Effects of NaCl on ion relations and carbohydrate status of roots and on osmotic regulation of roots and shoots of Atriplex amnicola

Abstract Atriplex amnicola was grown at 25, 200 or 400 mol m³ NaCl. Root tissues at different stages of development were investigated for concentrations of K⁺, Na⁺ and Mg²⁺, and in some cases for Cl^?. Sugar and starch concentrations were measured for plants grown at 25 or 400 mol m³ NaCl. In the ‘slightly vaeuolated’ root tips, Na⁺ was only 40 mol m^?3 at an external concentration of 400 mol m^?3 NaCl. The concentrations of K⁺ were not affected substantially by external NaCl between 25 mol m^?3 and 400 mol m^?3. The ‘highly vacuolated’ root tissues had substantially higher concentrations of K⁺, Na⁺ and Cl^? in plants grown at 200 and 400 mol m ³ NaCl than in plants grown at 25 mol m^?3 NaCl. Concentrations of Cr and of the sum of the cations in recently expanded tissue were similar to those in the bulk of the roots, consisting mainly of old cells. However, the K⁺: Na⁺ decreased with age; at 400 mol m^?3 external NaCl with a K⁺: Na⁺ of 0.012, the K⁺: Na⁺ in recently expanded 12 mm root tips was as high as 1.6, compared with 0.7 for the bulk of the roots. These ion data were used to estimate cytoplasmic and vacuolar concentrations of K⁺ and Na ⁺. Such calculations indicated that between 25 mol m³ and 400 mol m^?3 external NaCl the concentration of the sum of (Na⁺+K⁺) in the cytoplasm was maintained at about 180–200 mol m^?3 (cell water basis). In contrast, the (Na⁺+ K⁺) concentration in the vacuole was 170 mol m^?3 for plants grown at 25 mol m^?3 NaCl and 420 mol 400 mol m^?3 NaCl. The expanding root (issues exhibited greatly decreased soluble sugars and starch between dusk and dawn. Ai both times, sugar and starch concentrations in these tissues were 2.5–4.0 times greater in plants grown at 400 mol m^?3 NaCl compared with plants grown at 25 mol m^?3 NaCl. In contrast, carbohydrate concentrations in expanded root tissues were very similar at 25 and 400 mol m^?3 and showed little diurnal fluctuation. This paper considers the causes for the slower growth of A. amnicola at 400 than at 25 mol m”³ NaCl, using the data for the roots described here, and those for the shoots presented in the preceding paper (Aslam et al., 1986). There is no support for possible adverse effects by high internal ion concentrations. Instead, there may be deficiencies in supply of organic solutes for osmotic regulation; during part of the night a limited supply of such solutes may well restrict the rate of expansion of cells in plants growing at 400 mol m^?3 NaCl. There is insufficient evidence to decide whether this limitation in the expanding tissues is particularly prominent for the roots or for the shoots.     

Influence of summer rainfall on root and shoot growth of a cold-winter desert shrub,Atriplex confertifolia

Summary The influence of irrigation and nitrogen fertilization in early summer on root and shoot growth of Atriplex confertifolia, a C₄ shrub species, was examined in a cold-winter desert community in northern Utah. Soil water and xylem pressure potentials were monitored during the summer period.At the time of watering the surface soil (0–30 cm) was dry but there were turgid fine roots in this horizon. Watering of the soil reduced plant water stress from-30 to-15 bars (dawn values) indicating that roots near the surface were capable of absorbing water, and induced root growth in the 0–30 cm zone. The addition of N to the water treatment did not further increase root production. However, watering and watering +N fertilizer failed to stimulate shoot elongation or any dry weight increase of shoots. This shoot dormancy during summer is not typical of C₄ plants and is probably associated with adaptation to the cool arid environment.This work was carried out while the senior author was on study leave from CSIRO     

Effect of high salinity on Atriplex portulacoides: Growth,leaf water relations and solute accumulation in relation with osmotic adjustment

