Apical Meristems

Dennis Francis, Cardiff University, Cardiff, UK

The cells in the meristem do not all divide at the same rate; the control of the cell cycle in meristems is the subject of current research.

Secondary article
Article Contents
. Apical Meristems . Cell Cycle Control in Meristems

Apical Meristems
The true vegetative shoot apical meristem (SAM) comprises a relatively small population of cells above the youngest primordium varying from as few as 50 cells in Arabidopsis thaliana to over 1000 in Chrysanthemum segetum. This cell population conforms to an apical dome, which can be pronounced in some species (e.g. Lolium temulentum) but attened in others (e.g. Helianthus annuus). Apical dome size varies between species (Table 1); the basal diameter of the dome can vary from 50 to 3000 mm (Mauseth, 1991). However, in the 250 000 angiosperms worldwide, there will probably be some that exhibit larger apical domes than indicated in Table 1. A vegetative SAM changes size constantly. Through cell division, it increases to a threshold size large enough to partition itself into a smaller dome, primordia and subapical tissue (Figure 1). The interval between the initiation of successive primordia is dened as the plastochron. The process will be repeated in a semiiterative fashion because, during vegetative growth, the plastochron tends to lengthen; this, in turn, is reected in dierent growth rates between successive plastochrons (Lyndon, 1977). Notably, the apical dome cells exhibit exponential cell cycle kinetics so that dome growth during successive plastochrons can be expressed as sawtooth plots when temporal increases in cell number are plotted on a loge scale. Vegetative SAMs are described in at least three dierent ways: according to planes of division (Tunica Corpus theory), according to cell fates (L1, L2, L3) or according to cell size/rates of cell division (zonation). The Tunica Corpus theory (Schu epp, 1917) supports the partitioning of cells into domains according to their plane of cell division. In the tunica, cells divide with the plane of chromosome separation at anaphase parallel to the surface
Table 1 Sample of vegetative apical domes ranked in order of increasing size (for a review, see Francis, 1997) Plant Arabidopsis thaliana Helianthus annuus Silene coeli-rosa Chrysanthemum segetum Diameter of vegetative apical dome (mm) 50 70 100 1400

(anticlinal). Typically the tunica is conned to the outer cell layers. In the corpus, cells divide in any plane and this domain is inside the tunica. Most dicotyledonous apical domes exhibit a tunica of two to four layers. When there are two tunica layers they are often described as L1 and L2, with L3 representing the outermost layer of corpus. As mentioned above, leaves are initiated from the apical dome. Fate maps of leaves often indicate plasticity because cells in a particular leaf tissue may be derived from all three layers, but sometimes only from L1 and L2. In other words, cell layers of the apical dome might be best regarded as sources of cells, while fate is imposed on them through positional controls during development (Lyndon, 1998). On the basis of cell size and rates of cell division, dierent regions of the dome can be classied in dierent zones: a central zone (CZ) of large slowly dividing cells, a peripheral zone (PZ) of smaller, faster dividing cells and a pith-rib meristem (PRM) of cells with intermediate size and an intermediate rate of cell division relative to the CZ and PZ ` de, 1967). Leaf primordia arise on the side of the (Nougare dome from cells of the PZ, which, as explained above, can include L1, L2 and L3 layers. In the garden pea, the primordium begins as a cohort of periclinal cell divisions (plane of anaphase separation at 908 to the surface) within the PZ occurring about one-third of the way through a plastochron (Lyndon, 1970). In general terms, leaf initiation is a function of both the plane of cell division in the PZ coupled with changes in the elasticity of the cell walls of the epidermal cells at the point of primordium outgrowth (Green, 1994). However, the primary signal for leaf primordium initiation has yet to be discovered. At the end of each plastochron, part of the PRM is partitioned into the subapical region. Within this region are the progenitor cells for both the internode and node although it is impossible to resolve these tissues at this early stage (Figure 1). Recent studies of mutants of Arabidopsis, which are decient in normal SAM function, have led to the discovery of genes that regulate vegetative development. For example, the shootmeristemless (stm) mutant of Arabidopsis is virtually unable to form a shoot meristem and is regarded as a more severe mutation than the socalled topless (tpl) mutant, which is also unable to form a shoot meristem (Evans and Barton, 1997). SHOOTMERISTEMLESS (STM) encodes a KNOTTED1-type of homeodomain protein, which is rst detected in one to two
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Apical Meristems

