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.     

Leaf responses to soil water deficits: Comparative sensitivity of leaf expansion rate and leaf conductance in field-grown sunflower (Helianthus annuus L.)

The relative importance of changes in leaf expansion rate (LER) and leaf conductance (g₁) in the control of crop transpiration depends primarily on their sensitivity to soil water deficits. The aim of this paper was to quantify the responses of LER and g₁ to soil water deficits in sunflower (Helianthus annuus L.) under conditions of moderate (spring) and high (summer) evaporative demand. Soil water content, g₁, and LER were measured in dryland (DRY) and daily-irrigated (WET) crops established on a deep sandy-loam (Typic Xerofluvent) in a Mediterranean environment. There was no difference between g₁ of DRY and WET plants (p>0.20) in contrast with a highly significant difference in LER (p<0.001). Even under the harsh conditions of the summer experiment, g₁ did not respond to water deficit in a ten-day period in which LER of DRY plants was reduced to approx. 30% of that measured in WET controls. This field study indicates that g₁ plays at most a minor role in the control of sunflower transpiration in the pre-anthesis period and confirms the importance of leaf expansion in the regulation of gas exchange of expanding canopies subjected to soil water deficits.     

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.     

Leaf expansion in sunflower as influenced by salinity and short-term changes in carbon fixation

Abstract With a view to defining factors regulating the growth responses of sunflower to salinity, plants were grown in solution culture (0, 50 or 100 mol m⁻³ NaCl) and under natural light, and the areas of every leaf measured once or twice daily from 22 until 38 d after germination. During this period, carbon availability for growth was manipulated by changing light levels and by the use of a photosynthesis inhibitor, DCMU. Salinity reduced relative leaf expansion rates per plant (RLER) by an average of 0.04 (50 mol m⁻³) and 0.08 (100 mol m⁻³) m² m⁻² d⁻¹ compared with control plants of equivalent leaf area: the effects were found in expanding leaves regardless of age or size. Control plants expanded faster during the day than the night, but plants grown in salt had an almost constant RLER throughout the 24 h, indicating that salt influences the rate of utilization of assimilates independently of their production. DCMU reduced RLER considerably in both control and salt-treated plants and reduced the advantage of control plants during the day. Conditions of low light also reduced the differences in RLER between control and salt-treated plants. When salt was removed from the root medium of non-DCMU plants, the expansion rates equalled that of the controls within 24 h and remained at the same levels for the following 3 d measurement period: this recovery applied to leaves of all ages. Salt-grown plants with no photosynthesis (DCMU treatments) also increased their expansion rates upon removal of salt from the root medium, thus providing further evidence that growth was not limited by carbohydrate status, i.e. that salt influences growth primarily via its effects on the rate of utilization of stored assimilates.     

The role of auxin-binding protein 1 in the expansion of tobacco leaf cells

Tobacco leaf was used to investigate the mechanism of action of auxin-binding protein 1 (ABP1). The distributions of free auxin, ABP1, percentage of leaf nuclei in G2 and the amount of auxin-inducible growth were each determined in control tobacco leaves and leaves over-expressing Arabidopsis ABP1. These parameters were compared with growth of tobacco leaves, measured both spatially and temporally throughout the entire expansion phase. Within a defined window of leaf development, juvenile leaf cells that inducibly expressed Arabidopsis ABP1 prematurely advanced nuclei to the G2 phase. The ABP1-induced increase in cell expansion occured before the advance to the G2 phase, indicating that the ABP1-induced G2 phase advance is an indirect effect of cell expansion. The level of ABP1 was highest at the position of maximum cell expansion, maximum auxin-inducible growth and where the free auxin level was the lowest. In contrast, the position of maximum cell division correlated with higher auxin levels and lower ABP1 levels. Consistent with the correlations observed in leaves, tobacco cells (BY-2) in culture displayed two dose-dependent responses to auxin. At a low auxin concentration, cells expanded, while at a relatively higher concentration, cells divided and incorporated [3H]-thymidine. Antisense suppression of ABP1 in these cells dramatically reduced cell expansion with negligible effect on cell division. Taken together, the data suggest that ABP1 acts at a relatively low level of auxin to mediate cell expansion, whereas high auxin levels stimulate cell division via an unidentified receptor.     

