Effects of mucilage on seed germination of the desert ephemeral plant Plantago minuta Pall. under osmotic stress and cycles of wet and dry conditions

To test the role of the seed mucilage of Plantago minuta Pall. in regulating germination under osmotic stress and cycles of hydration and dehydration, two experiments were carried out using seeds with intact mucilage and mucilage‐free seeds. In Experiment 1 seeds were immersed in a range of iso‐osmotic polyethylene glycol solutions (?1.15 to 0 MPa) for 14 days; any ungerminated seeds were transferred to deionized water to investigate the recovery germination. In Experiment 2 seeds were immersed in deionized water for 24 h, and were then incubated on filter paper for an additional 13 days to ensure complete desiccation before reimbibition to test the germination recovery percentage. Under mild osmotic stress (?0.73 to 0 MPa), the intact seeds with mucilage were shown to have higher germination rates than the mucilage‐free seeds, indicating that the mucilage led to a “fast sprouting” germination strategy under mild osmotic stress. However, when seeds were exposed to high osmotic stress (?1.15 MPa), the mucilage apparently slowed the germination rate, resulting in a “risk‐balancing” germination strategy. Extreme drought induced by polyethylene glycol solution and the desiccation pretreatment accelerated germination rates compared to non‐pretreated seeds; both germination potential and recovery percentage of the mucilage seeds were significantly higher than that of the mucilage‐free seeds. Our results revealed that the seed mucilage of P. minuta plays a crucial role in regulating seed germination rates and the germination strategies adopted by controlling seed water absorption when the seeds experience different osmotic stresses or alternating wet and dry conditions.     

Probing short-term root exudation in Zea mays

Exudation of maize roots was studied using a microdrop recorder. The high-resolution measurements of relatively short-term changes in exudation seems to be one of the most useful and unproblematic applications of the microdrop recorder. When mannitol, polyethylene glycol (PEG) and kinetin were supplied to the medium bathing, the surfaces of excised maize roots, a marked decrease in root exudation was observed. The action of fusicoccin and that of abscisic acid (ABA) showed a sharp and then a slower decline on root exudation, though, enhanced exudation was sustained over a much longer period, in comparison to that recorded for mannitol and polyethylene glycol. A decline in the volume of exudates is related to an increase in the water deficit, in coincidence to changes in the osmotic gradient between root cells and the bathing medium generated by expelling exudates.     

INFLUENCE OF MANNITOL ON ABSORPTION AND RETENTION OF RUBIDIUM BY EXCISED CORN ROOTS

Excised root segments of corn were subjected to osmotic stress by immersion in solutions of mannitol before, during, or after a period of Rb absorption. Both the time course of uptake (or loss) and selectivity of uptake were studied. Stress before or during the absorption period reduced Rb absorption to 20 % or less of that of controls, whereas it had no detectable influence on the constancy and selectivity of uptake. Stress imposed following a period of Rb accumulation caused root segments to lose only slightly more Rb than controls during the first 30 min, after which rates of change were insignificant in both. Segments stressed after the Rbabsorption period retained from 70–90 % as much Rb as did the controls, even after two hours. The comparatively great quantitative difference between effect of stress on uptake and its effect on loss is interpreted to mean that the mechanism of stress-induced reduction of ion absorption cannot be adequately explained on the sole basis of increased efflux of ions.     