Figure 1 Median longitudinal section of a shoot apical meristem of the grass, Dactylis glomerata. A is the apical dome; B is the subapical region containing the new leaf primordium as a bump on the left side adjacent to a central region in which the progenitor cells of the node and internode are located. Bar, 100 mm.

cells of the late globular stage of embryogenesis. First discovered in maize, mutations in KN1 result in knots of tissue forming around veins of the leaf blades (reviewed by Evans and Barton, 1997). A consensus view is that STM is a transcription factor regulating the expression of other genes, including TPL. Another target gene is WUCHSEL, which is required for cell identity in the centre of the SAM. This gene cascade is essential for normal SAM morphogenesis (Evans and Barton, 1997). STM also has important interactions with CLAVATA1(CLV1), the expression of which can regulate the number of cell layers within the meristem. One idea was that STM:CLV1 could regulate cell fates in the meristem (Laux and Shoof, 1997). Root apical meristems are located in the tips of roots and can comprise between 125 000 and 250 000 cells spanning approximately 2 mm of apical tissue. Root apical meristems exhibit specic tissue domains. At the centre is the central stele, surrounded by the cortex, which is in turn surrounded by the epidermal layer. Classically, these three layers correspond to Hansteins (1870) histogens: the periblem, plerome and dermatogen, which were perceived as discrete domains of founder cells for each tissue. These terms were originally applied to shoot apices but the plerome and periblem are not easily distinguishable in SAMs. Strictly speaking, the RAM is subapical in that it is protected by a root cap, thus invoking Hansteins fourth histogen, the calypterogen. Hence, the RAM gives rise to cells of the various tissue domains, including the root cap. The histogens were shown to be less specic in terms of
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tissue-organizing ability by F. A. L. Clowes at Oxford, who predicted, and subsequently discovered, a population of cells at the pole of the various tissue domains that divided much more slowly than surrounding cells (Clowes, 1954, 1956). This was the quiescent centre (QC), which led Clowes to develop a theory of root growth based on apical initials displaced from the QC by cell division, which then surround the quiescent centre. Central to this theory was that the apical initials themselves have no set fate but give rise to descendants, which then establish the tissue domains (Clowes, 1967). The cells of the QC are stimulated into more rapid cell division by decapping, tissue damage or by radiation leading to regeneration of the lost parts (Clowes, 1970). Hence, QC cells would conform to a population of founder cells (Barlow, 1997). More recently, Scheres and colleagues in Utrecht, using laser ablation on roots of Arabidopsis, have demonstrated that QC cells exert a negative regulation of cell dierentiation on surface contacting apical initials. These observations conformed to a hypothesis whereby QC cells repress the dierentiation of contacting apical initials, enabling the latter to divide (van den Berg et al., 1997). Clearly, the frequency of cell division is highest in the RAM and SAM of the plant. At the margins of the RAM, cells are left behind to elongate and dierentiate. Thus, at this boundary region, the meristem is exhibiting steadystate cell cycle kinetics, where a cell division at the periphery of the meristem results in one cell elongating while the other remains part of the proliferative pool. This

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Apical Meristems

helps to explain how the RAM remains fairly constant in size during root growth, whereas the SAM is constantly changing size (see above).

