ULTRASTRUCTURAL EVIDENCE FOR SECRETORY PHASES IN THE LIFE HISTORY OF STIGMATIC HAIRS OF COTTON: A DRY TYPE STIGMA

Stigmatic hairs of the cotton flower were studied through their developmental stages up to anthesis. Stigmatic hairs invariably develop from a densely straining band of epidermal cells opposite the transmitting tissue cells. At anthesis, these are single cell structures measuring up to 300 μm long. At the 5-mm stage of stylar length (7–10 days before anthesis), some stigmatic hair cells begin to accumulate an osmiophilic substance between the plasmalemma and the cell wall, possibly synthesized in the endoplasmic reticulum. This material is apparently never secreted outside the cell wall. Immediately following this secretory phase in some stigmatic hair cells a second secretory phase starts. A dense osmiophilic substance, different in appearance from the previous phase, accumulates in the vacuoles of each hair cell. Concomitantly, dimorphism develops in the cytoplasmic densities of stigmatic hair. Some stigmatic hair cytoplasm appears very dense and shows signs of degeneration while other cytoplasm appears normal. A third secretory phase, which begins at anthesis, occurs in the normal hair cells. This phase is characterized by enhanced activity in the cytoplasm of the endoplasmic reticulum and Golgi apparatus. Large vesicles containing granular material are seen fusing with the plasmalemma. Coincident with this activity there is dissolution of the middle layers of the cell wall and the cuticle is ruptured at various points. The dense osmiophilic substance that had accumulated in the vacuole breaks down into fine granular material. Significance of these changes is discussed in relation to the pollen germination mechanism on the dry type stigma of cotton.     

The Ultracytochemical Localization of ATPase Activity in the Ovules of Sunflower

The ultracytochemical localization of ATPase activity was determined employing the method of lead precipitation in the ovules of sunflower (Helianthus annuus L.). No ATPase activity is observed in the egg and synergids except some at the filiform apparatus. Much ATPase activity is localized on the plasma membrane and wall of the central cell. In the antipodal cells, ATPase activity is also found on the plasma membranes, but only a little in their walls. In the integumentary tapetum, besides the plasma membranes, most of the nuclei are rich in ATPase. Between the integumentary tapetum and uncontinuous cuticle surrounding the embryo sac, there is a gap where a lot of ATPase are found. These ATPases are continuously linked with those in the central cell wall throuth the intervals of the cuticle. At the sites of the wall ingrowths of the central celT, abundant vesicles and other structures with high ATPase activity aggregate noticeably in the gap region. According to the ATPase distribution in the ovules, we propose that the whole surface of embryo sac functions in absorbing nutrients directly from the apoplast outside the cuticle, especially via the wall-membrane apparatus of 'he central cell.     

The ultrastructure of a pheromone-secreting gland in the male scorpion-fly Harpobittacus Australis (Bittacidae: Mecoptera)

The eversible vesicles of the sex pheromone glands consist of three cell types, secreting the cuticle, the duct and the pheromone, respectively. Each pheromone-secreting cell has an end apparatus, bounded by microvilli and lined by a granular mass, that is penetrated by remarkable elongate filaments extending far up into the secretory duct. Vesicles containing secretion or secretion precursors abound. The possible role is discussed of the various organelles in the elaboration and transport of the pheromone.     

Microspore ofSecale cereale as a transfer cell type

Summary In the mature microspore ofSecale cereale, a set of wall ingrowths deposited as the first (outer) intine layer between exine and the microspore plasma membrane, are revealed by electron microscopy. The wall ingrowths form a girdle in the vicinity of the apertural region at the external pole of microspore which is in contact with the tapetum, so the microspore can be considered as a transfer cell which is polarized. After microspore division the second (inner) intine layer is deposited by the vegetative cell and forms a labyrinth of branched wall ingrowths. As a result, the periphery of a vegetative cell is also irregular and appears as very thin plasmatubules or evaginations delimited by plasma membrane and penetrating the pollen wall.The possible functions of the microspore as a transfer cell and the wall-membrane system of the vegetative cell are discussed.     

