Quantitative Analyses of Cell Division in Plants

At the molecular level regulatory interactions between cell cycle genes are being uncovered rapidly, but less progress is made in unravelling how these molecular events regulate growth processes at the level of cells and of the whole organism. The main obstacle is the absence of a set of analytical tools that are powerful enough to determine pertinent parameters and, at the same time, relatively easy to use by non-specialized laboratories. Appropriate methodology to obtain this type of data has been pioneered in the first half of the last century and is now commonly defined as ‘kinematic analysis’. Unfortunately, the laborious nature of these analyses and the relatively complex numerical methods used, have limited their use to only a handful of specialized research groups. In this article we attempt to present an accessible entry to this methodology, particularly in terms of the mathematical framework. We start describing the simplest possible system, i.e., a virtually homogenous cell suspension culture. Then, we outline the analysis of dicotyledonous leaves, root tips, monocotyledonous leaves, and finally shoot apical meristems. For each of these systems we discuss the details of the calculation of cell division parameters such as cell cycle duration, size of the meristem and number of cells contained in it, which enables answering fundamental questions about the relative contribution of differences in cell production and cell size to variation in growth. In addition, we discuss the assumptions and limitations of these and alternative methodologies with the aim to facilitate the choice of appropriate analyses depending on the specific research question.

[1]  C. Bergounioux,et al.  Molecular and biochemical characterization of the involvement of cyclin-dependent kinase A during the early development of tomato fruit. , 1999, Plant physiology.

[2]  U. Schurr,et al.  A cellular growth model for root tips. , 2004, Journal of theoretical biology.

[3]  H. Tsukaya,et al.  Cell cycling and cell enlargement in developing leaves of Arabidopsis. , 1999, Developmental biology.

[4]  K. Kiêu,et al.  Cellular Parameters of the Shoot Apical Meristem in Arabidopsis , 1998, Plant Cell.

[5]  F. Sala,et al.  Specific and reversible inhibition by aphidicolin in the alpha-like DNA polymerase of plant cells. , 1980, FEBS letters.

[6]  J. Turner The Quantitative Relation Between Plant and Bacterial Cells Involved in the Hypersensitive Reaction , 1974 .

[7]  Stephane Rombauts,et al.  Transcriptome analysis during cell division in plants , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[8]  J. Masle,et al.  The effects of elevated CO(2) concentrations on cell division rates, growth patterns, and blade anatomy in young wheat plants are modulated by factors related to leaf position, vernalization, and genotype. , 2000, Plant physiology.

[9]  T. Nagata,et al.  Role of cytoskeletal components in the migration of nuclei during the cell cycle transition from G1 phase to S phase of tobacco BY-2 cells , 1997 .

[10]  D. Inzé,et al.  The Plant-Specific Cyclin-Dependent Kinase CDKB1;1 and Transcription Factor E2Fa-DPa Control the Balance of Mitotically Dividing and Endoreduplicating Cells in Arabidopsis , 2004, The Plant Cell Online.

[11]  F. Miglietta,et al.  Spatial and Temporal Effects of Free-Air CO2Enrichment (POPFACE) on Leaf Growth, Cell Expansion, and Cell Production in a Closed Canopy of Poplar1 , 2003, Plant Physiology.

[12]  Lucila Ohno-Machado,et al.  Analysis of matched mRNA measurements from two different microarray technologies , 2002, Bioinform..

[13]  F. Fiorani,et al.  Control of Leaf Growth and its Role in Determining Variation in Plant Growth Rate from an Ecological Perspective , 1999 .

[14]  D. Inzé,et al.  CKS1At overexpression in Arabidopsis thaliana inhibits growth by reducing meristem size and inhibiting cell-cycle progression. , 2001, The Plant journal : for cell and molecular biology.

[15]  P. Burman,et al.  Effect of Water Stress on Cortical Cell Division Rates within the Apical Meristem of Primary Roots of Maize , 1997, Plant physiology.

[16]  Janet K. Thompson,et al.  The Quiescent Centre and Rates of Mitosis in the Root Meristem of Allium sativum , 1968 .

[17]  R. O. Erickson Modeling of Plant Growth , 1976 .

[18]  J. Doonan,et al.  Cellular basis of shoot apical meristem development. , 2001, International review of cytology.

[19]  R. O. Erickson,et al.  Kinematics of plant growth. , 1979, Journal of theoretical biology.

