Water stress induced changes in concentrations of proline and total soluble sugars in nodulated alfalfa (Medicago sativd) plants

Susceptibility of alfalfa ( Medicago saliva L. cv. Aragón) nodules and leaves to water stress has been investigated. Nodule acetylene reduction activity (ARA), leaf CO₂ exchange rate (CER) as well as soluble protein, proline and total soluble sugar (TSS) contents were determined during drought. Water status was estimated as water potential (Ψ_w) and Relative water content (RWC) of the respective tissues. Maximum rates of ARA required higher Ψ_w than CER. Nodules had lower RWC for a given Ψ_w than leaves. Water stress reduced soluble protein content in both tissues; however, the decline in soluble protein content was detected at greater Ψ_w in nodules than in leaves. Proline and TSS increased in leaves and nodules, and again the threshold Ψ_w triggering such accumulation was higher in nodule tissues. Oior results suggest that alfalfa nodules are more susceptible to water shortage than leaves. Effects of accumulated TSS and proline upon leaf and nodule physiology are discussed in relation to protein stability (proline), pH control (proline) and osmotic adjustment (proiine and TSS). The TSS accumulation induced by water stress suggests that substrate shortage would not be the primary effect of drought on nodule activity.     

The Extent and Pattern of Osmotic Adjustment in White Clover (Trifolium repens L.) During the Development of Water Stress

White clover plants were subjected to water stress followingthe cessation of watering. As a water deficit developed, waterand osmotic potentials were measured in stolon tips, leavesfrom the stolon tip and leaves from the plant crown. Pressurepotentials were calculated. Pressure potential was maintainedin stolon tips even when water potential fell to around –2·0MPa. In contrast, pressure potential in leaves fell rapidlyas water stress developed. Total amino acid and potassium levels were largely unaffectedin both stolon tips and leaves. Water-soluble carbohydratesand proline accumulated during water stress. The increase inproline level in leaves did not follow the same pattern as thatin stolon tips, although toward the end of the water stressperiod the level had increased by a similar extent in both partsof the plant. Additionally, pressure potential and osmotic potentialappeared to be significantly related to proline content in stolontips. No such relationship was found for leaves. The role ofproline in osmotic adjustment is discussed. Trifolium repens L. cv. Olwen, white clover, water stress, osmotic adjustment, proline     

Photosynthetic, Physiological and Biochemical Responses of Tomato Plants to Polyethylene Glycol-Induced Water Deficit

Polyethylene glycol （PEG 6000）-induced water deficit causes physiological as well as biochemical changes in plants. The present study reports on the results of such changes in hydroponically grown tomato plants （Lycopersicon esculentum Mill. cv. Nikita）. Plants were subjected to moderate and severe levels of water stress （i.e. water potentials in the nutrient solution of- 0.51 and -1.22 MPa, respectively）. Water stress markedly affected the parameters of gas exchange. Net photosynthetic rate （Pn） decreased with the induction of water stress. Accordingly, a decrease in the transpiration rate （E） was observed. The ratio of both （Pn/E） resulted in a decrease in water use efficiency. One of the possible reasons for the reduction in Pn is structural damage to the thylakoids, which affects the photosynthetic transport of electrons. This was indicated by an increase in non-photochemical quenching and a reduction in the quantum yield of photosystem Ⅱ. Furthermore, a decrease in both leaf water potential and leaf osmotic potential was observed, which resulted in a significant osmotic adjustment during stress conditions. Analysis of the physiological responses was complemented with a study on changes in proline content. In stressed plants, a 10-fold increase in proline content was detected compared with control plants. It is clear that water stress tolerance is the result of a cumulative action of various physiological and biochemical processes, all of which were affected by PEG 6000-induced water stress.     