Atriplex (Halimione) portulacoides is a halophyte with potential interest for saline soil reclamation and phytoremediation. Here, we assess the impact of salinity reaching up to two-fold seawater concentration (0–1000 mM NaCl) on the plant growth, leaf water status and ion uptake and we evaluate the contribution of inorganic and organic solutes to the osmotic adjustment process. A. portulacoides growth was optimal at 200 mM NaCl but higher salinities (especially 800 and 1000 mM NaCl) significantly reduced plant growth. Na⁺ and Cl⁻ contents increased upon salt exposure especially in the leaves compared to the roots. Interestingly, no salt-induced toxicity symptoms were observed and leaf water content was maintained even at the highest salinity level. Furthermore, leaf succulence and high instantaneous water use efficiency (WUE_i) under high salinity significantly contributed to maintain leaf water status of this species. Leaf pressure–volume curves showed that salt-challenged plants adjusted osmotically by lowering osmotic potential at full turgor (Ψ_π¹⁰⁰) along with a decrease in leaf cell elasticity (values of volumetric modulus elasticity (ε) increased). As a whole, our findings indicate that A. portulacoides is characterized by a high plasticity in terms of salt-response. Preserving leaf hydration and efficiently using Na⁺ for the osmotic adjustment especially at high salinities (800–1000 mM NaCl), likely through its compartmentalization in leaf vacuoles, are key determinants of such a performance. The selective absorption of K⁺ over Na⁺ in concomitance with an increase in the K⁺ use efficiency also accounted for the overall plant salt tolerance.     

The effects of salt stress on growth,water relations and ion accumulation in two halophyte Atriplex species

Atriplex halimus is found in the Mediterranean Basin along the coastal areas of Sardinia, but few data are available on its adaptability to salinity. The effects of drought and salinity under controlled conditions on two clones of A. halimus, designated MOR2 and SOR4, originating from southern and northern Sardinia, respectively, were compared with those of seedlings of A. nummularia, an Australian species widely used in the restoration of arid areas. The effects of increasing NaCl salinity above seawater concentrations and of increasing the KCl concentration gradient were tested. Plants were harvested and analysed after 10 and 20 days of NaCl and KCl treatments. All plants remained alive until the end of treatment, although growth was strongly reduced, mainly for the A. halimus MOR2 clone, under increasing concentrations of KCl. The leaves and roots of both species responded positively to increasing NaCl concentrations up to 600 mM NaCl for A. halimus, whereas the optimal growth of A. nummularia was recorded at 300 mM NaCl. SOR4 was more sensitive to KCl toxicity. The Na⁺ concentration in the plants increased with increased salinity and was higher in A. halimus than in A. nummularia, suggesting that A. halimus is an ion accumulator and may be used for phytoremediation. The sodium accumulation in the roots of the A. halimus MOR2 clone was far greater than in its leaves. This suggests that MOR2 is an Na⁺ excluder, either by minimising the entry of salt into the plant or by an excretion mechanism via the vesiculated hairs that play a significant role in the removal of salt from the remainder of the leaf, thereby preventing its accumulation to toxic levels in the leaves, whereas SOR4 acted as an Na⁺ includer. Higher levels of proline were detected in the MOR2 clone under NaCl treatments, suggesting a more developed adaptative mechanism for the selection of this characteristic in the southern part of the island, which is more exposed to abiotic stresses, particularly water stress that is either generated by salinity or by other causes.     

Effects of external NaCl on the growth of Atriplex amnicola and the ion relations and carbohydrate status of the leaves