Day length affects the dynamics of leaf expansion and cellular development in Arabidopsis thaliana partially through floral transition timing

Background and Aims: Plant aerial development is well known to be affected by daylength in terms of the timing and developmental stage of floraltransition. Arabidopsis thaliana is a ‘long day’plant in which the time to flower is delayed by short days andleaf number is increased. The aim of the work presented herewas to determine the effects of different day lengths on individualleaf area expansion. The effect of flower emergence per se onthe regulation of leaf expansion was also tested in this study. Methods: Care was taken to ensure that day length was the only sourceof micro-meteorological variation. The dynamics of individualleaf expansion were analysed in Ler and Col-0 plants grown underfive day lengths in five independent experiments. Responsesat cellular level were analysed in Ler plants grown under variousday lengths and treatments to alter the onset of flowering. Key Results: When the same leaf position was compared, the final leaf areaand both the relative and absolute rates of leaf expansion weredecreased by short days, whereas the duration of leaf expansionwas increased. Epidermal cell number and cell area were alsoaltered by day-length treatments and some of these responsescould be mimicked by manipulating the date of flowering. Conclusions: Both the dynamics and cellular bases of leaf development arealtered by differences in day length even when visible phenotypesare absent. To some extent, cell area and its response to daylength are controlled by whole plant control mechanisms associatedwith the onset of flowering.     

The CURLY LEAF gene controls both division and elongation of cells during the expansion of the leaf blade in Arabidopsis thaliana

The CURLY LEAF (CLF ) gene in Arabidopsis thaliana (L.) Heynh. is required for stable repression of a floral homeotic gene, AGAMOUS in leaves and stems To clarify the function of CLF in organ development, we characterized clf mutants using an anatomical and genetic approach. The clf mutants had normal roots, hypocotyls, and cotyledons, but the foliage leaves and the stems had reduced dimensions. A decrease both in the extent of cell elongation and in the number of cells was evident in the clf mutant leaves, suggesting that the CLF gene might be involved in the division and elongation of cells during leaf morphogenesis. An analysis of the development of clf mutant leaves revealed that the period during which cell division or cell elongation occurred was of normal duration, while the rates of both cell production and cell elongation were lower than in the wild type. Two phases in the elongation of cells were also recognized from this analysis. From analysis of an angustifolia clf double mutant, we found that the two phases of elongation of leaf cells were regulated independently by each gene. Thus, the CLF gene appears to affect cell division at an earlier stage and cell elongation throughout the development of leaf primordia. Received: 19 February 1998 / Accepted: 24 March 1998     

Kinematic evidence that atmospheric nitrogen dioxide increases the rates of cell proliferation and enlargement to stimulate leaf expansion in Arabidopsis

To elucidate the stimulation of leaf growth by atmospheric nitrogen dioxide (NO₂), we performed a kinematic analysis of the eighth leaves of Arabidopsis thaliana (accession C24) plants grown for 17–35 days after sowing in the presence or absence of 50 ppb NO₂ (designated +NO₂ plants and –NO₂ plants, respectively). We found that the peak and mean values of the relative rates of leaf expansion, cell division and cell expansion were always greater in +NO₂ plants than in –NO₂ plants. No evidence for prolonged duration was obtained. Thus, NO₂ treatment increased the rates of both cell proliferation and enlargement to increase leaf size. Furthermore, a fold-change analysis showed that cell proliferation and enlargement differentially regulated NO₂-induced leaf expansion.     

Spatial distributions of tissue expansion and cell division rates are related to irradiance and to sugar content in the growing zone of maize roots

We have investigated the way in which the radiation absorbed by leaves affects the rate of elongation of maize ( Zea mays L.) roots. In five repeated growth chamber experiments, plants previously grown at a photon irradiance of 23 mol m^–2 d^–1 received either 7 or 34 mol m^–2 d^–1 from day 10 to day 20 after germination. The elongation rate of primary roots steadily decreased for 4 d after reduction in irradiance and then stabilized at 60% of that in plants at high irradiance. The elongating zone was slightly shorter after 2 d at low irradiance, and was further reduced after 8 d. The concentrations of sucrose and glucose in the elongating zone were greatly decreased after 2 d at low irradiance and the gradient of both sugars was suppressed. The longer period at low irradiance affected neither sugar content nor gradient. In the same way, cell production rate was reduced after 2 d at low irradiance and was not appreciably decreased afterwards. The root zone with cell division was shorter in plants at low irradiance, but cell division rate remained nearly constant temporally and spatially, and was unaffected by the irradiance treatment. Our results suggest that primary events after a reduction in irradiance were a change in cell flux and sugar content in the elongating zone. Change in elongation rate was slower and probably the result of a time-related developmental effect, which may be related to the change in cell production.     