Water stress and indol-3yl-acetic acid content of maize roots

Water-stress conditions were applied to the apical 12 mm of intact or excised roots ofZea mays L. (cv. LG 11) using mannitol solutions (0 to 0.66 M) and changes in weight, water content, growth and IAA level of these roots were investigated. With increasing stress a decrease in growth, correlated with an increased IAA level, was observed. The largest increase in IAA (about 2.7-fold) was found in the apical 5 mm of the root and was obtained under a stress corresponding to an osmotic potential of −1.39 MPa in the solution. This stress led to an isotonic state in the cells after 1 h. When the duration of water stress (−1.09 MPa) was increased to 2 or 3 h, no further increase in the IAA content was observed in the root segments. This indicated that there was no correlation between a hypothetical passive penetration of mannitol in the cells and IAA content. Indol-3yl-acetic acid rose to the same level in excised as in intact roots. In both cases, IAA accumulation was apparently independent of the hydrolysis of the conjugated form. The caryopsis and shoot seem not to be necessary to induce the increase of the IAA level in the roots during water stress (−1.09 MPa). Therefore, there seems to be a high rate of IAA biosynthesis in excised maize roots under water-stress conditions. Exodiffusion of IAA was observed during an immersion in either buffer or stress (−1.09 MPa) solution. In both cases, this IAA efflux into the medium represented about 50% of the endogenous level. Considering the present results, IAA appears to play an important part in the regulation of maize root metabolism and growth under water deficiency.     

Effects of rapidly and slowly permeating osmotica on metabolism

Zea mays was exposed to solutions of low water potentials by addition of ethylene glycol or mannitol. Intact seedlings were treated for 1 hr at potentials between −10 and −20 atmospheres and then returned to high water potentials. Subsequent root extension was slow after mannitol treatment, but rapid when ethylene glycol had been used as the osmoticum. Cellular activity of excised roots was also affected much less by ethylene glycol than by mannitol. Processes studied included respiration, glucose uptake, and synthesis of methanol-insoluble compounds. These differences in response to various osmotica applied both during and after treatment at low water potentials.     

Enhanced Maturation and Desiccation Tolerance of White Spruce [Picea glauca (Moench) Voss] Somatic Embryos: Effects of a Non-plasmolysing Water Stress and Abscisic Acid

A non-plasmolysing moisture stress effected by polyethyleneglycol (PEG) was beneficial when applied to maturing white spruce(Picea glauca) somatic embryos for the following reasons. Anosmotic treatment of 5.0–7.5% PEG stimulated a threefoldincrease in the maturation frequency. The osmotically treatedsomatic embryos displayed higher dry weights and lower moisturecontents than the controls, indicating a greater accumulationof storage reserves. Moisture contents of mature, osmotically-treated,hydrated somatic embryos were 40–45%, in contrast to 57%for the non-osmotically treated controls. Desiccation was achievedby placing the somatic embryos in a range of relative-humidityenvironments. No clear trend for the effect of PEG on survivalof desiccated somatic embryos was observed; mean survival valuesranged from 34 to 62% when somatic embryos from all osmotictreatments were desiccated for 14 d at 81% relative humidity.Following this desiccation treatment, somatic embryos from allosmotic concentrations had moisture contents of 26–31%,similar to the 32% recorded for unimbibed zygotic embryos. Afterimbibition, moisture contents for these zygotic and somaticembryos were in the order of 60%. Somatic embryos matured withPEG remained quiescent during desiccation due to their low initialmoisture contents, and gave rise to plantlets of normal appearance.Gradual desiccation of the somatic embryos directly followingmaturation with abscisic acid (ABA) was crucial to survivalduring desiccation. A plasmolysing water stress effected bysucrose at osmotic potentials similar to PEG was detrimentalto somatic embryo maturation, thereby emphasizing the importanceof the choice of osmoticum. Desiccation, maturation, osmotic potential, Picea glauca, polyethylene glycol, somatic embryo, water stress, white spruce     

Measurement of free space and sorption of large molecules by cereal roots

Abstract. Large molecular weight solutes that do not penetrate the root have been used to correct for the surface film in measurements with mannitol of the volume of the Apparent Free Space (FS) in bailey roots. The results are compared with those obtained using other correction techniques for elimination of the surface film. Large molecules seem to be adsorbed on the root surface and the kinetics of adsorption differ between the polyhydric alcohol mannitol or the polysaccharide dextran on the one hand, and the polyether polyethylene glycol (PEG-4000) on the other. The significance of this difference in kinetics is discussed in relation to the use of PEG as an osmoticum in studies on root water relations and its effect on ion uptake. Although smaller molecular weight PEG's penetrate the FS and diminish sodium uptake from 10 mol m⁻³ NaCl, more dilute solutions of mannitol and larger PEG polymers are unlikely to affect ion uptake from dilute nutrient solutions. Use of these substances along with labelled nutrients in kinetic studies of the compartmentation of ions in roots can help to distinguish between ions associated with the surface film, those in the FS and those that have crossed the cell membranes into the protoplast.     