Abstract Atriplex amnicola, was grown in nutrient solution cultures with concentrations of NaCl up to 750 mol m^?3. The growth optimum was at 25–50 mol m^?3 NaCl and growth was 10–15% of that value at 750 mol m^?3 NaCl. Sodium chloride at 200 mol m^?3 and higher reduced the rate of leaf extension and increased the time taken for a leaf to reach its maximal length. Concentrations of Na⁺, K⁺ and Mg²⁺ in leaves of different ages were investigated for plants grown at 25, 200 and 400 mol m^?3 NaCl. Although leaves of plants grown at 200 and 400 mol m^?3 NaCl had high Na⁺ concentrations at young developmental stages, much of this Na⁺ was located in the salt bladders. Leaves excluding bladders had low Na⁺ concentrations when young, but very high in Na⁺ when old. In contrast to Na⁺, K⁺ concentrations were similar in bladders and leaves excluding bladders. Concentrations of K⁺ were higher in the rapidly expanding than in the old leaves. At 400 mol m^?3 NaCl, the K⁺:Na⁺ ratios of the leaves excluding bladders were 0.4–0.6 and 0.1 for rapidly expanding and oldest leaves, respectively. The Na⁺ content in moles per leaf, excluding bladders, increased linearly with the age of the leaves; concurrent increases in succulence were closely correlated with the Na ⁺ concentration in the leaves excluding the bladders. Soluble sugars and starch in leaves, stems and buds were determined at dusk and dawn. There was a pronounced diurnal fluctation in concentrations of carbohydrates. During the night, most plant parts showed large decreases in starch and sugar. Concentrations of carbohydrates in most plant organs were similar for plants grown at 25 and 400 mol m^?3 NaCl. One notable exception was buds at dusk, where sugar and starch concentrations were 30–35% less in plants grown at 400 mol m^?3 NaCl than in plants grown at 25 mol m^?3 NaCl. The data indicate that the growth of A. amnicola at 400 mol m^?3 NaCl is not limited by the availability of photosynthate in the plant as a whole. However, there could have been a growth limitation due to inadequate organic solutes for osmotic regulation.     

Germination of winterfat (Eurotia lanata (Pursh) Moq.) seeds at reduced water potentials: testing assumptions of hydrothermal time model

The hydrothermal time model not only quantifies seed germination progress as affected by temperature and water potential, but also has ecological and biological significance. Assumptions of the hydrothermal time model were tested using two non-dormant seed collections of winterfat (Eurotia lanata) with two seed size classes. Winterfat is a native shrub with superior forage quality. Germination rates (GR) of subpopulations were estimated from germination time courses over a water potential range from 0 to −1.33 MPa at 2, 5, 10, 15, 20, and 25 °C. Parameters of the hydrothermal time model were estimated from the relationships between GR and temperature at various water potentials and between GR and ψ at various temperatures. Model assumptions were tested using these estimated parameters. Results indicate that base temperature (T_b(50)) and base water potential (ψ_b(50)) of the 50% subpopulation were not independent of temperature and water potential. The ψ_b(50) was lowest at intermediate temperatures between 10 and 15 °C, while T_b decreased linearly with increasing water availability. The estimated shift rates of T_b(50) with ψ were between 2.18 and 3.81 °C MPa⁻¹ for the two collections and large seeds had a greater shift rate than small seeds. Hydro time (θ_H) was constant among subpopulations only at optimal temperatures. A linear increase of θ_H with subpopulation was found at lower temperatures, especially at 2 °C. There were no significant differences in ψ_b(50) between large and small seeds, but significant differences were observed in hydrothermal time requirement (θ_HT(50)), which was lower at intermediate temperatures than at either lower or higher temperatures. The predictability of the hydrothermal time model was improved especially at low temperatures when θ_HT(50) was allowed to change with temperature as measured by a modified R² value. Changes in other parameters with temperature or water potential did not further improve the predictability of the hydrothermal time model. Therefore, further efforts in improving the hydrothermal time model should focus on variations in θ_HT.     

Winterfat (Eurotia lanata(Pursh) Moq.) Seedbed Ecology: Low Temperature Exotherms and Cold Hardiness in Hydrated Seeds as Influenced by Imbibition Temperature

