Growth of the maize primary root at low water potentials : I. Spatial distribution of expansive growth

Seedlings of maize (Zea mays L. cv WF9 × Mo 17) were grown in vermiculite at various water potentials. The primary root continued slow rates of elongation at water potentials which completely inhibited shoot growth. To gain an increased understanding of the root growth response, we examined the spatial distribution of growth at various water potentials. Time lapse photography of the growth of marked roots revealed that inhibition of root elongation at low water potentials was not explained by a proportional decrease in growth along the length of the growing zone. Instead, longitudinal growth was insensitive to water potentials as low as − 1.6 megapascal close to the root apex, but was inhibited increasingly in more basal locations such that the length of the growing zone decreased progressively as the water potential decreased. Cessation of longitudinal growth occurred in tissue of approximately the same age regardless of spatial location or water status, however. Roots growing at low water potentials were also thinner, and analysis revealed that radial growth rates were decreased throughout the elongation zone, resulting in greatly decreased rates of volume expansion.     

Turgor and growth at low water potentials

Turgor affects cell enlargement but has not been measured in enlarging tissue of intact plants when growth is inhibited by inadequate water. Mature or excised tissue can be problematic for these measurements because turgor may not be the same as in intact enlarging cells. Therefore, we measured the average turgor in the elongating region of intact stems of soybean (Glycine max [L.] Merr.) while the seedlings were exposed to low water potentials by transplanting to vermiculite of low water content. Stem growth was completely inhibited by the transplanting, and the average turgor decreased in the mature stem tissue. However, it did not decrease in the elongating region whether measured in intact or excised tissue (total of four methods). At the cellular level, turgor was uniform in the elongating tissue except at transplanting, when turgor decreased in a small number of cortical cells near the xylem. The reduced turgor in these cells, but constant turgor in most of the cells, confirmed that no general turgor loss had occurred but indicated that gradients in water potential extending from the xylem into the enlarging tissue were reduced, thus decreasing the movement of water into the tissue for cell enlargement. A modest growth recovery occurred after 2 days and was preceded by a recovery of the gradient. This suggests that under these conditions, growth initially was inhibited not by turgor loss but by a collapse of the water potential gradient necessary for the growth process.     

Complete turgor maintenance at low water potentials in the elongating region of maize leaves

Leaf elongation rate, water potential, and osmotic potential were measured in the fifth leaf of maize (Zea mays L.) plants growing in soil from which water was withheld for varying times. Elongation occurred in the basal region, which was enclosed by other leaf sheaths. When water was withheld from the soil, leaf elongation decreased and eventually ceased even though enough solutes accumulated in the elongating region to maintain turgor virtually constant. In the exposed blade, however, turgor was lost and wilt symptoms developed. If the night was prolonged, the elongating region lost much of its ability to accumulate solute, which suggests that the accumulating solutes were of recent photosynthetic origin. Under these conditions, leaf elongation was restricted to higher water potentials than under the usual photoperiodic regime.     

Increased endogenous abscisic Acid maintains primary root growth and inhibits shoot growth of maize seedlings at low water potentials

Roots of maize (Zea mays L.) seedlings continue to grow at low water potentials that cause complete inhibition of shoot growth. In this study, we have investigated the role of abscisic acid (ABA) in this differential growth sensitivity by manipulating endogenous ABA levels as an alternative to external applications of the hormone. An inhibitor of carotenoid biosynthesis (fluridone) and a mutant deficient in carotenoid biosynthesis (vp 5) were used to reduce the endogenous ABA content in the growing zones of the primary root and shoot at low water potentials. Experiments were performed on 30 to 60 hour old seedlings that were transplanted into vermiculite which had been preadjusted to water potentials of approximately −1.6 megapascals (roots) or −0.3 megapascals (shoots). Growth occurred in the dark at near-saturation humidity. Results of experiments using the inhibitor and mutant approaches were very similar. Reduced ABA content by either method was associated with inhibition of root elongation and promotion of shoot elongation at low water potentials, compared to untreated and wild-type seedlings at the same water potential. Elongation rates and ABA contents at high water potential were little affected. The inhibition of shoot elongation at low water potential was completely prevented in fluridone-treated seedlings during the first five hours after transplanting. The results indicate that ABA accumulation plays direct roles in both the maintenance of primary root elongation and the inhibition of shoot elongation at low water potentials.     

