Rate Of Cell Enlargement Research Articles

What drives forest carbon storage? The ramifications of source-sink decoupling.

This article is a Commentary on Martínez‐Sancho et al. (2022), 236: 58–70.

Modelling the spatial crosstalk between two biochemical signals explains wood formation dynamics and tree-ring structure.

In conifers, xylogenesis during a growing season produces a very characteristic tree-ring structure: large, thin-walled earlywood cells followed by narrow, thick-walled latewood cells. Although many factors influence the dynamics of differentiation and the final dimensions of xylem cells, the associated patterns of variation remain very stable from one year to the next. While radial growth is characterized by an S-shaped curve, the widths of xylem differentiation zones exhibit characteristic skewed bell-shaped curves. These elements suggest a strong internal control of xylogenesis. It has long been hypothesized that much of this regulation relies on a morphogenetic gradient of auxin. However, recent modelling studies have shown that while this hypothesis could account for the dynamics of stem radial growth and the zonation of the developing xylem, it failed to reproduce the characteristic tree-ring structure. Here, we investigated the hypothesis of regulation by a crosstalk between auxin and a second biochemical signal, by using computational morphodynamics. We found that, in conifers, such a crosstalk is sufficient to simulate the characteristic features of wood formation dynamics, as well as the resulting tree-ring structure. In this model, auxin controls cell enlargement rates while another signal (e.g. cytokinin, tracheary element differentiation inhibitory factor) drives cell division and auxin polar transport.

Tree physiological monitoring of the 2018 larch budmoth outbreak: preference for leaf recovery and carbon storage over stem wood formation in Larix decidua.

Insect defoliation impacts forest productivity worldwide, highlighting the relevance of plant-insect interactions. The larch budmoth (Zeiraphera griseana Hübner) is one of the most extensively studied defoliators, where numerous tree ring-based analyses on its host (Larix decidua Mill.) have aided in identifying outbreak dynamics over the past millennia. Yet, outbreaks have been widely absent after the early 1980s, and little is known about the in situ tree physiological responses and the allocation of carbon resources during and after defoliation. In summer 2018, we tracked an ongoing larch budmoth outbreak in a well-studied larch forest in the Swiss Alps. We performed biweekly monitoring on an affected and unaffected site using a unique combination of xylogenesis observations, measurements of non-structural carbohydrates, isotopic analysis of needle assimilates and ground-based and remote-sensed leaf trait observations. The budmoth induced a defoliation that lasted 40days and could be detected by satellite observations. Soluble sugars significantly decreased in needles and stem phloem of the defoliated trees, while starch levels remained stable in the stem and root xylem compared to the control. Carbon and oxygen isotope ratios in needle assimilates indicated that neither photosynthetic assimilation rates nor stomatal conductance was different between sites before, during and after the outbreak. Defoliated trees ceased cell wall thickening 17days earlier than unaffected trees, showing the earliest halt of ring formation recorded from 2007 untill 2013 and causing significant thinner cell walls, particularly in the latewood. No significant differences were found for cell enlargement rates and ring width. Our study revealed that an outbreak causes a downregulation of cell wall thickening first, while no starch is mobilized or leaf physiology is adjusted to compensate for the reduced carbon source due to defoliation. Our observations suggest that affected larch trees prioritize leaf recovery and carbon storage over wood biomass development.

Involvement of cell proliferation and cell enlargement in increasing the fruit size of Malus species

Cell number and cell size in the fruit flesh of five Malus cultivars (two crabapples and three domesticated apples) were analyzed from soon after blossom until maturity. M. floribunda, whose fruit size is one of the smallest among Malus, exhibited absence of cell proliferation throughout fruit ontogeny. Furthermore, the rate of cell enlargement, which was almost uniform until the mature stage in other species, slowed down from about 45 days after full blossom (DAFB), resulting in a cell size approximately half that of the other cultivars. The fruit of M. coronaria, which is of medium size, possessed approximately half the cell number of M. domestica cv. ‘Fuji’, due to a slower proliferation rate. The cv. ‘Sekaiichi’ of M. domestica had the largest fruit among the cultivars investigated, and showed a faster increase in cell number. These results indicate that a combination of greater cell division capacity and an enhanced degree of cell enlargement are involved in the increase of Malus fruit size.

