Salinity and drought tolerance of mannitol-accumulating transgenic tobacco

Tobacco plants (Nicotiana tabacum L.) were transformed with a mannitol-1-phosphate dehydrogenase gene resulting in mannitol accumulation. Experiments were conducted to determine whether mannitol provides salt and/or drought stress protection through osmotic adjustment. Non-stressed transgenic plants were 20–25% smaller than non-stressed, non-transformed (wild-type) plants in both salinity and drought experiments. However, salt stress reduced dry weight in wild-type plants by 44%, but did not reduce the dry weight of transgenic plants. Transgenic plants adjusted osmotically by 0.57 MPa, whereas wild-type plants did not adjust osmotically in response to salt stress. Calculations of solute contribution to osmotic adjustment showed that mannitol contributed only 0-003-0-004 MPa to the 0.2 MPa difference in full turgor osmotic potential (π_o) between salt-stressed transgenic and wild-type plants. Assuming a cytoplasmic location for mannitol and that the cytoplasm constituted 5% of the total water volume, mannitol accounted for only 30–40% of the change in π_o of the cytoplasm. Inositol, a naturally occurring polyol in tobacco, accumulated in response to salt stress in both transgenic and wild-type plants, and was 3-fold more abundant than mannitol in transgenic plants. Drought stress reduced the leaf relative water content, leaf expansion, and dry weight of transgenic and wild-type plants. However, π_o was not significantly reduced by drought stress in transgenic or wild-type plants, despite an increase in non-structural carbohydrates and mannitol in droughted plants. We conclude that (1) mannitol was a relatively minor osmolyte in transgenic tobacco, but may have indirectly enhanced osmotic adjustment and salt tolerance; (2) inositol cannot substitute for mannitol in this role; (3) slower growth of the transgenic plants, and not the presence of mannitol per se, may have been the cause of greater salt tolerance, and (4) mannitol accumulation was enhanced by drought stress but did not affect π_o or drought tolerance.     

Control of leaf cell elongation in barley. Generation rates of osmotic pressure and turgor, and growth-associated water potential gradients

In a previous study on the effects of N-supply on leaf cell elongation, the spatial distribution of relative cell elongation rates (RCER), epidermal cell turgor, osmotic pressure (OP) and water potential (Ψ) along the elongation zone of the third leaf of barley was determined (W. Fricke et al. 1997, Planta 202: 522–530). The results suggested that in plants receiving N at fixed relative addition rates (N-supply limitation of growth), cell elongation was rate-limited by the rate of solute provision, whereas in plants growing on complete nutrient solution containing excessive amounts of N (N-demand limitation), cell elongation was rate-limited by the rate of water supply or wall yielding. In the present paper, these suggestions were tested further. The generation rates of cell OP, turgor and Ψ along the elongation zone were calculated by applying the continuity equation of fluid dynamics to the previous data. To allow a more conclusive interpretation of results, anatomical data were collected and bulk solute concentrations determined. The rate of OP generation generally exceeded the rate of turgor generation. As a result, negative values of cell Ψ were created, particularly in demand-limited plants. These plants showed highest RCER along the elongation zone and a Ψ gradient of at least −0.15 MPa between water source (xylem) and expanding epidermal cells. The latter was similar to a theoretically predicted value (−0.18 MPa). Highest rates of OP generation were observed in demand-limited plants, with a maximum rate of 0.112 MPa · h⁻¹ at 16–20 mm from the leaf base. This was almost twice the rate in N-supply-limited plants and implied that the cells in the leaf elongation zone were capable of importing (or synthesising) every minute almost 1 mM of osmolytes. Potassium, Cl⁻ and NO₃ ⁻ were the main inorganic osmolytes (only determined for demand-limited plants). Their concentrations suggest that, unlike the situation in fully expanded epidermal cells, sugars are used to generate OP and turgor. Anatomical data revealed that the zone of lateral cell expansion extended distally beyond the zone of cell elongation. It is concluded that leaf cell expansion in barley relies on high rates of water and solute supply, rates that may not be sustainable during periods of sufficient N-supply (limitation by water supply: Ψ gradients) or limiting N-supply (limitation by solute provision: reduced OP-generation rates). To minimise the possibility of growth limitation by water and osmolyte provision, longitudinal and lateral cell expansion peak at different locations along the growth zone. Received: 15 October 1997 / Accepted: 12 March 1998     

