Changes in cell length and mitotic index in vascular pattern-related pericycle cell types along the apical meristem and elongation zone of the onion root

Summary Three pericycle cell types (opposite xylem, opposite phloem and intervening) distinguished by their location in relation to different elements of the vascular system were studied in the adventitious root ofAllium cepa L. Changes in cell length and mitotic index were analysed in these cells along the apical meristem and elongation zone of the root. The opposite phloem and intervening pericycle cells are significantly shorter than the opposite xylem pericycle cells in the apical half of the meristem. Between 1,200 and 1,400 m behind the tip, length became similar in all three pericycle cell types, while in more proximal zones the opposite phloem cells were significantly longer. These results suggest that the number of transverse divisions is different in the three types of pericycle cells. In the apical half of the meristem, mitotic index increased in intervening and opposite xylem cells but remained unchanged in opposite phloem cells, a fact likely to account for the relative lengthening of the latter. In the proximal half of the meristem, mitotic index fell in all three cell types until cell division had ceased. However, mitotic index in opposite xylem cells remained high for longer than in the other two cell types, implying that increase of the mean cell length in the former was slower. These results suggest that differences in mean cell length between the three pericycle cell types are due to different rates of proliferation.     

Pre-prophase bands of microtubules in all categories of formative and proliferative cell division in Azolla roots

Pre-prophase bands of microtubules were found in every category of cell division, symmetrical and asymmetrical, in the cell lineages of the root apex of Azolla pinnata R.Br. and A. filiculoides Lam., and in the transverse divisions in the cell files of the roots. They are also found in the asymmetrical cell division that gives rise to trichoblasts in roots of Hydrocharis dubia (B1). Backer. It is possible, in a variety of cell types in roots of Azolla, to predict within a fraction of a micrometre where a new cell wall will be located. In every such case the midline of the 1.5–3-m-wide pre-prophase band anticipates this location. Each of the daughter cells thus inherits approximately half of the former pre-prophase band site. Images interpreted as stages of formation of the band were obtained, its microtubules replacing the interphase cortical arrays. In one highly asymmetrical division, band formation precedes migration of the nucleus to the site of mitosis. The asymmetrical division that gives rise to root hairs passes acropetally along every cell in the dermatogen layer, and preprophase bands were seen up to 8 cells in advance of the last completed division. Here, and in the zone of formative divisions, the band is present for much longer than the duration of mitosis. The ubiquity of the band in the Azolla root tip is discussed in relation to the literature, and a working hypothesis is presented that takes into account current knowledge of occurrence, development and function of the band.     

Autoreproductive cells and plant meristem construction: The case of the tomato cap meristem

Summary Root apical meristems are composed of two zones in which either formative or proliferative cell divisions occur. Within the formative zone, autoreproductive initial cells (a-cells) occupy distinctive locations. By means of graph-L-systems, the behavior of one such type of a-cells has been investigated, with particular reference to root caps within the developing primordia of lateral roots ofLycopersicon esculentum cultivated in vitro. Here, the a-cells constitute the protoderm initials, cells which are found also in the root cap of many angiosperm species. A set of cuboidal (i.e., six-sided) acells develops early in the ontogeny of a lateral-root primordium. Then, according to both anatomical observations and theoretical simulations obtained by the application of graph-L-systems, sequential production of descendents from each a-cell leads to the formation of a new autoreproductive cell (a), a cap columella initial (c), and two mother cells (e and f) whose respective descendents differentiate as root epidermis and cap flank cells. In this graph-L-system, there is specification of the location of sister cells with respect to the three orthogonal directions of a cuboidal. In the early stage of root cap formation, only a few rounds of these formative cell divisions by each a-cell and its four types of descendents are required to provide the basic set of cells necessary for full cap development. After the lateral root emerges from the parent root, there may be a temporary cessation of the formative divisions of the a-cells which give rise to columella initials. Columella production is then supported entirely by its own independent set of autoreproductive c-initials. At the same time, division of the autoreproductive protoderm initial cell is directed towards maintaining the cap flank and the epidermal cell files. The regulation of the types of formative division by the a-cell may be represented by means of a division counter which may be specific for a given species.Dedicated to Professor Brian E. S. Gunning on the occasion of his 65th birthday     

