On the constancy of cell division rate in the root meristem

This review examines under what circumstances the rate of cell division among cells of the root meristem is known to vary. First, methods are compared that have been used to quantify cell division rate. These can be grouped as being either cytological, in which the rate of accumulation of cells in a particular phase of the cell cycle is determined based on some form of cytological labeling, or kinematic, in which the rate of cell accumulation is determined from the net movement of cells. Then, evidence is reviewed as to whether cell division rates vary between different tissues or cell types, between different positions in the root, or finally between different environments. The evidence is consistent with cells dividing at a constant rate, and well documented examples where cell division rate changes substantially are rare. The constancy of cell division rate contrasts with the number of dividing cells, which varies extensively, and implies that a major point for cell cycle control is governing the exit from the proliferative state at the basal boundary of the meristem.     

Quantitative analysis of venation patterns of Arabidopsis leaves by supervised image analysis

The study of transgenic Arabidopsis lines with altered vascular patterns has revealed key players in the venation process, but details of the vascularization process are still unclear, partly because most lines have only been assessed qualitatively. Therefore, quantitative analyses are required to identify subtle perturbations in the pattern and to test dynamic modeling hypotheses using biological measurements. We developed an online framework, designated Leaf Image Analysis Interface (LIMANI), in which venation patterns are automatically segmented and measured on dark-field images. Image segmentation may be manually corrected through use of an interactive interface, allowing supervision and rectification steps in the automated image analysis pipeline and ensuring high-fidelity analysis. This online approach is advantageous for the user in terms of installation, software updates, computer load and data storage. The framework was used to study vascular differentiation during leaf development and to analyze the venation pattern in transgenic lines with contrasting cellular and leaf size traits. The results show the evolution of vascular traits during leaf development, suggest a self-organizing mechanism for leaf venation patterning, and reveal a tight balance between the number of end-points and branching points within the leaf vascular network that does not depend on the leaf developmental stage and cellular content, but on the leaf position on the rosette. These findings indicate that development of LIMANI improves understanding of the interaction between vascular patterning and leaf growth.     

Developmental control of cell division patterns in the shoot apex

The shoot apical meristem is a group of rapidly dividing cells that generate all aerial parts of the plant. It is a highly organised structure, which can be divided into functionally distinct domains, characterised by specific proliferation rates of the individual cells. Genetic studies have enabled the identification of regulators of meristem function. These factors are involved in the formation and maintenance of the meristem, as well as in the formation of the primordia. Somehow, they must also govern cell proliferation rates within the shoot apex. Possible links between meristem regulators and the cell cycle machinery will be discussed. In order to analyse the role of cell proliferation in development, cell cycle gene expression has been perturbed using transgenic approaches and mutation. The effect of these alterations on growth and development at the shoot apex will be presented. Together, these studies give a first insight into the regulatory networks controlling the cell cycle and into the significance of cell proliferation in plant development.     

水热条件与烟叶钾素吸收的关系研究

在恒温光照培养条件下,研究了土壤水分和温度对烟叶钾素吸收的影响。结果表明：土壤含水量在14％～18％范围,烟叶钾含量随水分增加而增加,而在20％～25％范围,烟叶钾含量随水分增加而减少,烟叶钾素吸收较适宜的水分条件为18％;温度对烟叶钾素吸收的影响表现为15～25℃范围随温度升高而增加,30～35℃范围随温度升高而减少,较适宜的温度条件为25℃;水热在高温和土壤含水量较少时对烟叶钾素吸收有互补效应,而在高温高湿时又对烟叶钾素吸收具有抑制效应。     

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.     

Spatial and temporal quantitative analysis of cell division and elongation rate in growing wheat leaves under saline conditions

Leaf growth in grasses is determined by the cell division and elongation rates, with the duration of cell elongation being one of the processes that is the most sensitive to salinity. Our objective was to investigate the distribution profiles of cell production, cell length and the duration of cell elongation in the growing zone of the wheat leaf during the steady growth phase. Plants were grown in loamy soil with or without 120 mmol/L NaCl in a growth chamber, and harvested at day 3 after leaf 4 emerged. Results show that the elongation rate of leaf 4 was reduced by 120 mmol/L NaCl during the steady growth phase. The distribution profile of the lengths of abaxial epidermal cells of leaf 4 during the steady growth stage shows a sigmoidal pattern along the leaf axis for both treatments. Although salinity did not affect or even increased the length of the epidermal cells in some locations in the growth zone compared to the control treatment, the final length of the epidermal cells was reduced by 14% at 120 mmol/L NaCl. Thus, we concluded that the observed reduction in the leaf elongation rate derived in part from the reduced cell division rate and either the shortened cell elongation zone or shortened duration of cell elongation. This suggests that more attention should be paid to the effects of salinity on those properties of cell production and the period of cell maturation that are related to the properties of cell wall.     

