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

The spatial distributions of leaf expansion rate, cell division rate and cell size was examined under contrasting soil water conditions, evaporative demands and temperatures in a series of experiments carried out in either constant or naturally fluctuating conditions. They were examined in the epidermis and all leaf tissues. (1) Meristem temperature affected relative elongation rate by a constant ratio at all positions in the leaf. If expressed per unit thermal time, the distribution of relative expansion rate was independent of temperature and was similar in all experiments with low evaporative demand and no water deficit. This provides a reference distribution, characteristic of the studied genotype, to which any distribution in stressed plants can be compared. (2) Evaporative demand and soil water deficit affected independently the distribution of relative elongation rate and had near-additive effects. For a given stress, a nearly constant difference was observed, at all positions of the leaf, between the relative elongation rates of stressed plants and those of control plants. This caused a reduction in the length of the zone with tissue elongation. (3) Methods for calculating cell division rate in the epidermis and in all leaf tissues are proposed and discussed. In control plants, the zone with cell division was 30 mm and 60 mm long in the epidermis and in whole tissues, respectively. Both this length and relative division rate were reduced by soil water deficit. The size of epidermal and of mesophyll cells was nearly unaffected in the leaf zone with both cell division and tissue expansion, suggesting that water deficit affects tissue expansion rate and cell division rate to the same extent. Conversely, cell size of epidermis and mesophyll were reduced by water deficit in mature parts of the leaf.     

Spatial and temporal analysis of non-steady elongation of rice leaves

A precise knowledge of the temporal and spatial distributions of cell division and tissue expansion is essential for appropriate leaf sampling in omics studies and for analyses of plant–environment relations. Elongating leaves of rice were studied during their whole development for elongation rate, distribution of cell length, cell production rate and spatial distribution of growth in the leaf. In seven genotypes, the pattern of leaf elongation rate followed three phases: (1) an exponential increase before leaf appearance; (2) a short phase (2–4 d at 20 °C) with a stable leaf elongation rate around leaf appearance; and (3) a phase of 8–10 d with a progressive decrease in elongation rate. The profile of cell length along the leaf changed with time during the first and last phases, but was time invariant around appearance. We propose a method adapted to non-steady elongation based on anatomical measurements, which was successfully tested by comparing it with the pricking method. It allowed analysis of the change with time in the spatial distribution of growth from initiation to end of leaf growth. The length of leaf zones with cell division and tissue elongation varied with time, with maximums of 21 and 60 mm respectively around leaf appearance.     

Temperature‐compensated cell production rate and elongation zone length in the root of Arabidopsis thaliana

To understand how root growth responds to temperature, we used kinematic analysis to quantify division and expansion parameters in the root of Arabidopsis thaliana. Plants were grown at temperatures from 15 to 30 °C, given continuously from germination. Over these temperatures, root length varies more than threefold in the wild type but by only twofold in a double mutant for phytochrome‐interacting factor 4 and 5. For kinematics, the spatial profile of velocity was obtained with new software, Stripflow. We find that 30 °C truncates the elongation zone and curtails cell production, responses that probably reflect the elicitation of a common pathway for handling severe stresses. Curiously, rates of cell division at all temperatures are closely correlated with rates of radial expansion. Between 15 to 25 °C, root growth rate, maximal elemental elongation rate, and final cell length scale positively with temperature whereas the length of the meristem scales negatively. Non‐linear temperature scaling characterizes meristem cell number, time to transit through either meristem or elongation zone, and average cell division rate. Surprisingly, the length of the elongation zone and the total rate of cell production are temperature invariant, constancies that have implications for our understanding of how the underlying cellular processes are integrated.     