Metabolic engineering using <Emphasis Type="Italic">mtlD</Emphasis> gene enhances tolerance to water deficit and salinity in sorghum

Sorghum bicolor L. Moench cv. SPV462 was transformed with the mtlD gene encoding for mannitol-1-phosphate dehydrogenase from E. coli with an aim to enhance tolerance to water deficit and NaCl stress. Transgene (pCAM mtlD) integration and expression were successfully confirmed by PCR, Southern, RT-PCR and Western analysis. Segregation analysis based on germination of T0 seed on hygromycin-supplemented medium revealed an expected Mendelian ratio 3:1 in lines 5, 72 and 75. Retention of leaf water content was remarkably higher in transgenic leaf segments when exposed to polyethylene glycol 8000 (−2.0 MPa), as compared to the untransformed controls. Another significant finding is that the transgenics maintained a 1.7 to 2.8 fold higher shoot and root growth, respectively, under NaCl stress (200 mM) when compared to untransformed controls. These results demonstrate that engineering mannitol biosynthetic pathway into sorghum can impart enhanced tolerance to water deficit and salinity.     

Inhibition of phosphorus and water passage across intact roots by polyethylene glycol and phenylmercuric acetate

Application of polyethylene glycol or phenylmercuric acetate to intact bean (Phaseolus vulgaris L., cv. Red Wade) roots inhibited passage of phosphorus across the roots to the xylem. The same results occurred for foliar application of phenylmercuric acetate when time was allowed for absorption and distribution of the chemical in the plant. For both chemicals the inhibition of phosphorus was proportional to or greater than any accompanying restriction on water flow across the root.     

Polyamine concentrations and arginine decarboxylase activity in wheat exposed to osmotic stress

The activity of L-arginine decarboxylase (ADC: EC 4.1.1.19)and polyamine content were examine in intact wheat plants ( Triticum aestivum L. cv. Sappo) exposed to osmotic stress (0.4 M mannitol) for 5 days. ADC activity was increased in first and second leaves and in roots of mannitol-stressed plants. Concentrations of putrescine, cadaverine and spermine were generally increased in leaves and roots of plants exposed to mannitol, whereas spermidine was reduced in first leaves and roots of these plants. In an attempt to determine the localization of mannitol in stressed wheat. ¹⁴C-mannitol was fed to plants grown in liquid culture. Most of the mannitol was detected in roots (84%), while small amounts were found in first (9%) and second (7%) leves.
Since it seemed possible that some of the effects on polyamine metabolism caused by exposure to mannitol could have been the result of water stress. polyamine metabolism was also studied in plants water stressed by exposure to 2% polyethylene glycol (PEG) 4000. ADC activity was not altered by exposure to PEG. but concentrations of putrescine, spermidine and spermine were generally reduced in leaves and roots of stressed plants. Cadaverine concentrations were not significantly affected by exposure to PEG. Spermidine and spermine concentrations were reduced in first and second leaves but remained unchanged in roots of plants exposed to PEG.     

The increasing desiccation sensitivity of recalcitrant Avicennia marina seeds with storage time

To test the hypothesis that desiccation sensitivity increases with storage time, recalcitrant seeds of Avicennia marina (Forssk.) Vierh. were dehydrated by soaking in polyethylene glycol solutions after inreasing periods of storage. Germination characteristics and the ultrastructure of root primorida were assessed before and after dehydration. Short-term storage enhanced the apparent rate of germination, consistent with the hypothesis that these seeds commence germination in storage. Root primordia of stored seeds initially showed enhanced subcellular activity, including cell division and vacuolation. Increased storage time resulted in the onset and progression of deleterious changes.
Newly shed seeds and seeds stored up to the stage of cell division were comparatively resistant to desiccation. As storage time increased, subsequent dehydration caused increasing subcellular damange and consequent reduction in the rates of germination relative to non-dehydrated controls. Ultrastructural results suggest that after the initiation of cell division of seeds in storage, there is a requirement for additional water for the germination process to continue. A model for the behaviour of recalcitrant seeds is proposed.     