Primary events regulating stem growth at low water potentials

Cell enlargement is inhibited by inadequate water. As a first step toward understanding the mechanism, all the physical parameters affecting enlargement were monitored to identify those that changed first, particularly in coincidence with the inhibition. The osmotic potential, turgor, yield threshold turgor, growth-induced water potential, wall extensibility, and conductance to water were measured in the elongating region, and the water potential was measured in the xylem of stems of dark-grown soybean (Glycine max [L.] Merr.) seedlings. A stepdown in water potential was achieved around the roots by transplanting the seedlings to vermiculite of low water content, and each of the parameters was measured simultaneously in the same plants while intact or within a few minutes of being intact using a newly developed guillotine psychrometer. The gradient of decreasing water potential from the xylem to the enlarging cells (growth-induced water potential) was the first of the parameters to decrease to a growth-limiting level. The kinetics were the same as for the inhibition of growth. The decreased gradient was caused mostly by a decreased water potential of the xylem. This was followed after 5 to 10 hours by a similar decrease in cell wall extensibility and tissue conductance for water. Later, the growth-induced water potential recovered as a result of osmotic adjustment and a rise in the water potential of the xylem. Still later, moderate growth resumed at a rate apparently determined by the low wall extensibility and tissue conductance for water. The turgor did not change significantly during the experiment. These results indicate that the primary event during the growth inhibition was the change in the growth-induced water potential. Because the growth limitation subsequently shifted to the low wall extensibility and tissue conductance for water, the initial change in potential may have set in motion subsequent metabolic changes that altered the characteristics of the wall and cell membranes.     

Chloroplast response to low leaf water potentials: I. Role of turgor

The effect of decreases in turgor on chloroplast activity was studied by measuring the photochemical activity of intact sunflower (Helianthus annuus L. cv. Russian Mammoth) leaves having low water potentials. Leaf turgor, calculated from leaf water potential and osmotic potential, was found to be affected by the dilution of cell contents by water in the cell walls, when osmotic potentials were measured with a thermocouple psychrometer. After the correction of measurements of leaf osmotic potential, both the thermocouple psychrometer and a pressure chamber indicated that turgor became zero in sunflower leaves at leaf water potentials of −10 bars. Since most of the loss in photochemical activity occurred at water potentials below −10 bars, it was concluded that turgor had little effect on the photochemical activity of the leaves.     

Osmotic adjustment and the inhibition of leaf,root, stem and silk growth at low water potentials in maize

The expansion growth of plant organs is inhibited at low water potentials ( _w), but the inhibition has not been compared in different organs of the same plant. Therefore, we determined elongation rates of the roots, stems, leaves, and styles (silks) of maize (Zea mays L.) as soil water was depleted. The _w was measured in the region of cell expansion of each organ. The complicating effects of transpiration were avoided by making measurements at the end of the dark period when the air had been saturated with water vapor for 10 h and transpiration was less than 1% of the rate in the light. Growth was inhibited as the _w in the region of cell expansion decreased in each organ. The _w required to stop growth was-0.50,-0.75, and-1.00 MPa, in this order, in the stem, silks, and leaves. However, the roots grew at these _w and ceased only when _w was lower than-1.4 MPa. The osmotic potential decreased in each region of cell expansion and, in leaves, roots and stems, the decrease was sufficient to maintain turgor fully. In the silks, the decrease was less and turgor fell. In the mature tissue, the _w of the stem, leaves and roots was similar to that of the soil when adequate water was supplied. This indicated that an equilibrium existed between these tissues, the vascular system, and the soil. At the same time, the _w was lower in the expanding regions than in the mature tissues, indicating that there was a _w disequilibrium between the growing tissue and the vascular system. The disequilibrium was interpreted as a _w gradient for supplying water to the enlarging cells. When water was withheld, this gradient disappeared in the leaf because _w decreased more in the xylem than in the soil, indicating that a high flow resistance had developed in the xylem. In the roots, the gradient did not decrease because vascular _w changed about the same amount as the soil _w. Therefore, the gradient in _w favored water uptake by roots but not leaves at low _w. The data show that expansion growth responds to low _w differently in different growing regions of the plant. Because growth depends on the maintenance of turgor for extending the cell walls and the presence of _w gradients for supplying water to the expanding cells, several factors could have been responsible for these differences. The decrease of turgor in the silks and the loss of the _w gradient in the leaves probably contributed to the high sensitivity of these organs. In the leaves, the gradient loss was so complete that it would have prevented growth regardless of other changes. In the roots, the maintenance of turgor and _w gradients probably allowed growth to continue. This difference in turgor and gradient maintenance could contribute to the increase in root/shoot ratios generally observed in water-limited conditions.Symbols _s osmotic potential - _w water potential     

