Leaf water potential of differentially salinized plants

Water and osmotic potential energies were measured with thermocouple psychrometers, at intervals during a 4-week period, in growing leaves of bean (Phaseolus vulgaris, var. Blue Lake) and barley (Hordeum vulgare, var. Liberty) plants having roots equally split between 2 differentially salinized nutrient solutions. The osmotic potentials of plants with half their roots in saline solutions were about halfway between the osmotic potentials of plants grown in nonsaline solutions and those grown in saline solutions. By the end of the 4-week measurement period, the beans and barley were almost mature. The final dry weights of shoots of plants with half their roots in saline solutions were about halfway between the dry weights of the shoots of plants grown in nonsaline solutions and the dry weights of those in saline solutions. The results obtained showed that the degree of osmotic adjustment and the rate of growth were functions of the proportion of the root system exposed to saline conditions.     

Effects of waterlogging and salinity on plant-water relations and on the accumulation of solutes in three mangrove species

Dispersal units of Avicennia marina (Forsk.) Vierh., Rhizophora mucronata Lam. and Bruguiera gymnorrhiza (L.) Lam. were cultivated in mangrove sediment under greenhouse conditions. After 7 months plants were subjected to the following waterlogging and salinity treatments for 60 days: drained non-saline, drained saline, waterlogged non-saline and waterlogged saline. Measurements were made of stomatal resistance (r_s), tissue water potentials, relative water content, and the concentration of Na, K, Ca and Mg in plant organs. Measurements were made 4 days before the termination of waterlogging and continued for 12 days thereafter. Generally, the highest stomatal resistance occurred in the waterlogged saline treatments and the least stomatal resistance occurred in the drained non-saline treatments. Water potentials were usually lower in the saline treatments than in the non-saline treatments. Depression in water potential was greatest in Avicennia and least in Bruguiera. In all 3 species, the concentration of cations was high in the roots. Salinity had greater effects on the concentration of ions in Avicennia than in Rhizophora or Bruguiera. The low stomatal resistance, low tissue water potentials, high relative water content and high tissue cation concentration in Avicennia suggest that it is ideally suited as the pioneer in the mangrove association. Generally, Rhizophora was more tolerant of salinity than Bruguiera. The responses of the plants reveal the remarkable adaptability of mangroves to the saline environment. The adaptive features of each species determine its characteristic habitat in intertidal areas.     

Development of water conducting capacity in the root systems of young plants of corn and some other c4 grasses

Development of the primary and early nodal roots was studied in Zea mays L., Zea mexicana (Schrad.) Reeves & Mangelsd., Sorghum bicolor (L.) Moench., and Sorghum sudanese (Piper) Stapf. in relation to shoot development. In all the types studied all roots reached lengths of about 30 centimeters before the late metaxylem (LMX) was open, and young plants with total root lengths of around 100 centimeters had almost no open LMX. On average, corn seedlings with up to 36 square centimeters of leaf had no open LMX. The name “immature apices” is suggested for such long but not fully functional roots. In plants up to 50 days old a fairly constant proportion of less than half the total root length had open LMX. A pilot study of stomatal resistance on days of high evaporative demand suggested that young seedlings may show higher resistance than older plants in the afternoon. Estimates of longitudinal permeability of corn roots with only early metaxylem vessels open indicate very steep gradients of water potential would develop under such conditions.     

The role of arbuscular mycorrhizal fungi in alleviating salt stress in Medicago sativa L. var. icon