Polyploidy and cellular mechanisms changing leaf size: comparison of diploid and autotetraploid populations in two species of Lolium

BACKGROUND AND AIMS: Growth and development of plant organs, including leaves, depend on cell division and expansion. Leaf size is increased by greater cell ploidy, but the mechanism of this effect is poorly understood. Therefore, in this study, the role of cell division and expansion in the increase of leaf size caused by polyploidy was examined by comparing various cell parameters of the mesophyll layer of developing leaves of diploid and autotetraploid cultivars of two grass species, Lolium perenne and L. multiflorum. METHODS: Three cultivars of each ploidy level of both species were grown under pot conditions in a controlled growth chamber, and leaf elongation rate and the cell length profile at the leaf base were measured on six plants in each cultivar. Cell parameters related to division and elongation activities were calculated by a kinematic method. KEY RESULTS: Tetraploid cultivars had faster leaf elongation rates than did diploid cultivars in both species, resulting in longer leaves, mainly due to their longer mature cells. Epidermal and mesophyll cells differed 20-fold in length, but were both greater in the tetraploid cultivars of both species. The increase in cell length of the tetraploid cultivars was caused by a faster cell elongation rate, not by a longer period of cell elongation. There were no significant differences between cell division parameters, such as cell production rate and cell cycle time, in the diploid and tetraploid cultivars. CONCLUSION: The results demonstrated clearly that polyploidy increases leaf size mainly by increasing the cell elongation rate, but not the duration of the period of elongation, and thus increases final cell size.     

Can meristematic activity determine variation in leaf size and elongation rate among four Poa species? A kinematic study

We studied inherent variation in final leaf size among four Poa spp. that live at different elevations. The average final length of leaf 7 of the main stem of the smallest species (Poa alpina) was only one-half that of the largest species (Poa trivialis); it was correlated with leaf elongation rate, but not with the duration of leaf elongation. A faster rate of leaf elongation rate was associated with (a) larger size of the zone of cell expansion, and (b) faster rates of cell production (per cell file) in the meristem, which in turn were due to greater numbers of dividing cells, whereas average cell division rates were very similar for all species (except Poa annua). Also we found that the proliferative fraction equaled 1 throughout the meristem in all species. It was remarkable that rates of cell expansion tended to be somewhat higher in the species with slower growing leaves. We discuss the results by comparing the spatial and material viewpoints, which lead to different interpretations of the role of cell division. Although the presented data do not strictly prove it, they strongly suggest a regulatory role for cell division in determining differences in growth rate among the present four Poa spp.     

The decrease in growth of phosphorus-deficient maize leaves is related to a lower cell production

The spatial distribution of leaf elongation and adaxial epidermal cell production in leaf 6 of maize (Zea mays L. cv. Cecilia) plants grown in a growth chamber under two contrasting availabilities of P in the soil was investigated. Lower displacement velocities from 32.5 mm from leaf base and a shorter growth zone were found in low P (LP) leaves compared with control leaves. P deficiency significantly diminished maximum relative elemental growth rate and shifted its location closer to the leaf base. Cells were significantly longer in LP than in control leaves for all positions from the leaf base except at the end of the growth zone. For both treatments it took a similar time for a cell situated at the leaf base to reach the limit of the growth zone. The average length of the cell division zone was decreased by 21% in LP leaves. Significant differences were found in cell production and cell division rates from 12.5 mm from the leaf base although maximum values were similar between P treatments. A shorter zone of cell division with lower cell production rates along most of its length was the regulatory event that decreased cell production, and ultimately leaf elongation rates, in P‐deficient maize plants.     

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.     

Effect of N deficiency on leaf growth and cell wall peroxidase activity in contrasting maize genotypes