Alteration of division polarity and preprophase band orientation in stomatogenesis by light

We have developed an experimental system in which the irradiation with a red light pulse induces stomatal disorientation in the hypocotyl epidermis ofCucumis sativus L. In this system, the orientation of the division plane in guard mother cells was not defined correctly. Preprophase bands formed in these cells but their orientation was abnormal.     

A lysine-rich arabinogalactan protein in Arabidopsis is essential for plant growth and development, including cell division and expansion

Arabinogalactan proteins (AGPs), a family of hydroxyproline-rich glycoproteins, occur throughout the plant kingdom. The lysine-rich classical AGP subfamily in Arabidopsis consists of three members, AtAGP17, 18 and 19. In this study, AtAGP19 was examined in terms of its gene expression pattern and function. AtAGP19 mRNA was abundant in stems, with moderate levels in flowers and roots and low levels in leaves. AtAGP19 promoter-controlled GUS activity was high in the vasculature of leaves, roots, stems and flowers, as well as styles and siliques. A null T-DNA knockout mutant of AtAGP19 was obtained and compared to wild-type (WT) plants. The atagp19 mutant had: (i) smaller, rounder and flatter rosette leaves, (ii) lighter-green leaves containing less chlorophyll, (iii) delayed growth, (iv) shorter hypocotyls and inflorescence stems, and (v) fewer siliques and less seed production. Several abnormalities in cell size, number, shape and packing were also observed in the mutant. Complementation of this pleiotropic mutant with the WT AtAGP19 gene restored the WT phenotypes and confirmed that AtAGP19 functions in various aspects of plant growth and development, including cell division and expansion, leaf development and reproduction.     

Effect of light and gibberellic acid on cell division in the first foliage leaf of durum wheat (Triticum durum Desf.)

The present paper is part of a research program which aims at a quantitative analysis of the effects of light and gibberellic acid (GA₃) on growth of the first foliage leaf in durum wheat (Triticum durum Desf.). Since leaf growth is the combined result of the increase in cell number (cell division) and cell enlargement, the influence of light and GA₃ treatment on cell division in the basal meristem of the first leaf in two cultivars, Cappelli and Creso, was investigated. Creso is a short-strawed cultivar carrying the Gai 1 gene which influences both plant height and insensitivity to applied GA₃. Cell division, as measured by mitotic index, was similar in darkness, continuous red light and dichromatic irradiation (far-red plus red), while lower mitotic rates were observed under continuous far-red light: this indicates that the response of cell division is modulated by a high-irradiance reaction of phytochrome in both cultivars. The two cultivars showed different responses to blue light. In Cappelli, blue light and dichromatic irradiation (blue plus red) gave lower mitotic indices than the dark control, indicating the action of a specific blue-light-absorbing photoreceptor, whereas in Creso the response kinetics to all light regimes which included blue light were more complex. On the basis also of the results obtained with GA₃ application in Cappelli, it appears that (i) the hormonal treatment is able to change the pattern of mitotic index only in the presence of the action of a blue-light receptor and (ii) the different responses of the two cultivars could be the result of different endogenous hormonal levels. The importance of the observations in relation to the data for first-leaf longitudinal growth reported in a previous paper (Baroncelli et al. 1984, Planta 160, 298–304) is discussed.Abbreviations BL blue light - D darkness - FR far-red light - GA gibberellin - GA₃ gibberellic acid - m.i. mitotic index - Norflurazon 4-chloro-5-(methylamino)-2-(,,,-trifluoro-m-totyl-3(2H)) pyridazinone - R red light - WL white light - phytochrome photoequilibrium     

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.     

