Endosperm cell division in maize kernels cultured at three levels of water potential

The influence of osmoticum treatments on early kernel development of maize (Zea mays L.) was studied using an in vitro culture method. Kernels with subtending cob sections were placed in culture at 5 days after pollination. Sucrose (0.29, 0.44, or 0.58 molar) and sorbitol (0, 0.15, or 0.29 molar) were used to obtain six media with water potentials of −1.1, −1.6, or −2.0 megapascals. Kernel water potential declined in correspondence with the water potential of the medium; however, fresh weight growth was not significantly inhibited from 5 to 12 days after pollination. In stress treatments with media water potentials of −1.6 or −2.0 megapascals, endosperm tissue accumulated water and solutes from 10 and 12 days after pollination at a rate similar to or greater than that of the control (−1.1 megapascals). In contrast, endosperm cell division was inhibited in all treatments relative to control. At 10 days after pollination, endosperm sucrose concentration was greater in two of the −2.0 megapascal treatments with 0.44 or 0.58 molar media sucrose compared to control kernels cultured in 0.29 molar sucrose at −1.1 megapascals. Significant increases in abscisic acid content per gram of fresh weight were detected in two −2.0 megapascal treatments (0.29 molar sucrose plus 0.29 molar sorbitol and 0.58 molar sucrose) at 10 days after pollination. We conclude that in cultured maize kernels, endosperm cell division was more responsive than fresh weight accumulation to low water potential treatments. Data were consistent with mechanisms involving abscisic acid or lowered tissue water potential, or an interaction of the two factors.     

Water Deficit and Associated Changes in Some Photosynthetic Parameters in Leaves of ;Valencia' Orange (Citrus sinensis [L.] Osbeck)

Photosynthetic CO₂ assimilation, transpiration, ribulose-1,5-bisphosphate carboxylase (RuBPCase), and soluble protein were reduced in leaves of water-deficit (stress) `Valencia' orange (Citrus sinensis [L.] Osbeck). Maximum photosynthetic CO₂ assimilation and transpiration, which occurred before midday for both control and stressed plants, was 58 and 40%, respectively, for the stress (−2.0 megapascals leaf water potential) as compared to the control (−0.6 megapascals leaf water potential). As water deficit became more severe in the afternoon, with water potential of −3.1 megapascals for the stressed leaves vs. −1.1 megapascals for control leaves, stressed-leaf transpiration declined and photosynthetic CO₂ assimilation rapidly dropped to zero. Water deficit decreased both activation and total activity of RuBPCase. Activation of the enzyme was about 62% (of fully activated enzyme in vitro) for the stress, compared to 80% for the control. Water deficit reduced RuBPCase initial activity by 40% and HCO₃⁻/Mg²⁺-saturated activity by 22%. However, RuBPCase for both stressed and control leaves were similar in K_cat (25 moles CO₂ per mole enzyme per second) and K_m for CO₂ (18.9 micromolar). Concentrations of RuBPCase and soluble protein of stressed leaves averaged 80 and 85%, respectively, of control leaves. Thus, reductions in activation and concentration of RuBPCase in Valencia orange leaves contributed to reductions in enzyme activities during water-deficit periods. Declines in leaf photosynthesis, soluble protein, and RuBPCase activation and concentration due to water deficit were, however, recoverable at 5 days after rewatering.     

Water Transfer in an Alfalfa/Maize Association : Survival of Maize during Drought

We investigated the possibility of interspecific water transfer in an alfalfa (Medicago sativa L.) and maize (Zea mays L.) association. An alfalfa plant was grown through two vertically stacked plastic tubes. A 5 centimeter air gap between tubes was bridged by alfalfa roots. Five-week old maize plants with roots confined to the top tube were not watered, while associated alfalfa roots had free access to water in the bottom tube (the −/+ treatment). Additional treatments included: top and bottom tubes watered (+/+), top and bottom tubes droughted (−/−), and top tube droughted after removal of alfalfa root bridges and routine removal of alfalfa tillers (−^*). Predawn leaf water potential of maize in the −/+ treatment fell to −1.5 megapascals 13 days after the start of drought; thereafter, predawn and midday potentials were maintained near −1.9 megapascals. Leaf water potentials of maize in the −/− and −^* treatments declined steadily; all plants in these treatments were completely desiccated before day 50. High levels of tritium activity were detected in water extracted from both alfalfa and maize leaves after ³H₂O was injected into the bottom −/+ tube at day 70 or later. Maize in the −/+ treatment was able to survive an otherwise lethal period of drought by utilizing water lost by alfalfa roots.     