Two maize genotypes differing in leaf elongation rate (high-LER and low-LER) were used for the investigation of the effects of nitrogen deficiency on leaf growth and development and activity of enzyme cell wall peroxidase in the leaf growth zone. Plants were grown in a growth cabinet in perlite as a substrate and watered with complete N-NO₃ solution (+N) and N-NO₃ deficient solution (–N). Comparison between the investigated genotypes showed that final leaf length in both N treatments was related with LER, but not with the duration of leaf elongation. Faster leaf elongation rate in high-LER compared with low-LER genotype, was associated with longer growth zone, a bigger number of cells in it, and higher cell flux rate, although cell elongation rate was similar in both genotypes. These lines of evidence indirectly indicated that leaves of the faster growing genotype were characterized by higher meristematic activity. Nitrogen deficiency reduced the flux of cells and cell elongation rate, length of cell division zone and the number of cells in whole zone, significantly for both genotypes, although duration of cell elongation was increased and final epidermal cell length was unchanged. These results showed that N deficiency reduced both cell division and cell elongation, which in turn resulted in decreased leaf length and prolonged time for leaf development. Nitrogen deficiency significantly increased both bulk and segmental cell wall peroxidase activity in the growth zone of both investigated genotypes, thus showing an interaction between leaf growth cessation and enzyme activity.     

Is thermal time adequate for expressing the effects of temperature on sunflower leaf development?

We have tested whether the effects of temperature on sunflower leaf growth could be documented by using thermal time. The rates of leaf expansion and of cell division were analysed in leaves located at two positions on the stem, and a spatial analysis of expansion rate was carried out. Experiments were performed in growth chamber (stable conditions), in the field or in a greenhouse (fluctuating conditions). We compared three methods for characterizing the rate and the duration of expansion. Responses to leaf temperature were consistent only when expansion was characterized as a two-phase process — a period of exponential expansion (constant relative expansion rate, RER ) followed by a decrease in RER . RER and relative cell division rate ( RDR ) responded linearly to temperature with a common response curve for all studied conditions. This response curve was also common to all studied zones within a leaf and to leaves at two positions on the stem. The reciprocals of the durations of the periods of exponential expansion, non-zero expansion and non-zero division were also linearly related to leaf temperature with common response curves in a given leaf zone. The x -intercepts of all these response curves and of the response curve of leaf initiation rate to temperature did not significantly differ in an analysis of covariance, with a common value of 4·8 °C. The expression of time in cumulative degree days, with a base temperature of 4·8 °C, resulted in a unique time course of RER and cell division rate regardless of temperature. These results suggest that a powerful 'program' of leaf development exists in a sunflower plant.     

Day/night temperature environment affects cell elongation but not division in Lilium longiflorum Thunb

Lilium tongiflorum Thunb. cv. ‘Nellie White’ plantswere grown in different day/night temperature (DT/NT) environmentsto determine the anatomical basis for differential responsesof stem elongation to DT and NT. Lilium plants were forced in1986 and 1987 under 25 and 12 different DT/NT environments,respectively, with temperatures ranging from 14 to 30 °C.Parenchyma and epidermal cell length and width were measuredin stem tissue (1987) and epidermal cell length and width weremeasured in leaf tissue (1986). Total cell number per internodeand vertical cell number per internode were calculated. Stemparenchyma and stem and leaf epidermal cell length increasedlinearly as the difference (DIF) between DT and NT increased(DIF = DT —- NT), i.e. as DT increased relative to NT.DIF had no effect on stem parenchyma width, stem and leaf epidermalcell width, or cell number per internode. Data suggested thatstem elongation responses to DIF are elicited primarily througheffects on cell elongation and not division. Key words: Thermoperiodism, thermomorphogenesis, stem elongation, DIF, cell division, cell elongation, leaf expansion     