Hydraulic Conductivity and Aquaporins of Cortical Cells in Gravitropically Bending Roots of Pisum sativum L.

We examined the differential elongation of gravitropically bending roots of Pisum sativum L. in terms of cell enlargement and water uptake by cells in the growing tissue. Hydraulic conductivity between the elongating and mature tissues (Lp) was estimated from the equation G = A × Lp × Δψ, where G is the water-uptake rate, A is the surface area of a single cell and Δψ is the driving force. The rate of entry of water into a cell was estimated from the rate of increase in the volumes of cells in the outer cortex, which were calculated from longitudinal sections at given times. Gravitropic bending occurred 1 h after the application of gravi-stimulation and the curvature increased rapidly for the next 3 h. The biggest difference in the partial elongation rate between opposite sides of a root was found in the region 3 to 4 mm from the root tip at the start of stimulation. Cell enlargement rate was 2.8 to 3.8 times greater on the upper side of the root than on the lower side. The water potential and the osmotic potential, in both the elongating and mature tissues, were the same on both sides of the root. Therefore, there was no difference in the driving force for water flow. Hydraulic conductivity was 2.3 to 4.2 times greater on the upper side of the root than on the lower side. There was no difference between the upper and lower sides of the root in the amounts of 19-kD and 24-kD proteins in membrane fractions, which we assumed to be aquaporins (putative aquaporins), as estimated with two preparations of polyclonal antibodies. The differential elongation that occurred during root gravitropism was caused by a difference in Lp. However, the difference in Lp did not appear to be regulated by the concentration in cell membranes of the putative aquaporins.

Auxin-activated NADH oxidase activity of soybean plasma membranes is distinct from the constitutive plasma membrane NADH oxidase and exhibits prion-like properties.

The hormone-stimulated and growth-related cell surface hydroquinone (NADH) oxidase activity of etiolated hypocotyls of soybeans oscillates with a period of about 24 min or 60 times per 24-h day. Plasma membranes of soybean hypocotyls contain two such NADH oxidase activities that have been resolved by purification on concanavalin A columns. One in the apparent molecular weight range of 14-17 kDa is stimulated by the auxin herbicide 2,4-dichlorophenoxyacetic acid (2,4-D). The other is larger and unaffected by 2,4-D. The 2,4-D-stimulated activity absolutely requires 2,4-D for activity and exhibits a period length of about 24 min. Also exhibiting 24-min oscillations is the rate of cell enlargement induced by the addition of 2,4-D or the natural auxin indole-3-acetic acid (IAA). Immediately following 2,4-D or IAA addition, a very complex pattern of oscillations is frequently observed. However, after several hours a dominant 24-min period emerges at the expense of the constitutive activity. A recruitment process analogous to that exhibited by prions is postulated to explain this behavior.

Soybean cell enlargement oscillates with a temperature-compensated period length of ca. 24 min.

Rate of enlargement of epidermal cells from soybean, when measured at intervals of 1 min using a light microscope equipped with a video measurement system, oscillated with a period length of about 24 min. This oscillation parallels the 24-min periodicity observed for the oxidation of NADH by the external plasma membrane NADH oxidase. The increase in length was not only non-linear, but intervals of rapid increase in area alternated with intervals of rapid decrease in area. The length of the period was temperature compensated, and was approximately the same when measured at 14, 24 and 34 degrees C even though the rate of cell enlargement varied over this same range of temperatures. These observations represent the first demonstration of an oscillatory growth behavior correlated with a biochemical activity where the period length of both is independent of temperature (temperature compensated) as is the hallmark of clock-related biological phenomena.

CHO cell enlargement oscillates with a temperature-compensated period of 24 min

The rate of increase in cell area of CHO cells when measured at intervals of 1 min using a light microscope equipped with a video measurement system, oscillated with a minimum period of about 24 min. The pattern of oscillations paralleled those of the 24 min period observed with the oxidation of NADH by an external cell surface or plasma membrane NADH oxidase. The increase in cell area was non-linear. Intervals of rapid increase in area alternated with intervals of rapid decrease in area. The length of the 24 min period was temperature-compensated (approximately the same when measured at 14°C, 24°C or 34°C) while the rate of cell enlargement increased with temperature over this same range of temperatures.