The effect of sodium chloride on solute potential and proline accumulation in soybean leaves

Two cultivars of soybean (Glycine max [L.] Merr.) were grown in solution with up to 100 millimolar NaCl. Leaf solute potential was −1.1 to −1.2 megapascals in both cultivars without NaCl. At 100 millimolar NaCl leaf solute potential was −3.1 to −3.5 megapascals in Bragg and −1.7 megapascals in Ransom. The decrease in solute potential was essentially proportional to the concentration of NaCl. In both salt susceptible Bragg and salt semitolerant Ransom, leaf proline was no more than 0.4 micromole per gram fresh weight at or below 20 millimolar NaCl. At 40 and 60 millimolar NaCl, Bragg leaf proline levels were near 1.2 and 1.9 micromoles per gram fresh weight, respectively. Proline did not exceed 0.5 micromole per gram fresh weight in Ransom even at 100 millimolar NaCl. Proline accumulated in Bragg only after stress was severe enough to induce injury; therefore proline accumulation is not a sensitive indicator of salt stress in soybean plants.     

Water Relations of Seed Development and Germination in Muskmelon (Cucumis melo L.) : III. Sensitivity of Germination to Water Potential and Abscisic Acid during Development

Muskmelon (Cucumis melo L.) seeds are germinable 15 to 20 days before fruit maturity and are held at relatively high water content within the fruit, yet little precocious germination is observed. To investigate two possible factors preventing precocious germination, the inhibitory effects of abscisic acid and osmoticum on muskmelon seed germination were determined throughout development. Seeds were harvested at 5-day intervals from 30 to 65 days after anthesis (DAA) and incubated either fresh or after drying on factorial combinations of 0, 1, 3.3, 10, or 33 micromolar abscisic acid (ABA) and 0, −0.2, −0.4, −0.6, or −0.8 megapascals polyethylene glycol 8000 solutions at 30°C. Radicle emergence was scored at 12-hour intervals for 10 days. In the absence of ABA, the water potential (Ψ) required to inhibit fresh seed germination by 50% decreased from −0.3 to −0.8 megapascals between 30 and 60 DAA. The Ψ inside developing fruits was from 0.4 to 1.4 megapascals lower than that required for germination at all stages of development, indicating that the fruit Ψ is sufficiently low to prevent precocious germination. At 0 megapascal, the ABA concentration required to inhibit germination by 50% was approximately 10 micromolar up to 50 DAA and increased to >33 micromolar thereafter. Dehydration improved subsequent germination of immature seeds in ABA or low Ψ. There was a linear additive interaction between ABA and Ψ such that 10 micromolar ABA or −0.5 megapascal osmotic potential resulted in equivalent, and additive, reductions in germination rate and percentage of mature seeds. Abscisic acid had no effect on embryo solute potential or water content, but increased the apparent minimum turgor required for germination. ABA and osmoticum appear to influence germination rates and percentages by reducing the embryo growth potential (turgor in excess of a minimum threshold turgor) but via different mechanisms. Abscisic acid apparently increases the minimum turgor threshold, while low Ψ reduces turgor by reducing seed water content.     

