Peroxidase Activity in the Leaf Elongation Zone of Tall Fescue : II. Spatial Distribution of Apoplastic Peroxidase Activity in Genotypes Differing in Length of the Elongation Zone

Previous work suggested that cell wall peroxidase activity increased as cells were displaced through the elongation zone in leaf blades of tall fescue (Festuca arundinacea Schreb.). In this study, two genotypes that differ in length of the elongation zone were used to examine the relationship between peroxidase activity in apoplastic fluid of intact leaf blade segments and the spatial distribution of leaf growth. Apoplastic fluid was extracted by vacuum infiltration and centrifugation, and peroxidase activity was assayed spectrophotometrically. Isoelectric focusing was used to characterize the isoforms of apoplastic peroxidase within the region of elongation and in the region of secondary cell wall deposition, which is distal to the elongation zone. A striking correlation was found in each genotype between both the location and timing of increase in apoplastic peroxidase activity and the onset of growth deceleration. Only cationic isoforms of apoplastic peroxidase could be identified in the elongation zone, whereas additional anionic isoforms appeared in the region of secondary cell wall deposition. We conclude that cessation of elongation growth in tall fescue leaf blades is likely to be related to the secretion of cationic isoforms of peroxidase into the cell wall.     

Photosynthate partitioning in Basal zones of tall fescue leaf blades

Elongating grass leaves have successive zones of cell division, cell elongation, and cell maturation in the basal portion of the blade and are a strong sink for photosynthate. Our objective was to determine dry matter (DM) deposition and partitioning in basal zones of elongating tall fescue (Festuca arundinacea Schreb.) leaf blades. Vegetative tall fescue plants were grown in continuous light (350 micromoles per square meter per second photosynthetic photon flux density) to obtain a constant spatial distribution of elongation growth with time. Content and net deposition rates of water-soluble carbohydrates (WSC) and DM along elongating leaf blades were determined. These data were compared with accumulation of ¹⁴C in the basal zones following leaf-labeling with ¹⁴CO₂. Net deposition of DM was highest in the active cell elongation zone, due mainly to deposition of WSC. The maturation zone, just distal to the elongation zone, accounted for 22% of total net deposition of DM in elongating leaves. However, the spatial profile of ¹⁴C accumulation suggested that the elongation zone and the maturation zone were sinks of equal strength. WSC-free DM accounted for 55% of the total net DM deposition in elongating leaf blades, but only 10% of incoming ¹⁴C-photosynthate accumulated in the water-insoluble fraction (WIF ≈ WSC-free DM) after 2 hours. In the maturation zone, more WSC was used for synthesis of WSC-free DM than was imported as recent photosynthate.     

Measurement of a growth-induced water potential gradient in tall fescue leaves

Spatial distribution of cell turgor pressure, cell osmotic pressure and relative elemental growth rate were measured in growing tall fescue leaves ( Festuca arundinacea ). Cell turgor pressure (measured with a pressure probe) was c . 0.55 MPa in expanding cells but increased steeply (+0.3 MPa) in cells where elongation had stopped. However, cell osmotic pressure (measured with a picolitre osmometer) was almost constant at 0.85 MPa throughout the leaf. The water potential difference between the growth zone and the mature zone (0.3 MPa) was interpreted as a growth-induced water potential gradient. This and further implications for the mechanism of growth control are discussed.     

Regulation of leaf growth of grass by blue light

Changes in light quality occur naturally within a canopy when a plant grows from unshaded to shaded conditions, and the reverse occurs after a cut that reduces shading. These changes in light quality could be responsible for the variation in leaf elongation and appearance rates of grasses. The role of blue light in leaf growth was investigated in tall fescue (Festuca arundinacea Schreb.) and perennial ryegrass (Lolium perenne L.). Leaf length was measured daily following a decrease or an increase in blue light to evaluate effects on duration of leaf growth, leaf elongation and the rate of leaf appearance rate. A reduction in blue light increased sheath length by 8 to 14% and lamina length by 6 to 12% for both species. These increases could be reversed by enrichment of blue light. With low blue light treatment, final leaf length was increased due to a greater leaf elongation rate. In tall fescue, but not in perennial ryegrass, this effect was coupled with a greater phyllochron and a longer duration of leaf elongation. Development of successive leaves on a tall fescue tiller were co-ordinated. A decrease in blue light increased the duration of elongation in the oldest growing leaf and also delayed the appearance of a new leaf, maintaining this co-ordination. We conclude that final leaf size and phyllochron for tall fescue can be significantly modified by blue light. Perennial ryegrass appeared less responsive, except for displaying longer sheaths and laminae in low blue light, as also occurred for tall fescue. We hypothesize that leaf length could be regulated by the quality of the light reaching the growing region itself.     