Osmoregulation,solute distribution,and growth in soybean seedlings having low water potentials

Soybean (Glycine max (L.) Merr.) seedlings osmoregulate when the supply of water is limited around the roots. The osmoregulation involves solute accumulation (osmotic adjustment) by the elongating region of the hypocotyls. We investigated the relationship between growth, solute accumulation, and the partitioning of solutes during osmoregulation. Darkgrown seedlings were transplanted to vermiculite containing 1/8 (0.13 x) the water of the controls. Within 12–15 h, the osmotic potential of the elongating region had decreased to-12 bar, but it was-7 bar in the controls. This osmoregulation involved a true solute accumulation by the hypocotyls, since cell volume and turgor were virtually the same regardless of the water regime. The hypocotyls having low water potentials elongated slowly but, when deprived of their cotyledons, did not elongate or accumulate solute. This result indicated a cotyledonary origin for the solutes and a dependence of slow growth on osmotic adjustment. The translocation of nonrespired dry matter from the cotyledons to the seedling axis was unaffected by the availability of water, but partitioning was altered. In the first 12 h, dry matter accumulated in the elongating region of the 0.13 x hypocotyls, and osmotic adjustment occurred. The solutes involved were mostly free amino acids, glucose, fructose, and sucrose, and these accounted for most of the increased dry weight. After osmotic adjustment was complete, dry matter ceased to accumulate in the hypocotyls and bypassed them to accumulate in the roots, which grew faster than the control roots. The proliferation of the roots resulted in an increased root/shoot ratio, a common response of plants to dry conditions.Osmotic adjustment occurred in the elongating region of the hypocotyls because solute utilization for growth decreased while solute uptake continued. Adjustment was completed when solute uptake subsequently decreased, and uptake then balanced utilization. The control of osmotic adjustment was therefore the rate of solute utilization and, secondarily, the rate of solute uptake. Elongation was inhibited by unknown factors(s) despite the turgor and substrates associated with osmotic adjustment. The remaining slow elongation depended on osmotic adjustment and represented some optimum between the necessary inhibition for solute accumulation and the necessary growth for seedling establishment.     

Cell wall proteins at low water potentials

We investigated the proteins extractable from cell walls of stem tissues when plants were subjected to low water potentials (low ψ_w). Dark-grown soybean seedlings (Glycine max [L.] Merr.) showed decreased stem growth when the roots were exposed to vermiculite having low water content (ψ_w = −3 bar). After a time, growth resumed but at a reduced rate relative to the controls. The extractable protein increased in the cell walls as ψ_w decreased, especially a 28-kilodalton protein in the young tissue. In contrast, a 70 kilodalton protein, mainly extractable from mature cell walls, appeared to decrease slightly at low ψ_w. No hydroxyproline was present in either protein, which shows that neither protein is related to extensin. The level of the 28 kilodalton protein increased in the cell wall of the dividing region soon after the initial growth inhibition, and it appeared in the elongating tissue at about the time growth resumed. The correlation between growth and these protein changes suggests that the two events could be related.     

