Oblique Divisions Research Articles

Reproducibly oriented cell divisions pattern the prothallus to set up dorsoventrality and de novo meristem formation in Marchantia polymorpha

Land plant bodies develop from stem cells located in meristems. However, we know little about how meristems initiate from non-meristematic cells. The haploid body of bryophytes develops from unicellular spores in isolation from the parental plant, which allows all stages of development to be observed. We discovered that the Marchantia spore undergoes a series of reproducibly oriented cell divisions to generate a flat prothallus on which a meristem later develops de novo. The young sporeling comprises an early cell mass. One cell of the early cell mass elongates and undergoes a formative division that produces the prothalloblast, which initiates prothallus formation. A symmetric division of the prothalloblast followed by two transverse divisions generates a four-celled plate that expands into a flat disc through oblique divisions in three of the four plate-cell-derived quadrants. One quadrant gives rise to a flat flabellum. A notch with a meristem and apical stem cell develops at the margin of the flabellum. The transcription factor Marchantia class III homeodomain-leucine-zipper (MpC3HDZ) is a marker of the first flat prothallus structure and polarizes to the dorsal tissues of flabella and meristems. Mpc3hdz mutants are defective in setting up dorsoventrality and thallus body flatness. We report how a regular set of cell divisions forms the prothallus-the first dorsoventral structure-and how cells on the margin of the prothallus develop a dorsoventralized meristem de novo.

LGN loss randomizes spindle orientation and accelerates tumorigenesis in PTEN-deficient epidermis.

Loss of cell polarity and disruption of tissue organization are key features of tumorigenesis that are intrinsically linked to spindle orientation. Epithelial tumors are often characterized by spindle orientation defects, but how these defects impact tumor formation driven by common oncogenic mutations is not fully understood. Here, we examine the role of spindle orientation in adult epidermis by deleting a key spindle regulator, LGN, in normal tissue and in a PTEN-deficient mouse model. We report that LGN deficiency in PTEN mutant epidermis leads to a threefold increase in the likelihood of developing tumors on the snout, and an over 10-fold increase in tumor burden. In this tissue, loss of LGN alone increases perpendicular and oblique divisions of epidermal basal cells, at the expense of a planar orientation of division. PTEN loss alone does not significantly affect spindle orientation in these cells, but the combined loss of PTEN and LGN fully randomizes basal spindle orientation. A subset of LGN- and PTEN-deficient animals have increased amounts of proliferative spinous cells, which may be associated with tumorigenesis. These results indicate that loss of LGN impacts spindle orientation and accelerates epidermal tumorigenesis in a PTEN-deficient mouse model.

A non-canonical function for Centromere-associated protein-E controls centrosome integrity and orientation of cell division

Centromere-associated protein-E (CENP-E) is a kinesin motor localizing at kinetochores. Although its mitotic functions have been well studied, it has been challenging to investigate direct consequences of CENP-E removal using conventional methods because CENP-E depletion resulted in mitotic arrest. In this study, we harnessed an auxin-inducible degron system to achieve acute degradation of CENP-E. We revealed a kinetochore-independent role for CENP-E that removes pericentriolar material 1 (PCM1) from centrosomes in late S/early G2 phase. After acute loss of CENP-E, centrosomal Polo-like kinase 1 (Plk1) localization is abrogated through accumulation of PCM1, resulting in aberrant phosphorylation and destabilization of centrosomes, which triggers shortened astral microtubules and oblique cell divisions. Furthermore, we also observed centrosome and cell division defects in cells from a microcephaly patient with mutations in CENPE. Orientation of cell division is deregulated in some microcephalic patients, and our unanticipated findings provide additional insights into how microcephaly can result from centrosomal defects.

DNA Sequence Analyses Reveal Two New Species of Caloglossa (Delesseriaceae, Rhodophyta) from the Skin of West Indian Manatees

Epizoic macroalgae collected from the skin of West Indian manatees included specimens of the red algal family Delesseriaceae. Morphological and rbcL sequence analyses indicated that these specimens represented two novel species of Caloglossa. One species, described here as Caloglossa kamiyana Freshwater, Cath.E. Miller & Frankovich sp. nov., had been previously studied and recognized as part of the C. ogasawaraensis species complex. The rbcL sequence divergence between C. kamiyana and other taxa within the complex ranged from 4.6–5.3%, and tetrasporangial mother cells are cut off from the lateral pericentral cells by oblique divisions instead of transverse divisions as in C. ogasawaraensis. The second species was resolved as a closely related sister species to C. fluviatilis, with a minimum interspecific sequence divergence of 2.0%. It was morphologically indistinguishable from C. fluviatilis except for one potential character—mostly one, instead of multiple rhizoids, developing from rhizoid-bearing pericentral and marginal wing cells. It is herein described as Caloglossa manaticola Freshwater, Cath.E. Miller & Frankovich sp. nov.