Thermal analyses of freezing events in hydrated lettuce (LactucasativaL.) seeds show a correlation between low temperature exotherms(LTEs) (evidence of ice crystal formation) and seed death. Yet,weather patterns common to the Northern Great Plains of NorthAmerica regularly create conditions where non-dormant seedsof native plants hydrate with snow melt and are subsequentlyexposed to -30 °C or colder conditions. To determine ifsuch weather patterns decimate dispersed seeds, we measuredthe effects of freezing on fully hydrated winterfat (Eurotialanata(Pursh) Moq.) seeds harvested from the Northern Plainsat two USA and one Canadian location. Survival of hydrated seedsto -30 °C at a cooling rate of 2.5 °C h^-1was similarto that of seeds not subjected to cooling, even though botha high temperature exotherm (HTE) and an LTE were observed.Although the LTE was not related to winterfat seed survival,freeze-stressed seeds had reduced germination rates and reducedseedling vigour, particularly for the collection with the lightestseeds. The temperature of LTEs was similar among seed collectionswith a mean of -17.6 °C, but was warmer when the seeds wereimbibed at 0 °C compared to 5, 10 or 20 °C. We founda significant correlation between the HTE and LTE temperatures.The difference and the correlation may be due to the highermoisture content of seeds imbibed at 0 °C. After pericarpremoval, only one exotherm in the range of the LTE was observed.This was also true for the naked embryo. We conclude that anLTE indicates ice formation in the embryo, but that it doesnot signal the death of a winterfat seed.Copyright 1998 Annalsof Botany Company Eurotia lanata(Pursh) Moq.,Krascheninnikovia, Ceratoides,winterfat, exotherm, freezing tolerance, freezing avoidance, seedbed ecology, germination, D50, seedling vigour, seed collection     

Water diffusion,ion transport and lipid composition of the skin of Rana cyanophlyctis Schneider under osmotic stress

• 1.1. Analysis of the total lipid content (TL), components of the neutral lipid fraction (NLF), phospholipid fraction (PLF), recordings of electrical potential differences and diffusional permeability were carried out in the skin of the aquatic frog Rana cyanophlyctis subjected to in vivo salt stress (0.9 sodium chloride) for different durations (0, 1, 3 and 7 days).
• 2.2. A general decrease of skin TL and of components of the NEE and PEE was observed.
• 3.3. Stoichiometric ratios for skin PLF components under initial salt stress of different durations reveal an increase of the ratios of sphingomyelin and phosphatidyl choline after osmotic stress.
• 4.4. The diffusional permeability of water increased following exposure to salt stress of I, 3 and 7 days duration.
• 5.5. The transepithelial potential difference measured in vitro after a salt stress of 3 days was considerably higher than the controls.

     

Isolation and osmotic relations of developing megagametophytes of Torenia fournieri

A new method is reported to isolate and handle living megagametophytes of Torenia fournieri at any developmental stage. The stages were determined using light microscopy and delimited by correlating floral morphological traits. When significant changes in the osmotic pressure were found during development, enzyme solutions contained different concentrations of osmoticum. Osmotic pressure is lowest in the megaspore, increases until the four-nucleate stage and then gradually decreases until complete embryo sac formation. In enzymatic solutions containing appropriate concentrations of osmoticum, protoplasts of megaspores, two-, four-, eight-nucleate embryo sacs, egg cells, synergids and central cells were successfully isolated. The living protoplasts were collected by micromanipulator, transferred into microdroplets and tested for viability. Received: 1 June 1998 / Revision accepted: 20 May 1999     

Mineral nutrition and the water relations of plants

Summary When young tomato plants were transferred from nutrient solution to mineral-free water, reductions in transpiration, water content of the shoots and stomatal aperture were not accompanied by a reduction in the relative water content or an increase in the suction pressure of the leaves. The relative water content of the leaves was increased and the suction pressure was little affected.Following transfer of the plants to mineral-free water, the mineral content of the shoots and the osmotic pressure of expressed leaf sap were reduced. It was concluded that mineral salts were necessary for maintaining the osmotic pressure of the leaf cell sap and that this was achieved, at least in part, by maintaining the mineral concentration of the sap. The amount of water that could be taken up by leaves and their turgor pressure were related to the osmotic pressure of the sap and calculations of turgor pressure showed that it was less in the leaves of plants with their roots in mineral-free water than in the leaves of plants in nutrient solution.Evidence was obtained that in leaflets detached from plants with their roots in mineral-free water, stomatal closure could occur at a higher water content than in leaflets detached from plants in nutrient solution, indicating a further role of minerals in leaf water relations. It is suggested that this role may be related to the properties of the cell walls.     