SECRETORY CELLS OF LILY PISTILS. II. ELECTRON MICROSCOPE CYTOCHEMISTRY OF CANAL CELLS

The secretory cells which line the canal of Lilium longiflorum pistils possess, on the side facing the canal, an elaborate wall which, with associated structures, Rosen and Thomas (1970) termed the “secretion zone.” We examined the secretion zone in the electron microscope following treatment of excised pistil slices with extraction procedures which remove pectin, hemicellulose, cellulose, lipid, or protein. The outer, fibrillar wall (layer 1) of the secretion zone contains protein, pectin, and cellulose. Internal to layer 1 is a granular-fibrillar wall (layer 2) several microns thick. It consists of outer and inner regions which can be distinguished from each other cytochemically. The granular component is composed of pectin which is not esterified with methyl groups and which may be complexed with protein. The short, randomly dispersed microfibrils of layer 2 were sensitive to procedures which dissolve cellulose. The extraction procedures did not reveal the chemical nature of the “osmiophilic islands” of layer 2. Paramural body membranes appear to be composed of glycoprotein and may function in secretion by serving as sites of pinocytic interchange at the plasmalemma. The origin of stigmatic exudate and the release of canal cell secretion product are discussed.     

Ultrastructure of an exocrine dermal gland in the gills of the grass shrimp,Palaemonetes pugio: Occurrence of transitory ciliary axonemes associated with the sloughing and reformation of the ductule

Exocrine dermal glands, comparable to the class 3 glandular units of insects, are found in the gills of the grass shrimp, Palaemonetes pugio. The dermal glands are composed of three cells: secretory cell, hillock cell and canal cell. Originating as a complex invagination of the apical cytoplasm of the granular secretory cell, a duct ascends through the hillock and canal cells to the cuticular surface. The duct is divisible into four regions: the secretory apparatus in the granular secretory cell, the locular complex, the hillock region within the hillock cell and the canal within the canal cell. A tubular ductule is contained within the latter two regions. As the ductule ascends to the cuticular surface, its constitution gradually changes from one of a fibrous material to one which possesses layers of epicuticle. During the proecdysial period, the ductule is extruded into the ecdysial space and this is followed by the secretion of a new ductule. Temporary ciliary structures, located near the secretory apparatus of the secretory cell, are associated with the extrusion and reformation of the ductule. Characterized only by a basal body and rootlets throughout most of the intermolt cycle, the ciliary organelles give rise to temporary axonemic processes which ascend through the ductule toward the ecdysial space at the onset of proecdysis. Subsequently, the old ductule is sloughed off and a new ductule is reformed around the ciliary axonemes. Following this reformation, the ciliary axonemes degenerate. The function of cytoplasmic processes, derived from the apical cytoplasm of the secretory cell, is also discussed.     

Development and functions of seed transfer cells

In secretion or absorption processes, solutes are transported across the plasmalemma between the symplastic and apoplastic compartments. For this purpose, certain plant cells have developed a specialised transfer cell morphology characterised by wall ingrowths, which amplify the associated plasmalemma surface area up to 20-fold. Detailed studies on the function and development of transfer cells in the context of seed filling have been carried out mainly in cereal endosperm, and for the cotyledon and seed coat cells of legumes. The major solutes transferred are amino acids, sucrose and monosaccharides. The contributions of recently identified symporter proteins to solute transfer are reviewed here, as is the role of apoplastic invertases in promoting solute assimilation. Expression of invertase and monosaccharide transporters early in both cereal and legume seed development orchestrates the distribution of free sugars which play an important role in regulating transfer cell function and determining final endosperm or embryo cell number. Transfer cell differentiation is subject to developmental control, and may also be modulated by sugar levels. The most abundant genes specifically expressed in the transfer layer of maize endosperm encode small antipathogenic proteins, pointing to a role for these cells in protecting the developing endosperm against pathogen ingress. The functional characterisation of the corresponding transfer layer-specific promoters has provided a tool for dissecting transfer cell functions. Transfer cells are highly polar in their organisation, the characteristic cell wall ingrowths developing on one face only. The presence of cytoskeletal components bordering wall ingrowths is documented, but their role in establishing transfer cell morphology remains to be established.     

The transmitting tract in Trimezia fosteriana (Iridaceae). 1. Ultrastructure in the stigma, style and ovary

An ultrastructural investigation of the entire transmitting tract in Trimezia fosteriana (Iridaceae) was undertaken. The transmitting tissue is secretory but transfer cells do not occur at any level. With exception for the stigma papillae, the cells are covered with large amounts of secretory products. The papillae have a thick and ridged cuticle. The cuticle in the rest of the transmitting tract is thin and detached from the cell wall by the secretory products. It is more or less ruptured in the secretory parts of the stigma and ovary. In the stylar canal the major part of the cuticle is continuous and covers the secretory products. The occurence of a large amount of vesicles in the stigma transmitting tissue cells is interpreted as a result of high dictyosome activity. An electron opaque material is produced in the dictyosomes and appears in vesicles and vacuoles but also between the plasma membrane and the cell walls in the stigma. A small amount of such material is present in the cell walls. Corresponding material is also present in the style and the ovary but declines basipetally. Plastids with strongly electron opaque plastoglobules are present at all levels in the transmitting tract.     

