The plant cell cycle in context

Biological scientists are eagerly confronting the challenge of understanding the regulatory mechanisms that control the cell division cycle in eukaryotes. New information will have major implications for the treatment of growth-related diseases and cancer in animals. In plants, cell division has a key role in root and shoot growth as well as in the development of vegetative storage organs and reproductive tissues such as flowers and seeds. Many of the strategies for crop improvement, especially those aimed at increasing yield, involve the manipulation of cell division. This review describes, in some detail, the current status of our understanding of the regulation of cell division in eukaryotes and especially in plants. It also features an outline of some preliminary attempts to exploit transgenesis for manipulation of plant cell division.     

Opportunities for regulation of sugar beet storage root growth

The percentage of sucrose in sugar beet storage root fresh and dry matter is closely related to root structure. It has been suggested that the sucrose content might be increased by using plant growth regulators to modify storage root structure through control of cambial development, cell division and cell expansion. During storage root development correlations were found between the changing phytohormone profiles and the formation of secondary cambia and their subsequent cell division and expansion. Sugar beet root derived cell suspension cultures were used for detailed studies of the roles of endogenous phytohormones. The gibberellin synthesis inhibitor paclobutrazol was tested in cell cultures and whole plants. The observations provide a basis for development of plant growth regulator regimes to optimise sucrose yield from sugar beet.     

Strigolactone Positively Controls Crown Root Elongation in Rice

Strigolactones are recently identified plant hormones that inhibit shoot branching. Pleiotropic defects in strigolactone-deficient or -insensitive mutants indicate that strigolactones control various aspects of plant growth and development. However, our understanding of the hormonal function of strigolactones in plants is very limited. In this study we demonstrate that rice dwarf mutants that are strigolactone-deficient or -insensitive exhibit a short crown root phenotype. Exogenous application of GR24, a synthetic strigolactone analog, complemented the crown root defect in strigolactone-deficient mutants but not in strigolactone-insensitive mutants. These observations imply that strigolactones positively regulate the length of crown roots. Histological observations revealed that the meristematic zone is shorter in dwarf mutants than in wild type, suggesting that strigolactones may exert their effect on roots via the control of cell division. We also show that crown roots of wild type, but not dwarf mutants, become longer under phosphate starvation.     

The dynamic plant stem cell niches

Stem cells exist in specific locations called niches, where extracellular signals maintain stem cell division and prevent differentiation. In plants, the best characterised niches are within the shoot and root meristems. Networks of regulatory genes and intercellular signals maintain meristem structure in spite of constant cell displacement by division. Recent works have improved our understanding of how these networks function at the cellular and molecular levels, particularly in the control of the stem cell population in the shoot meristem. The meristem regulatory genes have been found to function partly through localised control of widely used signals such as cytokinin and auxin. The retinoblastoma protein has also emerged as a key regulator of cell differentiation in the meristems.     

Stem cells: The root of all cells

The plant basic body plan is laid down during embryogenesis. All post-embryonic development has its origin in the stem cells located in niches in the heart of the shoot and root meristems. Creating the root niche requires auxin dependent patterning cues that provide positional information in combination with parallel inputs to specify and maintain the root stem cell niche from embryogenesis onwards. Once established, the architecture of the root niche differs from that in the shoot but recent findings reveal a conserved module for stem cell control. Important for stem cell maintenance is the balance between cell division and differentiation. Dealing with the environment is the biggest challenge for plants and that includes complete regeneration of stem cell systems upon damage. Here we will address these issues as we follow the formation, function and maintenance of the root stem cell niche during development.     