Developmental anatomy of the fifth shoot-borne root in young sporophytes of<Emphasis Type="Italic"> Ceratopteris richardii</Emphasis>

Young sporophytes of the homosporous fern Ceratopteris richardii produce a single shoot-borne root below each leaf. The developmental anatomy of the fifth sporophyte root is described using scanning electron microscopy and histological techniques. Three merophyte orthostichies in the body of the root originate from three proximal division faces of a tetrahedral root apical cell. Eight or nine divisions occur in a relatively regular sequence within each merophyte and produce a characteristic radial anatomical pattern in the root. The exact number of early divisions within a merophyte depends on the merophytes position within the root as a whole. Predictable inter-merophyte differences arise because a 2-fold (diarch) anatomical symmetry that is characteristic of mature roots is superimposed on a 3-fold radial symmetry that originates behind the apical cell. Before early formative divisions within a merophyte are completed, additional proliferative divisions begin to increase the number of cells within previously established tissue zones. The cellular parameters of early fifth root development in C. richardii are relatively invariant, and are reminiscent of patterns previously described for the heterosporous fern Azolla. Young sporophytes of C. richardii provide a useful model to further investigate the genetic regulation of root development in a non-seed plant, where the anatomy of meristem organization differs from that seen in flowering plant species.Abbreviations SEM Scanning electron microscopy - RAC Root apical cell     

Age-related and origin-related control of the numbers of plasmodesmata in cell walls of developing Azolla roots

Plasmodesmata were counted in the longitudinal and transverse walls in developmental sequences of merophytes in roots of Azolla pinnata R.Br. The differences between certain categories of longitudinal wall were traced to factors that govern the surface area of the cell plates, the density of plasmodesmata (number per unit area of cell plate), and the amount by which each type of plate expands. No evidence for secondary augmentation of plasmodesmatal numbers after the cell-plate stage of development was found, but plasmodesmata are lost from the walls of sieve and xylem elements during their differentiation. Losses caused by cell separation occur in other tissues. The relatively high density of plasmodesmata in transverse walls is based not so much on a high density in the cell plates as on the relatively low expansion that these walls undergo. There appears to be a compensatory mechanism that relates initial plasmodesmatal density to the future expansion of the cell plate. The root shows determinate growth, the apical cell dividing about 55 times. Beginning at about the 35th division there is a progressive failure to maintain the plasmodesmatal frequencies that were developed in earlier cell divisions in the apical cell. The divisions that occur within the later-produced merophytes also show progressive diminution of plasmodesmatal numbers. The result is that the apex of the root, and particularly the apical cell, becomes more and more isolated symplastically, a phenomenon which could account for its limited lifespan and the determinate growth pattern of the root.     

Cell division patterns of the protoderm and root cap in the “closed” root apical meristem ofArabidopsis thaliana

Summary The peripheral root cap and protoderm inArabidopsis thaliana are organized into modular packets of cells derived from formative T-divisions of the root cap/protoderm (RCP) initials and subsequent proliferative divisions of their daughter cells. Each module consists of protoderm and peripheral root cap packets derived from the same periclinal T-division event of an RCP initial. Anatomical analyses are used to interpret the history of extensively coordinated cell divisions producing this modular construction. Within a given layer of root cap, the columella and RCP initials divided in a centrifugal sequence from the innermost columella initials toward the RCP initials. All RCP initials in the lineages around the circumference of the root divided nearly simultaneously in waves to form one module prior to the next wave of initial divisions forming a younger module. The peripheral root cap and protoderm packets within each module completed four rounds of proliferative divisions in the axial plane to produce, on average, 16 cells per packet in the basalmost modules in axial view. Peripheral root cap and protoderm cells predominantly in the T-type (trichoblast) lineages also underwent radial divisions as they were displaced basipetally. The regularity in the cellular pattern within the modules suggests a timing mechanism controlling highly coordinated cell division in the initials and their daughter cells.Abbreviations RAM root apical meristem - RCP root cap protoderm - prc peripheral root cap     