Leaf expansion and cell division are affected by reducing absorbed light before but not after the decline in cell division rate in the sunflower leaf

The effect of absorbed photosynthetic photon flux density (PPFD) on leaf expansion is a key issue for analysing the phenotypic variability between plants and for modelling feedback loops. Expansion and epidermal cell division in leaf 8 of sunflower were analysed in a series of five experiments where absorbed photosynthetic photon flux density (PPFD) was reduced either by shading or by covering part of the leaf area. These treatments were imposed at different times during leaf development. Expansion and cell division were affected by a reduction in absorbed PPFD only in the first part of leaf development, while the leaf area was less than 2% of its final value and while absolute expansion rate was slow. In contrast, it was not affected if imposed later when the leaf was visible and absolute expansion rate was at maximum. A reduction in absorbed PPFD caused the same reduction in expansion and in cell division whether it was due to a reduction in incident PPFD or to a reduction in photosynthetic leaf area, suggesting that carbon metabolism was involved. Relative expansion rate recovered to control levels when relative division rate began to decline, in all experiments and in all zones of a leaf. This was probably linked to the source–sink transition, after which the leaf had such a high priority in carbon allocation that it was largely insensitive to changes in absorbed PPFD. The final leaf area was therefore closely related to the cumulated PPFD absorbed by the plant from leaf initiation to the end of exponential cell division.     

Short-term responses of leaf growth rate to water deficit scale up to whole-plant and crop levels: an integrated modelling approach in maize

Physiological and genetic studies of leaf growth often focus on short-term responses, leaving a gap to whole-plant models that predict biomass accumulation, transpiration and yield at crop scale. To bridge this gap, we developed a model that combines an existing model of leaf 6 expansion in response to short-term environmental variations with a model coordinating the development of all leaves of a plant. The latter was based on: (1) rates of leaf initiation, appearance and end of elongation measured in field experiments; and (2) the hypothesis of an independence of the growth between leaves. The resulting whole-plant leaf model was integrated into the generic crop model APSIM which provided dynamic feedback of environmental conditions to the leaf model and allowed simulation of crop growth at canopy level. The model was tested in 12 field situations with contrasting temperature, evaporative demand and soil water status. In observed and simulated data, high evaporative demand reduced leaf area at the whole-plant level, and short water deficits affected only leaves developing during the stress, either visible or still hidden in the whorl. The model adequately simulated whole-plant profiles of leaf area with a single set of parameters that applied to the same hybrid in all experiments. It was also suitable to predict biomass accumulation and yield of a similar hybrid grown in different conditions. This model extends to field conditions existing knowledge of the environmental controls of leaf elongation, and can be used to simulate how their genetic controls flow through to yield.     

Effects of nitrogen deprivation on cell division and expansion in leaves of Ricinus communis L.

The effects of nitrogen deprivation on leaf extension, cell numbers and epidermal cell size were followed in leaves of Ricinus communis L. The extent to which reductions in final cell number or final epidermal cell size contributed to the reduction in final leaf size depended on the developmental stage of the leaf at the time of N deprivation. In leaves which already had their full complement of cells (leaf 2), the reduction in final leaf size following nitrogen deprivation was associated with a reduction in final cell size. In leaves that were at earlier stages of development at the onset of N deprivation (leaves 3 and 4), the reduction in final leaf size was greater than in leaf 2. In these younger leaves, the final cell size was even smaller than in leaf 2, but the greatest contribution to reduced final leaf size was a reduction in the number of cells produced. This accounted for approximately 80% of the reduction in final leaf size in leaf 4. During leaf development, the contribution from different tissue layers to the total cell number changed. In the smallest leaf sizes, the contribution from upper and lower epidermis and spongy parenchyma was greater than that from palisade parenchyma. As the leaf size increased, cells in the palisade parenchyma continued to divide for longer than in the other layers. At final leaf size, the contribution from the different tissue layers to total cell number was the same for leaves 2, 3 and 4, irrespective of N treatment. In these final leaf structures, palisade parenchyma contributed 60% of the total cell number. Thus, although nitrogen deprivation affected leaf size variously through cell division and cell expansion, depending on leaf developmental stage at the time of nitrogen deprivation, the ratio of cell numbers and sizes in different tissue layers, at final leaf size, was unaffected.     