Peroxidase activity in the leaf elongation zone of tall fescue : I. Spatial distribution of ionically bound peroxidase activity in genotypes differing in length of the elongation zone

Cessation of cell expansion has been associated with cell wall cross-linking reactions catalyzed by peroxidase. This study utilized two genotypes of tall fescue (Festuca arundinacea Schreb.) that differ in length of the leaf elongation zone to investigate the relationship between ionically bound peroxidase activity and the spatial distribution of leaf elongation. Peroxidase activity was also localized histochemically in transverse sections of the leaf blade using 3,3′ -diaminobenzidine. Soluble or soluble plus ionically bound peroxidase activities were extracted from homogenized segments of the elongating leaf blade and assayed spectrophotometrically. Activity of the ionically bound fraction, expressed per milligram fresh weight or per microgram protein, increased as cells were displaced through the distal half of the elongation zone, corresponding to the region in which the elongation rate declined. In both genotypes, the initial increase in activity preceded the onset of growth deceleration by about 10 hours. In the basal region where elongation began, histochemical localization showed that peroxidase activity was found only in vascular tissues. As cells were displaced farther through the elongation zone, peroxidase activity appeared in walls of other longitudinally continuous tissues such as the epidermis and bundle sheaths. Increase in ionically bound peroxidase activity and changes in localization of peroxidase activity occurred at comparable developmental stages in the two genotypes. The results indicate that cessation of elongation followed an increase in cell wall peroxidase activity.     

Characterization of fructan from mature leaf blades and elongation zones of developing leaf blades of wheat, tall fescue, and timothy

Water-soluble carbohydrate composition of mature (ceased expanding) leaf blades and the elongation zone of developing leaf blades was characterized in wheat (Triticum aestivum L.), tall fescue (Festuca arundinacea Schreb.), and timothy (Phleum pratense L.). These species were chosen because they differ in mean degree of polymerization (DP) of fructan in the mature leaf blade. Our objective was to compare the nature and DP of the fructan. Vegetative plants were grown with a 14-hour photoperiod and constant 21°C at the leaf base. Gel permeation chromatography of leaf blade extracts showed that the apparent mean fructan DP increased in the order wheat < tall fescue < timothy. Apparent mean DP of elongation zone fructan was higher than that of leaf blade fructan in wheat and timothy, but the reverse occurred for tall fescue. Low DP (≤10) and high DP (>10) pools were found in both tissues of tall fescue and wheat, but concentration of low DP fructan was very low in either tissue of timothy. All three species have high DP fructan. Comigration with standards on thin-layer chromotography showed that wheat contained 1-kestose and a noninulin fructan oligomer series. Tall fescue contained neokestose, 1-kestose and higher oligosaccharides that comigrated with neokestose-based compounds and inulins. Thin-layer chromatography showed that small amounts of fructose-containing oligosaccharides were present in timothy.     

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%).     

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.     

Nitrogen Effects on Leaf Anatomy within the Intercalary Meristems of Tall Fescue Leaf Blades

Longitudinal elongation contributes most to leaf area expansionof grasses and its rate is known to be strongly affected byN. Our objective was to determine the effect of two N regimes(N₀and N+) on the gradient of leaf tissue formation in meristemsof two contrasting tall fescue (Festuca arundinacea Schreb.)genotypes. Proportions of epidermal, mesophyll and vasculartissue as well as intercellular air space were determined throughoutthe base of actively elongating leaves. The area of leaf transversesections nearly doubled between the ligule and the distal endof the growth zone (about 30 mm), and was mainly associatedwith lateral epidermal and mesophyll cell division in the proximal5.0–7.5 mm. Further increase in transverse area was dueto the formation of intercellular airspace and transverse expansionof epidermal cells. Depending on genotype and N treatment themesophyll, epidermis, vascular bundles and air space comprised45–54%, 20–28%, 6–9%, and 17–21%, respectively,of transverse leaf area in the distal part of the growth zone.After a slight increase close to the leaf base, the area ofvascular tissue remained constant throughout the growth zone.The proportion of air space to mesophyll space was higher atN₀than at N+ because mesophyll area was enhanced by N+ to agreater degree than by N₀. In the genotype with slow leaf elongation,the increase in cross-sectional leaf area was due to an increasein both leaf width and leaf thickness. In the genotype whichhad faster leaf elongation and wider leaves, only leaf thicknesswas enhanced by N+. Copyright 2001 Annals of Botany Company Festuca arundinacea(Schreb.), tall fescue, leaf anatomy, growth zone, nitrogen     