Emerging role of cytokinin as a regulator of cellular differentiation

Perhaps the most amazing feature of plants is their ability to grow and regenerate for years, sometimes even centuries. This fascinating characteristic is achieved thanks to the activity of stem cells, which reside in the shoot and root apical meristems. Stem cells function as a reserve of undifferentiated cells to replace organs and sustain postembryonic plant growth. To maintain meristem function, stem cells have to generate new cells at a rate similar to that of cells leaving the meristem and differentiating, thus achieving a balance between cell division and cell differentiation. Recent findings have improved our knowledge on the molecular mechanisms necessary to establish this balance and reveal a fundamental signaling role for the plant hormone cytokinin. Evidence has been provided to show that in the root meristem cytokinin acts in defined developmental domains to control cell differentiation rate, thus controlling root meristem size.     

The organization of roots of dicotyledonous plants and the positions of control points

BACKGROUND: The structure of roots has been studied for many years, but despite their importance to the growth and well-being of plants, most researchers tend to ignore them. This is unfortunate, because their simple body plan makes it possible to study complex developmental pathways without the complications sometimes found in the shoot. In this illustrated essay, my objective is to describe the body plan of the root and the root apical meristem (RAM) and point out the control points where differentiation and cell cycle decisions are made. Hopefully this outline will assist plant biologists in identifying the structural context for their observations. SCOPE AND CONCLUSIONS: This short paper outlines the types of RAM, i.e. basic-open, intermediate-open and closed, shows how they are similar and different, and makes the point that the structure and shape of the RAM are not static, but changes in shape, size and organization occur depending on root growth rate and development stage. RAMs with a closed organization lose their outer root cap layers in sheets of dead cells, while those with an open organization release living border cells from the outer surfaces of the root cap. This observation suggests a possible difference in the mechanisms whereby roots with different RAM types communicate with soil-borne micro-organisms. The root body is organized in cylinders, sectors (xylem and phloem in the vascular cylinder), cell files, packets and modules, and individual cells. The differentiation in these root development units is regulated at control points where genetic regulation is needed, and the location of these tissue-specific control points can be modulated as a function of root growth rate. In Arabidopsis thaliana the epidermis and peripheral root cap develop through a highly regulated series of steps starting with a periclinal division of an initial cell, the root cap/protoderm (RCP) initial. The derivative cells from the RCP initial divide into two cells, the inner cell divides again to renew the RCP and the other cell divides through four cycles to form 16 epidermal cells in a packet; the outer cell divides through four cycles to form the 16 cells making up the peripheral root cap packet. Together, the epidermal packet and the peripheral root cap packet make up a module of cells which are clonally related.     

dhm1, an Arabidopsis mutant with increased sensitivity to alkamides shows tumorous shoot development and enhanced lateral root formation

The control of cell division by growth regulators is critical to proper shoot and root development. Alkamides belong to a class of small lipid amides involved in plant morphogenetic processes, from which N-isobutyl decanamide is one of the most active compounds identified. This work describes the isolation and characterization of an N-isobutyl decanamide-hypersensitive (dhm1) mutant of Arabidopsis (Arabidopsis thaliana). dhm1 seedlings grown in vitro develop disorganized tumorous tissue in petioles, leaves and stems. N-isobutyl decanamide treatment exacerbates the dhm1 phenotype resulting in widespread production of callus-like structures in the mutant. Together with these morphological alterations in shoot, dhm1 seedlings sustained increased lateral root formation and greater sensitivity to alkamides in the inhibition of primary root growth. The mutants also show reduced etiolation when grown in darkness. When grown in soil, adult dhm1 plants were characterized by reduced plant size, and decreased fertility. Genetic analysis indicated that the mutant phenotype segregates as a single recessive Mendelian trait. Developmental alterations in dhm1 were related to an enhanced expression of the cell division marker CycB1-uidA both in the shoot and root system, which correlated with altered expression of auxin and cytokinin responsive gene markers. Pharmacological inhibition of auxin transport decreased LR formation in WT and dhm1 seedlings in a similar manner, indicating that auxin transport is involved in the dhm1 root phenotype. These data show an important role of alkamide signaling in cell proliferation and plant architecture remodeling likely acting through the DHM1 protein.     