Plant Morphological and Biochemical Responses to Field Water Deficits : II. Responses of Leaf Glycerolipid Composition in Cotton

The effects of water deficits on leaf glycerolipid composition were analyzed in two photoperiodic strains of field grown cotton (Gossypium kirsutum L.) that differ in sensitivity to drought. Leaves from plants grown under dryland conditions exhibited increased dry weight and specific leaf weight. The average midday leaf water potential in the dryland treatment decreased to −1.9 and −2.4 megapascals, respectively, for the T25 and T185 genotypes. Total leaf lipid content of plants exposed to dryland conditions was 5.9 and 7.5% of leaf dry weight for strain T25 and T185, respectively. The difference in leaf lipid content between these genotypes was caused by water deficits and was attributed to loss of both phospholipids and glycolipids in strain T25. There was no apparent loss of phospholipids due to water deficits in the T185 genotype; however, a significant loss of glycolipids was partially compensated by a 2-fold increase in triacylglycerol. No change in triacylglycerol was found between treatments in T25 leaves. Water deficit caused a significant decline in the relative degree of acylunsaturation in phospholipids and glycolipids from both genotypes; however, the double bond index for triacylglycerol increased in both genotypes. It is believed that the observed responses of leaf lipid composition to dryland conditions may be an additional criterion for characterization and selection of new drought-tolerant cotton genotypes.     

Osmotic Response of Sugar Beet Source Leaves at CO(2) Compensation Point

As sugar beet source leaves lowered the CO₂ concentration to compensation point in a closed atmosphere, leaf thickness and relative water content decreased. Leaf water potential declined rapidly from −0.5 to −1.4 megapascals. At 340 microliters CO₂ per liter, water potential and sucrose, glucose, and fructose contents were steady in photosynthesizing source leaves. Within 90 minutes after leaves were exposed to a CO₂ concentration at the compensation point, leaf sucrose content declined to 60% of the preteatment level, rapidly in the first 30 minutes and then more slowly. During the subsequent 200 minutes, sucrose content increased to 180% of pretreatment level. Glucose and fructose remained unchanged during the treatment. Degradation of starch was sufficient to account for the additional sucrose that accumulated. Labeled carbon lost from starch appeared in sucrose and several other compounds that likely contributed to the recovery in leaf water content.     

Heterogenous stomatal closure in response to leaf water deficits is not a universal phenomenon

The extent and occurrence of water stress-induced “patchy” CO₂ uptake across the surface of leaves was evaluated in a number of plant species. Leaves, while still attached to a plant, were illuminated and exposed to air containing [¹⁴C]CO₂ before autoradiographs were developed. Plant water deficits that caused leaf water potential depression to −1.1 megapascals during a 4-day period did result in heterogenous CO₂ assimilation patterns in bean (Phaseolus vulgaris). However, when the same level of stress was imposed more gradually (during 17 days), no patchy stomatal closure was evident. The patchy CO₂ assimilation pattern that occurs when bean plants are subjected to a rapidly imposed stress could induce artifacts in gas exchange studies such that an effect of stress on chloroplast metabolism is incorrectly deduced. This problem was characterized by examining the relationship between photosynthesis and internal [CO₂] in stressed bean leaves. When extent of heterogenous CO₂ uptake was estimated and accounted for, there appeared to be little difference in this relationship between control and stressed leaves. Subjecting spinach (Spinacea oleracea) plants to stress (leaf water potential depression to −1.5 megapascals) did not appear to cause patchy stomatal closure. Wheat (Triticum aestivum) plants also showed homogenous CO₂ assimilation patterns when stressed to a leaf water potential of −2.6 megapascals. It was concluded that water stress-induced patchy stomatal closure can occur to an extent that could influence the analysis of gas exchange studies. However, this phenomenon was not found to be a general response. Not all stress regimens will induce patchiness; nor will all plant species demonstrate this response to water deficits.     

Fluorescence Quenching and Gas Exchange in a Water Stressed C(3) Plant, Digitalis lanata

A leaf cuvette has been adapted for use with a pulse-modulation fluorometer and an open gas exchange system. Leaf water potential (ψ) was decreased by withholding watering from Digitalis lanata EHRH. plants. At different stages of water deficiency the photochemical (q_Q) and nonphotochemical (q_E) fluorescence quenching was determined during the transition between darkness and light-induced steady state photosynthesis of the attached leaves. In addition, the steady state CO₂ and H₂O gas exchange was recorded. Following a decrease of leaf water potential with increasing water deficiency, the transition of photochemical quenching was almost unaffected, whereas nonphotochemical quenching increased. This is indicative of an enhanced thylakoid membrane energization during the transition and is interpreted as a partial inhibition of either the ATP generating or the ATP consuming reaction sequences. Complete reversion of the stress induced changes was achieved within 6 hours after rewatering. In contrast to the variations during transition, the final steady state values of q_Q and q_E remained unchanged over the entire stress range from −0.7 to −2.5 megapascals. From these results we conclude that, once established, electron transport via photosystem II and the transmembrane proton gradient remain unaffected by water stress. These data are indicative of a protective mechanism against photoinhibition during stress, when net CO₂ uptake is limited.     