Role of proline and leaf expansion rate in the recovery of stressed white clover leaves with increased phosphorus concentration

Osmotic adjustment (OA) and increased cell-wall extensibility required for expansive leaf growth are well defined components of adaptation to water stress in dry soil, which might interact with soil phosphorus (P) concentration and defoliation frequency for intensively grazed white clover in legume-based pastures. Experiments were conducted with frequently and infrequently defoliated mini-swards of white clover growing in dry soil with low and high P concentrations. The higher yielding high-P plants were able to dry the soil to greater soil water suctions; their leaves had lower water potential values, yet they showed fewer water stress symptoms and underwent a more complete recovery from the water stress symptoms on rewatering, than the low-P plants. High- P plants had greater OA, proline concentration and leaf expansion rate. On the other hand, low-P plants showed an increased osmotic concentration when there was no change in the total solute content per unit of leaf d. wt, indicating more loss of water from the leaf tissue. The key measures that appeared to be directly associated with plant recovery over a short period following water stress were increased proline concentration and leaf expansion rate, probably resulting from increased cell-wall extensibility rather than increased production of cells for the high-P plants.     

Arv1 promotes cell division by recruiting IQGAP1 and myosin to the cleavage furrow

Cell division is strictly regulated by a diversity of proteins and lipids to ensure proper duplication and segregation of genetic material and organelles. Here we report a novel role of the putative lipid transporter ACAT-related protein required for viability 1 (Arv1) during telophase. We observed that the subcellular localization of Arv1 changes according to cell cycle progression and that Arv1 is recruited to the cleavage furrow in early telophase by epithelial protein lost in neoplasm (EPLIN). At the cleavage furrow Arv1 recruits myosin heavy chain 9 (MYH9) and myosin light chain 9 (MYL9) by interacting with IQ-motif-containing GTPase-activating protein (IQGAP1). Consequently the lack of Arv1 delayed telophase-progression, and a strongly increased incidence of furrow regression and formation of multinuclear cells was observed both in human cells in culture and in follicle epithelial cells of egg chambers of Drosophila melanogaster in vivo. Interestingly, the cholesterol-status at the cleavage furrow did not affect the recruitment of either IQGAP1, MYH9 or MYL. These results identify a novel function for Arv1 in regulation of cell division through promotion of the contractile actomyosin ring, which is independent of its lipid transporter activity.     

Overexpression of OsKTN80a,a katanin P80ortholog,caused the repressed cell elongation and stalled cell division mediated by microtubule apparatus defects in primary root in Oryza sativa

Katanin, a microtubule‐severing enzyme, consists of two subunits: the catalytic subunit P60, and the regulatory subunit P80. In several species, P80 functions in meiotic spindle organization, the flagella biogenesis, the neuronal development, and the male gamete production. However,the P80 function in higher plants remains elusive. In this study, we found that there are three katanin P80 orthologs(OsKTN80a, OsKTN80b, and OsKTN80c) in Oryza sativa L.Overexpression of OsKTN80a caused the retarded root growth of rice seedlings. Further investigation indicates that the retained root growth was caused by the repressed cell elongation in the elongation zone and the stalled cytokinesis in the division zone in the root tip. The in vivo examination suggests that OsKTN80a acts as a microtubule stabilizer. We prove that OsKTN80a, possibly associated with OsKTN60, is involved in root growth via regulating the cell elongation and division.     

Overexpression of OsKTN80a,a katanin P80 ortholog,caused the repressed cell elongation and stalled cell division mediated by microtubule apparatus defects in primary root in Oryza sativa

Katanin, a microtubule-severing enzyme, consists of two subunits：the catalytic subunit P60, and the regulatory subunit P80. In several species, P80 functions in meiotic spindle organization, the flagella biogenesis, the neuronal development, and the male gamete production. However, the P80 function in higher plants remains elusive. In this study, we found that there are three katanin P80 orthologs （OsKTN80a, OsKTN80b, and OsKTN80c） in Oryza sativa L. Overexpression of OsKTN80a caused the retarded root growth of rice seedlings. Further investigation indicates that the retained root growth was caused by the repressed cell elongation in the elongation zone and the stalled cytokinesis in the division zone in the root tip. The in vivo examination suggests that OsKTN80a acts as a microtubule stabilizer. We＆amp;nbsp;prove that OsKTN80a, possibly associated with OsKTN60, is involved in root growth via regulating the cell elongation and division.