Chemical hormesis in cell growth: a molecular target at the cell surface

A multifunctional ubiquinol (NADH) oxidase with protein disulfide-thiol interchange activity of the cell surface, abbreviated as NOX, is described as a molecular target for chemical hormesis of cell growth. The activity of the NOX correlates with rate of cell enlargement, which helps to determine how rapidly cells will divide. When NOX activity is inhibited, cells fail to enlarge following division and the result is a population of small cells unable to reach the minimum size required for them to divide again. In plants, cells fail to enlarge when NOX activity is inhibited. When NOX activity is stimulated or constitutively activated, as in cancer, cells enlarge more rapidly and the rate of cell division also is enhanced. Both cell growth and NOX activity are sometimes stimulated by low concentrations of normally inhibitory molecules. These properties define chemical hormesis, making the NOX molecule a molecular target to explain hormetic growth responses and to utilize hormetic principles to increase, for example, crop yields with plants. The NOX activity at the cell surface oscillates with a temperature-compensated 24-min ultradian (<24 h) periodicity. The indicated function of the NOX protein as a time-keeping mechanism adds to its potential importance as a molecular target for chemical hormesis.

NADH oxidase activity of soybean plasma membranes inhibited by submicromolar concentrations of ATP.

The activity of an auxin-stimulated NADH oxidase activity from soybean hypocotyls was inhibited by submicromolar concentrations of ATP. Auxins are plant growth regulators that increase the rate of cell enlargement in plant stems. A synthetic auxin, 2,4-dichlorophenoxyacetic acid (2,4-D), was used. The inhibition was half maximal at 1 nM ATP and was not observed with other nucleotides and nucleosides. The inhibition was the result of an increase in the Km for NADH from about 60 microM to > 100 microM and was noncompetitive. The decrease in Km due to ATP was enhanced by the addition of 1 microM 2,4-D. The Vmax of the plasma membrane NADH oxidase was approximately doubled (1.5-2.8-fold) by ATP and by 1 microM 2,4-D. No further increase in the Vmax was observed by the combination of 1 nM to 0.1 mM ATP in the presence of 1 microM 2,4-D. The results demonstrate a response of the NADH oxidase activity of isolated vesicles of soybean plasma membranes to ATP distinct from that observed previously with other nucleotide di- and triphosphates. The results are suggestive either of control of the cell surface NADH oxidase by phosphorylation or a direct response to ATP binding at nanomolar concentrations of ATP.

Differences in cell proliferation and enlargement between seeded and seedless grape berries induced parthenocarpically by gibberellin

SummaryA comparative study was conducted of cell proliferation and enlargement in seeded and seedless ‘Delaware’ grape berries induced parthenocarpically by pre-bloom application of gibberellin A3. Using paraffin-celloidin thin sections taken from the equatorial region of berry, number of cell layers and cells, and cell size were measured in the inner wall, outer wall parenchyma, subepidermis and epidermis directly by microscope or using microphotography. In the seedless berry, cell division and enlargement proceeded simultaneously in all tissues after post-bloom application of gibberellin A3. By then, in the seeded berry, cell division in the tissues, with the exception of the epidermis, was nearly complete. The cell numbers in epidermis and subepidermis collenchyma were fewer in the seedless berry than in the seeded berry. On the other hand, the number of parenchymatous cells in the outer wall of the seedless berry was approximately 1.4 times that found in the seeded berry. Cell enlargement rate was higher in the seedless berry than in the seeded berry, in all tissues. The cells in the outer wall parenchyma and the inner wall of the seedless berry were larger tangentially and radially, respectively, than those in the seeded berry. These results indicate that development of the seedless berry induced by application of gibberellin A3 follows quite a different course of cell division and enlargement when compared with the seeded berry.