Timing cell-fate determination during asymmetric cell divisions

From invertebrates to mammals, cell-cycle progression during an asymmetric cell division is accompanied by precisely timed redistribution of cell-fate determinants so that they segregate asymmetrically to enable the two daughter cells to choose different fates. Interestingly, studies on how cell fates are specified in such divisions reveal that the same fate determinants can be reiteratively used to specify a variety of cell types through multiple rounds of cell divisions or to exert seemingly contradictory effects on cell proliferation and differentiation. Here I summarize the molecular mechanisms governing asymmetric cell division and review recent findings pointing to a novel mechanism for coupling intracellular signaling and cell-cycle progression. This mechanism uses changes in the morphology, subcellular distribution, and molecular composition of cellular organelles like the Golgi apparatus and centrosomes, which not only accompany the progression of cell cycle to activate but also temporally constrain the activity of fate determinants during asymmetric cell divisions.     

Cell division and cell enlargement in isolated Cucurbita cotyledons grown in darkness and in light

The spatial and temporal patterns of post-embryonal cell growth and cell division were characterised in excised cotyledons of vegetable marrow (Cucurbita pepo L. var. giromontia Alef.) incubated in water. The concurrent roles of these two processes in cotyledon growth were determined using paradermal sections of the first palisade layer of developing cotyledons. Tissue specificity was observed in the pattern of cell division. The daughter cells derived from an initial cell, which had already differentiated before imbibition of the seeds, were tightly packed in a cluster, which enabled us to monitor cell division during early cotyledon development. Heterogeneity of cell size was recognised during the process of cell proliferation in the cluster, suggesting that cell division is uncoupled from control of cell size. There was significantly more cell division in the marginal part of the cotyledons than in other parts, suggesting high activity of the marginal meristem. Light enhanced cell and cotyledon enlargement, but had no effect on the number of divisions. This study elucidated the cellular basis of post-germinative Cucurbita cotyledon morphogenesis and development. Electronic Publication     

Asymmetric cell division and cell fate in plants

A variety of approaches has recently been employed to investigate how sister cells adopt distinct fates following asymmetric divisions during plant development. Surgical and drug studies have been used to analyze asymmetric divisions during both early embryogenesis in brown algae and pollen development in tobacco. Genetic screens have been used to identify genes in Arabidopsis thaliana that are required for specific asymmetric cell divisions during pollen and root development. These studies indicate that cell polarity and division orientation are closely tied to the process of cell fate specification, and suggest that differential inheritance of determinants and positional information may both be involved in the specification of cell fates following asymmetric cell division.     

Root apical organization inArabidopsis thaliana ecotype ‘WS’ and a comment on root cap structure

Arabidopsis thaliana roots have closed apical organization with three initial tiers. The dermatogen/calyptrogen tier consists of two parts-the central initials form the columella root cap, and the peripheral initial cells form the protoderm (epidermis) and the peripheral root cap. These peripheral initials divide in a sequence to form a root cap consisting of interconnected cones. the periblem initial tier forms the ground meristem (cortex). For the first week after germination the periblem consists of one layer of initial cells. The peripheral cells of the tier divide periclinally and then anticlinally (a T-division) to form the two-layered cortex (outer cortex and endodermis). After about one week, all the peripheral cells have divided periclinally forming two initials; the outermost produces the outer cortex while the inner initial produces the endodermis and middle cortex layer. The latter two cells arise via a periclinal division. During this time, other cells within the tier divide periclinally to form a two-layered tier. The plerome forms the cells of the procambium (vascular cylinder) by simple anticlinal divisions followed by longitudinal divisions to fill out the cell files of the vascular cylinder. A survey (27 dicot species in 17 families) of roots with closed apical organization revealed that there are three different types of root cap-concentric cylinders of cells (e.g.Linum), interconnecting cones (e.g.Arabidopsis) or overlapping arcs (e.g.Gossypium). H Lambers Section editor     