Water-Stress-Induced Ethylene Production in Wheat : A Fact or Artifact?

Effects of water stress on ethylene evolution from excised leaf segments and intact plants of wheat (Triticum aestivum L. cv Katepwa) were studied. Excised leaf segments of 8 day or 6 week old plants were dried until they lost 8% of their fresh weight (water potential about −2.3 megapascals). These and nondried control leaf segments (water potential about −1.0 megapascal) were sealed in glass tubes, and their ethylene production rates were compared by head space analysis via gas-chromatography. The dried leaves of both ages produced significantly more ethylene than the corresponding controls. However, when 6 week old intact plants were water-stressed by withholding water supply, and their ethylene production measured using a continuousflow system, no increase in ethylene was deteceted despite a drop in water potential to −2.9 megapascals over 6 days. Even the leaf segments excised from plants that had been subjected to water stress for 2, 4, or 6 days produced no more ethylene (in sealed tubes) than the leaves from well-watered plants. In fact, the ethylene production by these segments decreased with the increase in the severity of stress experienced by the plants. The results show that the commonly reported overproduction of ethylene by excised leaves subjected to rapid drying represents an artifact, which has little relevance to the water stress responses of intact wheat plants.     

Drought-induced increases in abscisic Acid levels in the root apex of sunflower

Abscisic acid (ABA) levels in 3-mm apical root segments of slowly droughted sunflower plants (Helianthus annuus L. cv Russian Giant) were analyzed as the methyl ester by selected ion monitoring gas chromatography-mass spectrometry using characteristic ions. An internal standard, hexadeuterated ABA (d6ABA) was used for quantitative analysis. Sunflower seedlings, grown in aeroponic chambers, were slowly droughted over a 7-day period. Drought stress increased ABA levels in the root tips at 24, 72, and 168 hour sample times. Control plants had 57 to 106 nanograms per gram ABA dry weight in the root tips (leaf water potential, −0.35 to −0.42 megapascals). The greatest increase in ABA, about 20-fold, was found after 72 hours of drought (leaf water potential, −1.34 to −1.47 megapascals). Levels of ABA also increased (about 7− to 54-fold) in 3-mm apical root segments which were excised and then allowed to dessicate for 1 hour at room temperature.     

Effect of water stress on the reduction of nitrate and nitrite by soybean nodules

The effects of water stress on nitrate reductase and nitrite reductase activities in symbiotic nodules were examined in field-grown soybean plants (Glycine max L Merr. cv Clark). The in vitro assays of enzyme activity indicated that the nodule cytosol and bacteroids contained both nitrate reductase and nitrite reductase activities. The reduction of nitrate in bacteroids increased significantly as nodule water potential declined from −0.6 to −1.4 megapascals, and then decreased when −1.8 megapascals water potential was reached. On the contrary, the reduction of nitrate in nodule cytosol was inhibited as water stress progressed. Increases in water stress intensity also caused a significant inhibition in nitrite reductase activities of bacteroids and nodule cytosol within soybean nodules. The results show that nitrate reduction occurred both in the cytosol and bacteroids of water-stressed soybean nodules. The reduction of nitrate functioned at different physiological modes in these two fractions.     