Effect of modified carbon allocation on turgor, osmolality, sugar and potassium content, and membrane potential in the epidermis of transgenic potato (Solanum tuberosum L.) plants

The effects of modification in sugar concentrations on turgor pressure and membrane potential in epidermal leaf cells of transgenic potato (Solanum tuberosum cv. Desirée) plants were studied. Measurements of turgor pressure were performed by insertion of a micro pressure probe. Osmolality and sugar concentrations were determined by micro analysis of single cell extracts. Membrane potentials and cell diameters were calculated from repeated, computer-controlled scans with voltage-sensitive microelectrodes. Epidermal cells of sucrose transporter antisense plants showed a more than 100% elevation in osmolality and turgor pressure compared to the wild-type. As a consequence, cell diameters were enhanced in this transgenic line. However, membrane potentials were only slightly reduced in sucrose transporter antisense plants. In addition to sucrose transporter antisense lines, transgenic plants that were reduced in their capacity to accumulate starch due to antisense inhibition of the chloroplastic fructose-1,6-bisphosphatase (FBPase) were investigated. These antisense plants maintained membrane potential and turgor pressure comparable to the wild-type.     

Plumbing the depths: extracellular water storage in specialized leaf structures and its functional expression in a three‐domain pressure –volume relationship

A three‐domain pressure–volume relationship (PV curve) was studied in relation to leaf anatomical structure during dehydration in the grey mangrove, Avicennia marina. In domain 1, relative water content (RWC) declined 13% with 0.85 MPa decrease in leaf water potential, reflecting a decrease in extracellular water stored primarily in trichomes and petiolar cisternae. In domain 2, RWC decreased by another 12% with a further reduction in leaf water potential to ?5.1 MPa, the turgor loss point. Given the osmotic potential at full turgor (?4.2 MPa) and the effective modulus of elasticity (~40 MPa), domain 2 emphasized the role of cell wall elasticity in conserving cellular hydration during leaf water loss. Domain 3 was dominated by osmotic effects and characterized by plasmolysis in most tissues and cell types without cell wall collapse. Extracellular and cellular water storage could support an evaporation rate of 1 mmol m^?2s^?1 for up to 54 and 50 min, respectively, before turgor loss was reached. This study emphasized the importance of leaf anatomy for the interpretation of PV curves, and identified extracellular water storage sites that enable transient water use without substantive turgor loss when other factors, such as high soil salinity, constrain rates of water transport.     

A Simple Instrument for Measuring Leaf Extension in Grasses, and its Application in the Study of the Effects of Water Stress on Maize and Sorghum

The design of a simple instrument to monitor leaf expansionin grasses is described. The instrument was used to comparethe effects of water stress on leaf extension of two cultivarsof maize and sorghum. The effect of withholding water for 3days was an appreciable reduction in the rate of leaf expansionin both plants, particularly during the light period. In well-wateredplants of both species, leaf extension continued at a steadyrate even when leaf turgor fell to around 0.1 MPa. In water-stressedmaize plants, leaf turgor during the light period fell to zeroand leaf growth ceased. When turgor was restored, followingstomatal closure, leaf extension resumed at a slow rate. Inunwatered sorghum plants, leaf turgor remained at a value greaterthan 0.1 MPa but the rate of leaf extension was significantlyreduced. The reduction in leaf turgor in the unwanted plantsresulted partly from an increase in solute potential. Zea mays L, maize, Sorghum bicolor L, leaf expansion, leaf turgor, water stress     

Dynamic Relation between Expansion and Cellular Turgor in Growing Grape (Vitis vinifera L.) Leaves