TheCaesalpinia peltophoroides cell cycle under different aeration conditions

Summary Cell cycle parameters were studied inCaesalpinia peltophoroides meristems proliferating under different oxygen tensions. This species has been selected for mixed planting in degraded areas in Brazil, some of which are occasionally flooded. As the species’ adaptation to oxygen deprivation during flooding is not fully understood, the objective of this study was to characterize the meristematic activity of root cells under various oxygen regimes. Synchronous binucleate cells, induced by a pulse of caffeine, showed a cell-cycle time constant under both control (5.6 mg of O₂ per l) and oxygenated conditions (7.9 and 3.2 mg of O₂ per l). The whole cell cycle lasted 10 h, although the relative duration of metaphase and anaphase/early telophase increased in more hypoxic conditions. The species appeared to utilise oxygen diffusing from the shoot to the root system to maintain cell division and root growth.     

The simulation model of growth and cell divisions for the root apex with an apical cell in application to Azolla pinnata

In contrast to seed plants, the roots of most ferns have a single apical cell which is the ultimate source of all cells in the root. The apical cell has a tetrahedral shape and divides asymmetrically. The root cap derives from the distal division face, while merophytes derived from three proximal division faces contribute to the root proper. The merophytes are produced sequentially forming three sectors along a helix around the root axis. During development, they divide and differentiate in a predictable pattern. Such growth causes cell pattern of the root apex to be remarkably regular and self-perpetuating. The nature of this regularity remains unknown. This paper shows the 2D simulation model for growth of the root apex with the apical cell in application to Azolla pinnata. The field of growth rates of the organ, prescribed by the model, is of a tensor type (symplastic growth) and cells divide taking principal growth directions into account. The simulations show how the cell pattern in a longitudinal section of the apex develops in time. The virtual root apex grows realistically and its cell pattern is similar to that observed in anatomical sections. The simulations indicate that the cell pattern regularity results from cell divisions which are oriented with respect to principal growth directions. Such divisions are essential for maintenance of peri-anticlinal arrangement of cell walls and coordinated growth of merophytes during the development. The highly specific division program that takes place in merophytes prior to differentiation seems to be regulated at the cellular level.     

Arabidopsis MPK6 is involved in cell division plane control during early root development,and localizes to the pre‐prophase band,phragmoplast, trans‐Golgi network and plasma membrane

The proper spatial and temporal expression and localization of mitogen‐activated protein kinases (MAPKs) is essential for developmental and cellular signalling in all eukaryotes. Here, we analysed expression, subcellular localization and function of MPK6 in roots of Arabidopsis thaliana using wild‐type plants and three mpk6 knock‐out mutant lines. The MPK6 promoter showed two expression maxima in the most apical part of the root meristem and in the root transition zone. This expression pattern was highly consistent with ‘no root’ and ‘short root’ phenotypes, as well as with ectopic cell divisions and aberrant cell division planes, resulting in disordered cell files in the roots of these mpk6 knock‐out mutants. In dividing root cells, MPK6 was localized on the subcellular level to distinct fine spots in the pre‐prophase band and phragmoplast, representing the two most important cytoskeletal structures controlling the cell division plane. By combining subcellular fractionation and microscopic in situ and in vivo co‐localization methods, MPK6 was localized to the plasma membrane (PM) and the trans‐Golgi network (TGN). In summary, these data suggest that MPK6 localizing to mitotic microtubules, secretory TGN vesicles and the PM is involved in cell division plane control and root development in Arabidopsis.     

