Growth, Water Relations, and Accumulation of Organic and Inorganic Solutes in Roots of Maize Seedlings during Salt Stress.

Seedlings of maize (Zea mays L. cv Pioneer 3906), hydroponically grown in the dark, were exposed to NaCl either gradually (salt acclimation) or in one step (salt shock). In the salt-acclimation treatment, root extension was indistinguishable from that of unsalinized controls for at least 6 d at concentrations up to 100 mM NaCl. By contrast, salt shock rapidly inhibited extension, followed by a gradual recovery, so that by 24 h extension rates were the same as for controls, even at 150 mM NaCl. Salt shock caused a rapid decrease in root water and solute potentials for the apical zones, and the estimated turgor potential showed only a small decline; similar but more gradual changes occurred with salt acclimation. The 5-bar decrease in root solute potential with salt shock (150 mM NaCl) during the initial 10 min of exposure could not be accounted for by dehydration, indicating that substantial osmotic adjustment occurred rapidly. Changes in concentration of inorganic solutes (Na+, K+, and Cl-) and organic solutes (proline, sucrose, fructose, and glucose) were measured during salt shock. The contribution of these solutes to changes in root solute potential with salinization was estimated.

Ethylene Biosynthesis during Aerenchyma Formation in Roots of Maize Subjected to Mechanical Impedance and Hypoxia.

Germinated maize (Zea mays L.) seedlings were enclosed in modified triaxial cells in an artificial substrate and exposed to oxygen deficiency stress (4% oxygen, hypoxia) or to mechanical resistance to elongation growth (mechanical impedance) achieved by external pressure on the artificial substrate, or to both hypoxia and impedance simultaneously. Compared with controls, seedlings that received either hypoxia or mechanical impedance exhibited increased rates of ethylene evolution, greater activities of 1-aminocyclopropane-1-carboxylic acid (ACC) synthase, ACC oxidase, and cellulase, and more cell death and aerenchyma formation in the root cortex. Effects of hypoxia plus mechanical impedance were strongly synergistic on ethylene evolution and ACC synthase activity; cellulase activity, ACC oxidase activity, or aerenchyma formation did not exhibit this synergism. In addition, the lag between the onset of stress and increases in both ACC synthase activity and ethylene production was shortened by 2 to 3 h when mechanical impedance or impedance plus hypoxia was applied compared with hypoxia alone. The synergistic effects of hypoxia and mechanical impedance and the earlier responses to mechanical impedance than to hypoxia suggest that different mechanisms are involved in the promotive effects of these stresses on maize root ethylene biosynthesis.

Shielding from solar particle event exposures in deep space.

The physical composition and intensities of solar particle event exposures of sensitive astronaut tissues are examined under conditions approximating an astronaut in deep space. Response functions for conversion of particle fluence into dose and dose equivalent averaged over organ tissues are used to establish significant fluence levels and the expected dose and dose rates of the most important events from past observations. The BRYNTRN transport code is used to evaluate the local environment experienced by sensitive tissues and used to evaluate bioresponse models developed for use in tactical nuclear warfare. The present results will help to clarify the biophysical aspects of such exposure in the assessment of RBE and dose rate effects and their impact on design of protection systems for the astronauts. The use of polymers as shielding material in place of an equal mass of aluminum would provide a large safety factor without increasing the vehicle mass. This safety factor is sufficient to provide adequate protection if a factor of two larger event than has ever been observed in fact occurs during the mission.

Influence of soil water stress on evaporation, root absorption, and internal water status of cotton.

Diurnal variations in leaf water potential, diffusion resistance, relative water content, stem diameter, leaf temperature, and energy balance components were measured in cotton (Gossypium hirsutum L. var. Lankart 57) during drought stress under field conditions. A plot of leaf water potential against either relative water content or stem diameter during the 24-hour period yielded a closed hysteresis loop. The relation between cell hydration and evaporation is discussed.Despite low soil water potential in the main root zone, significant plant evaporation rates were maintained. Root absorption rates as a function of soil depth were calculated from water content profiles measured with a neutron probe. The maximal root absorption rate of 3.5 x 10(-3) day(-1) occurred at the 75-centimeter depth, well below the main root zone.Stomatal resistance of individual leaves during the daylight hours remained nearly constant at 2.5 seconds centimeter(-1) even though leaf water potentials approached -30 bars. A growth chamber study indicated stomatal closure occurred at potentials near -16 bars. Possible implications of high soil water stress in relation to stomatal function and growth are discussed. Based on an energy balance method, the actual to potential plant evapotranspiration ratio was 0.43 for the 24-hour period, indicating partial stomatal closure. A surface resistance, r(s), of 4.0 seconds centimeter(-1) was calculated for the incomplete canopy with the use of the energy balance data. Alternatively, a canopy resistance of 1.3 seconds centimeter(-1) was attained from a relationship between leaf area and stomatal resistance of individual leaves. If the soil resistance was assumed to be very large and the canopy resistance was weighted for the fractional ground cover of the crop, the calculated surface resistance was 4.3 seconds centimeter(-1). Under these conditions, the two independent estimates of r(s) were in essential agreement.