A cytochemical and immunocytochemical analysis of the wall labyrinth apparatus in leaf transfer cells in Elodea canadensis

Background and Aims

Transfer cells are plant cells specialized in apoplast/symplast transport and characterized by a distinctive wall labyrinth apparatus. The molecular architecture and biochemistry of the labyrinth apparatus are poorly known. The leaf lamina in the aquatic angiosperm Elodea canadensis consists of only two cell layers, with the abaxial cells developing as transfer cells. The present study investigated biochemical properties of wall ingrowths and associated plasmalemma in these cells.

Methods

Leaves of Elodea were examined by light and electron microscopy and ATPase activity was localized cytochemically. Immunogold electron microscopy was employed to localize carbohydrate epitopes associated with major cell wall polysaccharides and glycoproteins.

Key Results

The plasmalemma associated with the wall labyrinth is strongly enriched in light-dependent ATPase activity. The wall ingrowths and an underlying wall layer share an LM11 epitope probably associated with glucuronoarabinoxylan and a CCRC-M7 epitope typically associated with rhamnogalacturonan I. No labelling was observed with LM10, an antibody that recognizes low-substituted and unsubstituted xylan, a polysaccharide consistently associated with secondary cell walls. The JIM5 and JIM7 epitopes, associated with homogalacturonan with different degrees of methylation, appear to be absent in the wall labyrinth but present in the rest of cell walls.

Conclusions

The wall labyrinth apparatus of leaf transfer cells in Elodea is a specialized structure with distinctive biochemical properties. The high level of light-dependent ATPase activity in the plasmalemma lining the wall labyrinth is consistent with a formerly suggested role of leaf transfer cells in enhancing inorganic carbon inflow. The wall labyrinth is a part of the primary cell wall. The discovery that the wall ingrowths in Elodea have an antibody-binding pattern divergent, in part, from that of the rest of cell wall suggests that their carbohydrate composition is modulated in relation to transfer cell functioning.     

Histogenesis of the transmitting tract in Strelitzia reginae

Organisation and development of the stigmatic, stylar and ovarian parts of the transmitting tract in Strelitzia reginae were evaluated. They were characterised by 1) cell shape, 2) appearance of distal cell wall, 3) type of plastid, 4) and vacuolar system. The long stigmatic trichomes have a secondary irregular wall layer separated from the primary wall. Cell structures include pleomorphic plastids with vesiculated thylakoids and frequently a crystal or a lipidic globule. In early stages of bud development the extensive endoplasmic reticulum (ER) is smooth, whereas it is mainly rough in older buds. Coated vesicles are frequent, as are dictyosomes. Prominent invaginations along the plasma membrane contain floccular deposits in the older flower buds. These deposits are similar in structural appearance to material in the large vacuoles. The basal part of the stigma has wedge-shaped cells with wall ingrowths. Three stages of stigmatic secretion during the development were characterised. The stylar canal is initially narrow but widens subsequently. The cuticle is detached and the apical cell walls show a fringed surface; from this wall inbuddings develop shortly before anthesis. At this stage sheets of rough ER are evident in the cell cortex. The plastids have a few vesiculated thylakoids, proteinaceous crystals and starch grains. The epithelial cells of young buds have numerous vacuoles, the volume of which decreases in more mature cells. The stylar canal is filled with a secretion at all stages of bud development. The face of the ovarian transmitting tract, lining the placenta, is smooth in young buds but lobed in older ones due to the division pattern of the epithelial cells. These cells are large, elongated and culumnar as young but narrow and wedge-shaped when more mature. Cell wall inbuddings are formed late during bud development.     

Unusual transfer cells in the epithelium of the nectaries ofAsclepias curassavica L.