Proline accumulation and the adaptation of cultured plant cells to water stress

The transfer of cultured tomato cells (Lycopersicon esculentum cv VFNT-Cherry) to a low water potential environment resulted in an increased dry weight to fresh weight ratio accompanied by a rapid accumulation of proline. Proline content continued to increase as osmotic adjustment and growth occurred. The initial increase in proline concentration was accompanied by a drop in turgor. However, proline levels continued to increase with a gain in turgor during osmotic adjustment. Thus, the accumulation of proline depended not only on cell water potential, or on the initial loss of turgor but more closely on cell osmotic potential. The ultimate level of proline depended on the level of adaptation. Proline levels remained high after more than 100 cell generations in low water potential media, but declined rapidly after transfer to media with a less negative water potential. Addition of exogenous proline to the medium during water stress and during osmotic downshock alleviated the normally resulting inhibition of growth. The results suggest a positive role for proline accumulation in adaptation of cells to changing external water potentials.     

Growth and osmoregulation in Pisum sativum subhooks as affected by gibberellic acid and cotyledon excision

Growth stimulation by gibberellic acid (GA) of the Alaska pea ( Pisum sativum L.) subhook was observed within 6 h after its application; the stimulation being larger in cuttings with cotyledons than in decotylized ones. The osmotic potential in the subhook increased as it grew, the rate of its increase being faster in cuttings without than in cuttings with cotyledons. GA had no effect on the change in the osmotic potential until 8 h after GA application, but afterwards it suppressed the increase in cuttings with cotyledons. This GA effect was not observed in decotylized cuttings. Changes in the osmotic potential were well correlated with changes in the concentration of soluble sugars, but not with changes in amino acids and K⁺, Soluble sugars accumulated in the subhook of cuttings with and without cotyledons in proportion to growth, irrespective of the presence or absence of GA. Cotyledon excision suppressed sugar accumulation, and GA promoted it in cuttings with cotyledons but not in decotylized ones. These results suggest that GA stimulates the translocation of sugars from the cotyledons to the subhook and, thereby, maintains the osmotic potential low, resulting in enhanced growth.     

Water use efficiency by alfalfa: Mechanisms involving anti-oxidation and osmotic adjustment under drought

Water use and mechanisms relating to osmotic adjustment and anti-oxidation were investigated in alfalfa (Medicago sativa L.) plants under reduced water availability. Water use efficiency (WUE), MDA and proline contents, and antioxidant enzyme activities were measured in three alfalfa cultivars under three levels of soil water availability in a greenhouse pot experiment. WUE was determined indirectly using discriminating carbon isotope composition. WUE increased with the severity of water deficit. Under all water regimes examined, cv. Longdong showed the greatest WUE values and the least reduction in biomass production under a 50% soil field water capacity. Stomatal density increased with increasing water deficit, but stomatal conductance decreased. This suggests that water stress can increase WUE by modifying stomatal regulation of the balance between the rates of CO₂ assimilation and water loss. The maintenance of leaf physiological function and leaf water status suggests that alfalfa has some mechanisms to maintain cell function when the plant is subjected to water deficit. The increase in the MDA content under drought conditions indicates that some degree of damage to cell membranes is unavoidable, whereas other results showing increases in the contents of proline and soluble sugars and activities of superoxide dismutase, peroxide dismutase, and catalase indicate how cell function may be to some extent maintained to result in the higher WUE. Alfalfa is shown to exhibit cultivar-specific differences in WUE with the maintenance of cell function under water deficit being related to anti-oxidation and osmotic adjustment.     

Response of French bean cultivars to water deficits: Changes in endogenous hormones, proline and chlorophyll

Effects of water stress at different stages of plant growth on leaf relative water content (RWC), osmotic potential (Ψos) and changes in contents of chlorophyll, abscisic acid (ABA), zeatin riboside (t-ZR), ethylene and proline in six cultivars of French bean (Phaseolus vulgaris L.) were studied. Under water stress, Ψos and RWC were highest in cv. Contender and lowest in cvs. IIHR-909 and Sel-2. The increase in contents of ABA and proline was marked in cv. Contender followed by cv. UPF-626. Decrease in t-ZR and chlorophyll contents was prominent in cv. IIHR-909. Ethylene production surged in all the cultivars under 4- and 8-d stress and declined under 12-d stress. This revised version was published online in August 2006 with corrections to the Cover Date.     