Medicago sativa L. is the most important forage crop in arid and semi-arid areas, where increased salinity is a major factor limiting plant growth and crop productivity. The role of arbuscular mycorrhizal (AM) fungus Glomus viscosum H.T. Nicolson strain A6 in protecting alfalfa plants from salt stress, induced by sodium chloride (NaCl), was studied in two ways. Firstly, the root systems of 3-month old M. sativa plants, both mycorrhizal (AM+) and non-mycorrhizal (non-AM) (M. sativa L. var. icon), were placed in solutions of increasing salt concentrations (0, 50, 100, 150, 200 mM NaCl) to study the wilting response. G. viscosum improved the tolerance to salinity stress and the benefit was expressed in terms of the time required to reach the T4 stage in the wilting experiment. Secondly, to evaluate the ability of the Glomus-alfalfa symbiosis to tolerate salt, a pot experiment was set up in a glasshouse in which 3-month old alfalfa plants (M. sativa var. icon) were grown in a peat substratum at three salinity levels (0, 100, 150 mM NaCl). The AM symbiosis stimulated plant height, leaf area, root density, fresh and dry plant weight under saline conditions. Furthermore, proline accumulation was higher in mycorrhizal M. sativa plants than in non-mycorrhizal plants under conditions of salt stress. These and other results indicated that the micropropagated selected clone of M. sativa var. icon, when in symbiosis with G. viscosum H.T. Nicolson strain A6, exhibited better growth and physiological activities under saline conditions than non-AM plants. The AM+ plants also had lower sodium and chloride concentrations in tissues than non-AM plants.     

Effect of Nitrate and Ammonium Nutrition of Nonnodulated Phaseolus vulgaris L. on Phosphoenolpyruvate Carboxylase and Pyruvate Kinase Activity

Young bean plants (Phaseolus vulgaris L. var Saxa) were fed with 3.5 or 10 millimolar N in either the form of NO₃⁻ or NH₄⁺, after being grown on N-free nutrient solution for 8 days. The pH of the nutrient solutions was either 6 or 4. The cell sap pH and the extractable activities of phosphoenolpyruvate carboxylase and of pyruvate kinase from roots and primary leaves were measured over several days.

The extractable activity of phosphoenolpyruvate carboxylase (based on soluble protein) from primary leaves increased with NO₃⁻ nutrition, whereas with NH₄⁺ nutrition and on N-free nutrient solution the activity remained at a low level. Phosphoenopyruvate carboxylase activity from the roots of NH₄⁺-fed plants at pH 4 was finally somewhat higher than from the roots of plants grown on NO₃⁻ at the same pH. There was no difference in activity from the root between the N treatments when pH in the nutrient solutions was 6. The extractable activity of pyruvate kinase from roots and primary leaves seemed not to be influenced by the N nutrition of the plants.

The results are discussed in relation to the physiological function of both enzymes with special regard to the postulated functions of phosphoenolpyruvate carboxylase in C3 plants as an anaplerotic enzyme and as part of a cellular pH stat.

     

Continuous monitoring of plant water potential

Plant water potential was monitored continuously with a Wescor HR-33T dewpoint hygrometer in conjunction with a L51 chamber. This commercial instrument was modified by replacing the AC-DC mains power converter with one stabilized by zener diode controlled transistors. The thermocouple sensor and electrical lead needed to be thermally insulated to prevent spurious signals. For rapid response and faithful tracking a low resistance for water vapor movement between leaf and sensor had to be provided. This could be effected by removing the epidermis either by peeling or abrasion with fine carborundum cloth. A variety of rapid plant water potential responses to external stimuli could be followed in a range of crop plants (sunflower (Helianthus annuus L., var. Hysun 30); safflower (Carthamus tinctorious L., var. Gila); soybean (Glycine max L., var. Clark); wheat (Triticum aestivum L., var. Egret). These included light dark changes, leaf excision, applied pressure to or anaerobiosis of the root system. Water uptake by the plant (safflower, soybean) mirrored that for water potential changes including times when plant water status (soybean) was undergoing cyclical changes.     

Temperature dependence of photosynthesis in cotton

Cotton plants (Gossypium hirsutum L., var. Deltapine Smooth Leaf) were grown under controlled environmental conditions over a range of day/night temperatures from 20/15 to 40/35 C. Their photosynthetic characteristics were then measured over a comparable temperature range. Net photosynthesis tended stongly to be greatest, and intracellular resistance to CO₂ transport to be lowest, when the measurement temperature corresponded to the daytime growth temperature, suggesting pronounced acclimation of the plants to the growth temperature. The preferred growth temperature was close to the 25/20 C regime, since net photosynthesis of these plants, regardless of measurement temperature, was higher and intracellular resistance lower, than in plants from any other regime.     