Turgor and osmotic relations of the desert shrub Larrea tridentata

Abstract Leaf water relations characteristics of creosote bush, Larrea tridentata, were studied in view of previous reports that its leaves commonly experience zero or negative turgor under dry conditions. Leaf turgor loss point () was determined by a pressure-volume method for samples subjected to a hydration procedure and for untreated samples. Hydration caused to increase by as much as 3 M Pa. Hydration of samples also caused changes in other leaf water relations characteristics such as symplastic solute content, tissue elasticity and symplasmic water fraction, but total leaf solute content was unchanged. Comparison of our field plant water potential data with values of obtained by the two methods resulted in predictions of turgor loss during part or all of a diurnal cycle based on hydrated samples, and turgor maintenance (at least 0.3 MPa) based on untreated samples. Pooled data for obtained from both partially hydrated and untreated samples showed that L. tridentata maintains fairly constant levels of turgor over a wide range of leaf water potential. Dilution of cell contents by apoplastic water introduced significant errors in psychrometric determinations of osmotic potential in both frozen and thawed leaf tissue and expressed cell sap. Use of these values of osmotic potential resulted in predictions of zero turgor at all plant water potentials measured in the field.     

Response to non-uniform salinity in the root zone of the halophyte Atriplex nummularia: growth, photosynthesis, water relations and tissue ion concentrations

Background and Aims

Soil salinity is often heterogeneous, yet the physiology of halophytes has typically been studied with uniform salinity treatments. An evaluation was made of the growth, net photosynthesis, water use, water relations and tissue ions in the halophytic shrub Atriplex nummularia in response to non-uniform NaCl concentrations in a split-root system.

Methods

Atriplex nummularia was grown in a split-root system for 21 d, with either the same or two different NaCl concentrations (ranging from 10 to 670 mm), in aerated nutrient solution bathing each root half.

Key Results

Non-uniform salinity, with high NaCl in one root half (up to 670 mm) and 10 mm in the other half, had no effect on shoot ethanol-insoluble dry mass, net photosynthesis or shoot pre-dawn water potential. In contrast, a modest effect occurred for leaf osmotic potential (up to 30 % more solutes compared with uniform 10 mm NaCl treatment). With non-uniform NaCl concentrations (10/670 mm), 90 % of water was absorbed from the low salinity side, and the reduction in water use from the high salinity side caused whole-plant water use to decrease by about 30 %; there was no compensatory water uptake from the low salinity side. Leaf Na⁺ and Cl⁻ concentrations were 1·9- to 2·3-fold higher in the uniform 670 mm treatment than in the 10/670 mm treatment, whereas leaf K⁺ concentrations were 1·2- to 2·0-fold higher in the non-uniform treatment.

Conclusions

Atriplex nummularia with one root half in 10 mm NaCl maintained net photosynthesis, shoot growth and shoot water potential even when the other root half was exposed to 670 mm NaCl, a concentration that inhibits growth by 65 % when uniform in the root zone. Given the likelihood of non-uniform salinity in many field situations, this situation would presumably benefit halophyte growth and physiology in saline environments.Key words: Split-root system, salinity heterogeneity, root zone heterogeneity, water potential, water use, stomatal conductance, saltbush, leaf ions, photosynthesis, NaCl     

Water relations of drought hardy shrubs: osmotic potential and stomatal reactivity

Abstract. Diurnal measurements of total water potential and stomatal opening were made at six sites. Pressure-volume curves were established on parallel leaf samples. In eastern Austria, the species investigated were Cornus mas L., Cornus sanguinea L., Crateagus monogyna Jacq., Sorbus aria (L.) Crantz and Viburnum lantana L. in southern France Crateagus monogyna , and in southern Turkey Crateagus monogyna and Olea europaea L. Osmotic adjustment, defined as a change in osmotic potential larger than the passive change resulting from the loss of cell water, was relatively small from day to day or week to week in mature, non-senescing leaves. Cornus sanguinea was an exception. A recently suggested method for the demonstration of diurnal active osmotic adjustment seems not to be reliable without further independent corroboration. Changes in the leaf water potential threshold for stomatal closure were either insignificant when the pressure-volume characteristics of the plant material were stable, or significant when shifts in such parameters as the turgor loss point occurred ( Cornus sanguinea ).     