Water deficit enhancement of proline and α-amino nitrogen accumulation in potato plants and its association with susceptibility to drought

Potato plants ( Solanum tuberosum L. cvs 'Up-to-Date', 'Desiree', 'Alpha', 'Spunta', 'Elvira' and 'Troubadour') were exposed to cycles of water stress and relief during growth. Severe water deficit induced increased proline content 6- to 7-fold in nonturgid leaves which just started to wilt, and 8- to 27-fold in fully wilted leaves of potatoes. However, proline content was not affected during the early stages of stress development over a range of osmotic potentials in the leaves. The rising proline content was related to turgor loss of leaves independent of changes in the osmotic potentials, which indicates that proline involvement in osmoregulation of potato leaves is unlikely.
Repeated cycles of water stress and relief resulted in increased proline and α-amino nitrogen content in the tuber tissue of some of the cultivars. The smallest increase in proline content was obtained in 'Alpha' tubers and the content of α-amino nitrogen remained unaffected by the water stress. Concomitantly, 'Alpha' was the most drought-tolerant cultivar, as determined by its capacity to accumulate dry matter in tubers under stress conditions. On the other hand, in tubers of cultivars which were more susceptible to drought, a marked increase in proline and α-amino nitrogen was observed in response to water stress. The possible association of these findings with tolerance of potatoes to repeated short periods of drought is discussed.     

Implication of ABA and proline on cell membrane injury of water deficit stressed barley seedlings

The aim of this work was to examine the ability of ABA and proline to counteract the deleterious effect of water deficit stress on cell membrane injuries. Six-day-old seedlings of two barley genotypes (cv. Aramir, line R567) were treated with ABA (2·10⁻⁴ M) or proline (0.1 M) for 24 h, and then subjected to osmotic stress for 24h, by immersing their roots in polyethylene glycol (PEG 6000) solution of osmotic potential of −1.0 MPa and −1.5 MPa or by submerging the leaf pieces in PEG solution of osmotic potential of −1.6 MPa. Pretreatment of plants with ABA and proline caused an increase of free proline level in the leaves. Plants treated with ABA exhibited a lower membrane injury index under water stress conditions than those untreated even when no effect of this hormone on RWC in the leaves of stressed plants was observed. Pretreatment of plants with proline prevented to some extent membrane damage in leaves of the stressed seedlings, but only in the case when stress was imposed to roots. Improvement in water status of leaves was also observed in seedlings pretreatment with proline. The protective effect of both ABA and proline was more pronounced in line R567 that exhibited higher membrane injury under water deficit stress conditions.     

Contrasting physiological responses of six eucalyptus species to water deficit

BACKGROUND AND AIMS: The genus Eucalyptus occupies a broad ecological range, forming the dominant canopy in many Australian ecosystems. Many Eucalyptus species are renowned for tolerance to aridity, yet inter-specific variation in physiological traits, particularly water relations parameters, contributing to this tolerance is weakly characterized only in a limited taxonomic range. The study tests the hypothesis that differences in the distribution of Eucalyptus species is related to cellular water relations. METHODS: Six eucalypt species originating from (1) contrasting environments for aridity and (2) diverse taxonomic groups were grown in pots and subjected to the effects of water deficit over a 10-week period. Water potential, relative water content and osmotic parameters were analysed by using pressure-volume curves and related to gas exchange, photosynthesis and biomass. KEY RESULTS: The six eucalypt species differed in response to water deficit. Most significantly, species from high rainfall environments (E. obliqua, E. rubida) and the phreatophyte (E. camaldulensis) had lower osmotic potential under water deficit via accumulation of cellular osmotica (osmotic adjustment). In contrast, species from low rainfall environments (E. cladocalyx, E. polyanthemos and E. tricarpa) had lower osmotic potential through a combination of both constitutive solutes and osmotic adjustment, combined with reductions in leaf water content. CONCLUSIONS: It is demonstrated that osmotic adjustment is a common response to water deficit in six eucalypt species. In addition, significant inter-specific variation in osmotic potential correlates with species distribution in environments where water is scarce. This provides a physiological explanation for aridity tolerance and emphasizes the need to identify osmolytes that accumulate under stress in the genus Eucalyptus.     