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

We have investigated the spatial distributions of cell division rate, p34(cdc2) kinase activity, and amount of p34(cdc2a) in maize (Zea mays) leaves grown at contrasting temperatures and soil water conditions. An original method for calculating cell division rate in all leaf tissues is proposed. In all studied conditions, cell division rate was stable and maximum in the first 2 cm beyond the leaf insertion point, declined afterward, and reached zero at 7 cm from the insertion point. The spatial distribution of p34(cdc2) kinase activity, expressed on a per cell basis, followed the same pattern. In contrast, the amount of p34(cdc2a) was maximum in the first centimeter of the leaf, declined afterward, but remained at 20% of maximum in more distal zones with a near-zero cell division rate. A mild water deficit caused a reduction in cell division rate and p34(cdc2) kinase activity by approximately 45% in all leaf zones, but did not affect the amount of p34(cdc2a). Growth temperature affected to the same extent cell division rate and p34(cdc2) kinase activity, but only if p34(cdc2) kinase activity was assayed at growth temperature, and not if a standard temperature was used in all assays. A common linear relationship between cell division rate and p34(cdc2) kinase activity applied to all causes of changes in cell division rate, i.e. cell aging, water deficit, or changes in temperature. It is shown that temperature has two distinct and additive effects on p34(cdc2) kinase activity; first, an effect on the rate of the reaction, and second, an effect on the amount of p34(cdc2a).     

The Roles of Cell Division and Cell Expansion in the Growth of Alfalfa Leaf Mesophyll

Leaf mesophyll of Medicago sativa (L.) was investigated to determinethe roles of cell division and cell expansion in tissue growth.Samples of leaf tissue were macerated, stained, and squashed.The slides were studied under a phase microscope to determinethe percentage of recently divided cells and the average celldiameter for leaflets of varying lengths. Cell division wasgreatest in young leaflets and virtually ceased as a leaf lengthof 12 mm was attained. For leaflets less than 12 mm in length,the rate of increase in cell size appeared to be inversely associatedto the degree of cell division. For alfalfa leaflets greaterthan 12 mm in length, the mean cell size increased in proportionto leaf length since cell division had virtually ceased.     

Spatial distribution of growth rates and of epidermal cell lengths in the elongation zone during leaf development in Lolium perenne L.

Relative elemental growth rates (REGR) and lengths of epidermal cells along the elongation zone of Lolium perenne L. leaves were determined at four developmental stages ranging from shortly after emergence of the leaf tip to shortly before cessation of leaf growth. Plants were grown at constant light and temperature. At all developmental stages the length of epidermal cells in the elongation zone of both the blade and sheath increased from 12 m at the leaf base to about 550 m at the distal end of the elongation zone, whereas the length of epidermal cells within the joint region only increased from 12 to 40 m. Throughout the developmental stages elongation was confined to the basal 20 to 30 mm of the leaf with maximum REGR occurring near the center of the elongation zone. Leaf elongation rate (LER) and the spatial distributions of REGR and epidermal cell lengths were steady to a first approximation between emergence of the leaf tip and transition from blade to sheath growth. Elongation of epidermal cells in the sheath started immediately after the onset of elongation of the most proximal blade epidermal cells. During transition from blade to sheath growth the length of the blade and sheath portion of the elongation zone decreased and increased, respectively, with the total length of the elongation zone and the spatial distribution of REGR staying near constant, with exception of the joint region which elongated little during displacement through the elongation zone. Leaf elongation rate decreased rapidly during the phase when only the sheath was growing. This was associated with decreasing REGR and only a small decrease in the length of the elongation zone. Data on the spatial distributions of growth rates and of epidermal cell lengths during blade elongation were used to derive the temporal pattern of epidermal cell elongation. These data demonstrate that the elongation rate of an epidermal cell increased for days and that cessation of epidermal cell elongation was an abrupt event with cell elongation rate declining from maximum to zero within less than 10 h.Abbreviations LER leaf elongation rate - REGR relative elemental growth rates     

Ultraviolet-B radiation reduces the rates of cell division and elongation in the primary leaf of wheat (Triticum aestivum L. cv Maris Huntsman)

The primary leaf of wheat (Triticum aestivum L. cv Maris Huntsman) was used as a model system to examine how elevated ultraviolet‐B (UV‐B; λ= 280–320 nm) radiation affected growth. A reduction in the rate and duration of growth of the primary leaf, in response to UV‐B, was the result of changes in both the rate and extent of cell division and elongation. UV‐B reduced the proportion of mitotically active cells (mitotic index) and increased the time taken for cell division (cell doubling time). Thus the supply of cells into the elongation zone was reduced, and this, coupled to a reduction in the rate of elongation, resulted in reduced leaf growth. This analysis of the spatial distribution of growth provided a means of calculating the age of cells within the leaves. Cells of UV‐B‐treated leaves were found to age more quickly than those of the controls. This analysis will enable future studies to take account of age‐related changes when interpreting the response of plants to any number of environmental stresses that affect leaf development.     