Fructan Content and Synthesis in Leaf Tissues of Festuca arundinacea

The concentration of fructan in tall fescue (Festuca arundinacea Schreb.) changes during growth and in response to environment. The objective of this research was to compare the fructan concentration and fructosyl-transferase activity of tall fescue leaf tissues. Expanding leaves, inner and outer sheaths, and expanded blades of greenhouse-grown tall fescue plants were assayed for fructan concentration and fructosyl-transferase activity. Leaf sheaths contained significantly more nonstructural carbohydrate than did the expanded blade. Sheaths also contained a greater percentage of fructan with more than six sugar residues (long chain fructan), than either the expanded blade or expanding leaf. Expanding leaves contained a greater concentration of fructose and oligosaccharides than did sheath or blade tissues. Expanding leaves also had the greatest fructosyl-transferase activity measured either as radiolabel incorporated into fructans in tissue pieces or protein extracts. Activity of fructosyl-transferase was greater in expanding leaf tissue than in sheath tissues.     

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     

Kinematics and Dynamics of Sorghum (Sorghum bicolor L.) Leaf Development at Various Na/Ca Salinities (I. Elongation Growth)

In many salt-sensitive species, elevated concentrations of Ca in the root growth media ameliorate part of the shoot growth reduction caused by NaCl stress. The physiological mechanisms by which Ca exerts protective effects on leaf growth are still not understood. Understanding growth inhibition caused by a stress necessitates locating the leaf expansion region and quantifying the profile of the growth reduction. This will enable comparisons and correlations with spatial gradients of probable physiologically inhibiting factors. In this work we applied the methods of growth kinematics to analyze the effects of elevated Ca concentrations on the spatial and temporal distributions of growth within the intercalary expanding region of salinized sorghum (Sorghum bicolor [L.] Moench, cv NK 265) leaves. NaCl (100 mM) caused a decrease in leaf elongation rate by shortening the leaf growing zone by 20%, as well as reducing the peak value of the longitudinal relative elemental growth rate (REG rate). Increasing the Ca concentrations from 1 to 10 mM restored the length of the growing zone of both emerged and unemerged salinized leaves and increased the peak value of the REG rate. The beneficial effects of supplemental Ca were, however, more pronounced in leaves after their appearance above the whorl of encircling older leaf sheaths. Elevated Ca then resulted in a peak value of REG rate higher than in the salinized leaves. The peak value of unemerged leaves was not increased, although it was maintained over a longer distance. The duration of elongation growth associated with a cell during its displacement from the leaf base was longer in salinized than control leaves, despite the fact that the elongation zone was shorter in salinity. Although partially restoring the length of the elongation region, supplemental Ca had no effect on the age of cessation of growth. Elongation of a tissue element, therefore, ceased when a cellular element reached a certain age and not a specific distance from the leaf base.     