Effect of gibberellic acid on epicotyl growth and carbohydrate distribution in derooted Pisum sativum cuttings with or without cotyledons

The effect of gibberellic acid (GA) on subhook growth in derooted cuttings of pea ( Pisum sativum L. cv. Alaska) grown in the dark was studied in relation to the distribution of sugar-related compounds in the epicotyl and cotyledons. GA stimulated subhook growth of cuttings with or without cotyledons. In cuttings with cotyledons, the net inflow of sugar-related compounds (soluble sugars, starch, cell wall polysaccharides and sugars consumed by respiration) to the epicoiyl balanced with the net outflow from the cotyledons. GA stimulated the net inflow of sugar-related compounds to the epicotyl and the net outflow from cotyledons. Among these compounds, GA substantially increased the amount of soluble sugars, starch and cell wall polysaccharides in the subhook. In cuttings without cotyledons, on the other hand, the net inflow of sugar-related compounds to the subhook almost balanced with the net outflow from the epicotyl below the subhook. GA stimulated the net inflow of sugar-related compounds to the subhook and the net outflow from the epicotyl below the subhook. Among these compounds, GA substantially increased the amount of soluble sugars and cell wall polysaccharides in the subhook. These results suggest that GA stimulates an increase in the net inflow of sugar-related compounds to the subhook, thereby preventing an increase in osmotic potential and stimulating cell wall polysaccharide synthesis, when pea subhook growth is stimulated.     

Nonrecirculating hydroponic system suitable for uptake studies at very low nutrient concentrations

Seedlings of maize (Zea mays L. cv WF9 × Mo 17) growing at low water potentials in vermiculite contained greatly increased proline concentrations in the primary root growth zone. Proline levels were particularly high toward the apex, where elongation rates have been shown to be completely maintained over a wide range of water potentials. Proline concentration increased even in quite mild treatments and reached 120 millimolal in the apical millimeter of roots growing at a water potential of −1.6 megapascal. This accounted for almost half of the osmotic adjustment in this region. Increases in concentration of other amino acids and glycinebetaine were comparatively small. We have assessed the relative contributions of increased rates of proline deposition and decreased tissue volume expansion to the increases in proline concentration. Proline content profiles were combined with published growth velocity distributions to calculate net proline deposition rate profiles using the continuity equation. At low water potential, proline deposition per unit length increased by up to 10-fold in the apical region of the growth zone compared to roots at high water potential. This response accounted for most of the increase in proline concentration in this region. The results suggest that osmotic adjustment due to increased proline deposition plays an important role in the maintenance of root elongation at low water potentials.     

Changes at panicle emergence in the water relations of a wetland and a dryland Japonica rice cultivar under wetland conditions

The diurnal and seasonal changes in plant water relations of two Japonica rice ( Oryza sativa L.) cultivars, Nipponbare and Tachiminori, were studied under flooded conditions at Kyoto University. The dryland cv. Tachiminori maintained higher predawn and midday leaf osmotic potentials relative to the wetland cv. Nipponbare during the vegetative stage, but the ranking was reversed after flowering. The relationship between leaf water potential and leaf osmotic potential showed that prior to panicle emergence Nipponbare was able to adjust osmotically to maintain turgor, whereas after heading there was little turgor maintenance. Tachiminori showed little difference in osmotic adjustment before and after panicle emergence. Fertilizer treatment during panicle development also helped to maintain the degree of osmotic adjustment in both cultivars.     

Some physiological and growth responses of kiwi fruit (Actinidia chinensis) to flooding

The effects of long-term flooding on the growth of six-month-old Actinidia chinensis Planch cv. Abbot plants and some effects on stomatal behaviour and leaf water relations were examined under controlled conditions for 28 days. Flooding caused stomatal closure and decreases in transpiration rate, xylem water potential, osmotic potential and turgor potential. Flooding also caused inhibition of the dry weight increase of leaves plus stems and of roots, chlorosis and necrosis of leaves, production of hypertrophied lenticels and the appearance of a small number of adventitious roots on the submerged portions of the stems. Rapid and partial stomatal closure by flooding may not only be due to the passive mechanical response which follows leaf dehydration, since flooded plants showed an increase in xylem water potential and osmotic potential during the first days of the experiment. The marked intolerance of Actinidia chinensis to flooding has been a serious barrier to its culture in poorly drained soils, hence careful irrigation management is required.     