Cytokinin-deficient transgenic Arabidopsis plants show multiple developmental alterations indicating opposite functions of cytokinins in the regulation of shoot and root meristem activity

Cytokinins are hormones that regulate cell division and development. As a result of a lack of specific mutants and biochemical tools, it has not been possible to study the consequences of cytokinin deficiency. Cytokinin-deficient plants are expected to yield information about processes in which cytokinins are limiting and that, therefore, they might regulate. We have engineered transgenic Arabidopsis plants that overexpress individually six different members of the cytokinin oxidase/dehydrogenase (AtCKX) gene family and have undertaken a detailed phenotypic analysis. Transgenic plants had increased cytokinin breakdown (30 to 45% of wild-type cytokinin content) and reduced expression of the cytokinin reporter gene ARR5:GUS (beta-glucuronidase). Cytokinin deficiency resulted in diminished activity of the vegetative and floral shoot apical meristems and leaf primordia, indicating an absolute requirement for the hormone. By contrast, cytokinins are negative regulators of root growth and lateral root formation. We show that the increased growth of the primary root is linked to an enhanced meristematic cell number, suggesting that cytokinins control the exit of cells from the root meristem. Different AtCKX-green fluorescent protein fusion proteins were localized to the vacuoles or the endoplasmic reticulum and possibly to the extracellular space, indicating that subcellular compartmentation plays an important role in cytokinin biology. Analyses of promoter:GUS fusion genes showed differential expression of AtCKX genes during plant development, the activity being confined predominantly to zones of active growth. Our results are consistent with the hypothesis that cytokinins have central, but opposite, regulatory functions in root and shoot meristems and indicate that a fine-tuned control of catabolism plays an important role in ensuring the proper regulation of cytokinin functions.     

Identification of novel meristem factors involved in shoot regeneration through the analysis of temperature-sensitive mutants of Arabidopsis

Adventitious organogenesis in plant tissue culture involves de novo formation of apical meristems and should therefore provide important information about the fundamentals of meristem gene networks. We identified novel factors required for neoformation of the shoot apical meristem (SAM) through an analysis of shoot regeneration in root initiation defective3 ( rid3 ) and root growth defective3 ( rgd3 ) temperature-sensitive mutants of Arabidopsis. After induction of callus to regenerate shoots, cell division soon ceased and was then reactivated locally in the surface region, resulting in formation of mounds of dense cells in which adventitious-bud SAMs were eventually constructed. The rgd3 mutation inhibited reactivation of cell division and suppressed expression of CUP-SHAPED COTYLEDON1 ( CUC1 ), CUC2 and SHOOT MERISTEMLESS ( STM ). In contrast, the rid3 mutation caused excess ill-controlled cell division on the callus surface. This was intimately related to enhanced and broadened expression of CUC1 . Positional cloning revealed that the RGD3 and RID3 genes encode BTAF1 (a kind of TATA-binding protein-associated factor) and an uncharacterized WD-40 repeat protein, respectively. In the early stages of shoot regeneration, RGD3 was expressed (as was CUC1 ) in the developing cell mounds, whereas RID3 was expressed outside the cell mounds. When RID3 was over-expressed artificially, the expression levels of CUC1 and STM were significantly reduced. Taken together, these findings show that both negative regulation by RID3 and positive regulation by RGD3 of the CUC–STM pathway participate in proper control of cell division as a prerequisite for SAM neoformation.     