Summary The secretory cells of the nectaries ofAsclepias curassavica form a glandular epithelium in the inner parts of the stigmatic chambers. They resemble transfer cells in having many infoldings of the plasmalemma. The wall protuberances, however, are poorly developed and often lacking. The plasmalemma is highly convoluted and forms, in places, external compound membranes where the extracytoplasmic space is collapsed completely. Active glands contain dilated cisternae of the ER and large vesicles which are mainly associated with the cis face of the dictyosomes. In addition, small vesicles are observed in high number. It is discussed whether the secretion is granulocrine or eccrine and whether the enlargement of the plasmalemma is the cause or the consequence of the high secretory activity. After the secretory phase the outer peripheral part of the cytoplasm disintegrates. The remaining part of the protoplast is covered by a new plasmalemma.     

Ultrastructural investigation of a salivary gland in a centipede: structure and origin of the maxilla I-gland of Scutigera coleoptrata (Chilopoda, Notostigmophora)

The maxilla I-gland of Scutigera coleoptrata was investigated using light and electron microscopy methods. This is the first ultrastructural investigation of a salivary gland in Chilopoda. The paired gland opens via the hypopharynx into the foregut and extends up to the third trunk segment. The gland is of irregular shape and consists of numerous acini consisting of several gland units. The secretion is released into an arborescent duct system. Each acinus consists of multiple of glandular units. The units are composed of three cell types: secretory cells, a single intermediary cell, and canal cells. The pear-shaped secretory cell is invaginated distally, forming an extracellular reservoir lined with microvilli, into which the secretion is released. The intermediary cell forms a conducting canal and connects the secretory cell with the canal cell. Proximally, the intermediary cell bears microvilli, whereas the distal part is covered with a distinct cuticle. The cuticle is a continuation of the cuticle of the canal cells. This investigation shows that the structure of the glandular units of the salivary maxilla I-gland is comparable to that of the glandular units of epidermal glands. Thus, it is likely that in Chilopoda salivary glands and epidermal glands share the same ground pattern. It is likely that in compound acinar glands a multiplication of secretory and duct cells has taken place, whereas the number of intermediary cells remains constant. The increase in the number of salivary acini leads to a shifting of the secretory elements away from the epidermis, deep into the head. Comparative investigations of the different head glands provide important characters for the reconstruction of myriapod phylogeny and the relationships of Myriapoda and Hexapoda.     

Ultrastructural analysis of the functional copulatory organ of the male rotifer,Asplanchna brightwelli

The male rotifer copulatory organ is composed of a urethral canal extending from the tip of the copulatory organ internally to a layer of microvilli. The microvilli project from two different cell types, referred to as the internal and peripheral microvillar cells according to their location. At this microvillar junction a second canal, the vas deferens, continues posteriorly and enters the sperm duct region of the testis. The channel of the vas deferens is formed from the inner wall of three separate cells; the cap, intermediate and basal cells. Peripheral to these cells and parallel to them for their entire length, cross sections of seven prostate gland cells can be observed. Anteriorly, these gland cells are connected to the basal end of the microvillar layer via a short neck region, through which glandular secretion occurs only during copulation. The mechanism of secretion appears to be a form of exocytosis whereby the secretory granule membrane fuses with the cell plasmalemma so that rupturing at the point of fusion will release the granule content into the neck region. The prostate gland cells contain an abundance of autophagic vacuoles while most of the other cells of the copulatory organ contain primary lysosomes and cytolosomes. These organelles may be associated with the aging process in rotifers, or, as in the case of the prostate gland-autophagic vacuoles, with a fast organelle turnover during secretion.     

Ultrastructural Differentiation of the Ovarian Transmitting Tissue in Lilium regale

The cells of the ovarian transmitting tissue of Lilium regaleare papilla shaped and form and epithelium on the placenta.Their ultrastructural organization and differentiation from1 d before to 7 d after anthesis is presented. These placentacells are typical transfer cells with a prominent secretionzone similar to that known from stylar canal cells. After anthesisthe secretion zone continues to grow by addition of vesiclefrom the numerous dictyosomes. Maximum depth of this zone isreached by day 4 after anthesis. The outer surface of the cellwall is distinctly rugged on cell maturation and the outermostlayer is corroded. The ER system undergoes transition from asmooth to a granular condition. Before anthesis there is a centralvacuole which at anthesis is reduced to a system of small vauoles.These are supplemented by autophagic vacuoles formed from theER. Such vacuoles are found near the secretion zone and mayalso fuse with the plasmalemma. The cuticle is sloughed andsecretion commences before anthesis. Accumulations of vesiclesfound in the nucleus and occasional connections between suchvesicles and the inner membrane of the nuclear envelope indicatethe presence of a nuclear network. Protein crystals are presentin the cytoplasm and the nucleus. The starch grains in the plastidsare digested after anthesis, but new ones are formed by days6 and 7.Copyright 1995, 1999 Academic Press Lilium regale, transmitting tissue, placenta, secretion, nuclear reticulum, transfer cells     