Salt tolerance of soybean cultivars

Soybean (Glycine max) cv. Clark tolerated NaCl salinity up to osmotic potential in soil -1800 kPa, cv. Forest to -1500 kPa and cv. Kint to -1200 kPa. The tolerance of cvs. Clark and Forest was found to be related to the accumulation of soluble proteins, amino acids, proline, K and Ca. The sensitivity of cv. Kint was found to be associated with decrease in water, saccharide, protein, K and Ca contents. The leaf osmotic potential decreased in cvs. Clark and Forest while remained unchanged in cv. Kint. This revised version was published online in July 2006 with corrections to the Cover Date.     

The role of organic and inorganic solutes in the osmotic adjustment of drought-stressed Jatropha curcas plants

This study aimed to assess the accumulation of organic and inorganic solutes and their relative contribution to osmotic adjustment in roots and leaves of Jatropha curcas subjected to different water deficit intensity. Plants were grown in vermiculite 50% (control), 40%, 30%, 20% and 10% expressed in gravimetric water content. The water potential, osmotic potential and turgor potential of leaves decreased progressively in parallel to CO₂ photosynthetic assimilation, transpiration and stomatal conductance, as the water deficit increased. However, the relative water content, succulence and water content in the leaves did not show differences between the control and stressed plants, indicating osmotic adjustment associated with an efficient mechanisms to prevent water loss by transpiration through stomatal closure. The K⁺ ions had greater quantitative participation in the osmotic adjustment in both leaves and roots followed by Na⁺ and Cl⁻, while the NO₃⁻ ion only showed minor involvement. Of the organic solutes studied, the total soluble sugars showed the highest relative contribution to the osmotic adjustment in both organs and its concentration positively increased with more severe water deficit. The free amino acids and glycinebetaine also effectively contributed to the osmotic potential reduction of both the root and leaves. The role of proline was quantitatively insignificant in terms of osmotic adjustment, in both the control and stressed roots and leaves. Our data reveal that roots and leaves of J. curcas young plants display osmotic adjustment in response to drought stress linked with mechanisms to prevent water loss by transpiration by means of the participation of inorganic and organic solutes and stomatal closure. Of all the solutes studied, soluble sugars uniquely display a prominent drought-induced synthesis and/or accumulation in both roots and leaves.     

Growth characteristics and proline content in relation to water status in twoZea mays L. Cultivars during rehydration

Growth behaviour, proline changes and water saturation deficit (WSD) changes were studied in two maize cultivars, Agati-76, a drought resistant oultivar, and cv. Vijay, a susceptible one after stress conditions were removed. Plants of both the cultivars showed a tendency to recover during rehydration. Phenotypically the plants appeared just normal on 7th day of rehydration, although recovery was never complete. Significant positive correlation existed between free proline content and concomitant change in water saturation deficit. Significantly higher proline content in cv. Agati-76 was reoorded in cv. Vijay, immediately after the stress was released, thereafter continuous decline was observed up to 7th day of rehydration in both the cultivars. Proline changes in relation to recovery of plants from stress conditions are discussed.     