Endfoot regeneration restricts radial glial state and prevents translocation into the outer subventricular zone in early mammalian brain development.

Neural stem cells, called radial glia, maintain epithelial structure during the early neocortical development. The prevailing view claims that when radial glia first proliferate, their symmetric divisions require strict spindle orientation; its perturbation causes precocious neurogenesis and apoptosis. Here, we show that despite this conventional view, radial glia at the proliferative stage undergo normal symmetric divisions by regenerating an apical endfoot even if it is lost by oblique divisions. We found that the Notch-R-Ras-integrin β1 pathway promotes the regeneration of endfeet, whose leading edge bears ectopic adherens junctions and the Par-polarity complex. However, this regeneration ability gradually declines during the subsequent neurogenic stage and hence oblique divisions induce basal translocation of radial glia to form the outer subventricular zone, a hallmark of the development of the convoluted brain. Our study reveals that endfoot regeneration is a temporally changing cryptic property, which controls the radial glial state and its shift is essential for mammalian brain size expansion.

Telophase correction refines division orientation in stratified epithelia.

During organogenesis, precise control of spindle orientation balances proliferation and differentiation. In the developing murine epidermis, planar and perpendicular divisions yield symmetric and asymmetric fate outcomes, respectively. Classically, division axis specification involves centrosome migration and spindle rotation, events occurring early in mitosis. Here, we identify a novel orientation mechanism which corrects erroneous anaphase orientations during telophase. The directionality of reorientation correlates with the maintenance or loss of basal contact by the apical daughter. While the scaffolding protein LGN is known to determine initial spindle positioning, we show that LGN also functions during telophase to reorient oblique divisions toward perpendicular. The fidelity of telophase correction also relies on the tension-sensitive adherens junction proteins vinculin, α-E-catenin, and afadin. Failure of this corrective mechanism impacts tissue architecture, as persistent oblique divisions induce precocious, sustained differentiation. The division orientation plasticity provided by telophase correction may enable progenitors to adapt to local tissue needs.

Lzts1 controls both neuronal delamination and outer radial glial-like cell generation during mammalian cerebral development

In the developing central nervous system, cell departure from the apical surface is the initial and fundamental step to form the 3D, organized architecture. Both delamination of differentiating cells and repositioning of progenitors to generate outer radial glial cells (oRGs) contribute to mammalian neocortical expansion; however, a comprehensive understanding of their mechanisms is lacking. Here, we demonstrate that Lzts1, a molecule associated with microtubule components, promotes both cell departure events. In neuronally committed cells, Lzts1 functions in apical delamination by altering apical junctional organization. In apical RGs (aRGs), Lzts1 expression is variable, depending on Hes1 expression levels. According to its differential levels, Lzts1 induces diverse RG behaviors: planar division, oblique divisions of aRGs that generate oRGs, and their mitotic somal translocation. Loss-of-function of lzts1 impairs all these cell departure processes. Thus, Lzts1 functions as a master modulator of cellular dynamics, contributing to increasing complexity of the cerebral architecture during evolution.

Three-Dimensional Analysis of Cell Division Orientation in Epidermal Basal Layer Using Intravital Two-Photon Microscopy.

Epidermal structures are different among body sites, and proliferative keratinocytes in the epidermis play an important role in the maintenance of the epidermal structures. In recent years, intravital skin imaging has been used in mammalian skin research for the investigation of cell behaviors, but most of these experiments were performed with rodent ears. Here, we established a non-invasive intravital imaging approach for dorsal, ear, hind paw, or tail skin using R26H2BEGFP hairless mice. Using four-dimensional (x, y, z, and time) imaging, we successfully visualized mitotic cell division in epidermal basal cells. A comparison of cell division orientation relative to the basement membrane in each body site revealed that most divisions in dorsal and ear epidermis occurred in parallel, whereas the cell divisions in hind paw and tail epidermis occurred both in parallel and oblique orientations. Based on the quantitative analysis of the four-dimensional images, we showed that the epidermal thickness correlated with the basal cell density and the rate of the oblique divisions.