Measurements of the growth and water relations of expanding grape (Vitis vinifera L.) leaves have been used to determine the relationship between leaf expansion rate and leaf cell turgor. Direct measurement of turgor on the small (approximately 15 micrometer diameter) epidermal cells over the midvein of expanding grape leaves was made possible by improvements in the pressure probe technique. Leaf expansion rate and leaf water status were perturbed by environmentally induced changes in plant transpiration. After establishing a steady state growth rate, a step decrease in plant transpiration resulted in a rapid and large increase in leaf cell turgor (0.25 megapascal in 5 minutes), and leaf expansion rate. Subsequently, leaf expansion rate returned to the original steady state rate with no change in cell turgor. These results indicate that the expansion rate of leaves may not be strongly related to the turgor of the leaf cells, and that substantial control of leaf expansion rate, despite changes in turgor, may be part of normal plant function. It is suggested that a strictly physical interpretation of the parameters most commonly used to describe the relationship between turgor and growth in plant cells (cell wall extensibility and yield threshold) may be inappropriate when considering the process of plant cell expansion.     

Replicase proteins as determinants of phloem-dependent long-distance movement of tobamoviruses in tobacco

Summary The turgor pressure of growing pollen tubes of the lily (Lilium longiflorum Thunb.) has been recorded using a turgor pressure probe. Insertion of the probe's micropipette was routinely accomplished, providing recording periods of 20 to 30 min. Probe insertion did not affect tube growth. The stable turgor values ranged between 0.1 and 0.4 MPa, the mean value being 0.209 ± 0.064 MPa (n=106). A brief increase in turgor, generated by injection of oil through the pressure probe, caused the tube to burst at its tip. Burst pressures ranged between 0.19 and 0.58 MPa, that is, individual lily pollen tubes do not withstand turgor pressure approaching twice their regular turgor pressure. In contrast, parallel experiments using the incipient plasmolysis technique yielded a mean putative turgor pressure of 0.79 MPa either using sucrose (n=24) or mannitol (n=25). Surprisingly, turgor pressure was not significantly correlated with tube growth rate which ranged from zero to 13 m/min. Nor did it correlate with tube length over the tested range of 100 to 1600 m. In addition the influence of the medium's osmolality was surprisingly low: raising the external osmotic pressure from 0.36 to 1.08 MPa, with sucrose or mannitol, only caused mean turgor pressure to decline from 0.27 to 0.18 MPa. We conclude that growing lily pollen regulates its turgor pressure remarkably well despite substantial variation in tube growth rate, tube length, and osmotic milieu.     

Salinity Stress Inhibits Bean Leaf Expansion by Reducing Turgor, Not Wall Extensibility

Treatment of bean (Phaseolus vulgaris L.) seedlings with low levels of salinity (50 or 100 millimolar NaCl) decreased the rate of light-induced leaf cell expansion in the primary leaves over a 3 day period. This decrease could be due to a reduction in one or both of the primary cellular growth parameters: wall extensibility and cell turgor. Wall extensibility was assessed by the Instron technique. Salinity did not decrease extensibility and caused small increases relative to the controls after 72 hours. On the other hand, 50 millimolar NaCl caused a significant reduction in leaf bulk turgor at 24 hours; adaptive decreases in leaf osmotic potential (osmotic adjustment) were more than compensated by parallel decreases in the xylem tension potential and the leaf apoplastic solute potential, resulting in a decreased leaf water potential. It is concluded that in bean seedlings, mild salinity initially affects leaf growth rate by a decrease in turgor rather than by a reduction in wall extensibility. Moreover, longterm salinization (10 days) resulted in an apparent mechanical adjustment, i.e. an increase in wall extensibility, which may help counteract reductions in turgor and maintain leaf growth rates.     

Nitrate Supply and the Biophysics of Leaf Growth in Salix viminalis

The influence of nitrogen on leaf area development and the biophysicsof leaf growth was studied using clonal plants of the shrubwillow, Salix viminalis grown with either optimal (High N) orsub-optimal (Low N) supplies of nitrate. Leaf growth rate andfinal leaf size were reduced in the sub-optimal treatment andthe data suggest that in young rapidly growing leaves, thiswas primarily due to changes in cell wall properties, sincecell wall extensibility (% plasticity) was reduced in the LowN plants. The biophysical regulation of leaf cell expansion also differedwith nitrogen treatment as leaves aged. In the High N leaves,leaf cell turgor pressure (P) increased with age whilst in theLow N leaves P declined with age, again suggesting that foryoung leaves, cell wall plasticity limited expansion in theLow N plants. Measurements of cell wall properties showed thatcell wall elasticity (%E) was not influenced by nitrogen treatmentand remained constant regardless of leaf age. Key words: Salix, cell wall extensibility, nitrogen nutrition, biophysics of leaf growth     