Water Potential and Stomatal Resistance of Sunflower and Soybean Subjected to Water Stress during Various Growth Stages

Plants of two varieties of soybean (Glycine max (L.) Merr.) and two varieties of sunflower (Helianthus annuus L.) were grown in controlled environments and subjected to water stress at various stages of growth. Leaf resistances and leaf water potentials were measured as stress developed. In soybeans the upper leaf surface had a higher resistance than the lower surface at all leaf water potentials and growth stages. Resistance of the upper surface began to increase at a higher water potential and increased more than the resistance of the lower surface. Resistances returned to prestress values 4 days after rewatering. In sunflowers upper and lower leaf surfaces had similar resistances at all water potentials and growth stages. Leaf resistances were higher in sunflower plants stressed before flowering than in those stressed later. Sunflower plants stressed to −16 bars recovered their prestress leaf resistance and water potential a few days after rewatering, but leaves of sunflower plants stressed to −23 bars died. Leaves of soybean and sunflower plants stressed before flowering suffered less injury than those of older plants and sunflowers stressed after flowering suffered more injury than soybeans.     

Characterization of water stress and low temperature effects on flower induction in citrus

Experiments were conducted with containerized `Tahiti' lime (Citrus latifolia Tan.) trees in order to define conditions needed to induce flowering. Cyclical or continuous water stress for 4 to 5 weeks induced flowering. Moderate (−2.25 megapascals, midday) or severe (−3.5 megapascals, midday) water stress as measured by leaf xylem pressure potential, for as little as 2 weeks induced flowering, but the response was more significant in severely stressed trees. Low temperature (18°C day/10°C night) induced a time dependent flowering response much like that of moderate water stress. Significantly negative leaf xylem pressure potentials as compared to controls were found only under water stress treatment, suggesting that a common stress-linked event, separate from low plant water potential is involved in floral induction. Leafless, immature cuttings from mature, field-grown trees were induced to flower by water stress treatment, suggesting that leaves are not essential for a flower inductive response.     

Mild Water Stress of Phaseolus vulgaris Plants Leads to Reduced Starch Synthesis and Extractable Sucrose Phosphate Synthase Activity

Mild water stress, on the order of −1.0 megapascals xylem water potential, can reduce the rate of photosynthesis and eliminate the inhibition of photosynthesis caused by O₂ in water-stress-sensitive plants such as Phaseolus vulgaris. To investigate the lack of O₂ inhibition of photosynthesis, we measured stromal and cytosolic fructose-1,6-bisphosphatase, sucrose phosphate synthase, and partitioning of newly fixed carbon between starch and sucrose before, during, and after mild water stress. The extractable activity of the fructose bisphosphatases was unaffected by mild water stress. The extractable activity of SPS was inhibited by more than 60% in plants stressed to water potentials of −0.9 megapascals. Water stress caused a decline in the starch/sucrose partitioning ratio indicating that starch synthesis was inhibited more than sucrose synthesis. We conclude that the reduced rate of photosynthesis during water stress is caused by stomatal closure, and that the restriction of CO₂ supply caused by stomatal closure leads to a reduction in the capacity for both starch and sucrose synthesis. This causes the reduced O₂ inhibition and abrupt CO₂ saturation of photosynthesis.     

Relationship of Xylem Embolism to Xylem Pressure Potential, Stomatal Closure, and Shoot Morphology in the Palm Rhapis excelsa

Xylem failure via gas embolism (cavitation) induced by water stress was investigated in the palm Rhapis excelsa (Thumb.) Henry. Xylem embolism in excised stems and petioles was detected using measurements of xylem flow resistance: a decrease in resistance after the removal of flow-impeding embolisms by a pressure treatment indicated their previous presence in the axis. Results supported the validity of the method because increased resistance in an axis corresponded with: (a) induction of embolism by dehydration, (b) increased numbers of cavitations as detected by acoustic means, (c) presence of bubbles in xylem vessels. The method was used to determine how Rhapis accommodates embolism; results suggested four ways. (a) Embolism was relatively rare because pressure potentials reach the embolism-inducing value of about −2.90 megapascals only during prolonged drought. (b) When embolism did occur in nature, it was confined to the relatively expendable leaf xylem; the stem xylem, which is critical for shoot survival, remained fully functional. (c) Even during prolonged drought, the extent of embolism is limited by complete stomatal closure, which occurred at the xylem pressure potential of −3.20 ± 0.18 megapascals. (d) Embolism is potentially reversible during prolonged rains, since embolisms dissolved within 5 h at a pressure potential of 0.00 megapascals (atmospheric), and xylem sap can approach this pressure during rain.     