Starch Synthesis in the Kernel of Wheat Under High Temperature Conditions

As temperature rises above 18-22�C, the observed decrease in the duration of deposition of dry matter in the kernel is not accompanied by a compensating increase in the rate of grain filling with the result that grain weight (and yield) is diminished at high temperature. Reduced starch content accounts for most of the reduction in grain dry matter at high temperature. Responses to temperature in the low temperature range, 20-30�C (the LTR), could possibly be ascribed to the temperature response characteristics of the reaction catalysed by soluble starch synthase (SSS), the enzyme synthesising starch. However, the rate of cell enlargement and the rate of accumulation of nitrogen in the grain also do not increase much as temperature rises, so other explanations are conceivable for the temperature responses in the LTR. Variation amongst cultivars of wheat in tolerance of high temperature is evident in the LTR. At temperatures above 30�C (in the high temperature range (HTR) between 30 and 40�C), even for short periods, the rate of starch deposition is slower than that observed at lower temperatures, an effect which is carried over after transfer from high to lower temperatures. This response is attributable to a reduction in the activity, possibly due to thermal denaturation, of SSS. Several forms of SSS are found in cereal endosperm, and some forms may be more tolerant of high temperature than others. Loss of enzyme activity at high temperature is swift, but is partly restored some time after transfer from hot to cool conditions. There appear to be two distinct mechanisms of response to elevated temperature, both resulting in a reduced grain weight through reduced starch deposition, but one of them is important only in the range of temperature above 30�C.

Loss of arabinogalactan-proteins from the plasma membrane of NaCl-adapted tobacco cells

Cultured cells of tobacco (Nicotiana tabacum L.) adapted to 428 mM NaCl exhibited a reduced rate of cell enlargement, which is probably due to decreased cell-wall extensibility. Arabinogalactan-protein (AGP) has been implicated as a cell-wall-loosening factor (Schopfer 1990). Levels of plasma membrane and extracellular AGPs that react with Yariv reagent were measured and compared between NaCl-adapted and unadapted tobacco cells. Unadapted cells contained a very high level of AGPs on the plasma membrane, which amounted to 0.16 μg·μg−1 membrane protein. In contrast, AGPs were virtually undetectable on the plasma membrane of NaCl-adapted cells. Accumulation of AGPs was also decreased in culture media of NaCl-adapted cells. These data support the hypothesis that AGPs participate in cell expansion. Possible mechanisms of the proposed cell-expansion role of AGPs are discussed.

In vivo creep and stress relaxation experiments to determine the wall extensibility and yield threshold for the sporangiophores of phycomyces

The pressure probe was used to conduct in vivo creep and in vivo stress relaxation experiments on the sporangiophores of Phycomyces blakesleeanus. The in vivo creep and in vivo stress relaxation methods are compared with respect to their utility for determining the irreversible wall extensibility and the yield threshold. The results of the in vivo stress relaxation experiments demonstrate that the growth usually does not cease when the external water supply is removed, and the turgor pressure does not decay for hours afterwards. A successful stress relaxation experiment requires that the cell enlargement rate (growth rate) be zero during the turgor pressure decay. In a few experiments, the growth rate was zero during the turgor pressure decay. However, in general only the yield threshold could be determined.In vivo creep experiments proved to be easier to conduct and more useful in determining values for both the irreversible wall extensibility and the yield threshold. The results of the in vivo creep experiments demonstrate that small steps-up in turgor pressure, generally <0.02 MPa, elicit increases in growth rate as predicted by the growth equations and the augmented growth equations. The irreversible wall extensibility and the yield threshold were determined from these results. The results also demonstrate that steps-up in turgor pressure larger than 0.02 MPa, produce a different response; a decrease in growth rate. The decreased growth rate behavior is related to the magnitude of the step-up, and in general, larger steps-up in turgor pressure produce larger decreases in growth rate and longer periods of decreased growth rate. Qualitatively, this growth behavior is very similar to the "stretch response" previously reported by Dennison and Roth (1967).