Tolerance of mannitol-accumulating transgenic wheat to water stress and salinity

Previous work with model transgenic plants has demonstrated that cellular accumulation of mannitol can alleviate abiotic stress. Here, we show that ectopic expression of the mtlD gene for the biosynthesis of mannitol in wheat improves tolerance to water stress and salinity. Wheat (Triticum aestivum L. cv Bobwhite) was transformed with the mtlD gene of Escherichia coli. Tolerance to water stress and salinity was evaluated using calli and T(2) plants transformed with (+mtlD) or without (-mtlD) mtlD. Calli were exposed to -1.0 MPa of polyethylene glycol 8,000 or 100 mM NaCl. T(2) plants were stressed by withholding water or by adding 150 mM NaCl to the nutrient medium. Fresh weight of -mtlD calli was reduced by 40% in the presence of polyethylene glycol and 37% under NaCl stress. Growth of +mtlD calli was not affected by stress. In -mtlD plants, fresh weight, dry weight, plant height, and flag leaf length were reduced by 70%, 56%, 40%, and 45% compared with 40%, 8%, 18%, and 29%, respectively, in +mtlD plants. Salt stress reduced shoot fresh weight, dry weight, plant height, and flag leaf length by 77%, 73%, 25%, and 36% in -mtlD plants, respectively, compared with 50%, 30%, 12%, and 20% in +mtlD plants. However, the amount of mannitol accumulated in the callus and mature fifth leaf (1.7-3.7 micromol g(-1) fresh weight in the callus and 0.6-2.0 micromol g(-1) fresh weight in the leaf) was too small to protect against stress through osmotic adjustment. We conclude that the improved growth performance of mannitol-accumulating calli and mature leaves was due to other stress-protective functions of mannitol, although this study cannot rule out possible osmotic effects in growing regions of the plant.     

Evaluation of water stress control with polyethylene glycols by analysis of guttation

The water relations of pepper plants (Capsicum frutescens L.) under conditions conducive to guttation were studied to evaluate the control of plant water stress with polyethylene glycols. The addition of polyethylene glycol 6000 to the nutrient solution resulted in water relations similar to those expected in soil at the same water potentials. Specifically, xylem pressure potential in the root and leaf became more negative during a 24-hour treatment period, while osmotic potential of the root xylem sap remained constant. The decrease in pressure potential was closely correlated with the decrease in osmotic potential of the nutrient solution. In contrast, the addition of polyethylene glycol 400 to the nutrient medium resulted in a reduction of osmotic potential in the root xylem sap; this osmotic adjustment in the xylem was large enough to establish an osmotic gradient for entry of water and cause guttation at a nutrient solution osmotic potential of −4.8 bars. Pressure potential in the root and leaf xylem became negative only at nutrient solution osmotic potentials lower than −4.8 bars. About half of the xylem osmotic adjustment in the presence of polyethylene glycol 400 was caused by increased accumulation of K⁺, Na⁺, Ca²⁺, and Mg²⁺ in the root xylem. These studies indicate that larger polyethylene glycol molecules such as polyethylene glycol 6000 are more useful for simulating soil water stress than smaller molecules such as polyethylene glycol 400.     

The Effects of Cryoprotective Substances on the Mechanical Stability and Geometric Parameters of Human Erythrocytes

The aim of this study was to examine the effects of sucrose, dextran, polyethylene glycol, glycerol, and mannitol, which possess cryoprotective properties to various degrees, on the mechanical stability and the geometric parameters of human erythrocytes. All substances, except mannitol, contributed to a decrease in hemolysis, which was caused by the movement of small beads. Glycerol and polyethylene glycol, which provide the highest level of protection of erythrocytes during cryopreservation, also showed the maximum efficiency under mechanical stress. Changes in cell resistance may be associated with the transformation of their geometric parameters. According to the cytometry data, 5% solutions of all the substances, except mannitol, caused similar changes in the geometric parameters of the cell. The relationship between changes in the mechanical stability and the geometric parameters of erythrocytes under the influence of cryoprotective agents may be caused by the modification of the membrane?cytoskeleton protein complex, which controls the mechanoelastic properties and morphology of erythrocytes.     