Cerebrospinal fluid-derived Semaphorin3B orients neuroepithelial cell divisions in the apicobasal axis

The spatial orientation of cell divisions is fundamental for tissue architecture and homeostasis. Here we analysed neuroepithelial progenitors in the developing mouse spinal cord to determine whether extracellular signals orient the mitotic spindle. We report that Semaphorin3B (Sema3B) released from the floor plate and the nascent choroid plexus in the cerebrospinal fluid (CSF) controls progenitor division orientation. Delivery of exogenous Sema3B to neural progenitors after neural tube opening in living embryos promotes planar orientation of their division. Preventing progenitor access to cues present in the CSF by genetically engineered canal obstruction affects the proportion of planar and oblique divisions. Sema3B knockout phenocopies the loss of progenitor access to the CSF. Sema3B binds to the apical surface of mitotic progenitors and exerts its effect via Neuropilin receptors, GSK3 activation and subsequent inhibition of the microtubule stabilizer CRMP2. Thus, extrinsic control mediated by the Semaphorin signalling orients progenitor divisions in neurogenic zones.

Deletion of RIC8A in neural precursor cells leads to altered neurogenesis and neonatal lethality of mouse.

RIC8A is a noncanonical guanine nucleotide exchange factor for a subset of Gα subunits. RIC8A has been reported in different model organisms to participate in the control of mitotic cell division, cell signalling, development and cell migration. Still, the function of RIC8A in the mammalian nervous system has not been sufficiently analysed yet. Adult mice express RIC8A in the brain regions involved in the regulation of memory and emotional behaviour. To elucidate the role of RIC8A in mammalian neurogenesis we have inactivated Ric8a in neural precursor cells using Cre/Lox system. As a result, the conditional knockout mice were born at expected Mendelian ratio, but died or were cannibalized by their mother within 12 h after birth. The cerebral cortex of the newborn Nes;Ric8a(CKO) mice was thinner compared to littermates and the basement membrane was discontinuous, enabling migrating neurons to invade to the marginal zone. In addition, the balance between the planar and oblique cell divisions was altered, influencing the neuron production. Taken together, RIC8A has an essential role in the development of mammalian nervous system by maintaining the integrity of pial basement membrane and modulating cell division.

Size correlations among cambial initials and their derivatives in Polyalthia longifolia Thw.

The length and breadth of the cambial initials and their derivatives have been examined in <i>Polyalthia longifolia</i>, a tropical tree possessing non-storied cambium. Taking the average size of the initials and the elements originating from them, most of the sieve-tube elements have been found to be slightly shorter in length than the fusiform initials. On the other hand, a few of these are still shorter - almost half of the fusiform initials, due to transverse or somewhat oblique divisions in the sieve element mother cells. The vessel elements are slightly shorter but 5-6 times wider than the fusiform initials. The parenchyma strands, in phloem comprising cells storing starch or tannin (pps), in xylem accumulating starch only (ssps), are more or less equal to fusiform initials indicating that the xylem and phloem mother cells forming parenchyma cells have not undergone any major change except for transverse divisions. The individual vessel-associated parenchyma cells (v.a.p. cells) are wider but much shorter in length as compared to the starch-storing parenchyma cells (s.s.p. cells) indicating that more transverse divisions have occured in the strands of the former than those of the latter. Among all the cambial derivatives, the fibers exhibit maximum increase in length, due to intrusive growth. The ray parenchyma cells are slightly longer than the ray initials possibly due to the elongation of these initials during their transformation into vascular ray cells.