Cell Cycle Control in Meristems

Clearly, there are many cell divisions that occur in meristems during the development of the plant. However, not all meristem cells divide at the same time. Indeed, there is evidence showing that substantial proportions of cells are nondividing in RAMs (e.g. cells within the quiescent centre see above) and are scattered throughout SAMs (Gonthier et al., 1987; Kinsman et al., 1997). Cells achieve competence for cell division during the cell cycle, which consists of four distinct phases: G1 (postmitotic interphase), S phase (period of DNA synthesis), G2 (postsynthetic interphase) and mitosis (M). Major checkpoints of the eukaryote cell cycle, at G1/S and G2/M, are governed by the expression of cell division cycle (cdc) genes or the closely related cyclin-dependent kinase (cdk) genes. At G2/M a protein kinase encoded by cdc2 binds to a cyclin (cyclins A/B), so called because its concentration reaches a peak at a specic point in the cell cycle. This binding contributes to the activation of the Cdc2 kinase, which drives the cell into mitosis (Norbury and Nurse, 1992). In vertebrates, another class of cyclins, the D types, are critical for the entry of cells from a noncycling condition (G0) into the cell cycle. In the presence of serum growth factor, cyclin D1 is upregulated, which in turn leads to transcriptional activation of cyclins D2 and D3 (Scherr, 1996). Homologues to these D cyclins have been cloned in Arabidopsis. The so-called cycD2At is upregulated by sucrose whereas cycD3At is activated by cytokinin, provided that sucrose is present (Soni et al., 1995). It seems likely that this is a major control regulating the proportion of cycling cells in meristems. Remarkably, the overexpression of cycD3At in tobacco led to a more rapid growth rate in these transgenic plants, which is consistent with the idea that the number of dividing cells in meristems has an impact on plant growth (Cockcroft et al., 2000). In Arabidopsis, cdc2b is expressed at the G2/M transition together with cyclin B and cyclin A. The expression of all of these genes is essential, but exactly what cyclin binds to which Cdk is a subject of current research (Mironov et al., 1999). Working on Arabidopsis, Inze and his colleagues at Ghent spliced the promoter of cdc2 to the b-glucuronidase (GUS) reporter gene and studied the spatial expression of the gene during root development. The GUS staining patterns obtained were consistent with high Cdk activity in the RAM but also in the pericycle, the outermost layer of the stele from which laterals arise (Hemerly et al., 1993). These observations were consistent in revealing that cdc2 expression was indicative of division competence. Very recent work from Inzes laboratory has indicated that

lateral root initiation requires the expression of a newly discovered cyclin, cycd4At (De Veylder et al., 1999). SAMs exhibit gradients of cell division, slowest in the CZ, fastest in the PZ and intermediate in the PRM. For example, in Chrysanthemum at 208C, cell cycle times for the CZ, PZ and PRM were 139, 48 and 70 h, respectively ` de, 1977). Clearly, Cdk activity is (Rembur and Nougare high in SAMs but the mechanism for spatial regulation of cell division is unknown. However, it is anticipated that there will be links between genes that regulate development and those that control the cell cycle. One clue in this direction was the nding that a gene that is essential for megagametophyte and embryo development, PROLIFERA, encodes a minichromosome maintenance (MCM)like protein; MCMs are important regulators of DNA replication (Springer et al., 1995). Integration of the data is shedding light on the way that extracellular growth factors impinge on the cell cycle. For example, gibberellic acid treatment can lead to the upregulation of Cdc2 in rice and cytokinins can inuence the activation of Cdc2 in the garden pea (for a review, see Francis and Sorrell, 2000). Over the next few years it is anticipated that Cdkcyclin interactions will be veried in plants and that signal transduction chains mediated by plant growth regulators that interface with cell cycle control will be resolved, as will some of the links between developmental genes and cell cycle genes. Integration of the data on such interactions will be important in understanding cell cycle regulation in meristems during higher plant development.

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Further Reading
Francis D, Dudits D and Inze D (1998) Plant Cell Division. London: Portland Press. Lyndon RF (1998) The Shoot Apical Meristem. Cambridge: Cambridge University Press. Meyerowitz EM (1997) Genetic control of cell division patterns in developing plants. Cell 88: 299308. Taiz L and Zeiger E (1998) Growth development and dierentiation. In: Plant Physiology, 2nd edn, chap. 16. Sunderland, MA: Sinauer Associates. Wolpert L (1998) Plant development. In: Principles of Development, chap. 7. Oxford: Oxford University Press.

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