Carbon allocation to shoots and roots in relation to nitrogen supply is mediated by cytokinins and sucrose: Opinion

In this paper we firstly show some general responses of biomass partitioning upon nitrogen deprivation. Secondly, these responses are explained in terms of allocation of carbon and nitrogen, photosynthesis and respiration, using a simulation model. Thirdly, we present a hypothesis for the regulation of biomass partitioning to shoots and roots.Shortly after nitrogen deprivation, the relative growth rate (RGR) of the roots generally increases and thereafter decreases, whereas that of the shoot decreases immediately. The increased RGR of the root and decreased RGR of the shoot shortly after a reduction in the nitrogen supply, cause the root weight ratio (root weight per unit plant weight) to increase rapidly.We showed previously that allocation of carbon and nitrogen to shoots and roots can satisfactorily be described as a function of the internal organic plant nitrogen concentration. Using these functions in a simulation model, we analyzed why the relative growth rate of the roots increases shortly after a reduction in nitrogen supply. The model predicts that upon nitrogen deprivation, the plant nitrogen concentration and the rate of photosynthesis per unit plant weight rapidly decrease, and the allocation of recently assimilated carbon and nitrogen to roots rapidly increases. Simulations show that the increased relative growth rate of the root upon nitrogen deprivation is explained by decreased use of carbon for root respiration, due to decreased carbon costs for nitrogen uptake. The stimulation of the relative growth rate of the root is further amplified by the increased allocation of carbon and nitrogen to roots. Using the simple relation between the plant nitrogen concentration and allocation, the model describes plant responses quite realistically.Based on information in the literature and on our own experiments we hypothesize that allocation of carbon is mediated by sucrose and cytokinins. We propose that nitrogen deprivation leads to a reduced cytokinin production, a decreased rate of cytokinin export from the roots to the shoot, and decreased cytokinin concentrations. A reduced cytokinin concentration in the shoot represses cell division in leaves, whereas a low cytokinin concentration in roots neutralizes the inhibitory effect of cytokinins on cell division. A reduced rate of cell division in the leaves leads to a reduced unloading of sucrose from the phloem into the expanding cells. Consequently, the sucrose concentration in the phloem nearby the expanding cells increases, leading to an increase in turgor pressure in the phloem nearby the leaf's division zone. In the roots, cell division continues and no accumulation of sugars occurs in dividing cells, leading to only marginal changes in osmotic potential and turgor pressure in the phloem nearby the root's cell division zone. These changes in turgor pressure in the phloem of roots and sink leaves affect the turgor pressure gradients between source leaf-sink leaf and source leaf-root in such a way that relatively more carbohydrates are exported to the roots. As a consequence RWR increases after nitrogen deprivation. This hypothesis also explains the strong relationship between allocation and the plant nitrogen status.     

The role of microbial signals in plant growth and development

Plant growth and development involves a tight coordination of the spatial and temporal organization of cell division, cell expansion and cell differentiation. Orchestration of these events requires the exchange of signaling molecules between the root and shoot, which can be affected by both biotic and abiotic factors. The interactions that occur between plants and their associated microorganisms have long been of interest, as knowledge of these processes could lead to the development of novel agricultural applications. Plants produce a wide range of organic compounds including sugars, organic acids and vitamins, which can be used as nutrients or signals by microbial populations. On the other hand, microorganisms release phytohormones, small molecules or volatile compounds, which may act directly or indirectly to activate plant immunity or regulate plant growth and morphogenesis. In this review, we focus on recent developments in the identification of signals from free-living bacteria and fungi that interact with plants in a beneficial way. Evidence has accumulated indicating that classic plant signals such as auxins and cytokinins can be produced by microorganisms to efficiently colonize the root and modulate root system architecture. Other classes of signals, including N-acyl-L-homoserine lactones, which are used by bacteria for cell-to-cell communication, can be perceived by plants to modulate gene expression, metabolism and growth. Finally, we discuss the role played by volatile organic compounds released by certain plant growth-promoting rhizobacteria in plant immunity and developmental processes. The picture that emerges is one in which plants and microbes communicate themselves through transkingdom signaling systems involving classic and novel signals.Key words: Arabidopsis, alkamides, auxins, quorum-sensing, cytokinins     