若干种子处理方法对偃松种子生活力的影响

为了提高偃松种子生活力,采用水浸、NaOH溶液浸泡、PEG6000（PEG,聚乙二醇）溶液浸泡、低温冷冻、吸湿/回干5种方法对偃松种子进行处理,研究不同处理中各因素（如溶液浓度、处理时间与温度）对偃松种子生活力的影响,并采用靛蓝染色法测定偃松种子的生活力。结果表明：10%PEG6000溶液、初始温度60℃、浸泡36 h时,偃松种子生活力最强;水浸、低温处理和吸湿/回干处理方法也能提高偃松种子生活力;然而4%~12%NaOH溶液浸泡处理会导致偃松种子生活力下降。因此可将PEG6000溶液处理种子的方法应用于偃松种子萌发及人工种苗培育的实践中,使偃松资源得到更好的可持续开发与利用。     

Oscillatory Transpiration and Water Uptake of Avena Plants

The oscillatory transpiration of 6 days old Avena plants was investigated with respect to the water potential of the root medium. The desired water potential was obtained by means of mannitol solutions. When the water potential was lowered (“mannitol step”), the amplitude of the oscillations decreased. Below –3.0 bars no oscillations persisted. A detailed study was made of the phase changes of the oscillations caused by a short time decrease of the water potential of the root medium (“mannitol pulse”). The duration of these short term treatments was either 9.0, 3.0 or J.5 min. The experimental results are discussed on the basis of an electric analogue previously presented in the literature. Published simulations based on the model were in clear contrast to the present experimental results as well as to earlier results in the literature. However, simulations in the present paper showed that the model could explain the experimental results if suitable parameter values were chosen.     

Differential Sensitivity of Macrocarpa and Microcarpa Types of Chickpea （Cicer arietinum L.） to Water Stress： Association of Contrasting Stress Response with Oxidative Injury

Chickpea (Cicer arietinum L.) is particularly sensitive to water stress at its reproductive phase and, under conditions of water stress, will abort flowers and pods, thus reducing yield potential. There are two types of chickpea: (i) Macrocarpa (“Kabuli”), which has large, rams head‐shaped, light brown seeds; and (ii) Microcarpa (“Desi”), which has small, angular and dark‐brown seeds. Relatively speaking, “Kabuli” has been reported to be more sensitive to water stress than “Desi”. The underlying mechanisms associated with contrasting sensitivity to water stress at the metabolic level are not well understood. We hypothesized that one of the reasons for contrasting water stress sensitivity in the two types of chickpea may be a variation in oxidative injury. In the present study, plants of both types were water stressed at the reproductive stage for 14 d. As a result of the stress, the “Kabuli” type exhibited an 80% reduction in seed yield over control compared with a 64% reduction observed for the “Desi” type. The decrease in leaf water potential (Ψ_w) was faster in the “Kabuli” compared with the “Desi” type. At the end of the water stress period, Ψ_w was reduced to ?2.9 and ?3.1 MPa in the “Desi” and “Kabuli” types, respectively, without any significant difference between them. On the last day of stress, “Kabuli” experienced 20% more membrane injury than “Desi”. The chlorophyll content and photosynthetic rate were significantly greater in “Desi” compared with “Kabuli”. The malondialdehyde and H₂O₂ content were markedly higher at the end of the water stress in “Kabuli” compared with “Desi”, indicating greater oxidative stress in the former. Levels of anti‐oxidants, such as ascorbic acid and glutathione, were significantly higher in “Desi” than “Kabuli”. Superoxide dismutase and catalase activity did not differ significantly between the two types of chickpea, whereas on the 10th day, the activities of ascorbate peroxidase, dehydroascorbate reductase, and glutathione reductase were higher in “Desi”. These findings indicate that the greater stress tolerance in the “Desi” type may be ascribed to its superior ability to maintain better water status, which results in less oxidative damage. In addition, laboratory studies conducted by subjecting both types of chickpea to similar levels of polyethylene glycol‐induced water stress and to 10 μ.mol/L abscisic acid indicated a greater capacity of the “Desi” type to deal with oxidative stress than the “Kabuli” type. (Managing editor: Ping He)     