[20]  B. Scheres,et al.  Cellular organisation of the Arabidopsis thaliana root. , 1993, Development.

[21]  T. Baskin,et al.  STUNTED PLANT 1, A Gene Required for Expansion in Rapidly Elongating but Not in Dividing Cells and Mediating Root Growth Responses to Applied Cytokinin , 1995, Plant physiology.

[22]  D. Inzé,et al.  Molecular dissection of plant cytokinesis and phragmoplast structure: a survey of GFP-tagged proteins. , 2004, The Plant journal : for cell and molecular biology.

[23]  N. Bernstein,et al.  The Determination of Relative Elemental Growth Rate Profiles from Segmental Growth Rates (A Methodological Evaluation) , 1997, Plant physiology.

[24]  F. Fiorani,et al.  Can meristematic activity determine variation in leaf size and elongation rate among four Poa species? A kinematic study. , 2000, Plant physiology.

[25]  Gerrit T.S. Beemster,et al.  Variation in Growth Rate between Arabidopsis Ecotypes Is Correlated with Cell Division and A-Type Cyclin-Dependent Kinase Activity1 , 2002, Plant Physiology.

[26]  F. Tardieu,et al.  Temperature Affects Expansion Rate of Maize Leaves without Change in Spatial Distribution of Cell Length (Analysis of the Coordination between Cell Division and Cell Expansion) , 1995, Plant physiology.

[27]  W. Silk,et al.  Growth Patterns Inferred from Anatomical Records : Empirical Tests Using Longisections of Roots of Zea mays L. , 1989, Plant physiology.

[28]  S. Pongor,et al.  Cell cycle phase specificity of putative cyclin-dependent kinase variants in synchronized alfalfa cells. , 1997, The Plant cell.

[29]  D. Inzé,et al.  Specific checkpoints regulate plant cell cycle progression in response to oxidative stress , 1999 .

[30]  V. B. Ivanov,et al.  Estimation of the Cell-Cycle Duration in the Root Apical Meristem: A Model of Linkage between Cell-Cycle Duration, Rate of Cell Production, and Rate of Root Growth , 1997, International Journal of Plant Sciences.

[31]  T. Vernoux,et al.  Developmental control of cell division patterns in the shoot apex , 2000, Plant Molecular Biology.

[32]  F. Tardieu,et al.  Co-Ordination of Cell Division and Tissue Expansion in Sunflower, Tobacco, and Pea Leaves: Dependence or Independence of Both Processes? , 2000, Journal of Plant Growth Regulation.

[33]  D. Inzé,et al.  Genome-Wide Analysis of Gene Expression Profiles Associated with Cell Cycle Transitions in Growing Organs of Arabidopsis1[w] , 2005, Plant Physiology.

[34]  L. Hennig,et al.  Genome-wide gene expression in an Arabidopsis cell suspension , 2003, Plant Molecular Biology.

[35]  R. H. Goodwin,et al.  GROWTH AND DIFFERENTIATION IN THE ROOT TIP OF PHLEUM PRATENSE , 1945 .

[36]  P. Barlow,et al.  Cellular Growth in Roots of a Gibberellin-Deficient Mutant of Tomato (Lycopersicon esculentum Mill.) and its Wild-type , 1991 .

[37]  F. Tardieu,et al.  Spatial distributions of expansion rate, cell division rate and cell size in maize leaves: a synthesis of the effects of soil water status, evaporative demand and temperature. , 2000, Journal of experimental botany.

[38]  M. Menges,et al.  Cell cycle regulation of cyclin-dependent kinases in tobacco cultivar Bright Yellow-2 cells. , 2001, Plant physiology.

[39]  F. Fiorani,et al.  Epidermal cell division and cell elongation in two Aegilops species with contrasting leaf elongation rates. , 2003, Functional plant biology : FPB.

[40]  C. J. Nelson,et al.  Responses of Tall Fescue Leaf Meristems to N Fertilization and Harvest Frequency 1 , 1983 .

[41]  A. Meister Calculation of binding length of base-specific DNA dyes by comparison of sequence and flow cytometric data. Application to Oryza sativa and Arabidopsis thaliana. , 2005, Journal of theoretical biology.

[42]  D. Francis,et al.  Elevated CO2 stimulates cells to divide in grass meristems: a differential effect in two natural populations of Dactylis glomerata , 1997 .