Tissue water relations and leaf growth of tropical corn cultivars under water deficits

Abstract This study reports on the effect of water deficit on the tissue water relations and leaf growth of six corn cultivars, growing in glasshouse conditions, in order to understand growth responses to drought of tropical corn. A mild water-stress treatment was imposed slowly; plants reached a minimum pre-dawn leaf water potential of about –1.5 MPa by day 12 after watering was withheld. Analysis of the water relation characteristics of growing leaves using the pressure–volume technique demonstrated that under water deficits all the cultivars changed their moisture-release curves compared with irrigated plants. Osmotic potential at full turgor was lowered in water-stressed plants of all the genotypes and the degree of such change was between 0.34 MPa and 0.58 MPa. Thus, turgor pressure was lost at a lower water potential in water-stressed plants than in irrigated plants of all the varieties. Volumetric elastic moduli were also increased under water deficits and the increase ranged between 10% and 141% among the cultivars. In all the genotypes, the stress imposed led to a reduction of leaf area and dry matter accumulation. Leaf expansion was very sensitive to low turgor pressure and it ceased when turgor reached 0.2 MPa. Thus, varieties able to maintain a higher degree of turgor pressure (i.e. by osmotic adjustment) under water deficits may be able to prolong leaf growth.     

High CO2 levels reduce ethylene production in kiwifruit

We examined the roles of turgor potential and osmotic adjustment in plant growth by comparing the growth of spring wheat ( Triticum aestivum cv. Siete cerrors) and sudangrass ( Sorghum vulgare var. Piper) seedlings in response to soil water and temperature stresses. The rates of leaf area expansion, leaf water potential and osmotic potential were measured at combinations of 5 soil water potentials ranging from −0.03 to −0.25 MPa and 6 soil temperatures ranging from 14 to 36°C. Spring wheat exhibited little osmotic adjustment while sudangrass exhibited a high degree of osmotic adjustment. However, the rate of leaf area growth for sudangrass was more sensitive to water stress than that of spring wheat. These results were used to evaluate the relationship between growth and turgor potential. The modified Arrhenius equation based on thermodynamic considerations of the growth process was evaluated. This equation obtains growth rate as a function of activation energy, enthalpy difference between active and inactive states of enzymes, base growth rate and optimum temperature. Analyses indicate that the modified Arrhenius equation is consistent with the Lockhart equation with a metabolically controlled cell wall extensibility.     

Regulation of Turgor Pressure by Suspension-Cultured Tobacco Cells

Turgor regulation and effects of high NaCl and water deficiton growth and internal solutes were studied after transferringtobacco cells from control culture medium (osmotic pressure= 0.13–0.15 MPa at time of transfer) to culture mediumcontaining either 82 mol m^–3 NaCl or 150 mol m^–3melibiose (osmotic pressure of media = 0.62 MPa). Followingtransfer to media with higher osmotic pressure, expansion rateand turgor pressure were reduced. Within 24 h of imposing thewater deficit, expansion rate had returned to that of cellsin control culture medium. However, by 24 h, turgor pressurehad only risen from 0.2 MPa to 0.65 MPa in the NaCl treatmentand to 0.53 MPa in the melibiose treatment, while it was 0.73MPa in the control treatment. Furthermore, turgor pressure remainedwithin 0.05 MPa of these respective values for the rest of the(75 h) experiment. These results suggest differences in bothcell wall properties (extensibility and/or threshold turgor)and the level at which turgor is maintained for cells in thevarious treatments. Solutes contributing nearly all (82–97%) of the osmoticpressure in cells were identified. The initial (up to 24 h)increases in turgor pressure were mainly due to increases insolute concentrations caused by relatively slow expansion rates.However, increased Na⁺ and Cl ^– uptake contributed toincreased turgor pressure in the NaCl treatment and caused turgorpressure of cells in this treatment to increase faster thanin the melibiose treatment. Likewise, expansion rate rose morequickly in the NaCl than in the melibiose treatment. After 24h, maximum expansion rate was reached and concentrations ofmost internal solutes began to decrease. Nevertheless, turgorpressure remained relatively constant. The constancy of turgorpressure was due to increased glucose uptake rates relativeto controls, with consequent increases in concentrations ofsucrose, glucose and fructose and, in cells in the melibiosetreatment, of organic acids. Glucose uptake was slower in theNaCl than in the melibiose treatment but higher turgor pressurewas maintained in the NaCl treatment due to high uptake of Na⁺and Cl^–. Glucose uptake appears to respond to a systemof turgor regulation, but further experiments are required toconfirm this and to determine whether Na⁺ and Cl^– uptakealso respond to a system of turgor regulation. Key words: Salinity, water deficit, growth     