Effects of water stress on the water relations of Phaseolus vulgaris and the drought resistant Phaseolus acutifolius

The tepary bean ( Phaseolus acutifolius Gray var. latifolius ), a drought resistant species, was compared under water stress conditions with the more drought susceptible P. vulgaris L. cvs Pinto and White Half Runner (WHR). In order to better understand the basis for the superior drought resistance of tepary, this study was designed to determine the relationships among leaf water potential, osmotic potential, turgor potential, and relative water content (RWC).
Plants were prestressed by withholding irrigation water. These stress pretreatments changed the relation between leaf water potential and relative water content of both species so that prestressed plants had lower water potentials than controls at the same leaf RWC. Tepary had lower water potentials at given RWC levels than Pinto or WHR; this can account for part of the superior resistance of tepary. In all genotypes, prestressed plants maintained osmotic potentials approximately 0.2 MPa lower than controls. Tepary reached osmotic potentials that were significantly lower (0.15 to 0.25 MPa) than Pinto or WHR. Both control and prestressed tepary plants had 0.05 to 0.25 MPa more turgor than Pinto or WHR at RWC values between 65 and 80%. Both prestressed and control tepary plants had greater elasticity (a lower elastic modulus) than Pinto or WHR. This greater turgor of tepary at low RWC values could be caused by several factors including greater tissue elasticity, active accumulation of solutes, or greater solute concentration.
Tepary had significantly lower osmotic potentials than the P. vulgaris cultivars, but there was little difference in osmotic potential between Pinto and WHR. Knowledge of differences in osmotic and turgor potentials among and within species could be useful in breeding for drought resistance in Phaseolus.     

Physiological responses to water stress following a conditioning period in berseem clover

Berseem clover (Trifolium alexandrinum L.) is an important crop in semi-arid regions; its herbage and seed yields are often reduced by water stress. Our objectives were (i) to determine the effect of water stress, applied after a conditioning period, on water relations, proline accumulation and plant dry weight, and (ii) to investigate if some physiological responses differed in varieties of berseem. Five cultivars (Axi, Bigbee, Lilibeo, Sacromonte and Saniros) were grown in a controlled environment, and subjected to four irrigation treatments (T1, T2, T3 and T4 referring to plants irrigated to field capacity every 1, 2, 3 or 4 d, respectively) during a conditioning period (12 d). T1 treatment indicated the well-watered control, whereas T2, T3 and T4 treatments represented the conditioned plants. Leaf water potential (Ψ), osmotic potential (Ψπ), relative water content (RWC), gravimetric soil water content (GSWC) and leaf proline concentration were recorded during the conditioning period and a subsequent water deficit period (3 d) applied at early flowering growth stage. The conditioned plants subjected to subsequent water deficit maintained higher values of Ψ, Ψπ, RWC and GSWC, and lower values of leaf proline concentration. Reductions in parameter values were inversely related to the water stress severity that plants had previously experienced. At the end of the experiment, T1 showed 42%, 58% and 31% lower values for Ψ, Ψπ and RWC, respectively, than those of T4. Conditioned plants were also shorter and accumulated less leaf, stem and total dry weight. The conditioning treatments did not affect the relation between Ψ and Ψπ since conditioned plants show similar values of Ψπ as the control at the same Ψ value. Thus, drought acclimation in berseem clover contributed to water stress tolerance by the maintenance of tissue hydration. The berseem cultivars examined showed differences in plant growth parameters, but they were very similar for physiological responses to water deficit. The main genetic difference was recorded for turgor maintenance capacity. This revised version was published online in June 2006 with corrections to the Cover Date.     

Roles of leaf water potential and soil-to-leaf hydraulic conductance in water use by understorey woody plants

Diurnal changes of leaf water potential and stomatal conductance were measured for 12 deciduous shrubs and tree saplings in the understorey of a temperate forest. Sunflecks raised the leaf temperature by 4°C, and vapor pressure deficit to 2 kPa. Although the duration of the sunflecks was only 17% of daytime, the photon flux density (PFD) of sunflecks was 52% of total PFD on a sunny summer day. Leaf osmotic potential at full turgor decreased in summer, except in some species that have low osmotic potential in the spring. Plants that endured low leaf water potential had rigid cell walls and low osmotic potential at full turgor. These plants did not have lower relative water content and turgor potential than plants with higher leaf water potential. There were three different responses to an increase in transpiration rate: (i) plants had low leaf water potential and slightly increased soil-to-leaf hydraulic conductance; (ii) plants decreased leaf water potential and increased the hydraulic conductance; and (iii) plants had high leaf water potential and largely increased the hydraulic conductance.