Abscisic acid accumulation maintains maize primary root elongation at low water potentials by restricting ethylene production

Previous work showed that primary root elongation in maize (Zea mays L.) seedlings at low water potentials (psi(w)) requires the accumulation of abscisic acid (ABA) (R.E. Sharp, Y. Wu, G.S. Voetberg, I.N. Saab, M.E. LeNoble [1994] J Exp Bot 45: 1743-1751). The objective of the present study was to determine whether the inhibition of elongation in ABA-deficient roots is attributable to ethylene. At a psi(w) of -1.6 MPa, inhibition of root elongation in dark-grown seedlings treated with fluridone to impose ABA deficiency was largely prevented with two inhibitors of ethylene synthesis (aminooxyacetic acid and aminoethoxyvinylglycine) and one inhibitor of ethylene action (silver thiosulfate). The fluridone treatment caused an increase in the rate of ethylene evolution from intact seedlings. This effect was completely prevented with aminooxyacetic acid and also when ABA was supplied at a concentration that restored the ABA content of the root elongation zone and the root elongation rate. Consistent results were obtained when ABA deficiency was imposed using the vp5 mutant. Both fluridone-treated and vp5 roots exhibited additional morphological symptoms of excess ethylene. The results demonstrate that an important role of ABA accumulation in the maintenance of root elongation at low psi(w) is to restrict ethylene production.     

Plant water relations and control of cell elongation at low water potentials

Recent developments in water status measurement techniques using the psychrometer, the pressure probe, the osmometer and pressure chamber are reviewed, and the process of cell elongation from the viewpoint of plant-water relations is discussed for plants subjected to various environmental stress conditions. Under water-deficient conditions, cell elongation of higher plants can be inhibited by interruption of water flow from the xylem to the surrounding elongating cells. The process of growth inhibition at low water potentials could be reversed by increasing the xylem water potential by means of pressure application in the root region, allowing water to flow from the xylem to the surrounding cells. This finding confirmed that a water potential field associated with growth process,i.e., the growth-induced water potential, is an important regulating factor for cell elongation other than metabolic factors. The concept of the growth-induced water potential was found to be applicable for growth retardation caused by cold stress, heat stress, nutrient deficiency and salinity stress conditions. In the present review, the fact that the cell elongation rate is primarily associated with how much water can be absorbed by elongating cells under water-deficiency, nutrient deficiency, salt stress, cold stress and heat stress conditions is suggested.     

Effect of inhibition of abscisic Acid accumulation on the spatial distribution of elongation in the primary root and mesocotyl of maize at low water potentials

Previous work showed that accumulation of endogenous abscisic acid (ABA) acts both to maintain primary root growth and inhibit shoot growth in maize seedlings at low water potentials (ψ_w) (IN Saab, RE Sharp, J Pritchard, GS Voetberg [1990] Plant Physiol 93: 1329-1336). In this study, we have characterized the growth responses of the primary root and mesocotyl of maize (Zea mays L. cv FR27 × FRMo 17) to manipulation of ABA levels at low ψ_w with a high degree of spatial resolution to provide the basis for studies of the mechanism(s) of ABA action. In seedlings growing at low ψ_w and treated with fluridone to inhibit carotenoid (and ABA) biosynthesis, ABA levels were decreased in all locations of the root and mesocotyl growing zones compared with untreated seedlings growing at the same ψ_w. In the root, low ψ_w (−1.6 megapascals) caused a shortening of the growing zone, as reported previously. The fluridone treatment was associated with severe inhibition of root elongation rate, which resulted from further shortening of the growing zone. In the mesocotyl, low ψ_w (−0.3 megapascal) also resulted in a shortened growing zone. In contrast with the primary root, however, fluridone treatment prevented most of the inhibition of elongation and the shortening of the growing zone. Final cell length measurements indicated that the responses of both root and mesocotyl elongation to ABA manipulation at low ψ_w involve large effects on cell expansion. Measurements of the relative changes in root and shoot water contents and dry weights after transplanting to a ψ_w of −0.3 megapascal showed that the maintenance of shoot elongation in fluridone-treated seedlings was not attributable to increased water or seed-reserve availability resulting from inhibition of root growth. The results suggest a developmental gradient in tissue responsiveness to endogenous ABA in both the root and mesocotyl growing zones. In the root, the capacity for ABA to protect cell expansion at low ψ_w appears to decrease with increasing distance from the apex. In the mesocotyl, in contrast, the accumulation of ABA at low ψ_w appears to become increasingly inhibitory to expansion as cells are displaced away from the meristematic region.     