Separation of cell enlargement and division in bean leaves

A method is presented for inducing cell enlargement in intact leaves and leaf strips of Phaseolus vulgaris L. without the complication of cell division. Primary bean leaves complete cell division and stop growing after 10 d in dim red light. Transfer to white light induces expansion (50% in 24 h) which is entirely the consequence of cell enlargement. Leaf strips from red-light-grown seedlings placed in white light and provided external solutes (10 mM KCl+10 mM sucrose) expand at the same rate as intact leaves in the light. This system makes possible future investigation of the mechanism of leaf cell enlargement.     

Blue light-regulation of cell division in Adiantum protonemata: an approach with pulse stimulation

Abstract Cell division of the single-celled Adiantum protonemata produced by red-light (RL) incubation of germinated spores is induced by transfer to darkness and is stimulated by blue light (BL). It is known that the cellular process leading to this cell division includes one cell cycle and the BL response results from shortening of the Gl phase. The authors studied this BL regulation of cell cycle by giving a pulse of BL after RL termination and measuring changes in the proportion of divided cells. To minimize phytochrome responses arising from BL irradiation, the plants were kept in continuous far-red light instead of total darkness after the RL incubation. The response to a pulse (10–100 s) approached saturation with increasing rluences in a manner that reciprocity is valid. The sensitivity to BL, investigated by measuring the response to a saturating pulse, showed an increase in the first several hours after RL termination, followed by a sustained sensitivity for 20 h. Time courses of the pulse-induced responses showed a lag of about 12 h, which was considerably shorter than in the non-stimulated control; the lag was approximately independent of the strength of BL stimulation or the timing of BL application after RL termination, and the major difference occurred in the slope. It is concluded that the sensitivity to BL is retained during the time span in which the dark-dependent Gl phase progresses, and that the BL response is initiated independently of the reactions involved in the dark-dependent Gl phase. A minimal reaction model of Gl phase is suggested to unify the results.     

Comparison of allometric relationships and morphological differences in Tilia cordata Mill. outer leaves exposed to different environmental conditions

The study is based on four leaf parameters: leaf width (LW), lobe length (LL), leaf size (LS) and leaf shape which is calculated as LW to leaf length (LW/LL) ratio. Under different environmental conditions, LL is an isometric character, LW shows positive allometry, whereas LW/LL shows negative allometry. Regression analysis results indicated that there is no significant difference either in slopes or in regression coefficients between investigated sites. Thus, in this study, we found that allometric relationships between leaf parameters and LS are character specific and that they tended not to differ significantly between Tilia cordata Mill. outer leaves exposed to different environmental conditions. Also, there are no significant interpopulation differences for both principal component PC1 and PC2 scores. The stepwise discriminant functional analysis results allowed us to identify a set of two leaf parameters (LS and LL) with a moderate discriminating ability (59.8%). T. cordata outer leaves are significantly larger and broader in the reference area (R-leaves) than leaves from polluted (P-leaves) site. The data also indicated that there is a relatively larger petiole size in R-leaves than in P-leaves. We found that in P-leaves, LW increased faster with increasing LS than in R-leaves.     

Response of cassava leaf area expansion to water deficit: cell proliferation, cell expansion and delayed development

BACKGROUND AND AIMS: Cassava (Manihot esculenta) is an important food crop in the tropics that has a high growth rate in optimal conditions, but also performs well in drought-prone climates. The objectives of this work were to determine the effects of water deficit and rewatering on the rate of expansion of leaves at different developmental stages and to evaluate the extent to which decreases in cell proliferation, expansion, and delay in development are responsible for reduced growth. METHODS: Glasshouse-grown cassava plants were subjected to 8 d of water deficit followed by rewatering. Leaves at 15 developmental stages from nearly full size to meristematic were sampled, and epidermal cell size and number were measured on leaves at four developmental stages. KEY RESULTS: Leaf expansion and development were nearly halted during stress but resumed vigorously after rewatering. In advanced-stage leaves (Group 1) in which development was solely by cell expansion, expansion resumed after rewatering, but not sufficiently for cell size to equal that of controls at maturity. In Group 2 (cell proliferation), relative expansion rate and cell proliferation were delayed until rewatering, but then recovered partially, so that loss of leaf area was due to decreased cell numbers per leaf. In Group 3 (early meristematic development) final leaf area was not affected by stress, but development was delayed by 4-6 d. On a plant basis, the proportion of loss of leaf area over 26 d attributed to leaves at each developmental stage was 29, 50 and 21 % in Group 1, 2 and 3, respectively. CONCLUSIONS: Although cell growth processes were sensitive to mild water deficit, they recovered to a large extent, and much of the reduction in leaf area was caused by developmental delay and a reduction in cell division in the youngest, meristematic leaves.     