ARABIDOPSIS HOMOLOG of TRITHORAX1 (ATX1) is required for cell production, patterning, and morphogenesis in root development

Arabidopsis homolog of trithorax1 (ATX1/SDG27), a known regulator of flower development, encodes a H3K4histone methyltransferase that maintains a number of genes in an active state. In this study, the role of ATX1 in root development was evaluated. The loss-of-function mutant atx1-1 was impaired in primary root growth. The data suggest that ATX1 controls root growth by regulating cell cycle duration, cell production, and the transition from cell proliferation in the root apical meristem (RAM) to cell elongation. In atx1-1, the quiescent centre (QC) cells were irregular in shape and more expanded than those of the wild type. This feature, together with the atypical distribution of T-divisions, the presence of oblique divisions, and the abnormal cell patterning in the RAM, suggests a lack of coordination between cell division and cell growth in the mutant. The expression domain of QC-specific markers was expanded both in the primary RAM and in the developing lateral root primordia of atx1-1 plants. These abnormalities were independent of auxin-response gradients. ATX1 was also found to be required for lateral root initiation, morphogenesis, and emergence. The time from lateral root initiation to emergence was significantly extended in the atx1-1 mutant. Overall, these data suggest that ATX1 is involved in the timing of root development, stem cell niche maintenance, and cell patterning during primary and lateral root development. Thus, ATX1 emerges as an important player in root system architecture.

Compared Anatomy of Young Leaves of Prunus persica (L.) Batsch with Different Degrees of Susceptibility to Taphrina deformans (Berk.) Tul

AbstractAnatomical observations of leaves infected by Taphrina deformans were studied in tolerant peach trees (TPT) and in very susceptible (VSPT) ones. Leaves from the first sampling (2nd April) showed hyphae penetrating through the stomata or into the cuticle of the host tissue; anatomical structures of leaf sections were similar for both TPT and VSPT. The ultrastructure of the leaves of TPT showed seemingly normal mesophyll cells. In contrast, mesophyll cells of the VSPT showed important signs of degradation. Cells were organelle‐free and the middle lamella was expanded and invaded by hyphae of T. deformans. In some samples, the leaves of TPT showed deformed epidermal cells, loss of some spongy cells and increase of the intercellular spaces and division of the palisade cells. The pathogen proliferation in the leaves of the VSPT was considerably superior. In this case, stimulation of cell division occurred in the abaxial epidermis. Cells showed periclinal and oblique divisions, with an increased number of plasmodesmata; palisade or spongy cells were not differentiable. Leaves from TPT collected on 26th April showed hyphae with a non‐cylindrical section and with a squashed aspect. The hyphae were very evident in the intercellular spaces, showing abundant endoplasmic reticulum of rough type (RER) in the cytoplasm. On the other hand, epidermis of the leaves of the VSPT had numerous hyphae under the cuticle, which were growing in a thick pectin matrix. Leaves from TPT and VSPT collected on 6th May showed relevant differences. The leaves of TPT had a palisade mesophyll with fewer cells but with active chloroplasts. In contrast, the leaves from VSPT showed empty mesophyll cells, the cytoplasm was collapsed and the adaxial epidermis was covered with the fungus fructification. The observed anatomical and ultrastructural differences of leaves from TPT and VSPT confirm a different behaviour in plant‐host reaction at early stages of infection.

Early cleavage in Phoronis muelleri (Phoronida) displays spiral features

The view that early cleavage in Phoronida follows a radial pattern is widely accepted. However, data supporting this characterization are ambiguous. Studies have been repeatedly reporting variation between individual embryos, and the occurrence of embryos exhibiting oblique divisions or nonradial cell arrangements. Such embryos were often considered to represent variation within radial cleavage, or artificial appearances. Cleavage in Phoronis muelleri was previously characterized as "derived radial," but also oblique spindles and cell elongations, and shifted cell arrangements were observed. We studied the early cleavage in P. muelleri applying 4D microscopy, fluorescent staining, and confocal laser scanning microscopy. To deal with the problem of variation we provide statistical evaluations of our data. These show that oblique divisions do not represent variational abnormalities. In fact, they reveal that most cells divide obliquely from the third cleavage onwards. What is more, in almost all cells the axis of the third cleavage is inclined dextrally. The fourth cleavage is even stronger sinistrally pronounced. Subsequently, the pattern of alternating cleavage orientation is largely restricted to animal and vegetal blastomeres. As a result of the obliqueness of divisions, four cells encircle the poles in most embryos. Cross furrows are occasionally present. We found no indications for radial cleavage in P. muelleri. In contrast, the observed cleavage displays several characters consistent with the pattern of spiral cleavage. A close relation of phoronid and spiralian cleavage is also suggested by molecular phylogenies, allying both groups in the Lophotrochozoa. We suggest our findings to represent morphological support for this lophotrochozoan/spiralian affinity of Phoronida.