Effects of the growth regulator 4PU-30 on growth, K+ content, and alkaloid production in tobacco callus cultures

Growth, K+ content, and alkaloid production were compared in nonorganogenetic callus cultures ofNicotiana tabacum cv. Burley 21 grown at 25°C in the dark on two different media: a basal medium with 1 μM α-naphthaleneacetic acid and 1 μM kinetin, and one with 1 μM α-naphthaleneacetic acid and 1 μM 4PU-30 (N-(2-chloro-4-pyridyl)-N′-phenylurea). These callus tissues behaved differently not only in growth and K+ content but also in alkaloid production. In comparison to cultures grown with kinetin, those grown with 4PU-30 showed a significantly higher fresh weight and dry weight and K+ content during the growth period studied. The data clearly indicate a positive correlation between K+ uptake rate stimulated by 4PU-30 and cell enlargement rate. However, the alkaloid biosynthesis in the callus tissues was activated by the supply of kinetin and diminished by that of 4PU-30. It thus appears that cellular enlargement of meristematic tissue stimulated by 4PU-30 limited alkaloid production.

Wall yield threshold and effective turgor in growing bean leaves

The rate of cell enlargement depends on cell-wall extensibility (m) and on the amount of turgor pressure (P) which exceeds the wall yield threshold (Y). The difference (P-Y) is the growth-effective turgor (P e). Values of P, Y and P ehave been measured in growing bean (Phaseolus vulgaris L.) leaves with an isopiestic psychrometer, using the stress-relaxation method to derive Y. When rapid leaf growth is initiated by light, P, Y and P eall decrease. Thereafter, while the growth rate declines in maturing leaves, Y continues to decrease and P eactually increases. These data confirm earlier results indicating that the changes in light-stimulated leaf growth rate are primarily controlled by changes in m, and not by changes in P e. Seedlings incubated at 100% relative humidity have increased P, but this treatment does not increase growth rate. In some cases Y changes in parallel with P, so that P eremains unchanged. These data point out the importance of determining P e, rather than just P, when relating cell turgor to the growth rate.

The Role of Hormones in Wall Loosening and Plant Growth

The rate at which any plant cell enlarges is determined by the product of two cellular parameters; the wall extensibility (m) and the effective turgor (�*p - Y), where �*p is the turgor pressure and Y is the wall yield threshold. When hormones modulate the rate of cell enlargement, they do so by altering one or both of these parameters. To determine whether the affected parameter is m, one must be able to measure it. There are four methods for assessing m; the Instron technique; stress relaxation; turgor relaxation; and the growth rate v. turgor method. Each has advantages and disadvantages. Ideally, at least two methods should be used and the results should agree, at least qualitatively. In every case so far examined, wherever auxin promotes cell elongation, it also increases m. Gibberellins and cytokinins, on the other hand, sometimes increase m and sometimes do not. Abscisic acid, in the few cases tested, decreases m, while the effects of ethylene are mixed. A change in m can occur only if the cell exports (or takes up) a wall loosening factor (WLF) in response to the hormone. In some cases the WLF must be, at least in part, protons, but in other cases it is clear that it is something different. In dicotyledonous stems it could even be the uptake of Ca�+ from the walls. Turgor can be influenced by hormones but, with the exception of gibberellins, the responses appear to be secondary, induced by the growth processes rather than by the hormone. However, the ability of cells to take up osmotic solutes and maintain �*p may be the most important factor, in nature, modulating the growth rate of plants on an hour-to-hour basis.

Control of the rate of cell enlargement: Excision, wall relaxation, and growth-induced water potentials