Leaf Age as a Determinant in Stomatal Control of Water Loss from Cotton during Water Stress.

The stomatal resistance of individual leaves of young cotton plants (Gossypium hirsutum L. var. Stoneville 213) was measured during a period of soil moisture stress under conditions of constant evaporative demand. When plants were subjected to increasing soil water stress, increases in stomatal resistance occurred first on the lower leaves and the stomata on the upper surfaces were the most sensitive to decreasing leaf-water potential. Stomatal closure proceeded from the oldest leaves to the youngest as the stress became more severe. This apparent effect of leaf age was not due to radiation differences during the stress period. Radiation adjustments on individual leaves during their development altered the stomatal closure potential for all leaves, but did not change the within-plant pattern. Our data indicate that no single value of leaf water potential will adequately represent a threshold for stomatal closure in cotton. Rather, the stomatal resistance of each leaf is uniquely related to its own water potential as modified by age and radiation regime during development. The effect of age on stress-induced stomatal closure was not associated with a loss of potassium from older leaves. Increases in both the free and bound forms of abscisic acid were observed in water-stressed plants, but the largest accumulations occurred in the youngest leaves. Thus, the pattern of abscisic acid accumulation in response to water stress did not parallel the pattern of stomatal closure induced by water stress.

Factors Involved in in Vitro Stabilization of Nitrate Reductase from Cotton (Gossypium hirsutum L.) Cotyledons.

Experiments were conducted to determine if pretreatment of cotton (Gossypium hirsutum L.) plants resulted in differential in vitro stabilities of nitrate reductase (NR) activity. Although NR activity declines markedly during the second half of the daily light period, in vitro NR stability is not modified by time of harvest. Phenylmethylsulfonylfluoride, iodoacetamide, and N-ethylmaleimide do not influence in vitro NR stability, suggesting that serine or sulfhydryl proteases are not responsible for in vitro lability of NR from cotton cotyledons.Imposition of water stress or artificial extension of the dark period lead to significant reductions in NR activity, but do not change in vitro NR stability.Dilution of a crude extract leads to increasing lability of NR; hence the marked instability of NR cannot be attributed to an inactivator which follows simple enzyme kinetics. Since in vitro NR activity is much more stable in presence of both NADH and NO(3) (-), substrate availability must be considered as a possible factor influencing in vivo NR stability.

Auxin Transport as Related to Leaf Abscission during Water Stress in Cotton.

Plant water deficits reduced the basipetal transport of auxin in cotyledonary petiole sections taken from cotton (Gossypium hirsutum L.) seedings. A pulse-labeling technique was employed to eliminate complications of uptake or exit of (14)C-indoleacetic acid from the tissue. The transport capacity or the relative amount of radioactivity in a 30-minute pulse which was basipetally translocated was approximately 30% per hour in petioles excised from well watered seedlings (plant water potentials of approximately -4 to -8 bars). No cotyledonary leaf abscission took place in well watered seedlings. Plant water potentials from -8 to -12 bars reduced the transport capacity from 30 to 15% per hour, and although the leaves were wilted, cotyledonary abscission did not increase appreciably at these levels of stress. The threshold water potential sufficient to induce leaf abscission was approximately -13 bars and abscission increased with increasing stress while the auxin transport capacity of the petioles remained relatively constant (15% per hour). The basipetal transport capacity of well watered petioles tested under anaerobic conditions and acropetal transport tested under all conditions were typically less than basipetal transport under the most severe stress conditions. Cotyledonary abscission took place during and 24 hours after relief of stress with little or no abscission taking place 48 hours after relief of stress. Although the water potential returned to -4 bars within hours after rewatering the stressed plants, partial recovery of the basipetal transport capacity of the petioles was not apparent until 48 hours after rewatering, and at least 72 hours was required to return the transport capacity to near normal values. These data support the view that decreased levels of auxin reaching the abscission zone from the leaf blade influence the abscission process and further suggest that the length of time that the auxin supply is maximally reduced is more critical than the degree of reduction.

Movement and Endogenous Levels of Abscisic Acid during Water-Stress-induced Abscission in Cotton Seedlings.