Mouse Inscuteable Induces Apical-Basal Spindle Orientation to Facilitate Intermediate Progenitor Generation in the Developing Neocortex

SummaryNeurons in the mammalian neocortex arise from asymmetric divisions of progenitors residing in the ventricular zone. While in most progenitor divisions, the mitotic spindle is parallel to the ventricular surface, some progenitors reorient the spindle and divide in oblique orientations. Here, we use conditional deletion and overexpression of mouse Inscuteable (mInsc) to analyze the relevance of spindle reorientation in cortical progenitors. Mutating mInsc almost abolishes oblique and vertical mitotic spindles, while mInsc overexpression has the opposite effect. Our data suggest that oblique divisions are essential for generating the correct numbers of neurons in all cortical layers. Using clonal analysis, we demonstrate that spindle orientation affects the rate of indirect neurogenesis, a process where progenitors give rise to basal progenitors, which in turn divide symmetrically into two differentiating neurons. Our results indicate that the orientation of progenitor cell divisions is important for correct lineage specification in the developing mammalian brain.

Effect of mechanical stress on Zea root apex. I. Mechanical stress leads to the switch from closed to open meristem organization.

The effect of mechanical stress on the root apical meristem (RAM) organization of Zea mays was investigated. In the experiment performed, root apices were grown through a narrowing of either circular (variant I) or elliptical (variant II) shape. This caused a mechanical impedance distributed circumferentially or from the opposite sides in variant I and II, respectively. The maximal force exerted by the growing root in response to the impedance reached the value of 0.15 N for variant I and 0.08 N for variant II. Significant morphological and anatomical changes were observed. The changes in morphology depended on the variant and concerned diminishing and/or deformation of the cross-section of the root apex, and buckling and swelling of the root. Anatomical changes, similar in both variants, concerned transformation of the meristem from closed to open, an increase in the number of the cell layers at the pole of the root proper, and atypical oblique divisions of the root cap cells. After leaving the narrowing, a return to both typical cellular organization and morphology of the apex was observed. The results are discussed in terms of three aspects: the morphological response, the RAM reorganization, and mechanical factors. Assuming that the orientation of division walls is affected by directional cues of a tensor nature, the changes mentioned may indicate that a pattern of such cues is modified when the root apex passes through the narrowing, but its primary mode is finally restored.

Oblique Radial Glial Divisions in the Developing Mouse Neocortex Induce Self-Renewing Progenitors outside the Germinal Zone That Resemble Primate Outer Subventricular Zone Progenitors

Radial glia cells function as neural stem cells in the developing brain and generate self-renewing and differentiating daughter cells by asymmetric cell divisions. During these divisions, the apical process or basal process of the elongated epithelial structure is asymmetrically partitioned into daughter cells, depending on developmental contexts. However, in mammalian neurogenesis, the relationship between these subcellular structures and self-renewability is largely unknown. We induced oblique cleavages of radial glia cells to split the apical and basal processes into two daughters, and investigated the fate and morphology of the daughters in slice cultures. We observed that the more basal daughter cell that inherits the basal process self-renews outside of the ventricular zone (VZ), while the more apical daughter cell differentiates. These self-renewing progenitors, termed "outer VZ progenitors," retain the basal but not the apical process, as recently reported for the outer subventricular zone (OSVZ) progenitors in primates (Fietz et al., 2010; Hansen et al., 2010); to self-renew, they require clonal Notch signaling between sibling cells. We also found a small endogenous population of outer VZ progenitors in the mouse embryonic neocortex, consistent with a low frequency of oblique radial glia divisions. Our results describe the general role of the basal process in the self-renewal of neural progenitors and implicate the loss of the apical junctions during oblique divisions as a possible mechanism for generating OSVZ progenitors. We propose that mouse outer VZ progenitors, induced by oblique cleavages, provide a model to study both progenitor self-renewal and OSVZ progenitors.

