall regions between the flanges, but when in contact with a large host vessel the ingrowths also differentiate on top of the flanges. Cells with such a transfer cell labyrinth have not been previously reported in the endophytic system of a mistletoe. The cells are confined to the xylary portion of the primary haustorium and sinkers. InP. macrophyllum, however, the cells differ from ordinary transfer cells in that they have differentiated as part of a flange parenchyma cell. This arrangement represents a novel anatomical situation. The name flange-walled transfer cell is used for these cells. The xylem of primary haustorium and sinkers also contain numerous ordinary flange cells. In both flange-walled transfer cells and ordinary flange cells the flanges are lignified and form a reticulate pattern of thickenings, separated by rounded areas of primary pit fields. The extent of development of the flange wall can vary in different parts of a sinker. At the host interface, the existence of a flange-walled transfer cell in direct contact with a vessel reflects a site associated with high loading into the parasite. Similarly, a labyrinth against a sinker vessel indicates a site of unloading from surrounding sinker tissue into the vessel for subsequent longdistance transport within the parasite.Dedicated to the memory of Dr. Katherine Esau (1898–1997)     

Effects of de- and rehydration on food-conducting cells in the moss Polytrichum formosum: a cytological study

BACKGROUND AND AIMS: Moss food-conducting cells (leptoids and specialized parenchyma cells) have a highly distinctive cytology characterized by a polarized cytoplasmic organization and longitudinal alignment of plastids, mitochondria, endoplasmic reticulum and vesicles along endoplasmic microtubules. Previous studies on the desiccation biology of mosses have focused almost exclusively on photosynthetic tissues; the effects of desiccation on food-conducting cells are unknown. Reported here is a cytological study of the effects of de- and rehydration on food-conducting cells in the desiccation-tolerant moss Polytrichum formosum aimed at exploring whether the remarkable subcellular organization of these cells is related to the ability of mosses to survive desiccation. METHODS: Shoots of Polytrichum formosum were dehydrated under natural conditions and prepared for transmission and scanning electron microscopy using both standard and anhydrous chemical fixation protocols. Replicate samples were then fixed at intervals over a 24-h period following rehydration in either water or in a 10 microM solution of the microtubule-disrupting drug oryzalin. KEY RESULTS: Desiccation causes dramatic changes; the endoplasmic microtubules disappear; the nucleus, mitochondria and plastids become rounded and the longitudinal alignment of the organelles is lost, though cytoplasmic polarity is in part retained. Prominent stacks of endoplasmic reticulum, typical of the hydrated condition, are replaced with membranous tubules arranged at right angles to the main cellular axis. The internal cytoplasm becomes filled with small vacuoles and the plasmalemma forms labyrinthine tubular extensions outlining newly deposited ingrowths of cell wall material. Whereas plasmodesmata in meristematic cells at the shoot apex and in stem parenchyma cells appear to be unaffected by dehydration, those in leptoids become plugged with electron-opaque material. Starch deposits in parenchyma cells adjoining leptoids are depleted in desiccated plants. Rehydration sees complete reestablishment over a 12- to 24-h period of the cytology seen in the control plants. Oryzalin effectively prevents leptoid recovery. CONCLUSIONS: The results point to a key role of the microtubular cytoskeleton in the rapid re-establishment of the elaborate cytoplasmic architecture of leptoids during rehydration. The reassembly of the endoplasmic microtubule system appears to dictate the time frame for the recovery process. The failure of leptoids to recover normal cytology in the presence of oryzalin further underlines the key role of the microtubules in the control of leptoid cytological organization.     

Structural and ultrastructural characteristics of the vascular apparatus of the sensitive plant (Mimosa pudica L.)

Summary 1. In motor organs ofMimosa pudica xylem contains living fibriform elements limited by a thick lignified highly pitted wall, whereas in other parts of the plant (stem, petiole, rachis), xylem and protoxylem vessels are closely associated with parenchyma cells which possess wall ingrowths. These ingrowths, at the apex of which the plasmalemma and the tonoplast touch, are localized like those of transfer cells of C type described byGunning andPate. Nevertheless, xylem parenchyma cells differ from cells of C type in several characteristics. Moreover, in motor organs, phloem contains cells characterized by wall ingrowths, less abundant on the parts adjacent to the sieve tubes; these cells which are localized near collenchyma cells of primary phloem, look like transfer cells of A type defined byGunning andPate; they are absent from internodes, petioles and rachides. 2. In motor organs, three types of vascular cells (companion cells, living xylem fibriform elements and protoxylem parenchyma cells) are characterized by reduced vacuolar volumes and well developed membrane systems, as compared with homologuous cells belonging to other parts of the plant. 3. A symplastic continuity holds from the middle of motor organs to their cortex: it is provided by the presence, in xylem and phloem respectively, of living fibriform elements and collenchyma cells bearing numerous pit fields containing large numbers of plasmodesmata. Several ultrastructural features suggest that the vascular apparatus ofMimosa pudica would be the site of intensive lateral transfer at different levels, specially in motor organs. Possible functions of certain structures observed are discussed in relation to some hypotheses relative to excitatory conduction pathways.     