A new guillotine thermocouple psychrometer was used to make continuous measurements of water potential before and after the excision of elongating and mature regions of darkgrown soybean (Glycine max L. Merr.) stems. Transpiration could not occur, but growth took place during the measurement if the tissue was intact. Tests showed that the instrument measured the average water potential of the sampled tissue and responded rapidly to changes in water potential. By measuring tissue osmotic potential (Ψ s ), turgor pressure (Ψ p ) could be calculated. In the intact plant, Ψ s and Ψ p were essentially constant for the entire 22 h measurement, but Ψ s was lower and Ψ p higher in the elongating region than in the mature region. This caused the water potential in the elongating region to be lower than in the mature region. The mature tissue equilibrated with the water potential of the xylem. Therefore, the difference in water potential between mature and elongating tissue represented a difference between the xylem and the elongating region, reflecting a water potential gradient from the xylem to the epidermis that was involved in supplying water for elongation. When mature tissue was excised with the guillotine, Ψ s and Ψ p did not change. However, when elongating tissue was excised, water was absorbed from the xylem, whose water potential decreased. This collapsed the gradient and prevented further water uptake. Tissue Ψ p then decreased rapidly (5 min) by about 0.1 MPa in the elongating tissue. The Ψ p decreased because the cell walls relaxed as extension, caused by Ψ p , continued briefly without water uptake. The Ψ p decreased until the minimum for wall extension (Y) was reached, whereupon elongation ceased. This was followed by a slow further decrease in Y but no additional elongation. In elongating tissue excised with mature tissue attached, there was almost no effect on water potential or Ψ p for several hours. Nevertheless, growth was reduced immediately and continued at a decreasing rate. In this case, the mature tissue supplied water to the elongating tissue and the cell walls did not relax. Based on these measurements, a theory is presented for simultaneously evaluating the effects of water supply and water demand associated with growth. Because wall relaxation measured with the psychrometer provided a new method for determining Y and wall extensibility, all the factors required by the theory could be evaluated for the first time in a single sample. The analysis showed that water uptake and wall extension co-limited elongation in soybean stems under our conditions. This co-limitation explains why elongation responded immediately to a decrease in the water potential of the xylem and why excision with attached mature tissue caused an immediate decrease in growth rate without an immediate change in Ψ p.

Photocontrol of extension growth: a biophysical approach

The analysis of plant growth as a physical process is briefly reviewed. Growth requires the coordinated uptake of water and the irreversible expansion of the cell wall. Any agent that affects the growth rate must act on one or more of the parameters governing water absorption (e.g. the hydraulic conductivity or the difference in osmotic pressure of the cell contents and the water source) or cell wall expansion (e.g. wall extensibility or the yield threshold). When the hydraulic conductivity of the pathway for water transport is small enough to impede the rate of cell enlargement, a substantial gradient in water potential within the growing tissue will develop to sustain the absorption of water. In such a case, the analysis shows that turgor pressure is a key indicator for determining whether an agent acts predominantly on the osmotic properties of the tissue or on the cell wall properties. Furthermore, the dynamic response to a slight perturbation from steady-state conditions is shown to be a function of parameters for both the water relations and cell wall expansion of the tissue. Blue irradiation of etiolated seedlings causes a large inhibition of stem elongation with lag times as short as 30 s and half-times as short as 20 to 25 s. The biophysical mechanism of blue-light suppression of growth was studied in cucumber and sunflower seedlings by means of direct and indirect measurements of turgor pressure. The results indicate that (a) blue light suppresses growth by influencing the cell wall properties of the growing tissue, and (b) the hydraulic conductivity of the growing tissue is large enough for it not to limit the rate of cell enlargement.

Osmotic Properties of Pea Internodes in Relation to Growth and Auxin Action

The water transport properties of etiolated pea (Pisum sativum L.) internodes were studied using both dynamic and steady-state methods to determine (a) whether water transport through the growing tissue limits the rate of cell enlargement, and (b) whether auxin stimulates growth in part by increasing the hydraulic conductance of the growing tissue.Measurements using the pressure probe technique showed that the hydraulic conductivity of cortical cell membranes was the same for both slowly growing and auxin-induced rapidly growing cells (membrane hydraulic conductivity, about 1.5 x 10(-5) centimeters per second per bar). In a second technique which measured the rate of water movement through the entire pea internode, the half-time for radial water flow was about 60 seconds and was not altered by auxin application. These results indicate that auxin does not alter the hydraulic conductance of pea stem tissue, either at the cellular or the whole tissue level.Measurements of the turgor pressure of cortical cells, combined with osmotic pressure measurements of expressed cell sap, show that the water potential of growing pea stems was about -3 bars. When the growth rate was altered by various treatments, including decapitation, auxin application, cold temperature, and KCN treatment, the water potential was independent of the growth rate of the stem. We attribute the depression of the water potential in young pea stems to the presence of solutes in the cell wall free space of the tissue. This interpretation is supported by the results of infiltration and perfusion experiments.From the results of these dynamic and steady-state experiments, we conclude that the internal gradient in water potential (from the xylem to the epidermis) needed to sustain cell enlargement is small (no greater than 0.5 bar). Thus, the hydraulic conductance of the tissue is sufficiently large that it does not control or limit the rate of cell enlargement.