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.     

Epidermal Cell Division and the Coordination of Leaf and Tiller Development

Initiation and development of grass leaves and tillers are oftendescribed individually with little attention to possible interrelationshipsamong organs. In order to better understand these interrelationships,this research examined epidermal cell division during developmentaltransitions at the apical meristem of tall fescue (Festuca arundinaceaSchreb.). Ten seedlings were harvested each day for a 9-d period,and lengths of main shoot leaves and primary tillers were measured.In addition, numbers and lengths of epidermal cells were determinedfor 0·5 mm segments along the basal 3 mm of each leafand tiller. Primordia development and onset of rapid leaf elongationwere characterized by an increase in the number of cells perepidermal file with mean cell length remaining near 20 µmper cell. After the leaf had lengthened to 1-1·5 mm,cells near the leaf tip ceased dividing and increased in length,at which time leaf elongation rate increased rapidly. Liguleformation, marking the boundary between blade and sheath cells,occurred prior to leaf tip emergence above the whorl of oldersheaths, while the earliest differentiation between blade andsheath cells probably began when leaves were < 1 mm long.Major transitions in leaf and tiller development appeared tobe synchronized among at least three adjacent nodes. At theoldest node, cessation of cell division in the leaf sheath wasaccompanied by initiation of cell division and elongation inthe associated tiller bud. At the next younger node the ligulewas being initiated, while at the youngest node cell divisioncommenced in the leaf primordium, as elongation of a new leafblade began. This synchronization of events suggests a key rolefor the cell division process in regulating leaf and tillerdevelopment.Copyright 1994, 1999 Academic Press Festuca arundinacea Schreb., tall fescue, cell division, leaf initiation, tillering, ligule development     

Analysis of Cell Division and Elongation Underlying the Developmental Acceleration of Root Growth in Arabidopsis thaliana

To investigate the relation between cell division and expansion in the regulation of organ growth rate, we used Arabidopsis thaliana primary roots grown vertically at 20°C with an elongation rate that increased steadily during the first 14 d after germination. We measured spatial profiles of longitudinal velocity and cell length and calculated parameters of cell expansion and division, including rates of local cell production (cells mm⁻¹ h⁻¹) and cell division (cells cell⁻¹ h⁻¹). Data were obtained for the root cortex and also for the two types of epidermal cell, trichoblasts and atrichoblasts. Accelerating root elongation was caused by an increasingly longer growth zone, while maximal strain rates remained unchanged. The enlargement of the growth zone and, hence, the accelerating root elongation rate, were accompanied by a nearly proportionally increased cell production. This increased production was caused by increasingly numerous dividing cells, whereas their rates of division remained approximately constant. Additionally, the spatial profile of cell division rate was essentially constant. The meristem was longer than generally assumed, extending well into the region where cells elongated rapidly. In the two epidermal cell types, meristem length and cell division rate were both very similar to that of cortical cells, and differences in cell length between the two epidermal cell types originated at the apex of the meristem. These results highlight the importance of controlling the number of dividing cells, both to generate tissues with different cell lengths and to regulate the rate of organ enlargement.     