Leaf expansion of four sunflower (Helianthus annuus L.) cultivars in relation to water deficits. II. Diurnal patterns during stress and recovery

Abstract. Leaf expansion of four sunflower cultivars ( Helianthus annuus L. cvs. Hysun 31, Havasupai, Hopi and Seneca) was monitored continuously in a growth cabinet through the final stages of a drying cycle and then throughout the first 2 days after rewatering in order to study the responses of leaf expansion to water deficits. Comparable plants were also measured throughout a diurnal cycle in a glasshouse.
In the cabinet, leaf extension was faster in the dark than in the light, but an extended dark period suppressed leaf extension. At similar leaf water potentials, the rate of leaf extension was greater in the light than in the dark, but as the osmotic potential was lower in the light than in the dark, the relationship between turgor pressure and leaf extension rate was similar in both environments. Throughout the drying and recovery cycles turgor and leaf extension rate was positively correlated: no significant differences among cultivars were observed.
In the plants grown and measured in the glasshouse, leaf expansion occurred at lower leaf water potentials in stressed than in unstressed plants, but the relationship between leaf expansion and turgor was similar in both stressed and unstressed plants as a result of a lowering of the osmotic potential in the former. Diurnal turgor maintenance resulting from osmotic adjustment was almost half that occurring during a complete drying cycle. During the day, the leaf expansion rate increased linearly with turgor pressure in all cultivars: the expansion rate per unit turgor pressure was greater in the glasshouse than in the growth cabinet. Nocturnal leaf expansion in the stressed and unstressed plants was not, however, correlated with turgor pressure.     

Leaf expansion of four sunflower (Helianthus annuus L) cultivars in relation to water deficits. I. Patterns during plant development

Abstract. The influence of a slow stress and recovery cycle on the pattern of leaf expansion in four diverse sunflower cultivars ( Helianthus annuus L. cvs. Hysun 31, Havasupai, Hopi and Seneca) was studied in a glasshouse. Stress had no significant effect on the time of flower bud emergence and anthesis, or on final leaf number, but delayed the appearance of leaves at high insertions in all cultivars except Hysun 31.
Leaf expansion was markedly reduced as the predawn leaf water potential decreased from −0.35 to −0.60 MPa, and the predawn turgor pressure decreased from 0.3 to 0.2 MPa, and expansion ceased at a predawn leaf water potential of about −1.0 MPa, i.e. when the predawn turgor pressure reached zero.
The leaves most reduced in final size when water was withheld were those at the insertions which grew the most rapidly in unstressed plants. The maximum reduction in final leaf size of 25–35% was similar in all cultivars and was due to retardation of the rate of leaf expansion: the duration of leaf expansion was actually increased by stress. However, leaves that were initiated during stress, but emerged after rewatering, had final leaf areas at least equal to those in the unstressed plants: in the cultivar Seneca, the final size of leaves of high insertion was significantly greater in stressed than unstressed plants, whereas in the three other cultivars the final leaf sizes were similar in both treatments. All four cultivars examined adjusted osmotically to the same degree, but leaf water potentials in one, Seneca, increased more rapidly after rewatering than in the other three, and this may have contributed to the greater relative leaf size in the leaves of high insertion in this cultivar.     