Foliar application of ascorbic acid enhances salinity stress tolerance in barley (Hordeum vulgare L.) through modulation of morpho-physio-biochemical attributes,ions uptake,osmo-protectants and stress response genes expression

Barley (Hordeum vulgare L.) is a major cereal grain and is known as a halophyte (a halophyte is a salt-tolerant plant that grows in soil or waters of high salinity). We therefore conducted a pot experiment to explore plant growth and biomass, photosynthetic pigments, gas exchange attributes, stomatal properties, oxidative stress and antioxidant response and their associated gene expression and absorption of ions in H. Vulgare. The soil used for this analysis was artificially spiked at different salinity concentrations (0, 50, 100 and 150 mM) and different levels of ascorbic acid (AsA) were supplied to plants (0, 30 and 60 mM) shortly after germination of the seed. The results of the present study showed that plant growth and biomass, photosynthetic pigments, gas exchange parameters, stomatal properties and ion uptake were significantly (p < 0.05) reduced by salinity stress, whereas oxidative stress was induced in plants by generating the concentration of reactive oxygen species (ROS) in plant cells/tissues compared to plants grown in the control treatment. Initially, the activity of antioxidant enzymes and relative gene expression increased to a saline level of 100 mM, and then decreased significantly (P < 0.05) by increasing the saline level (150 mM) in the soil compared to plants grown at 0 mM of salinity. We also elucidated that negative impact of salt stress in H. vulgare plants can overcome by the exogenous application of AsA, which not only increased morpho-physiological traits but decreased oxidative stress in the plants by increasing activities of enzymatic antioxidants. We have also explained the negative effect of salt stress on H. vulgare can decrease by exogenous application of AsA, which not only improved morpho-physiological characteristics, ions accumulation in the roots and shoots of the plants, but decreased oxidative stress in plants by increasing antioxidant compounds (enzymatic and non-enzymatic). Taken together, recognizing AsA's role in nutrient uptake introduces new possibilities for agricultural use of this compound and provides a valuable basis for improving plant tolerance and adaptability to potential salinity stress adjustment.     

Root-shoot ratio in irrigated plants

Relationship between roots and shoots ofPimpinella saxifraga L.,Petroselinum hortense Hoffm.,Lactuca sativa L., andBeta vulgaris subsp.esculenta (Salisbury) Gürke var.altissima Roessig was studied in irrigated and non-irrigated plants. It was found that the irrigation water did not influence distinctly the root-shoot ratio of the experimental plants. From the statical point of view, this relationship could be expressed by the following quantitative ratios for irrigated and non-irrigated plants: 3∶1 forPimpinella saxifraga L., 2∶1 forPetroselinum hortense Hoffm., 9∶1 forLactuca sativa L., and 1∶1 for beetroot. From the dynamic point of view it is possible to state that this ratio changes in the course of individual development. With plants cultivated for their aerial parts, it increased with age, while with plants cultivated for their roots, it decreased with age. The irrigation water produced much greater morphological changes (the shape and the surface area) in the root and shoot system of experimental plants.     

Arbuscular mycorrhizal symbiosis alleviates detrimental effects of saline reclaimed water in lettuce plants

The present study evaluated the effects of inoculation with arbuscular mycorrhizal fungi (AMF; Glomus iranicum var. tenuihypharum sp. nova) on the physiological performance and production of lettuce plants grown under greenhouse conditions and supplied with reclaimed water (RW; urban-treated wastewater with high electrical conductivity; 4.19 dS m^?1). Four treatments, fresh water, fresh water plus AMF inoculation, RW and RW plus AMF inoculation, were applied and their effects, over time, analyzed. Root mycorrhizal colonization, plant biomass, leaf-ion content, stomatal conductance and net photosynthesis were assessed. Overall, our results highlight the significance of the AMF in alleviation of salt stress and their beneficial effects on plant growth and productivity. Inoculated plants increased the ability to acquire N, Ca, and K from both non-saline and saline media. Moreover, mycorrhization significantly reduced Na plant uptake. Under RW conditions, inoculated plants also showed a better performance of physiological parameters such as net photosynthesis, stomatal conductance and water-use efficiency than non-mycorrhizal plants. Additionally, the high concentration of nutrients already dissolved in reclaimed water suggested that adjustments in the calculation of the fertigation should be conducted by farmers. Finally, this experiment has proved that mycorrhization could be a suitable way to induce salt stress resistance in iceberg lettuce crops as plants supplied with reclaimed water satisfied minimum legal commercial size thresholds. Moreover, the maximum values of Escherichia coli in the reclaimed water were close to but never exceeded the international thresholds established (Spanish Royal Decree 1620/2007; Italian Decree, 2003) and hence lettuces were apt for sale.     