In an effort to investigate possible involvement of abscisic acid (ABA) in foliar abscission processes, its movement and endogenous levels were examined in cotyledons taken from cotton seedlings (Gossypium hirsutum L.) subjected to varying degrees of water deficit, a condition which initiates leaf abscission. Using a pulse-labeling technique to avoid complications of uptake and exit from the tissue, ABA-1-(14)C movement was observed in both basipetal and acropetal directions in cotyledonary petioles taken from well watered, stressed, and rewatered plants. The label distribution patterns obtained after 1 and 3 hours of transport under all situations of water supply were diffusive in nature and did not change when tested under anaerobic conditions. The transport capacity of the petioles ranged from 3.6 to 14.4% ABA-1-(14)C transported per hour at estimated velocities of 0 to 2 millimeters per hour. Comparison of basipetal and acropetal movement indicated a lack of polarity under all conditions tested. These low transport capacities and slow velocities of movement, when compared to the active transport systems associated with auxin movement, as well as the lack of anaerobic effects and polarity, suggest that ABA movement in cotton cotyledonary petiole sections is facilitated by passive diffusion. Increases in free and bound ABA in the lamina with increased water stress did not correlate with patterns of cotyledonary abscission. Thus, no evidence was found to suggest that ABA is directly involved in stress-induced abscission processes.

Kinetics of early time calcium oxalate nephrolithiasis.

This paper examines the kinetics of calcium deposition in rat kidneys after an intraperitoneal sodium oxalate injection. From the results we conclude that only a limited portion of tubular surface is available for adsorption of calcium oxalate crystals, that adsorpption of calcium oxalate crystals onto tubular epithelium is a process of greater than first order with regard to the dose, and that the washout of retained particles from the tubules is a first-order process as related to time. Also, we conclude that in these animals, which were subjected to a large oxalate challenge, the deposition of calcium oxalate crystals is virtually all intratubular.

Movement of Kinetin and Gibberellic Acid in Leaf Petioles during Water Stress-induced Abscission in Cotton.

Movement of [(14)C]kinetin and [(14)C]gibberellic acid was examined in cotton (Gossypium hirsutum L.) cotyledonary petiole sections independent of label uptake or exit from the tissue. Sections 20 millimeters in length were taken from well watered, stressed, and poststressed plants. Transport capacity was determined using a pulse-chase technique. Movement of both kinetin and gibberellic acid was found to be nonpolar with a velocity of 1 millimeter per hour or less, suggesting passive diffusion. Neither water stress nor anaerobic conditions during transport of labeled material affected the transport capacity of the petioles.Results suggested strong kinetin binding but weak gibberellic acid binding in the tissue sections. Apparent binding of both growth regulators was unaltered by the experimental conditions. Movement of these two growth regulators within cotton cotyledonary petioles plays a minor role in the stress-induced, foliar abscission process.

Ethylene Evolution from Maize (Zea mays L.) Seedling Roots and Shoots in Response to Mechanical Impedance.

The effect of mechanical impedance on ethylene evolution and growth of preemergent maize (Zea mays L.) seedlings was investigated by pressurizing the growth medium in triaxial cells in a controlled environment. Pressure increased the bulk density of the medium and thus the resistance to growth. The elongation of maize primary roots and preemergent shoots was severely hindered by applied pressures as low as 10 kilopascals. Following a steep decline in elongation at low pressures, both shoots and roots responded to additional pressure in a linear manner, but shoots were more severely affected than roots at higher pressures. Radial expansion was promoted in both organs by mechanical impedance. Primary roots typically became thinner during the experimental period when grown unimpeded. In contrast, pressures as low as 25 kilopascals caused a 25% increase in root tip diameter. Shoots showed a slight enhancement of radial expansion; however, in contrast to roots, the shoots increased in diameter even when growing unimpeded. Such morphological changes were not evident until at least 3 hours after initiation of treatment. All levels of applied pressure promoted ethylene evolution as early as 1 hour after application of pressure. After 1 hour, ethylene evolution rates had increased 10, 32, 70, and 255% at 25, 50, 75, and 100 kilopascals respectively, and continued to increase linearly for at least 10 hours. When intact corn seedlings were subjected to a series of hourly cycles of pressure, followed by relaxation, ethylene production rates increased or decreased rapidly, illustrating tight coupling between mechanical impedance and tissue response. Seedlings exposed to 1 microliter of ethylene per liter showed symptoms similar to those shown by plants grown under mechanical impedance. Root diameter increased 5 times as much as the shoot diameter. Pretreatment with 10 micromolar aminoethoxyvinyl glycine plus 1 micromolar silver thiosulfate maintained ethylene production rates of impeded seedlings at basal levels and restored shoot and root extension to 84 and 90% of unimpeded values, respectively. Our results support the hypothesis that ethylene plays a pivotal role in the regulation of plant tissue response to mechanical impedance.