Ultrastructure of the haustorial interface and apoplastic continuum between host and the root hemiparasiteOlax phyllanthi (Labill.) R. Br. (Olacaceae)

Summary Structural features of haustorial interface parenchyma of the root hemiparasiteOlax phyllanthi are described. Walls contacting host xylem are thickened non-uniformly with polysaccharides, not lignin, and show only a thin protective wall layer when abutting pits in walls of host xylem vessels or tracheids. Lateral walls of interface parenchyma exhibit an expanded middle layer of open fibrillar appearance, sometimes with, but mostly lacking adjoining layers of dense wall material. Free ribosomes and rough endoplasmic reticulum are prominent and occasional wall ingrowths present. Experiments involving transpirational feeding of the apoplast tracers lanthanum nitrate or uranyl acetate to host roots cut below haustorial connections, indicate effective apoplastic transfer from host to parasite root via the haustorium. Deposits of the tracers suggest a major pathway for water flow through host xylem pits, across the thin protective wall layer, and thence into the haustorium via the electronopaque regions of the terminal and lateral walls of the contact parenchyma. Graniferous tracheary elements and walls of parenchyma cells of the body of the haustorium appear to participate in tracer flow as do walls of cortical cells, stele parenchyma and xylem conducting elements of the parasite root, suggesting that both vascular and non-vascular routes are involved in extracytoplasmic transfer of xylem sap from host to parasite. The Casparian strip of the endodermis and the suberin lamella of the exodermis of theOlax root act as barriers to flow within the system.     

Polarized microtubule deposition coincides with wall ingrowth formation in transfer cells ofVicia faba L. cotyledons

Summary The epidermal transfer cells in developingVicia faba L. cotyledons are highly polarized. Extensive wall ingrowths occur on their outer periclinal walls and extend part way down both anticlinal walls. This ingrowth development serves to increase the surface area of the plasma membrane and thus maximize porter-dependent uptake of sugars from the seed apoplasm. In contrast, the inner periclinal walls of these transfer cells do not form wall ingrowths. We have commenced a study of the mechanisms responsible for establishing this polarity by first analysing the microtubule (MT) cytoskeleton in developing transfer cells. Thin sections of fixed cotyledons embedded in methacrylate resin were processed for immunofluorescence microscopy using monoclonal anti--tubulin and counterstained with Calcofluor White to visualize wall ingrowths. In epidermal cells of young cotyledons where wall ingrowths were yet to develop, MT labelling was detected around all cortical regions of the cell. However, in cells where wall ingrowths were clearly established, MT labelling was detected almost exclusively in cortical regions adjacent to the wall ingrowths. Little, if any, MT labelling was detected on the anticlinal or inner periclinal walls of these cells. This distribution of MTs was most prominent in cells with well developed wall ingrowths. In these cells, a subpopulation of MTs were also detected emanating from the subcortex and extending towards the wall ingrowth region. The possible role of MT distribution in establishing transfer cell polarity and wall ingrowth formation is discussed.Abbreviations MT microtubule     

Die submikroskopische Struktur der Assimilatleitbahnen von Polytrichum commune

Summary In the young part of the stem of Polytrichum commune the protoplasts of the two types of conducting cells, the leptoids and parenchyma cells, are nearly identically equipped with cell organelles and cytoplasmic structures. Both types contain a nucleus, chloroplasts, mitochondria, and dictyosomes. The endoplasmic reticulum builds characteristic cisterns in form of hollow cylinders extending from one end wall to the other. The cisterns are connected with many plasmodesmata, which occur only in the end walls. Leptoids have oblique end walls with 16 to 20 plasmodesmata per m², and parenchyma cells show cross walls perpendicular to the axis with 9 to 12 plasmodesmata per m².Since the leptoids are supposed to be the pathways for the longitudinal transport of assimilates (Eschrich and Steiner, 1967, 1968), it is of interest that early in their development these elements undergo a change in their protoplasmatic structure. Two to 3 cm below the apical cell the protoplasts degenerate and show lysosome-like structures. The endoplasmic reticulum and other structures are deformed or dissolved; the plasmodesmata are constricted by callose deposits. At the same level the parenchyma cells still retain the original structure of their protoplasts.Thus, assimilates moving upward in one row of leptoids may penetrate the whole lumen of the leptoids at lower levels, but they are restricted to the cisterns of the endoplasmic reticulum at higher levels of the stem.     