Drought effects on cellular and spatial parameters of leaf growth in tall fescue

The effect of drought and recovery on cellular and spatial parametersof the growth process in tall fescue leaves was studied in twoexperiments. In both experiments plants grown on vermiculiteand maintained in a controlled environment were submitted toa 7 d drought period generated by withholding water. Droughtwas followed by a 3 d recovery period in experiment II. As leafelongation rate (LER) decreased during developing drought boththe growth zone length (initially 40 mm) and the maximum relativeelemental growth rate (initially 0.09 mm mm^–1 h^–1during the dark period of diurnal cycles) within the growthzone declined. But the growth zone still exhibited a lengthof approximately 15 mm when LER approached 0 under severe drought(–2.0 MPa predawn leaf water potential). The growth potentialof the basal 15-mm-long portion of the leaf was conserved duringthe period when drought effected the complete arrest of leafelongation. A (retrospective) analysis of the position-timerelationships of epidermal cells identified on leaf replicas(experiment II) indicated that the cell flux out of the growthzone responded very sensitively to drought. Before drought theflux was maximum at approximately 3.2 cells (cell file h)^–1during the dark period. Flux decreased to 0 when leaf elongationstopped. Flux also varied diurnally both under well-wateredand droughted conditions. In well-watered conditions it wasabout 30% less during the light than the dark period. Cell elongationwas also sensitive to drought. Under well-watered conditionsepidermal cell elongation stopped when cells attained a lengthof approximately 480 µm. During developing drought cellsstopped elongating at progressively shorter lengths. When LERhad decreased to almost nil, cells stopped elongating at a lengthof approximately 250 µn. When drought was relieved followinga 2 d complete arrest of leaf elongation then cells shorterthan 250 µm were able to resume expansion. Following rewateringcell flux out of the growth zone increased rapidly to and abovethe pre-drought level, but there was only a slow increase overtime in the length at which cell elongation stopped. About 2d elapsed until the leaf growth zone produced cells of similarlength as before drought (i.e. approximately 480 µm). Key words: Epidermal cell length, cell flux, (leaf) growth zone, leaf elongation rate, relative elemental growth rate, position-time relationships (path line, growth trajectory), drought, water deficit     

Grass Leaf Elongation Rate as a Function of Developmental Stage and Temperature: Morphological Analysis and Modelling

Elongation of successive leaves was measured following defoliationof tall fescue plants in controlled environments. Measurementswere made under constant temperatures of 24 °C and 14 °C,and after temperature changes from 24 to 14 °C andvice versa.A morphological analysis of the growing leaf was made from thetime it was 1 mm long until it was fully elongated. The timeelapsed from initiation until the leaf was 1 mm long was estimated.Young leaves less than 1.5 mm long elongated slowly at a constantleaf elongation rate (LER). By extrapolating this LER back toleaf initiation from the apex it was calculated that elongationlasted 42.5 d at 24 °C and 51 d at 14 °C. Lengths ofthe division zone (DZ) and the extension-only zone (E-OZ) increasedto a maximum and then decreased during leaf development. Temperaturechange had an immediate effect on LER but the response varieddepending on the direction of the temperature change. To describethese different features, an empirical model of DZ and E-OZwas designed. Its five parameters were optimized at constanttemperature. The model was then used to simulate the LER ofplants subjected to temperature changes. Instant and lastingeffects of the initial temperature on mean LER in plants transferredfrom 14 to 24 °C andvice versawere well simulated. It wasconcluded that the major reason for differences was due to thegrowth stage (DZ and E-OZ lengths) at which the changes occurredat both temperatures.Copyright 1999 Annals of Botany Company Festuca arundinaceaSchreb., tall fescue, growth zone, division zone.     

An in vitro microplant bioassay using clonal white ash to test for tall fescue allelopathy

Axillary shoots from three selected white ash (Fraxinus americana L.) clones were harvested from in vitro shoot cultures. Roots were initiated by pulsing excised shoots for eight days in the dark in MS medium supplemented with 2% sucrose, 0.7% agar, 5 M NAA, and 1 M IBA. Pulsed shoots were transferred to a root elongation medium consisting of 25% MS macrosalts, full-strength microsalts and organics, 1% sucrose, 0.7% agar and no auxins. When roots were visible (6–10 days after transfer to root elongation medium), microplants were transferred to vessels containing the same minimal medium and tall fescue (Festuca elatior var. arundinacea (Schreb.) Wimm.) leaf extracts, leaf leachates, or soil leachates from plant boxes with and without tall fescue sod. After four weeks in vitro, primary adventitious and secondary root growth was reduced by extracts obtained from 5 and 10 g ground leaves per 100 ml of medium. Leachates obtained from 5 g soaked leaves per 100 ml of medium stimulated primary root growth. Soil leachates from bare soil also stimulated primary root growth. Variation was observed among the clones for root growth when plantlets were grown in extracts or leachates from tall fescue.     