Water Stress Enhances Ethylene-mediated Leaf Abscission in Cotton.

Abscission of cotyledonary leaves from cotton (Gossypium hirsutum L. cv. Stoneville 213) seedlings occurred following relief from water stress. The amount of abscission was related to the magnitude of the plant water deficit. Leaf abscission promoted by exogenous ethylene was enhanced in seedlings subjected to water stress. Treatment with ethylene (2.0 to 3.2 microliters of ethylene per liter of air for 24 hours) raised the threshold plant water potential required to induce abscission from -17 to -7 bar, indicating that the stress caused the tissue to become predisposed to ethylene action. Based on the abscission response curve for seedlings treated with ethylene while under water stress, this apparent predisposition was developed as the plant water potentials reached the -7 to -10 bar range. The abscission-promoting effects of ethylene in combination with water stress were reversed with 15% CO(2) at plant water potentials above -12 bar, but the CO(2) reversal was lost at lower water potentials. These results are compatible with the concept that ethylene plays a regulatory role in leaf abscission induced by water stress.

Effect of atmospheric pressure on maize root growth and ethylene production.

Maize (Zea mays) seedlings were exposed to elevated atmospheric pressures while growing in moist sand in open plastic envelopes to evaluate the effects of directly applied atmospheric pressure on ethylene production and root growth. Effects were evaluated after 24 h. The threshold pressures necessary to promote ethylene production and decrease root elongation were about 600 and 400 kPa, respectively. Direct atmospheric pressure, at levels up to 300 kPa, mimicked the control decrease in root diameter and increased diameter only slightly at 500 to 1200 kPa. In contrast, in previous work it was shown that physical impedance resulting from compression of the growth medium by external application of 100 kPa increased the ethylene production rate 4-fold and the root diameter 7-fold while reducing elongation 75% in 10 h. The relative insensitivity of roots to direct atmospheric pressure suggests that they perceive physical impedance, achieved experimentally by compressing the growth medium, via a surface mechanism rather than via a pressure-sensing mechanism.

Metabolism of 1-Aminocyclopropane-1-Carboxylic Acid in Etiolated Maize Seedlings Grown under Mechanical Impedance.

We investigated the metabolism of 1-aminocyclopropane-1-carboxylic acid (ACC) in etiolated maize (Zea mays L.) seedlings subjected to mechanical impedance by applying pressure to the growing medium. Total concentrations of ACC varied little in unimpeded seedlings, but impeded organs accumulated ACC. Roots had consistently higher concentrations of ACC than shoots or seeds, regardless of treatment. The concentration of ACC in the roots increased more than 100% during the first hour of treatment irrespective of the pressure applied; in shoots, total ACC concentration increased 46% at either low or high pressure during the first hour of treatment. The bulk of ACC synthesized under impeded and unimpeded conditions was present in a conjugated form, presumably, 1-(malonylamino)-cyclopropane-1-carboxylic acid. However, 1-(malonylamino)-cyclopropane-1-carboxylic acid increased 73% over controls after 10 hours at 25 kilopascals of pressure. Unimpeded tissue had about 77% ACC as the conjugate and 17% as free ACC, and less than 6% was used in ethylene production. Increased amounts of ACC were converted into ethylene under stress. In vivo ACC synthase activity in roots became six and seven times higher only 1 hour after initiation of treatment at 25 and 100 kilopascals of pressure, respectively, and remained high for at least 6 hours. However, the immediate and massive conjugation of mechanically induced ACC suggests that ACC N-malonyltransferase may play an important role in the regulation of mechanically induced ethylene production. After 8 hours, in vivo activity of the ethylene-forming enzyme complex increased 100 and 50% above normal level at 100 and 25 kilopascals, respectively. Furthermore, ethylene-forming enzyme complex activity was significantly greater at 100 kilopascals than in controls as early as 1 hour after treatment initiation. These data suggest that regulation of ethylene production under mechanical impedance involves the concerted action of ACC synthase, the ethylene-forming enzyme complex, and ACC N-malonyltransferase.

Structural Changes and Associated Reduction of Hydraulic Conductance in Roots of Sorghum bicolor L. following Exposure to Water Deficit.