Cell proliferation in ectodermal explants from Xenopus embryos

Summary Dissociated prospective ectoderm cells from Xenopus laevis embryos divide autonomously up to the 17th division cycle of the embryo. To examine the requirements for the further proliferation of these cells, the continuation of cell division in compact ectodermal explants beyond the 17th division cycle has been studied. Such explants develop into aggregates of epidermal cells, as can be shown immunohistochemically with an anti-serum against Xenopus epidermal cytokeratin. Cell division in these explants is comparable to the in vivo proliferation rate at least during the first 24 h of cultivation, that is, well beyond the 17th division cycle. Thus, epidermal cells are provided with all the factors necessary for continued proliferation, but these can be effective only when the cells form tight aggregates. The long-term changes in cell number are complex. Mitotic figures are present until the explants disintegrate after 3–4 days. However, the total cell number per explant does not increase during later development. The production of cells by mitotic divisions is likely to be countered by the loss of cells due to cell death, which is indicated by the presence of pyknotic nuclei.     

Division,growth, and branch formation in protonema of the mossPhyscomitrium turbinatum: Studies of sequential cytological changes in living cells

Summary Elongating caulonemal tip cells ofPhyscomitrium turbinatum were cultivated on mediumcoated cover slips and periodically observed with Nomarski differential interference contrast optics. Tip cells exhibit apical growth and an average growth rate of 27.5 m/h. During cell elongation the nucleus migrates forward in the tip cell, but this movement slowly decreases so that there is a gradual increase in the distance between the nucleus and cell tip. Minimum length cells contain small vacuoles adjacent to the basal wall which coalesce during subsequent cell elongation to form a solitary large basal vacuole.An increase in chloroplasts during cell elongation is due to the presence of a population of proliferating chloroplasts located between the cell tip and the nucleus resulting in a gradient in chloroplast number and shape. The zone of chloroplast proliferation shifts progressively forward during cell elongation from a peri-nuclear position to a region closer to the cell tip. During division of the apical cell a perpendicular metaphase plate is formed. Reorientation movements of the phragmoplast-cell plate during telophase, and early stages of the following interphase produce a 35–40° cross wall. This rotation of the spindle axis positions the daughter nuclei temporarily adjacent to the lateral walls on opposite sides of the cell with the sub-apical nucleus on the side nearest the light source. It subsequently migrates across the cell to become situated on the wall farthest from the light source. Sub-apical cells form branches at the distal (= apical) end of the cell on the lateral wall closest to the light source. Branch development is accompanied by changes in chloroplast shape, number, and position.     

Histochemical analysis of root meristem activity in Arabidopsis thaliana using a cyclin:GUS (β-glucuronidase) marker line

We used a transgenic Arabidopsis line expressing a translational fusion between a mitotic cyclin and the reporter gene -glucuronidase (GUS) to investigate cell divisions in postembryonic root meristems. The fusion protein contains the cyclin destruction box (CDB) and this leads to a rapid degradation of the chimeric GUS-protein after mitosis. Hence, the staining pattern of the meristem marks dividing cells. We observed that upon germination the first cell divisions occur in epidermis cells at the junction with the hypocotyl. Moreover, the accelerated root growth on media supplemented with sucrose correlates with an increased number of dividing cells and an enlargement of the root meristematic zone. The conditional root expansion mutants pom pom1 and procuste1 (quill) suppress this sugar effect leading to a smaller meristematic zone. Simultaneous visualisation of the nucleus revealed that the CYCAT1:CDB:GUS expression is subcellularly localised around the nucleus. This particular staining starts at prophase and disappears after the completion of the new cell wall. In metaphase the staining invades the cytoplasm whereas in the telophase it concentrates again around the nucleus. This cell cycle-dependent distribution was used to characterise the two root specific cytokinesis mutants pleiade1 and hyade1. In both mutants, cells which fail to develop a complete cell wall during cytokinesis divide synchronously in further cell divisions leading to multinucleate cells. These experiments demonstrate the usefulness of the CYCAT1:CDB:GUS marker line for studying cell division of wild-type and mutants. Furthermore, this line can be used to analyse the influence of biotic and abiotic signals on the rate and spatial distribution of cell divisions.     