FAK-mediated extracellular signals are essential for interkinetic nuclear migration and planar divisions in the neuroepithelium

Introduction During the morphogenesis of the neural tube, neural progenitors continuously divide, and post mitotic neurons then migrate to form complex neural networks. It is widely conserved among vertebrates that this mitotic process takes place near the lumen, the apical side of the neural tube (Gotz and Huttner, 2005). When viewed at the cellular level, this apically localized mitosis is the region of output for the cell-cycle-dependent oscillation of the nuclei within the neuroepithelium. Mitotic (M phase) nuclei are located in close proximity to the apical surfaces, whereas nuclei undergoing DNA synthesis (S phase) are displaced more basally. This characteristic movement of nuclei is known as interkinetic nuclear migration (INM) (Fujita, 1964; Sauer, 1935) and is thought to be important for maintaining a neural progenitor pool during neurogenesis (Baye and Link, 2007; Del Bene et al., 2008; Murciano et al., 2002; Xie et al., 2007). Furthermore, planar cell divisions near the apical surface are thought to be important to maintain the progenitor pool (Gotz and Huttner, 2005). Indeed, recent works suggest that most divisions during both proliferative and neurogenic stages are planar (that is, parallel to the apical surface), whereas vertical or oblique divisions are rarely seen, and daughter cells that inherit both the apical and basal components are suggested to remain in the progenitor pool (Konno et al., 2008). Although INM and planar cell divisions have been regarded as hallmarks of vertebrate neural progenitors, their cellular and molecular mechanisms are just beginning to be elucidated. Previous studies have suggested the involvement of several intracellular components such as the actin and microtubule cytoskeletons, molecules involved in adherens junctions, polarity complex and the centrosome (Cappello et al., 2006; Imai et al., 2006; Lien et al., 2006; Murciano et al., 2002; Siller and Doe, 2009; Xie et al., 2007). However, it remains unclear whether extracellular molecules, such as components of the extracellular matrix (ECM), have any role in INM and mitotic orientation of neural progenitors, and if so, how they interact with the intracellular machinery that drives this process. Indeed, the ECM has long been shown to participate in neurogenesis (see below), but there are no reports of vertebrate ECM mutants with defective INM and mitotic orientation. In this context, the medaka mutant tacobo (tab) used in the present study provides a unique framework. The medaka fish, Oryzias latipes, is an emerging vertebrate model and now has a high quality draft genome and a FAK-mediated extracellular signals are essential for interkinetic nuclear migration and planar divisions in the neuroepithelium

Ontogenesis of stomata in Velloziaceae: paracytic versus tetracytic?

In Velloziaceae, the number of subsidiary cells has been used to characterize species and support groups. Nevertheless, the homology of the stomatal types have not been scrutinized. Stomatal ontogenesis of Vellozia epidendroides and V. plicata, assigned to have tetracytic stomata, and of V. glauca and Barbacenia riparia, assigned to have paracytic stomata, were investigated. In the four species studied, stomata followed perigenic development. Subsidiary cells arise from oblique divisions of neighbouring cells of the guard mother cell (GMC). These cells are elongated and parallel to the longer axis of the stoma. Polar cells show wide variation, following the shape and size of the epidermal cells in the vicinity. Hence, these cells cannot be called subsidiary cells. This wide variation is due to a much higher density of stomata in some regions of the leaf blade. This distribution of stomata forces the development of short polar cells, leading to an apparently tetracytic stomata. In regions of low concentration of stomata, higher spatial availability between the GMCs allows the elongation of polar cells, leading to evident paracytic stomata. Therefore, the four studied species are considered braquiparacytic, questioning the classification of stomata into tetracytic and paracytic in Velloziaceae.

Embryology of the dioecious Australian endemic Lomandra longifolia (Lomandraceae)

Microsporogenesis, embryogeny and endosperm development of Lomandra longifolia Labill. are described in detail. The formation of the anther wall is the basic type composed of four cell layers, namely an epidermis, an endothecium, one middle layer and a tapetum. The tapetum layer has glandular, uninucleate cells. Successive cytokinesis follows meiosis, subsequently forming a tetrahedral tetrad of microspores. The ovule in each carpel is hemitropous, crassinucellate and bitegmic, with the micropyle formed by the inner integument. The archesporial cell divides periclinally to form the primary parietal and primary sporogenous cells. The sporogenous cell functions as the megaspore mother cell, whereas the parietal cell divides to give rise to two parietal layers. The mature megagametophyte, which has enlarged synergids and antipodals, is of the Polygonum type, with the normal complement of seven cells and eight nuclei. Nucellar tissue in the mature ovule consists of enlarged dermal cells and irregular subdermal cells surrounding a central strand of markedly smaller cells. Endosperm development is of the nuclear type. Embryo development is of the Graminad type, characterised by oblique zygotic and early pro-embryonic divisions.