Correlation between leaf growth variables suggest intrinsic and early controls of leaf size in Arabidopsis thaliana

Leaf development is affected by both internal (genetic) and external (environmental) regulatory factors. The aim of this work was to investigate how leaf growth variables are related to one another in a range of environments. The leaf growth variables of wild-type Arabidopsis thaliana and leaf development mutants (ang4, ron2-1, elo1, elo2 and elo4) were studied under different incident light treatments (light and shade). The leaves studied were altered in various leaf development variables, such as the duration of expansion, relative and absolute expansion rates, epidermal cell size, epidermal cell number and initiation rate. Final leaf area was correlated to maximal absolute leaf expansion rate and cell number, but not to duration of leaf expansion or cell size. These relationships were common to all studied genotypes and light conditions, suggesting that leaf size is determined early in development. In addition, the early variables involved in leaf development were correlated to one another, and initial relative expansion rate was negatively correlated to the duration of expansion. These relationships between the leaf development variables were used to construct a conceptual model of leaf size control.     

Ectopic expression of wheat expansin gene TaEXPA2 improved the salt tolerance of transgenic tobacco by regulating Na+/K+ and antioxidant competence

High salinity is one of the most serious environmental stresses that limit crop growth. Expansins are cell wall proteins that regulate plant development and abiotic stress tolerance by mediating cell wall expansion. We studied the function of a wheat expansin gene, TaEXPA2, in salt stress tolerance by overexpressing it in tobacco. Overexpression of TaEXPA2 enhanced the salt stress tolerance of transgenic tobacco plants as indicated by the presence of higher germination rates, longer root length, more lateral roots, higher survival rates and more green leaves under salt stress than in the wild type (WT). Further, when leaf disks of WT plants were incubated in cell wall protein extracts from the transgenic tobacco plants, their chlorophyll content was higher under salt stress, and this improvement from TaEXPA2 overexpression in transgenic tobacco was inhibited by TaEXPA2 protein antibody. The water status of transgenic tobacco plants was improved, perhaps by the accumulation of osmolytes such as proline and soluble sugar. TaEXPA2‐overexpressing tobacco lines exhibited lower Na⁺ but higher K⁺ accumulation than WT plants. Antioxidant competence increased in the transgenic plants because of the increased activity of antioxidant enzymes. TaEXPA2 protein abundance in wheat was induced by NaCl, and ABA signaling was involved. Gene expression regulation was involved in the enhanced salt stress tolerance of the TaEXPA2 transgenic plants. Our results suggest that TaEXPA2 overexpression confers salt stress tolerance on the transgenic plants, and this is associated with improved water status, Na⁺/K⁺ homeostasis, and antioxidant competence. ABA signaling participates in TaEXPA2‐regulated salt stress tolerance.     

Impaired growth in transgenic plants over-expressing an expansin isoform

Expansins are cell wall proteins characterised by their ability to stimulate wall loosening during cell expansion. The expression of some expansin isoforms is clearly correlated with growth and the external application of expansins can stimulate cell expansion in vivo in several systems. We report here the expression of a heterologous expansin coding sequence in transgenic tomato plants (Lycopersicon esculentum Mill.) under the control of a constitutive promoter. In some transgenic lines with high levels of expansin activity extractable from cell walls, we observed alterations of growth: mature plants were stunted, with shorter leaves and internodes, and dark-grown seedlings had shorter and wider hypocotyls than their wild-type counterparts. Examination of hypocotyl sections revealed similar differences at the cellular level: cortical and epidermal cells were shorter and wider than those from wild-type seedlings. The observed stimulation of radial expansion did not compensate for the decreased elongation, and overall growth was reduced in the transgenics. As this observation can seem paradoxical given the known effect of expansins on isolated cell walls, we examined the mechanical behaviour of transgenic tissue. We measured a decrease in hypocotyl elongation in response to acidic pH in the transformants. This result may account for the alterations in cell expansion, and could itself be explained by a reduced susceptibility of transgenic cell walls to expansin action.     