[43]  F. Sala,et al.  Specific and reversible inhibition by aphidicolin of the α‐like DNA polymerase of plant cells , 1980 .

[44]  W. Wernicke,et al.  The First Cell Division Cycle in Nicotiana Mesophyll Protoplasts Cultured in vitro II. Microtubules and the Plane of Division , 1991 .

[45]  F. Tardieu,et al.  The elongation rate at the base of a maize leaf shows an invariant pattern during both the steady-state elongation and the establishment of the elongation zone. , 2001, Journal of experimental botany.

[46]  F. Tardieu,et al.  Quantitative analysis of cell division in leaves: methods, developmental patterns and effects of environmental conditions , 2000, Plant Molecular Biology.

[47]  R. Macleod,et al.  Characteristics of root apical meristem cell population kinetics: A review of analyses and concepts , 1980 .

[48]  Richard H. Goodwin,et al.  STUDIES ON ROOTS. III. AN ANALYSIS OF ROOT GROWTH IN PHLEUM PRATENSE USING PHOTOMICROGRAPHIC RECORDS , 1956 .

[49]  R. Bino,et al.  Cell division and subsequent radicle protrusion in tomato seeds are inhibited by osmotic stress but DNA synthesis and formation of microtubular cytoskeleton are not. , 2000, Plant physiology.

[50]  Yasuyuki Nemoto,et al.  Tobacco BY-2 Cell Line as the “HeLa” Cell in the Cell Biology of Higher Plants , 1992 .

[51]  D. Inzé,et al.  Cell Cycle Modulation in the Response of the Primary Root of Arabidopsis to Salt Stress1 , 2004, Plant Physiology.

[52]  W. Wernicke,et al.  The first cell division cycle in Nicotiana mesophyll protoplasts cultured in vitro. I, Methods to determine cycle kinetics , 1991 .

[53]  J. Croxdale Stomatal patterning in monocotyledons: Tradescantia as a model system , 1998 .

[54]  P. Green,et al.  Analysing the changing cell cycle. , 1977, Journal of theoretical biology.

[55]  D. Galbraith,et al.  Systemic Endopolyploidy in Arabidopsis thaliana. , 1991, Plant physiology.

[56]  W. Silk Steady Form from Changing Cells , 1992, International Journal of Plant Sciences.

[57]  D. Inzé,et al.  Spatial distribution of cell division rate can be deduced from that of p34(cdc2) kinase activity in maize leaves grown at contrasting temperatures and soil water conditions. , 2000, Plant physiology.

[58]  H. Maurer Potential Pitfalls of [3H]Thymidine Techniques to Measure Cell Proliferation , 1981, Cell and tissue kinetics.

[59]  H QUASTLER,et al.  Cell population kinetics in the intestinal epithelium of the mouse. , 1959, Experimental cell research.

[60]  D. Inzé,et al.  Cell cycle: the key to plant growth control? , 2003, Trends in plant science.

[61]  C. J. Nelson,et al.  Assessment of spatial distribution of growth in the elongation zone of grass leaf blades. , 1987, Plant physiology.

[62]  Tobias I. Baskin,et al.  On the constancy of cell division rate in the root meristem , 2000, Plant Molecular Biology.

[63]  J. Dumais,et al.  Analysis of surface growth in shoot apices. , 2002, The Plant journal : for cell and molecular biology.

[64]  F. Clowes APICAL MERISTEMS OF ROOTS , 1959 .

[65]  Kannappan Palaniappan,et al.  A New Algorithm for Computational Image Analysis of Deformable Motion at High Spatial and Temporal Resolution Applied to Root Growth. Roughly Uniform Elongation in the Meristem and Also, after an Abrupt Acceleration, in the Elongation Zone1 , 2003, Plant Physiology.

[66]  J. Dumais,et al.  Growth and morphogenesis at the vegetative shoot apex of Anagallis arvensis L. , 2003, Journal of experimental botany.

[67]  R. E. Sharp,et al.  Growth of the Maize Primary Root at Low Water Potentials : II. Role of Growth and Deposition of Hexose and Potassium in Osmotic Adjustment. , 1990, Plant physiology.

[68]  P. Green Growth and Cell Pattern Formation on an Axis: Critique of Concepts, Terminology, and Modes of Study , 1976, Botanical Gazette.