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.     

Vascular transfer cells in Angiosperm leaves A taxonomic and morphological survey

Summary A study of the fine structure of minor veins of mature leaves of 975 species and 242 families of Angiosperms shows that transfer cells are widespread amongst herbaceous Dicotyledons, are much rarer in woody Dicotyledons, and are virtually absent from the Monocotyledons. The evolutionary significance of the distribution of the cells amongst and within orders, families and minor groupings is discussed.Four types of transfer cell are recognized in minor veins, all possessing irregular ingrowths of wall material protruding into their protoplasts, and all being regarded as modified parenchyma of the minor vein. Two types occur in phloem. One (the A-cell), with ingrowths distributed right round its periphery, is associated specifically with the sieve elements. The other (the B-cell) occurs more generally throughout the phloem and has zones of wall ingrowths oriented towards sieve elements and their associated companion cells or A-cells. Two other types (C- and D-cells) occur in xylem parenchyma and bundle sheath respectively, and have ingrowths only on walls in contact with or in close proximity to vessels or tracheids. Each species has a characteristic combination of types of transfer cell. The variations encountered in the survey are classified. Consistent differences in the frequency and form of ingrowths are to be found between the different types of transfer cell of a single species, and between different species in respect to a particular type of transfer cell.The functional significance of transfer cells in minor veins is discussed in relation to the loading and unloading of the conducting elements and to the retrieval of extra-cytoplasmic solutes from the mesophyll and the transpiration stream.     

Role of light and jasmonic acid signaling in regulating foliar phloem cell wall ingrowth development

Phloem cells adjacent to sieve elements can possess wall invaginations. The role of light and jasmonic acid signaling in wall ingrowth development was examined in pea companion cells (CCs), Arabidopsis thaliana phloem parenchyma cells (PCs), and in Senecio vulgaris (with ingrowths in both cell types). Features characterized included wall ingrowths (from electron microscopic images), foliar vein density and photosynthetic capacity. In Arabidopsis, wall ingrowths were bulky compared with finger-like invaginations in pea and S. vulgaris. Relative to low light (LL), wall invagination in both CCs and PCs was greater in high light (HL). Treatment with methyl jasmonate in LL had no effect on CCs, but increased PC wall ingrowths. LL-to-HL transfer resulted in significantly less wall ingrowth in the fad7-1 fad8-1 (jasmonate-deficient) Arabidopsis mutant relative to the wild type. These results suggest that chloroplast oxidative status, via chloroplast-derived jasmonates, may modulate phloem structure and function. While CC wall ingrowths facilitate phloem loading by expanding the membrane area available for active uptake, one can speculate that phloem PC ingrowths may have two potential roles: to increase the efflux of sugars and/or protons into the apoplast to augment phloem loading; and/or to protect the phloem against pathogens and/or insects.     

Zostera capensis setchell: III. Some aspects of wall ingrowth development in leaf blade epidermal cells

Summary The development of wall ingrowths in leaf blade epidermal cells of the marine angiospermZostera capensis was studied by electron microscopy. Prior to the appearance of ingrowths long profiles of endoplasmic reticulum cisternae become arranged peripherally closely following the contours of the walls. The plasmalemma assumes a wavy appearance and in regions where wall ingrowths first start forming (i.e., along the radial, inner tangential and transverse walls) the plasmalemma becomes separated from the walls by an undulating extracytoplasmic space. Small, irregular projections of secondary wall material make their appearance here. Paramural bodies, dictyosomes, endoplasmic reticulum (ER) and possibly also microtubules seem to be closely associated with the initiation and subsequent development of wall projections. As the cells mature, new ingrowths arise in a centrifugal direction along the radial and transverse walls. When wall ingrowths reach a certain stage of their development, mitochondria become strongly polarized towards them and become closely associated with the plasmalemma which ensheaths the ingrowths. There is often also a close association between ER cisternae and the involuted plasmalemma of the wall projections. Initially ingrowths are slender, curved structures, but become more complex as the cells mature. Ingrowths are most extensively developed along the inner tangential and transverse walls. As epidermal cells age there is a loss of wall material from the ingrowths. The probable significance of the formation of wall ingrowths in the epidermal cells is also discussed.