Cell growth and water relations of the halophyte,Atriplex nummularia L., in response to NaCl

Summary Growth reduction or cessation is an initial response of Atriplex nummularia L. cells to NaCl. However, A. nummularia L. cells that are adapted to 342 and 428 mM NaCl are capable of sustained growth in the presence of salt. Cells that are adapted to NaCl exhibit a reduced rate of division compared to unadapted cells. Unlike salt adapted cells of the glycophyte Nicotiana tabacum L., A. nummularia L. cells do not exhibit reduced rate of cell expansion after adaptation. However, the cell expansion rate of unadapted A. nummularia L. cells is considerably slower than that of unadapted glycophyte cells and this normally low rate of cell expansion may contribute to the enhanced capacity of the halophyte to tolerate salt. Turgor of NaCl adapted cells was equivalent to unadapted cells indicating that the cells of the halophyte do not respond to salt by osmotic over adjustment as reported for the glycophyte tobacco (Binzel et al. 1985, Plant Physiol. 79:118–125).     

Mineral composition of cultured Ginseng cells]

The contents of macroelements and microelements in ginseng roots and callus cultures was determined by atom absorption spectroscopy. Ginseng cells and tissues were shown to accumulate considerable amounts of microelements. The content of six of eleven mineral components studied (K, Ca, Na, Mo, Mn, and Cr) in callus cultures was higher than that in roots of agricultural ginseng plants. We revealed good correlations between the contents of microelements (K, Ca, and Mg), as well as between the concentrations of macroelements (Mo, Li, Cu, and Cr) in ginseng cultures. The ability to accumulate elements varied between ginseng species, which was probably related to their genetic features. Our findings indicate that cultured ginseng cells hold much promise as the source of microelements.     

Effects of Polyploidy on Photosynthetic Rates, Photosynthetic Enzymes, Contents of DNA, Chlorophyll, and Sizes and Numbers of Photosynthetic Cells in the C(4) Dicot Atriplex confertifolia

Photosynthetic rates, chlorophyll content, and activities of several photosynthetic enzymes were determined per cell, per unit DNA, and per unit leaf area in five ploidal levels of the C₄ dicot Atriplex confertifolia. Volumes of bundle sheath and mesophyll protoplasts were measured in enzymatic digestions of leaf tissue. Photosynthetic rates per cell, contents of DNA per cell, and activities of the bundle sheath enzymes ribulose 1,5-bisphosphate carboxylase (RuBPC) and NAD-malic enzyme per cell were correlated with ploidal level at 99% or 95% confidence levels, and the results suggested a near proportional relationship between gene dosage and gene products. There was also a high correlation between volume of mesophyll and bundle sheath cells and the ploidal level. Contents of DNA per cell, activity of RuBPC per cell, and volumes of cells were correlated with photosynthetic rate per cell at the 95% confidence level. The mesophyll cells did not respond to changes in ploidy like the bundle sheath cells. In the mesophyll cells the chlorophyll content per cell was constant at different ploidal levels, there was less increase in cell volume than in bundle sheath cells with an increase in ploidy, and there was not a significant correlation (at 95% level) of phosphoenolpyruvate carboxylase activity or content and pyruvate,Pi dikinase activity with increase in ploidy. The number of photosynthetic cells per unit leaf area progressively decreased with increasing ploidy from diploid to hexaploid, but thereafter remained constant in octaploid and decaploid plants. Numbers of cells per leaf area were not correlated with cell volumes. The mean photosynthetic rates per unit leaf area were lowest in the diploid, similar in 4×, 6×, and 8×, and highest in the decaploid. The photosynthetic rate per leaf area was highly correlated with the DNA content per leaf area.