The effects of a severe water deficit on total root (L(t)) and axial (L(x)) hydraulic conductances and on the development of the hypodermis, endodermis, and xylem were studied in sorghum (Sorghum bicolor L.). Water deficit was imposed in the upper rooting zone while the lower zones were kept moist. L(t) and L(x) were based on water flow rates obtained by applying suction to proximal xylem ends of excised roots. The development of the hypodermis, endodermis, and other tissues were examined by staining with fluorescent berberine hemisulfate and phloroglucinol-HCl. The L(t) value (x 10(-8) meters per second per megapascal) for unstressed control roots was 22.0 and only 5.9 for stressed roots. The low L(t) in stressed roots was attributed, in part, to accelerated deposition of lignin and suberin in the hypodermis and endodermis. Calcofluor, an apoplastic tracer that binds to cellulose, was blocked in stressed roots at the lignified and suberized outer tangential walls of the hypodermis but readily penetrated the cortical walls of similar root regions in controls where the casparian band was not developed. L(x) per unit root length was about 100 times lower in stressed roots than in controls because of the persistence of late metaxylem cross-walls and the smaller diameter and lower number of conductive protoxylem and early metaxylem vessels.

Evidence for an Inactivating System of Nitrate Reductase in Hordeum vulgare L. during Darkness That Requires Protein Synthesis.

The disappearance of nitrate reductase activity in leaves of Hordeum vulgare L. during darkness was inhibited by cycloheximide, actinomycin D, and low temperature. Thus, protein synthesis was probably required for the disappearance of nitrate reductase in the dark. Since chloramphenicol did not affect the rate of loss of activity, the degradation or inactivation apparently required protein synthesis by the cytoplasmic ribosomal system. Consistent with this observation, nitrate reductase is also reportedly located in the cytoplasm. Thus, the amount of nitrate reductase activity present in leaves of barley may be controlled by a balance between activating and inactivating systems.

Reduction of auxin transport capacity with age and internal water deficits in cotton petioles.

Auxin transport was examined in leaf petioles taken from the upper, middle, and lower leaf canopy of large cotton plants. The ability of petioles to transport auxin decreased with age (position) of the leaves. Plant water deficit reduced transport regardless of age. These correlations support the view that reduced transport capacity of petioles plays a significant role in the induction of abscission of lower or older leaves during water deficits.

Abscission responses to moisture stress, auxin transport inhibitors, and ethephon.

The three abscission-inducing agents - water stress, Ethephon, and auxin transport inhibitors-acted synergistically to promote leaf fall in cotton (Gossypium hirsutum L.). However, the synergism was primarily between stress and Ethephon. Auxin transport inhibitors did not promote the effect of stress alone, only promoted the effect of Ethephon in well watered plants and gave a very small promotion with stress and Ethephon together. Abscission was rapid in stressed plants treated with Ethephon and an auxin transport inhibitor, while leaves fell more slowly from well watered plants treated with Ethephon alone. This suggests that water stress or auxin transport inhibitors influence initial events in abscission; since an auxin transport inhibitor will replace the effect of stress but not Ethephon, an initial event in stress-induced abscission appears to be inhibition of auxin transport. Ethephon promoted lateral bud release, and auxin transport inhibitors did not duplicate that effect alone or promote it in combination with Ethephon.

Hemophilia and extracorporeal shock wave lithotripsy: a case report.

We report renal calculi fragmentation in a hemophiliac patient through the use of extracorporeal shock wave lithotripsy. The Dornier Human Model III lithotriptor was used with the patient under general anesthesia and high frequency intermittent positive pressure ventilation was delivered via a Siemen-Elema 900-D Ventilator. The pathological features and clinical course are discussed.

Gene expression induced by physical impedance in maize roots.

Two cDNA clones, pIIG1 and pIIG2, corresponding to mRNAs that accumulate in maize root tips subjected to 10 min of physical impedance, were isolated by differential screening of a cDNA library. The deduced proteins, based on DNA sequence analysis, have molecular masses of 13 and 23 kDa for pIIG1 and pIIG2, respectively. pIIG1 showed 97% similarity at the nucleic acid level to a maize root cortical cell delineating protein (pZRP3) and was also similar to some bimodular proteins that are developmentally or stress regulated in other plant species. In situ localization of pIIG1 showed some expression in cortical cells of control maize roots; however, after a 10 min physical impedance treatment, pIIG1 accumulation increased greatly in cortical cells and extended to include the procambial region. pIIG2 did not show sequence similarity with any identified gene of known function, but a bipartite nuclear targeting sequence occurs in its deduced amino acid sequence which indicates it may function in the nucleus. Thus, rapid accumulation of specific mRNAs occurs in maize roots in response to impedance stress, and these mRNAs may be responsible for some responses of the roots to physical impedance.