Water deficit in developing endosperm of maize: cell division and nuclear DMA endoreduplication

Water deficit severely decreases maize (Zea mays L.) kernel growth; the effect is most pronounced in apical regions of ears. The capacity for accumulation of storage material in endosperms is thought to he partially determined by the extent of cell division and endoreduplication (post-mitotic nuclear DNA synthesis). To gain a better understanding of the regulatory mechanisms involved, we have examined the effect of water deficit on cellular development during the post-fertilization period. Greenhouse-grown maize was subjected to water-limited treatments during rapid cell division [from 1 to 10days after pollination (DAP)] or rapid endoreduplication (9 to 15 DAP). The number of nuclei and the nuclear DNA content were determined with flow cytometry. Water deficit from 1 to 10 DAP substantially decreased the rate of endosperm cell division in apical-region kernels, but had little effect on middle-region endosperms. Rewatcring did not allow cell division to recover in apical-region endosperms. Water deficit from 9 to 15 DAP also decreased cell division in apical-region endosperms. Endoreduplication was not affected by the late treatment in either region of the car, but was inhibited by the early treatment in the apical region. In particular, the proportion of nuclei entering higher DN A-content size classes was reduced. We conclude that cell division is highly responsive to water deficit, whereas endoreduplication is less so. We also conclude that the reduced proportion of nuclei entering higher DNA-content size classes during endoreduplication is indicative of multiple control points in the mitotic and endoreduplication cycles.     

Developmental process of sun and shade leaves in Chenopodium album L.

The authors’ previous study of Chenopodium album L. revealed that the light signal for anatomical differentiation of sun and shade leaves is sensed by mature leaves, not by developing leaves. They suggested that the two‐cell‐layered palisade tissue of the sun leaves would be formed without a change in the total palisade tissue cell number. To verify that suggestion, a detailed study was made of the developmental processes of the sun and shade leaves of C. album with respect to the division of palisade tissue cells (PCs) and the data was expressed against developmental time (leaf plastochron index, LPI). The total number of PCs per leaf did not differ between the sun and shade leaves throughout leaf development (from LPI ?1 to 10). In both sun and shade leaves, anticlinal cell division of PCs occurred most frequently from LPI ?1 to 2. In sun leaves, periclinal division of PCs occurred synchronously with anticlinal division. The constancy of the total number of PCs indicates that periclinal divisions occur at the expense of anticlinal divisions. These results support the above suggestion that two‐cell‐layered palisade tissue is formed by a change of cell division direction without a change in the total number of PCs. PCs would be able to recognize the polarity or axis that is perpendicular to the leaf plane and thereby change the direction of their cell divisions in response to the light signal from mature leaves.     

Photosynthetic acclimation to simultaneous and interacting environmental stresses along natural light gradients: optimality and constraints

There is a strong natural light gradient from the top to the bottom in plant canopies and along gap-understorey continua. Leaf structure and photosynthetic capacities change close to proportionally along these gradients, leading to maximisation of whole canopy photosynthesis. However, other environmental factors also vary within the light gradients in a correlative manner. Specifically, the leaves exposed to higher irradiance suffer from more severe heat, water, and photoinhibition stresses. Research in tree canopies and across gap-understorey gradients demonstrates that plants have a large potential to acclimate to interacting environmental limitations. The optimum temperature for photosynthetic electron transport increases with increasing growth irradiance in the canopy, improving the resistance of photosynthetic apparatus to heat stress. Stomatal constraints on photosynthesis are also larger at higher irradiance because the leaves at greater evaporative demands regulate water use more efficiently. Furthermore, upper canopy leaves are more rigid and have lower leaf osmotic potentials to improve water extraction from drying soil. The current review highlights that such an array of complex interactions significantly modifies the potential and realized whole canopy photosynthetic productivity, but also that the interactive effects cannot be simply predicted as composites of additive partial environmental stresses. We hypothesize that plant photosynthetic capacities deviate from the theoretical optimum values because of the interacting stresses in plant canopies and evolutionary trade-offs between leaf- and canopy-level plastic adjustments in light capture and use.     