Rhythmic plant morphogenesis: Recurrent patterns of idioblast cell production

Most plants are constructed from repeating modular units such as phytomers, merophytes, and cell packets. Even an organism as simple as the filamentous cyanobacterium Anabaena shows recurrent patterns of differentiated cellular structures, notably with respect to its heterocysts. These examples reflect the inherent rhythms established within developmental processes of living organisms. In the present article, attention is paid to repetitious production of idioblasts—isolated cells, or clusters of cells, with an identity different to that of neighbouring cells from which they are derived. In higher plant root tissues, idioblasts are contained within cell packets that grow up from mother cells during the course of a number of cycles of cell production. The heterocysts of Anabaena are also discussed; they, too, are a type of idioblast. The idioblasts of root tissues originate as small cells which result from unequal cell divisions. Such divisions are usually the final ones within a cell packet which has already undergone a number of division cycles and are characteristically located at one or both ends of a packet. The packet end walls are suggested to have a role in regulating division asymmetry. Idioblastic systems discussed are root cortical trichosclereids and diaphragm cells; in their earliest stage, the cells from which lateral root primordia arise are also considered as clusters of idioblasts because they, too, are the products of asymmetric divisions of pericyclic mother cells. The division patterns of all these idioblastic systems were modelled in a consistent way using L-systems, with the assumption that the age of a cell-packet end wall plays a special role in cell determination. This article is dedicated to Vsevelod Ya. Brodsky, doyen of Russian studies of rhythms in cell division and development, who celebrates his 80th birthday on August 4, 2008 This article was presented in original.     

Studies on the behavior of organelles and their nucleoids in the root apical meristem of Arabidopsis thaliana (L.) Col.

The behavior of cell nuclei, mitochondrial nucleoids (mt-nucleoids) and plastid nucleoids (ptnucleoids) was studied in the root apical meristem of Arabidopsis thaliana. Samples were embedded in Technovit 7100 resin, cut into thin sections and stained with 4′-6-diamidino-2-phenylindole for light-microscopic autoradiography and microphotometry. Synthesis of cell nuclear DNA and cell division were both active in the root apical meristem between 0 μm and 300 μm from the central cells. It is estimated that the cells generated in the lower part of the root apical meristem enter the elongation zone after at least four divisions. Throughout the entire meristematic zone, individual cells had mitochondria which contained 1–5 mt-nucleoids. The number of mitochondria increased gradually from 65 to 200 in the meristem of the central cylinder. Therefore, throughout the meristem, individual mitochondria divided either once or twice per mitotic cycle. By contrast, based on the incorporation of [³H]thymidine into organelle nucleoids, syntheses of mitochondrial DNA (mtDNA) and plastid DNA (ptDNA) occurred independently of the mitotic cycle and mainly in a restricted region (i.e., the lower part of the root apical meristem). Fluorimetry, using a videointensified microscope photon-counting system, revealed that the amount of mtDNA per mt-nucleoid in the cells in the lower part of the meristem, where mtDNA synthesis was active, corresponded to more than 1 Mbp. By contrast, in the meristematic cells just below the elongation zone of the root tip, the amount of mtDNA per mt-nucleoid fell to approximately 170 kbp. These findings strongly indicate that the amount of mtDNA per mitochondrion, which has been synthesized in the lower part of the meristem, is gradually reduced as a result of continual mitochondrial divisions during low levels of mtDNA synthesis. This phenomenon would explain why differentiated cells in the elongation zone have mitochondria that contain only extremely small amounts of mtDNA. This work was supported by a Grant-in Aid (T.K.) for Special Research on Priority Areas (Project No. 02242102, Cellular and Molecular Basis for Reproduction Processes in Plants) from the Ministry of Education, Science and Culture of Japan and by a Grant-in Aid (T.K.) for Original and Creative Research Project on Biotechnology from the Research Council, Ministry of Agriculture, Forestry and Fisheries of Japan.