Osmotic Stress in Coleoptiles and Primary Leaves of Wheat: I. EFFECTS ON SIZE, WEIGHT, TOTAL PROTEIN, DNA, AND PHOSPHORUS

Seedlings of wheat (Triticum durum, cv. Balcarceño-INTA)were water-stressed in darkness with 20% polyethylene glycol(PEG) 6000 or 0.3 M mannitol added to the root medium. At differenttimes and up to a total of 36 h of treatment the coleoptileand primary leaves were cut and analysed. The height and freshweight of shoots were lower in treated plants than in controlplants. Dry weight was not significantly different between controland water-stressed plants. Total protein concentration decreasedsignificantly (P < 0.01) after 36 h of PEG 6000 treatment.Total DNA concentration decreased in controls but not significantly(P < 0.025) in treated seedlings. This result was interpretedas indicating that cell elongation prevailed over cell divisionin controls and that cell enlargement was affected in stressedplants. Total phosphorus concentration fell in control and treatedseedlings. However, phosphorus specific radioactivity increasedby 116% in control plants, 93% in mannitol-treated plants, and22% in PEG 6000-treated seedlings. These data suggest that anearly metabolic effect of water stress may be on phosphorusturnover in shoots.     

Stress-induced accumulation of wheat germ agglutinin and abscisic Acid in roots of wheat seedlings

Wheat germ agglutinin (WGA) levels in roots of 2-day-old wheat seedlings increased up to three-fold when stressed by air-drying. Similar results were obtained when seedling roots were incubated either in 0.5 molar mannitol or 180 grams per liter polyethylene glycol 6000, with a peak level of WGA after 5 hours of stress. Longer periods of osmotic treatment resulted in a gradual decline of WGA in the roots. Since excised wheat roots incorporate more [³⁵S]cysteine into WGA under stress conditions, the observed increase of lectin levels is due to de novo synthesis. Measurement of abscisic acid (ABA) levels in roots of control and stressed seedlings indicated a 10-fold increase upon air-drying. Similarly, a five- and seven-fold increase of ABA content of seedling roots was found after 2 hours of osmotic stress by polyethylene glycol 6000 and mannitol, respectively. Finally, the stress-induced increase of WGA in wheat roots could be inhibited by growing seedlings in the presence of fluridone, an inhibitor of ABA synthesis. These results indicate that roots of water-stressed wheat seedlings (a) contain more WGA as a result of an increased de novo synthesis of this lectin, and (b) exhibit higher ABA levels. The stress-induced increase of lectin accumulation seems to be under control of ABA.     

Hardening of root cell walls: a growth inhibitory response to salinity stress

Primary roots of intact maize plants (Zea mays L.) grown for several days in nutrient solutions containing 100 mol m⁻³ NaCl and additional calcium, had relatively inhibited rates of elongation. Possible physical restraints underlying this salt induced inhibition were investigated. The inhibition did not involve reductions in osmotic potential gradients and turgor in the tip tissues responsible for root elongation growth. The apparent yield threshold pressure, which is related to capacity of cell walls to undergo loosening by stress relaxation, was estimated psychrometrically in excised root tips. Salinity increased yield threshold values. Comparative root extensibility values were obtained for intact plants by determining the initial (1 min) increase in root elongation rate induced by an 0.1 MPa osmotic jump. Comparative extensibility was significantly reduced in the salinized root tips. Salinity did not reduce capacities for water efflux and associated elastic contraction in root tip tissues of intact plants exposed to hypertonic mannitol. We conclude that cell wall hardening in the elongating root tips is an important component of root growth inhibition induced by long-term salinization.