A comparison of the water relations characteristics of Helianthus annuus and Helianthus petiolaris when subjected to water deficits

The effect of water deficits on the water relations and stomatal responses of Helianthus annuus and Helianthus petiolaris were compared in plants growing in the glasshouse under controlled conditions. Unirrigated plants of both genotypes were subjected to two different stress rates in which predawn leaf water potentials declined steadily at either 0.15 MPa day^?1 or 0.50 MPa day^?1. In both genotypes water stress induced a gradual and similar decrease in leaf conductance from 1.6 to 0.3 cm s^?1 as water potential decreased from-0.5 to-2.0 MPa. The relationship between leaf conductance and leaf water potential was not affected by the rate of stress development. Development of predawn leaf water potentials of-1.3 MPa had no significant effect on the relative water content at zero turgor, the apoplastic water content or the volumetric elastic modulus of whole leaves in either species, but decreased the osmotic potential at full turgor and zero turgor by 0.22 MPa and decreased the turgid weight: dry weight ratio from 10.6 to 8.4 in H. annuus, but not in H. petiolaris. In H. annuus leaves expanded during stress development, changes in the osmotic potential at full turgor induced by water deficits did not disappear on rewatering.     

Control of leaf expansion in sunflower (Helianthus annuus L.) by nitrogen nutrition

Seedlings of Helianthus annuus L. were grown at an initiallyhigh relative nitrate supply rate (0.27 mol N mol N^–1d^–1). The supply was subsequently reduced to a low rate(0.04 mol N mol N^–1 d^–1). The response of leaf areadevelopment to this abrupt decrease in nitrate availabilitywas characterized by following the expansion of the primaryand secondary leaf pairs. The timing of the drop in nitratesupply was when cell division in the epidermis of the primaryleaf pair was largely complete. Reducing the availability ofnitrate had a strong effect on leaf area expansion. The finalleaf size of the primary leaf pair was affected indicating aneffect of nitrate availability on cell expansion. By the endof the experiment the secondary leaf pair was only one-thirdthe area of that on control seedlings. The role of epidermalcell turgor pressure in this growth response was assessed bydirect measurements with a miniature cell pressure probe. Noreduction in cell turgor pressure following the decrease innitrate availability was detected. It is concluded that a reductionin turgor pressure was not responsible for the reduction inleaf area expansion and it is suggested that reduced cell expansionwas due to changes in cell wall properties. Concentrations ofleaf and root abscisic acid increased following the reductionin nitrate availability. Key words: Abscisic acid, cell size, cell turgor pressure, nitrate, nitrogen, relative rate of nitrate supply     

Mechanism for increased leaf growth in elevated CO2

The effect of exposure to elevated CO₂ on the processes of leafcell production and leaf cell expansion was studied using primaryleaves of Phaseolus vulgaris L. Cell division and expansionwere separated temporally by exposing seedlings to dim red lightfor 10 d (when leaf cell division was completed) followed byexposure to bright white light for 14 d (when leaf growth wasentirely dependent on cell expansion). When plants were exposedto elevated CO₂ during the phase of cell expansion, epidermalcell size and leaf area development were stimulated. Three piecesof evidence suggest that this occurred as a result of increasedcell wall loosening and extensibility, (i) cell wall extensibility(WEx, measured as tensiometric extension using an Instron) wassignificantly increased, (ii) cell wall yield turgor (V, MPa)was reduced and (iii) xyloglucan endotransglycosylase (XET)enzyme activity was significantly increased. When plants wereexposed to elevated CO₂ during the phase of cell division, thenumber of epidermal cells was increased whilst final cell sizewas significantly reduced and this was associated with reducedfinal leaf area, WEx and XET activity. When plants were exposedto elevated CO₂ during both phases of cell division and expansion,leaf area development was not affected. For this treatment,however, the number of epidermal cells was increased, but cellexpansion was inhibited, despite exposure to elevated CO₂ duringthe expansion phase. Assessments were also made of the spatialpatterns of WEx across the expanding leaf lamina and the datasuggest that exposure to elevated CO₂ during the phase of leafexpansion may lead to enhanced extensibility particularly atbasal leaf margins which may result in altered leaf shape. The data show that both cell production and expansion were stimulatedby elevated CO₂, but that leaf growth was only enhanced by exposureto elevated CO₂ in the cell expansion phase of leaf development.Increased leaf cell expansion is, therefore, an important mechanismfor enhanced leaf growth in elevated CO₂, whilst the importanceof increased leaf cell production in elevated CO₂ remains tobe elucidated. Key words: Phaseolus vulgaris L., dwarf beans, elevated CO², biophysics of cell expansion, xyloglucan endotransglycosylase, XET, water relations