Influence of cadmium on water relations, stomatal resistance, and abscisic Acid content in expanding bean leaves

Ten day old bush bean plants (Phaseolus vulgaris L. cv Contender) were used to analyze the effects of 3 micromolar Cd on the time courses of expansion growth, dry weight, leaf water relations, stomatal resistance, and abscisic acid (ABA) levels in roots and leaves. Control and Cd-treated plants were grown for 144 hours in nutrient solution. Samples were taken at 24 hour intervals. At the 96 and 144 hour harvests, additional measurements were made on excised leaves which were allowed to dry for 2 hours. From the 48 hour harvest, Cd-treated plants showed lower leaf relative water contents and higher stomatal resistances than controls. At the same time, root and leaf expansion growth, but not dry weight, was significantly reduced. The turgor potentials of leaves from Cd-treated plants were nonsignificantly higher than those of control leaves. A significant increase (almost 400%) of the leaf ABA concentration was detected after 120 hours exposure to Cd. But Cd was found to inhibit ABA accumulation during drying of excised leaves. It is concluded that Cd-induced decrease of expansion growth is not due to turgor decrease. The possible mechanisms of Cd-induced stomatal closure are discussed.     

NMR imaging of root water distribution in intact Vicia faba L plants in elevated atmospheric CO2

The effect of elevated atmospheric CO₂ on water distribution in the intact roots of Vicia faba L. bean seedlings grown in natural soil was studied noninvasively with proton (¹H) nuclear magnetic resonance (NMR) imaging. Exposure of 24-d-old plants to atmospheric CO₂-enriched air at 650 cm³ m^?3 produced significant increases in water imaged in upper roots, hypogeal cotyledons and lower stems in response to a short-term drying-stress cycle. Above ground, drying produced negligible stem shrinkage and stomatal resistance was unchanged. In contrast, the same drying cycle caused significant depletion of water imaged in the same upper root structures in control plants subject to ambient CO₂ (350 m³ m^?3), and stem shrinkage and increased stomatal resistance. The results suggest that inhibition of transpiration caused by elevated CO₂ does not necessarily result in attenuation of water transport from lower root structures. Inhibition of water loss from upper roots and lower stem in elevated CO₂ environments may be a mitigating factor in assessing deleterious effects of greenhouse changes on crops during periods of dry climate.     

Nutrient status and mycorrhizal enhancement of water transport in soybean

Mycorrhizal soybean (Glycine max L. Merr. var. Harosoy-63) plants have lower resistances to water transport than nonmy-corrhizal plants after 4.5 weeks of growth. Although resistances of whole plants differ by 40%, there were no differences in the resistances of stems plus leaves, indicating that the major effect of the mycorrhizae was to reduce the resistance of the roots. Since the fungitoxicant, p-chloronitrobenzene, had no effect on resistances to water transport, reduced resistances were probably not caused by a direct modification of the transport pathway by the fungus. Differences in resistance between mycorrhizal and nonmycorrhizal soybean were essentialy eliminated by the application of nutrients to the soil. Thus, lowered resistances of mycorrhizal roots growing in soil with low levels of nutrients probably resulted from the enhanced nutrient status of the plant brought about by the fungus. Mycorrhizal infection increased growth at both low and high nutrient levels.     