Fine structure of the accessory glands of the female genital tract of the ichneumonid Pimpla turionellae (Insecta,Hymenoptera)

Summary The female accessory glands include the tubular poison gland, the paired, lemon-shaped uterus glands, and Dufour's gland, an unbranched tubular organ. They consist essentially of a single layer of epithelium cells surrounded by a basement membrane. The lumen is lined by cuticle. The proteinaceous secretion of the poison gland is released into intracellular ducts provided with microvilli, each connected to a channel lined with cuticle which leads to the central lumen of the gland. The channel is formed by special canal cells. Nerve endings are interspersed among the gland cells. The uterus gland consists of four cell types derived from a single type of precursor cell found in newly hatched wasps. Type I cells are covered by type II cells and are thus without contact to the luminal surface of the gland. They contain stacks or whorls of mitochondria and smooth cisternae in an alternating arrangement. Vesicles with a secretory product are found in cells of types II and III. Deep anastomosing infoldings of the plasmalemma, stabilized by microtubules and dense material at the branchings, are characteristic for type II cells. Most secretory vesicles are found in type III cells, the prevalent cell type which is thought to be the source of the lipoprotein secretion. Coated vesicles are present at deep infoldings of the plasmalemma. The greatly enlarged apical surface area of type IV cells and the presence of mitochondria in slender outgrowths is suggestive of an osmoregulatory function. In Dufour's gland, two cell types appear in succession, the first with a very dense cytoplasm, the second with dense inclusions and many seemingly empty vesicles of smooth endoplasmic reticulum. The secretion products, lecithin and a cholesterol ester, are thought to be formed by the second cell type. The dense inclusion might be lecithin, which reacts with osmium tetroxide. The cholesterol ester could have been washed out of the empty vesicles by the embedding procedure.     

ULTRASTRUCTURE OF NECTARIES IN RELATION TO NECTAR SECRETION

In the last 15–20 years, ultrastructural studies have added new important cytological information to the relatively rich literature on the morphology and light microscopy of nectaries. On the basis of these studies, the following main conclusions can be drawn regarding the relation between the ultrastructure of nectaries and the process of nectar secretion: (1) the transport of the pre-nectar in the nectariferous tissue is mainly via the symplast; (2) the ER alone or the ER and the Golgi apparatus are involved in the process of secretion; (3) the elimination of nectar from the protoplast of the secretory cells is by reverse pinocytosis; (4) the outer walls of the secretory cells of nectaries in many plants possess wall ingrowths.     

黑缘烟蟋螽丝腺结构

【目的】蟋螽是直翅目中唯一具有吐丝筑巢行为的类群。本研究旨在探讨蟋螽丝腺的结构特点。【方法】应用解剖学观察、免疫荧光、苏木精-伊红染色、PAS苏木精染色、扫描电镜和透射电镜等方法从细胞水平对黑缘烟蟋螽Capnogryllacris nigromarginata丝腺的显微与超微结构进行了观察。【结果】黑缘烟蟋螽丝腺由导管和腺泡构成。腺泡由鞘细胞延伸形成的结缔组织鞘包围。腺泡的主体有4种细胞,分别为Ⅰ型分泌细胞、Ⅱ型分泌细胞、围细胞和腔细胞。Ⅰ型和Ⅱ型分泌细胞为大的腺细胞,形状不规则。分泌细胞细胞核很大,胞质内有大量的内质网和分泌颗粒。Ⅰ型分泌细胞靠近腺泡中心,PAS-苏木精染色表明Ⅰ型分泌细胞内含糖蛋白,Ⅱ型分泌细胞在腺泡外周,位于Ⅰ型分泌细胞与围细胞或结缔组织鞘之间。腔细胞分散在分泌细胞之间,包围形成胞外运输分泌物的通道。围细胞与鞘细胞接触,具有由细胞膜内陷形成的微绒毛腔,胞质内有大量的线粒体。围细胞微绒毛腔与腔细胞包围的细胞外运输通道相连,分泌细胞分泌的颗粒聚集在分泌细胞和胞外运输通道之间的连接处,并将分泌物排出至胞外运输通道。多个腺泡的胞外运输通道汇集到由单层细胞组成的丝腺导管。单层导管细胞靠近管腔外围具有规则排列的质膜内陷和大量伸长的线粒体;靠近管腔的一侧具连续的细胞膜突起,在导管壁的表皮下紧密排列。【结论】黑缘烟蟋螽丝腺分泌细胞分为Ⅰ型分泌细胞和Ⅱ型分泌细胞。分泌物质产生及分泌过程依次经过分泌细胞、腔细胞包围的胞外通道、分支导管、总导管和唾窦。其中在腺泡细胞之间,分泌物向外运输过程中,围细胞微绒毛腔的微丝束可能对分泌物的外排提供推动力。     