Regulation of Shoot and Root Development through Mutual Signaling

Plants adjust their development in relation to the availability of nutrient sources. This necessitates signaling between root and shoot. Aside from the well-known systemic signaling processes mediated by auxin, cytokinin, and sugars, new pathways involving carotenoid-derived hormones have recently been identified. The auxin-responsive MAX pathway controls shoot branching through the biosynthesis of strigolactone in the roots. The BYPASS1 gene affects the production of an as-yet unknown carotenoid-derived substance in roots that promotes shoot development. Novel local and systemic mechanisms that control adaptive root development in response to nitrogen and phosphorus starvation were recently discovered. Notably, the ability of the NITRATE TRANSPORTER 1.1 to transport auxin drew for the first time a functional link between auxin, root development, and nitrate availability in soil. The study of plant response to phosphorus starvation allowed the identification of a systemic mobile miRNA. Deciphering and integrating these signaling pathways at the whole-plant level provide a new perspective for understanding how plants regulate their development in response to environmental cues.     

A Modeling Study on How Cell Division Affects Properties of Epithelial Tissues Under Isotropic Growth

Cell proliferation affects both cellular geometry and topology in a growing tissue, and hence rules for cell division are key to understanding multicellular development. Epithelial cell layers have for long times been used to investigate how cell proliferation leads to tissue-scale properties, including organism-independent distributions of cell areas and number of neighbors. We use a cell-based two-dimensional tissue growth model including mechanics to investigate how different cell division rules result in different statistical properties of the cells at the tissue level. We focus on isotropic growth and division rules suggested for plant cells, and compare the models with data from the Arabidopsis shoot. We find that several division rules can lead to the correct distribution of number of neighbors, as seen in recent studies. In addition we find that when also geometrical properties are taken into account other constraints on the cell division rules result. We find that division rules acting in favor of equally sized and symmetrically shaped daughter cells can best describe the statistical tissue properties.     

GEM,a Novel Factor in the Coordination of Cell Division to Cell Fate Decisions in the Arabidopsis Epidermis

Cell division and cell fate decisions are highly regulated processes that need to be coordinated both spatially and temporally for correct plant growth and development. Gaining a deeper molecular and cellular understanding of these links is especially relevant for plant biology since, unlike in animals, formation of new organs is a process that takes place after embryogenesis and continues throughout the entire plant lifespan. The recent identification of a novel factor, GEM, has provided a molecular framework that coordinates cell division to cell fate in the Arabidopsis epidermis. GEM is an inhibitor of cell division through interacting with CDT1, a DNA replication protein. It also inhibits the expression of the homeobox GLABRA2 (GL2) gene that determines the hair/non-hair fate and the pavement/trichome fate in the root and leaf epidermis, respectively. GEM seems to be crucial in controlling the balance of activating/repressing histone modifications at its target promoters.Key Words: cell division, cell cycle, cell fate, GEM, GLABRA2, CDT1, DNA replication, chromatin, histone methylation, gene expression, root hair, Arabidopsis, plant     

RML1 and RML2, Arabidopsis genes required for cell proliferation at the root tip.

New cells are produced from the meristematic tissues located at the shoot and root tip throughout the life of higher plants. To investigate the genetic mechanism regulating meristematic activity, we isolated and characterized four single-gene, recessive mutants in Arabidopsis thaliana called root meristemless (rml). Complementation tests identified two RML loci; RML1 maps to chromosome IV and RML2 maps to chromosome III. These mutants produce normal embryonic roots that either did not undergo or experienced limited cell division following germination, resulting in primary roots of less than 2.0 mm in length. Mutants can produce lateral and adventitious roots, which can grow to a length comparable to the embryonic root and arrest, indicating that the growth arrest is unrelated to the embryonic dormancy process. Neither the addition of growth regulators to the media nor the removal of shoots can rescue mutant roots from growth arrest, indicating that the mutant phenotype is not caused by a shortage of known growth regulators or by a transmissible shoot inhibitor. Normal cell division ability in mutant embryo, shoot, and callus cells indicates that the RML gene functions are not part of the general cell division processes; rather, they are involved specifically in activating the cell division cycle in the root apical cells.