Growth Rates and Carbohydrate Fluxes within the Elongation Zone of Tall Fescue Leaf Blades

Investigations were performed to better understand the carbon economy in the elongation zone of tall fescue leaf blades. Plants were grown at constant 21°C and continuous 300 micromoles per square meter per second photosynthetic photon flux density where leaf elongation was steady for several days. Elongation occurred in the basal 20 mm of the blade (0-20 millimeters above the ligule) and was maximum at 9 to 12 millimeters. Eight 3-millimeter long segments were sampled along the length of the elongation zone and analyzed for water-soluble carbohydrates. Sucrose concentration was high in the zone of cell division (0-6 millimeters) whereas monosaccharide concentration was high at and distal to the location where cell elongation terminated (20 millimeters). Fructan concentration increased in the basal part, then remained constant at about 85% of the total mass of water-soluble carbohydrates through the remainder of the elongation zone. Data on spatial distribution of growth velocities and substance contents (e.g. microgram fructan per millimeter leaf length) were used to calculate local net rates of substance deposition (i.e. excess rates of substance synthesis and/or import over substance degradation and/or export) and local rates of sucrose import. Rates of sucrose import and net deposition of fructan were positively associated with local elongation rate, whereas net rates of sucrose deposition were high in the zone of cell division and those of monosaccharide were high near the termination of elongation. At the location of most active elongation imported sucrose (29.5 milligrams per square decimeter per hour) was used largely for synthesis of structural components (52%) and fructan (41%).     

A comparative analysis of the temperature response of leaf elongation in Bromus stamineus and Lolium perenne plants in the field: intrinsic and size-mediated effects

BACKGROUND AND AIMS: Growth of grass species in temperate-humid regions is restricted by low temperatures. This study analyses the origin (intrinsic or size-mediated) and mechanisms (activity of individual meristems vs. number of active meristems) of differences between Bromus stamineus and Lolium perenne in the response of leaf elongation to moderately low temperatures. METHODS: Field experiments were conducted at Balcarce, Argentina over 2 years (2003 and 2004) using four cultivars, two of B. stamineus and two of L. perenne. Leaf elongation rate (LER) per tiller and of each growing leaf, number of growing leaves and total leaf length per tiller were measured on 15-20 tillers per cultivar, for 12 (2003) or 10 weeks (2004) during autumn and winter. KEY RESULTS: LER was faster in B. stamineus than in L. perenne. In part, this was related to size-mediated effects, as total leaf length per tiller correlated with LER and B. stamineus tillers were 71% larger than L. perenne tillers. However, accounting for size effects revealed intrinsic differences between species in their temperature response. These were based on the number of leaf meristems simultaneously active and not on the (maximum) rate at which individual leaves elongated. Species differences were greater at higher temperatures, being barely notable below 5 degrees C (air temperature). CONCLUSIONS: Bromus stamineus can sustain a higher LER per tiller than L. perenne at air temperatures > 6 degrees C. In the field, this effect would be compounded with time as higher elongation rates lead to greater tiller sizes.     

Cell growth analysis during steady and non-steady growth in leaves of perennial ryegrass (Lolium perenne L.) subject to defoliation

The effect of defoliation on leaf elongation rate (LER) and on the spatial distribution of epidermal cell lengths in the leaf growth zone was studied in vegetative main tillers of perennial ryegrass (Lolium perenne L. cv Modus) grown in a controlled environment. A new material approach was used to analyse the responses of epidermal cell expansion and production during the initial, non‐steady growth phase following defoliation. The analysis involved assigning an identity to individual expanding cells, assessing the displacement and estimating the expansion of cells with assigned identity during day 1 and day 2 after defoliation. LER decreased by 34% during the first 2 d after defoliation and did not recover to the pre‐defoliation rate within the 14 day regrowth period. Decreased LER on day 1 and day 2 after defoliation was associated with (i) a decrease in the length of the leaf growth zone; (ii) a decrease in the length at which epidermal cells stopped expanding; (iii) a reduced expansion of cells at intermediate growth stages; and (iv) a reduction in cell production (i.e. division) and an associated decrease in the number of expanding cells in the growth zone. However, defoliation had no effect on the expansion of cells located in the proximal part of the growth zone. Reduced LER at 14 d after defoliation was associated with a reduced cell production rate (27% lower than the pre‐defoliation rate) and decreased final cell size ( ? 28%).