Ultrastructural investigation of the vesicular glands in Scutigera coleoptrata (Chilopoda,Notostigmophora)

In the notostigmophoran centipedes, two pairs of vesicular glands have evolved. These paired glands are situated in the first and second trunk segment and open via cuticular ducts in the upper part of the particular pleura. The vesicular glands of Scutigera coleoptrata were investigated using light and, for the first time, electron microscopical methods. The glands consist of wide sac‐like cavities that often appear vesicular. The epithelia of both glands are identically structured and consist of numerous glandular units. Each of these units consists of four different cells: a single secretory cell, a small intermediary cell, and one proximal and one distal canal cell. The intermediary cell forms a conducting canal and connects the secretory cell with the canal cells. Proximally, the intermediary cell bears microvilli, whereas the distal part is covered with a distinct cuticle. The cuticle is a continuation of the cuticle of the canal cells. This investigation shows that the ultrastructure of glandular units of the vesicular glands is comparable to that of the glandular units of other epidermal glands in Chilopoda and Diplopoda, although the glands look completely different in the light microscope. Thus, it is likely that the vesicular glands and epidermal glands share the same ground pattern. With regard to specific differences in the cuticular lining of the intermediary cells, a common origin of epidermal glands in Myriapoda and Hexapoda is not supported. J. Morphol. 2009. © 2008 Wiley‐Liss, Inc.     

Ultrastructural organization of Alicyclobacillus tolerans strain K1T cells

Electron microscopy examinations of thin sections and freeze-fracture replicas revealed the specific ultrastructural features of Alicyclobacillus tolerans strain K1(T). In particular, the cell wall displayed an ultrastructure typical of gram-positive bacteria and consisted of a thin murein layer (50-60 A in thickness); cells exhibited a surface S-layer constituted by large hexagonally packed (p6-symmetry) rod-shaped subunits of 150-160 A in diameter and 200 A in height. In the cytoplasmic membrane, there were intramembrane vesicular structures that sometimes appeared as large leaflets in the central part. The cytoplasm contained numerous vesicular inclusions covered with a monolayered wall, dissimilar to bilamellar lipid membranes. Endospore coats displayed an intricate structure and consisted of three thick layers; the outer layer had an unusual fine structure; the exosporium was also found.     

ULTRASTRUCTURE OF THE CORALLINACEAE (RHODOPHYTA) II. VEGETATIVE CELLS OF LITHOTHRIX ASPERGILLUM1

The ultrastructure of the calcareous red coralline alga Lithothrix aspergillum Gray and the development of the various tissue types has been studied. The sub-apical meristematic tissue alternately produces genicular or intergenicular cells. The genicular cells rapidly elongate and their cell walls thicken and become denser as more fibrillar wall material is laid down within the cell wall. These cells contain little cytoplasm and few organelles. The inter genicular cells which elongate only slightly during development have a small vacuole and many free starch grains in the cytoplasm. The peripheral cells in each inter genicular layer remain meristematic and form a cortical cell layer over the genicular cells. These cortical cells and the apical meristematic cells are covered by small epidermal cells which have extensive cell wall ingrowths between the chloroplasts. The inter genicular cells are calcified. Although the CaCO₃ is laid down within the cell walls, there is always a thin layer of CaCO₃-free organic cell wall material between the plasmalemma and the CaCO₃ impregnated wall. Only the distal tips of the genicular cells are calcified. In old genicular tissues of Lithothrix, secondary deposits of CaCO₃ of unknown crystallography are also found in the spaces between the cell walls. Thus there appear to be at least two mechanisms of calcification in this alga.