[69]  R. E. Sharp,et al.  Growth of the maize primary root at low water potentials : I. Spatial distribution of expansive growth. , 1988, Plant physiology.

[70]  W. Kühbauch,et al.  Spatial distribution of growth rates and of epidermal cell lengths in the elongation zone during leaf development in Lolium perenne L. , 1990, Planta.

[71]  W. Silk,et al.  Quantitative Descriptions of Development , 1984 .

[72]  J. Doležel,et al.  Bivariate flow cytometry DNA/BrdUrd analysis of plant cell cycle. , 1999, Methods in cell science : an official journal of the Society for In Vitro Biology.

[73]  C. Granier,et al.  Spatial and temporal analyses of expansion and cell cycle in sunflower leaves. A common pattern of development for all zones of a leaf and different leaves of a plant , 1998, Plant physiology.

[74]  T. Baskin,et al.  Analysis of cell division and elongation underlying the developmental acceleration of root growth in Arabidopsis thaliana. , 1998, Plant physiology.

[75]  R. Lempicki,et al.  Evaluation of gene expression measurements from commercial microarray platforms. , 2003, Nucleic acids research.

[76]  J. Hof EXPERIMENTAL CONTROL OF DNA SYNTHESIZING AND DIVIDING CELLS IN EXCISED ROOT TIPS OF PISUM , 1966 .

[77]  F. Clowes Estimation of Growth Fractions in Meristems of Zea mays L. , 1976 .

[78]  Patrick Laufs,et al.  In Vivo Analysis of Cell Division, Cell Growth, and Differentiation at the Shoot Apical Meristem in Arabidopsis Article, publication date, and citation information can be found at www.plantcell.org/cgi/doi/10.1105/tpc.017962. , 2004, The Plant Cell Online.

[79]  M. Menges,et al.  Synchronous Arabidopsis suspension cultures for analysis of cell-cycle gene activity. , 2002, The Plant journal : for cell and molecular biology.

[80]  D. Inzé,et al.  Analysis of cell division parameters and cell cycle gene expression during the cultivation of Arabidopsis thaliana cell suspensions. , 2001, Journal of experimental botany.

[81]  R. F. Lyndon The shoot apical meristem , 1998 .

[82]  D. Vreugdenhil,et al.  Cell division and cell enlargement during potato tuber formation , 1998 .

[83]  P. J. King,et al.  Control of the cell cycle in cultured plant cells , 1984 .

[84]  Gerrit T.S. Beemster,et al.  Effects of soil resistance to root penetration on leaf expansion in wheat (Triticum aestivum L.): kinematic analysis of leaf elongation , 1996 .

[85]  B. Jähne,et al.  Quantitative analysis of the local rates of growth of dicot leaves at a high temporal and spatial resolution, using image sequence analysis , 1998 .

[86]  C. J. Nelson,et al.  Elongation of the Grass Leaf and its Relationship to the Phyllochron , 1995 .

[87]  D. Inzé,et al.  Control of proliferation, endoreduplication and differentiation by the Arabidopsis E2Fa–DPa transcription factor , 2002, The EMBO journal.

[88]  P. L. Webster Analysis of Heterogeneity of Relative Division Rates in Root Apical Meristems , 1980, Botanical Gazette.

[89]  D. Kwiatkowska Structural integration at the shoot apical meristem: models, measurements, and experiments. , 2004, American journal of botany.

[90]  P. Gandar The Analysis of Growth and Cell Production in Root Apices , 1980, Botanical Gazette.

[91]  N. Kirchgeßner,et al.  Spatio-temporal dynamics of expansion growth in roots: automatic quantification of diurnal course and temperature response by digital image sequence processing. , 2002, Journal of experimental botany.

[92]  T. Steeves,et al.  Patterns in plant development: Subject index , 1972 .

[93]  F. Tardieu,et al.  Control of Leaf Expansion Rate of Droughted Maize Plants under Fluctuating Evaporative Demand (A Superposition of Hydraulic and Chemical Messages?) , 1997, Plant physiology.

[94]  Franky R. G. Terras,et al.  Functional Analysis of Cyclin-Dependent Kinase Inhibitors of Arabidopsis , 2001, The Plant Cell Online.

[95]  R. E. Sharp,et al.  Growth of the Maize Primary Root at Low Water Potentials : III. Role of Increased Proline Deposition in Osmotic Adjustment. , 1991, Plant physiology.