The effect of low temperature on patterns of cell division in developing second leaves of wild-type and slender mutant barley (Hordeum vulgare L.)

This study reports on investigations into the effect of long-term growth at reduced temperatures on cell elongation and cell division in the wild type and a temperature-insensitive ( slender ) mutant of barley. Plants were grown under two temperature regimes (20 and 5 °C) and the mitotic index, cell doubling time and cell lengths over the division and elongation zone were monitored at several stages of development in the second leaf. Leaf length and leaf growth rates were characteristically greater in the slender mutant than in the wild type and this was greatly exaggerated by growth at low temperature. Cell length and the length of the division zone were also greater in the slender mutant than in the wild type, and growing the plants at reduced temperature (5 °C) shortened cell lengths only in the wild type. The slender mutant had a higher mitotic index than the wild type, although in neither genotype was change in the mitotic index observed following growth at reduced temperature. Cell doubling time, on the other hand, was reduced by growth at reduced temperature in the wild type but not in the slender mutant. Thus, the data suggest very different growth responses to low temperature in the two genotypes. The results are discussed in terms of the ability of plants to sense their environment and optimize their metabolism for future growth.     

Plasticity to soil water deficit in Arabidopsis thaliana: dissection of leaf development into underlying growth dynamic and cellular variables reveals invisible phenotypes

Genetic variability in the plasticity of leaf area expansion in response to water deficit has been reported in Arabidopsis thaliana. Here, the objective was to identify the underlying dynamic and cellular processes involved in this variability. Twenty-five accessions were subjected to identical soil water deficit treatments. In all accessions, the plasticity of leaf production was low compared with that of individual leaf expansion. A subset of accessions was selected for further dissection of individual leaf expansion into its underlying variables: the rate and duration of leaf expansion and epidermal cell number and area. In all accessions, water deficit had opposite effects on the rate and duration of leaf expansion. The accumulation of these effects was reflected in changes in final leaf area. At the cellular level, moderate water deficits had opposite effects on cell number and cell size, but more severe ones reduced both variables. The importance of these opposing effects is highlighted by the behaviour of the accession An-1, for which the compensation between the decrease in leaf expansion rate and the increase in the duration of expansion is total. This dynamic plasticity in response to water deficit is not detectable when only final measurements are done.     

Stomatal dynamics are limited by leaf hydraulics in ferns and conifers: results from simultaneous measurements of liquid and vapour fluxes in leaves

Stomatal responsiveness to vapour pressure deficit (VPD) results in continuous regulation of daytime gas‐exchange directly influencing leaf water status and carbon gain. Current models can reasonably predict steady‐state stomatal conductance (g_s) to changes in VPD but the g_s dynamics between steady‐states are poorly known. Here, we used a diverse sample of conifers and ferns to show that leaf hydraulic architecture, in particular leaf capacitance, has a major role in determining the g_s response time to perturbations in VPD. By using simultaneous measurements of liquid and vapour fluxes into and out of leaves, the in situ fluctuations in leaf water balance were calculated and appeared to be closely tracked by changes in g_s thus supporting a passive model of stomatal control. Indeed, good agreement was found between observed and predicted g_s when using a hydropassive model based on hydraulic traits. We contend that a simple passive hydraulic control of stomata in response to changes in leaf water status provides for efficient stomatal responses to VPD in ferns and conifers, leading to closure rates as fast or faster than those seen in most angiosperms.     

Blue light-regulation of cell division in Adiantum protonemata: kinetic properties of the photosystem

Abstract Blue-light-induced cell division in single-celled Adiantum protonemata was studied by using two or three pulses (pulse duration: 30 s) separated by various periods and by using relatively long irradiation (e.g. 30 min). The results showed: (1) that the response is saturated by a single pulse, (2) that after the application of a saturating pulse, the protonemata gradually become responsive to another pulse, showing time-dependent saturation to the second pulse, and (3) that although reciprocity holds in the pulse-induced response, it becomes invalid as the exposure duration extends in the range of minutes. These results were analysed in view of a reaction model in which a molecular component is considered to exist in two forms A and B. The response to a single pulse is considered to result from a light-dependent conversion of the component (A→B), and the restoration response measured by two pulses, from its dark reversion (B→A). The analyses yielded (1) the value of the constant which relates the fluence rate to the rate constant of the light-induced reaction, and (2) the rate constant of the dark reaction. The model was extended to formulate the responses to long irradiations as a function of the integrated concentration of B over time. The responses predicted by the formula by using the parameter values estimated from the pulse responses were able to explain the responses measured for long irradiation.