Recovery of photosynthesis in sunflower after a period of low leaf water potential

Photosynthesis was studied in sunflower plants subjected to 1 to 2 days of desiccation and then permitted to recover. The leaf water potential to which leaves returned after rewatering was dependent on the severity of desiccation and the evaporative conditions. Under moderately evaporative conditions, leaf water potential returned to predesiccation levels after 3 to 5 hours when desiccation was slight. Leaf water potentials remained below predesiccation levels for several days after rewatering when leaf water potentials decreased to −13 to −19 bars during desiccation. Leaf water potential showed no sign of recovery when leaf water potentials decreased to −20 bars or below during desiccation. The lack of full recovery of leaf water potential was attributable to increased resistance to water transport in the roots and stem. The resistance ultimately became large enough to result in death of the leaves because net water loss continued even after the soil had been rewatered.     

Cell turgor changes associated with ripening in tomato pericarp tissue

The pressure microprobe was used to determine whether the turgor pressure in tomato (Lycopersicon esculentum Mill., variety “Castelmart”) pericarp cells changed during fruit ripening. The turgor pressure of cells located 200 to 500 micrometers below the fruit epidermis was uniform within the same tissue (typically ± 0.02 megapascals), and the highest turgors observed (<0.2 megapascals) were much less than expected, based on tissue osmotic potential (−0.6 to −0.7 megapascals). These low turgor values may indicate the presence of apoplastic solutes. In both intact fruit and cultured discs of pericarp tissue, a small increase in turgor preceded the onset of ripening, and a decrease in turgor occurred during ripening. Differences in the turgor of individual intact fruit occurred 2 to 4 days before parallel differences in their ripening behavior were apparent, indicating that changes in turgor may reflect physiological changes at the cell level that precede expression of ripening at the tissue level.     

Evaluation of a system for the imposition of plant water stress

A system which imposes a range of water stress levels was developed and evaluated. Water stress was controlled by employing a screen to suspend roots above a water column of known height. Levels of water stress were imposed by changing water column height and/or hydraulic conductivity of the medium in the column. The system was evaluated in a series of growth chamber experiments in which sunflowers (Helianthus annuus L. cv NK894) were given three levels of water availability for a period of 3 weeks. Third leaf midday water potentials at the end of the trials ranged from −0.73 ± 0.04 to −2.35 ± 0.17 megapascals in waterstressed plants compared to −0.40 ± 0.02 megapascals for control plants. Repetition of experiments showed no statistical differences in leaf water potentials, plant leaf areas, or plastochron indices between trials. During the experiments, the severity and pattern of water stress developments was related to both water column height and conductivity of the medium. Control plants exhibited normal diurnal water relations and transpirational behavior. Use of this system avoids many problems associated with other techniques and provides a means for subjecting plants to reproducible water stress levels for extended periods of time.     

Compartmentation of solutes and water in developing sugarcane stalk tissue

Previous studies have suggested that the apoplast solution of sugarcane stalk tissue contains high concentrations of sucrose, but the accuracy of these reports has been questioned because sucrose leakage from damaged cells may have influenced the results. In this study, the solute potential of the apoplast and symplast of the second (immature), tenth, twentieth, thirtieth, and fortieth internodes of field-grown sugarcane (Saccharum spp. hybrid) stalk tissue was determined by two independent methods. Solute potential of the apoplast was measured either directly by osmometry from solution collected by centrifugation, or inferred from the initial water potential of fully hydrated tissue determined by thermocouple psychrometry before the tissue was progressively dehydrated for generation of water potential isotherms. Both methods produced nearly identical values ranging from −0.6 to −1.8 megapascals for immature and mature tissue, respectively. The solute potential of the symplast determined by either method ranged from −1.0 to approximately −2.2 megapascals for immature and mature internodes, respectively. Solute quantitation by HPLC agreed with concentrations inferred from osmometry. Washing thirtieth internode tissue in deionized water increased pressure potential from 0.29 to 1.96 megapascals. The apoplast of mature sugarcane stalk tissue is a significant storage compartment for sucrose containing as much as 25% of the total tissue water volume and as much as 21% of the stored sucrose.