A transient increase in apoplastic peroxidase activity precedes decrease in elongation rate of B73 maize (Zea mays) leaf blades

Peroxidase (EC 1.11.1.7) activity from homogenized tissue or in apoplastic fluid was analyzed along the developmental gradient of expanding B73 maize ( Zea mays L.) leaf blades. Soluble plus ionically bound peroxidase activity from homogenized tissue was present in high levels at the leaf base, which includes the region of cell division, and decreased as tissue was displaced away from the base by growth. A different pattern of change in peroxidase activity was seen in apoplastic fluid extracted from segments of intact tissue, where an increase in peroxidase activity preceded a rapid decrease in leaf elongation rate. Similar patterns in peroxidase activity from homogenized and intact tissue have been found in leaf blades of tall fescue ( Festuca arundinacea Schreb.), suggesting a common phenomenon. At the location within the elongation zone where the increase in apoplastic peroxidase activity occurred, the activities of neutral and acidic (pl 4.6) peroxidase isoforms were also elevated in both the homogenate and in apoplastic fluid. The coincidence of these isoforms with the decline in leaf elongation rate suggests they may contribute to cessation of growth. At the distal end of the elongation zone, the activities of other acidic peroxidases (pI 5.6 and 5.7) increased in the homogenate and in apoplastic fluid, and remained elevated as tissue was displaced into the maturation region. The location of their appearance and their relatively high activity in the maturation region suggest the involvement of these isoforms in lignification.     

Diurnal Growth of Tall Fescue Leaf Blades : II. Dry Matter Partitioning and Carbohydrate Metabolism in the Elongation Zone and Adjacent Expanded Tissue

The spatial distributions of net deposition rates of water soluble carbohydrate-free dry matter (WSC-free DM) and WSC were evaluated within and above the elongation zone of tall fescue (Festuca arundinacea Schreb.) leaf blades during light and darkness. Imported DM used for WSC-free DM synthesis during darkness (67% of the total in experiment I and 59% in experiment II) was greater than during light (47% in both experiments), suggesting that the 65% higher leaf elongation rate during darkness was accompanied by higher rates of synthesis of cellular structural components. Deposition rates of WSC in the basal and central part of the elongation zone (0-20 mm from the ligule) were similar during light and darkness, but above 20 millimeters WSC deposition occurred during light and WSC loss occurred during darkness. WSC deposition and loss throughout the elongation zone and the recently expanded tissue were mostly due to net synthesis and degradation of fructan. Fructan was predominantly low molecular weight and contributed about 50% of the total osmotic partial pressure of WSC. In the most actively growing region, where fructan synthesis was most rapid, no diurnal change occurred in molecular weight distribution of fructan. WSC solute concentrations were diluted in the most actively growing tissue during darkness because net monosaccharide and fructan deposition were unaltered and sucrose deposition was decreased, but growth-associated water deposition was increased by 77%. Net rates of fructan synthesis and degradation were not related to tissue sucrose concentration, but appeared to respond to the balance between assimilate import and assimilate use in synthesis of cellular structural components (i.e. WSC-free DM) and deposition of monosaccharides. Fructan synthesized in tissue during most active elongation was degraded when the respective tissue reached the distal limit of the elongation zone where assimilate import in darkness was insufficient to maintain synthetic processes associated with further differentiation of cells.     

Growth and Deposition of Inorganic Nutrient Elements in Developing Leaves of Zea mays L

Spatial distributions of growth and of the concentration of some inorganic nutrient elements were analyzed in developing leaves of maize (Zea mays L.). Growth was analyzed by pinprick experiments with numerical analysis to characterize fields of velocity and relative elemental elongation rate. Inductively coupled plasma and atomic emission spectroscopy were used to measure nutrients extracted from segments of leaf tissue collected by position. Leaves 7 and 8, both elongating 3 millimeters per hour had maximum relative elemental growth rates of 0.06 to 0.08 millimeters per hour with maximum rates 20 to 50 millimeters from the node and cessation of growth by 90 millimeters from the node. Spatial distribution of dry weight density revealed that the rate of biomass deposition was maximum in the most rapidly expanding region and continued beyond the elongation zone. The nutrient elements K, Cl, Ca, Mg, and P showed different distribution patterns of ion density (on a dry weight basis). K and Cl had minimal density in the leaf tips; K density was maximum in the growing region, whereas Cl density was maximum at the region of growth cessation. Ca, Mg, and P had relatively high densities at the base of the elongation zone near the node and also in the tip regions. Near the node, P and Mg densities were higher in the young, growing leaves, whereas Ca density near the node was higher in older leaves that had completed elongation. Deposition rates of all nutrients were greatest in the region of maximum elongation rate.