Resistance Responses of Potato to Vesicular-Arbuscular Mycorrhizal Fungi under Varying Abiotic Phosphorus Levels

In mycorrhizal symbioses, susceptibility of a host plant to infection by fungi is influenced by environmental factors, especially the availability of soil phosphorus. This study describes morphological and biochemical details of interactions between a vesicular-arbuscular mycorrhizal (VAM) fungus and potato (Solanum tuberosum L. cv Russet Burbank) plants, with a particular focus on the physiological basis for P-induced resistance of roots to infection. Root infection by the VAM fungus Glomus fasciculatum ([Thaxt. sensu Gerdemann] Gerdemann and Trappe) was extensive for plants grown with low abiotic P supply, and plant biomass accumulation was enhanced by the symbiosis. The capacity of excised roots from P-deficient plants to produce ethylene in the presence or absence of exogenous 1-amino cyclopropane-1-carboxylic acid (ACC) was markedly reduced by VAM infection. This apparent inhibition of ACC oxidase (ACC_ox) activity was localized to areas containing infected roots, as demonstrated in split-root studies. Furthermore, leachate from VAM roots contained a potent water-soluble inhibitor of ethylene generation from exogenous ACC by nonmycorrhizal (NM) roots. The leachate from VAM-infected roots had a higher concentration of phenolics, relative to that from NM roots. Moreover, the rates of ethylene formation and phenolic concentration in leachates from VAM roots were inversely correlated, suggesting that this inhibitor may be of a phenolic nature. The specific activity of extracellular peroxidase recovered in root leachates was not stimulated by VAM infection, although activity on a fresh weight basis was significantly enhanced, reflecting the fact that VAM roots had higher protein content than NM roots. Polyphenol oxidase activity of roots did not differ between NM and VAM roots. These results characterize the low resistance response of P-deficient plants to VAM infection. When plants were grown with higher abiotic P supply, the relative benefit of the VAM symbiosis to plant growth decreased and root infection was lower. The in vivo ACC_ox activity was also greater in roots of plants grown on high levels of P compared with those grown on low levels, although the influence of VAM infection was partially to counteract the nutritional effect of P on ACC_ox activity. Similar to ACC_ox activity, extracellular peroxidase activity of roots increased linearly with increasing abiotic P supply, thus indicating a greater potential for resistance to VAM infection. These findings suggest that VAM fungi may alter phenolic metabolism of roots so as to hinder ethylene production and the root's ability to invoke a defense response. Raising the abiotic P supply to plants at least partially restores the capacity of roots to produce ethylene and may, in this way, increase the root's resistance to VAM infection.     

Influx and efflux of p in roots of intact maize plants : double-labeling with p and p

Rates of P influx and efflux were determined in whole plants at ambient P concentrations comparable to those found in soil solutions. Maize (Zea mays L. var NC+59) seedlings were trimmed (endosperm and adventitious roots removed) and grown in a greenhouse in solution cultures at P concentrations of approximately 0.4 and 1.8 micromolar. Roots of intact plants previously exposed to ³²P-labeled solutions at 0.2 and 2.0 micromolar P for 48 hours were rinsed 10 minutes in P-free solution and exposed to ³³P solutions at 0.2 and 2.0 micromolar for 10 minutes. Net depletion of ³³P from and appearance of ³²P in the ambient solution were used to measure influx and efflux. The ration of ³²P efflux to ³³P influx was about 0.68 at 0.2 micromolar and 0.08 at 2.0 micromolar. When plants were allowed to deplete P from solutions, the P concentration in the medium dropped to about 0.15 micromolar within 24 hours and 0.05 micromolar within 60 hours. Results indicate that P efflux is a substantial component of net P accumulation at P concentrations normally found in soil solutions.     

Stomatal Response to Light of Solanum pennellii, Lycopersicon esculentum, and a Graft-induced Chimera

To learn how species differences in stomatal behavior are regulated, the response of epidermal and leaf diffusive resistance to light was investigated in Lycopersicon esculentum Mill., Solanum pennellii Corr., and a periclinal chimera having an S. pennellii epidermis and an L. esculentum mesophyll that was produced from a graft of the two species. S. pennellii has about 23% fewer stomata per square millimeter than does L. esculentum, and the two species have contrasting stomatal sensitivities to light. The abaxial stomata of L. esculentum open in dimmer light and to a greater extent than the adaxial stomata. The abaxial and adaxial stomata of S. pennellii respond similarly to light incident on the adaxial epidermis and are less open at all quantum flux densities than comparable stomata of L. esculentum. The patterns of response to light of the abaxial and adaxial stomata of the chimera were practically identical to those of L. esculentum, and quite unlike those of S. pennellii. Thus, the pattern of stomatal light response in the chimera was regulated by the L. esculentum mesophyll. The reduction in stomatal frequency of the chimera, which was regulated by the epidermis of S. pennellii, contributed to the 40% difference in leaf diffusive resistance between the plants in moderate light.     

Circadian Rhythms in Photosynthesis : Oscillations in Carbon Assimilation and Stomatal Conductance under Constant Conditions

Net carbon assimilation and stomatal conductance to water vapor oscillated repeatedly in red kidney bean, Phaseolus vulgaris L., plants transferred from a natural photoperiod to constant light. In a gas exchange system with automatic regulation of selected environmental and physiological variables, assimilation and conductance oscillated with a free-running period of approximately 24.5 hours. The rhythms in carbon assimilation and stomatal conductance were closely coupled and persisted for more than a week under constant conditions. A rhythm in assimilation occurred when either ambient or intercellular CO₂ partial pressure was held constant, demonstrating that the rhythm in assimilation was not entirely the result of stomatal effects on CO₂ diffusion. Rhythms in assimilation and conductance were not expressed in plants grown under constant light at a constant temperature, demonstrating that the rhythms did not occur spontaneously but were induced by an external stimulus. In plants grown under constant light with a temperature cycle, a rhythm was entrained in stomatal conductance but not in carbon assimilation, indicating that the oscillators driving the rhythms differed in their sensitivity to environmental stimuli.     

Evidence for the nitrate-dependent spatial regulation of the nitrate reductase gene in chicory roots

Young chicory plants (Cichorium intybus L. var. Witloof) show a tenfold higher nitrate reductase NR activity in roots compared to leaves. Northern analysis revealed, besides the nitrate inducibility of the nitrate reductase gene (nia), a higher level of expression in the roots. By modifying the external nitrate concentration the NR activity in the leaves remained negligible whereas a maximal activity was observed in the roots when grown in the presence of 5 mM nitrate. Surprisingly, variation of the external nitrate concentration induced changes in the spatial regulation of nia within the root. In-situ hybridization mainly localized nia mRNA in the cortical cells of roots grown at low nitrate concentrations (0.2 mM). At high nitrate concentrations (5 mM), nia mRNA was more abundant in the vascular tissues. The root apex revealed a strong signal under both conditions. The isolation and characterization of the NR structural gene from chicory is also presented. Southern blot analysis revealed the presence of a single nia gene per haploid genome of chicory.     

Stomatal Behavior and CO(2) Exchange Characteristics in Amphistomatous Leaves

The possibility that differences in stomatal conductance between upper and lower surfaces of amphistomatous leaves are adaptations to differences in CO₂ exchange characteristics for the two surfaces was investigated. The ratio of upper to lower stomatal conductance was found to change little in response to light and humidity for well-watered sunflower (Helianthus annuus L.) plants. Stressing the plants (ψ = −17 bars) and rewatering 1 day before gas exchange measurements reduced upper conductance more severely than lower in both indoor- and outdoor-grown plants, and caused small changes in conductance ratio with light and humidity. A similar pattern was found using outdoor grown sunflower and cocklebur (Xanthium strumarium L.) plants. Calculated intercellular CO₂ concentrations for upper and lower surfaces were always close to identical for a particular set of environmental conditions for both sunflower and cocklebur, indicating that no differences in CO₂ exchange characteristics exist between the two surfaces. By artificially creating a CO₂ gradient across the leaf, the resistance to CO₂ diffusion through the mesophyll was estimated and found to be so low that despite possible nonhomogeneity of the mesophyll, differences in CO₂ exchange characteristics for the two surfaces are unlikely. It is concluded that differences in conductance between upper and lower stomates are not adaptations to differences in CO₂ exchange characteristics.