Acclimation of leaf growth to low water potentials in sunflower

Abstract Leaf growth is one of the most sensitive of plant processes to water deficits and is frequently inhibited in field crops. Plants were acclimated for 2 weeks under a moderate soil water deficit to determine whether the sensitivity of leaf growth could be altered by sustained exposure to low water potentials. Leaf growth under these conditions was less than in the controls because expansion occurred more slowly and for less of the day than in control leaves. However, acclimated leaves were able to grow at leaf water potentials (Ψ₁) low enough to inhibit growth completely in control plants. This ability was associated with osmotic adjustment and maintenance of turgor in the acclimated leaves. Upon rewatering, the growth of acclimated leaves increased but was less than the growth of controls, despite higher concentrations of cell solute and greater turgor in the acclimated leaves than in controls. Therefore, factors other than turgor and osmotic adjustment limited the growth of acclimated leaves at high ψ₁ Four potentially controlling factors were investigated and the results showed that acclimated leaves were less extensible and required more turgor to initiate growth than control leaves. The slow growth of acclimated leaves was not due to a decrease in the water potential gradient for water uptake, although changes in the apparent hydraulic conductivity for water transport could have occurred. It was concluded that leaf growth acclimated to low ψ₁, by adjusting osmotically, and the concomitant maintenance of turgor permitted growth where none otherwise would occur. However, changes in the extensibility of the tissue and the turgor necessary to initiate growth caused generally slow growth in the acclimated leaves.     

Spatial variation in tree root distribution and growth associated with minirhizotrons

Four minirhizotrons were installed in each of three replicate plots in a deciduous forest dominated by Acer saccharum Marsh. The length growth of tree roots along the surface of the minirhizotrons was measured for a period of one year, and the resulting data were analyzed in nested, averaged and pooled arrangements. The analyses of nested data showed that spatial variation in root growth and abundance among minirhizotrons within plots was greater than variation among plots. Averaging data from minirhizotrons within plots prior to analysis reduced variation about plot means, but extensive intraplot variation invalidates this approach on statistical grounds. Both nested and averaged data failed to account for the contribution of individual roots to the mean, and root production rates were consequently overestimated. Pooling the data from the four minirhizotrons reduced variation about the means, and resulted in a more representative estimate of root production rates. The analysis of composited data can be used to incorporate small-scale variation into a single replicate sample in those circumstances where the activity of the root systems of plant communities is the object of study.     

Phosphorylation in crested wheatgrass seeds at low water potentials

Crested wheatgrass seeds [Agropyron desertorum (Fisch. ex Link) Schult.] were tested for their ability to carry on phosphorylation reactions at low water potentials. Seeds were treated with ³²P labeled sodium phosphate and incubated in air having different controlled relative humidities. Ion exchange chromatography and radioassay of phosphate esters indicated that some phosphorylation occurred at a water potential of −880 atmospheres. Seeds did not incorporate ³²P in nicotinamide adenine dinucleotide, adenosine triphosphate, and uridine diphosphate hexose until they were moistened to a water potential of −130 atmospheres.     

Amylase synthesis and stability in crested wheatgrass seeds at low water potentials

Drying of seeds of Agropyron desertorum (Fisch. ex Link) Schult. did not result in breakdown of α-amylase nor impair the ability of seeds to resume its synthesis when moistened again. β-Amylase activity did not change during 5 days of germination at a water potential of 0 atmosphere nor during 40 days of incubation at −40 atmospheres. Seeds synthesized α-amylase at 0, −20, and −40 atmospheres, but not at −60 atmospheres. At 0 and −20 atmospheres, the log of α-amylase activity was linearly related to hastening of germination. But at −40 atmospheres, seeds synthesized α-amylase during a time when there was little hastening of germination. Thus, it appears that other biochemical reactions are less drought-tolerant than synthesis of α-amylase. It is concluded that inhibition of α-amylase synthesis is not a controlling factor in the germination of these seeds at low water potentials.     

Influences of root distribution and growth on predicted water uptake and interspecific competition

Summary Root distribution and growth measured in the field were incorporated into a water uptake model for the CAM succulent Agave deserti and its nurse plant Hilaria rigida, a common desert bunchgrass. Agave deserti responds to the infrequent rainfalls of the Sonoran Desert by extending its existing established roots and by producing new roots. Most of such root growth was completed within one month after soil rewetting, total root length of A. deserti increasing by 84% for a seedling and by 58% for a mediumsized plant in the summer. Root growth in the winter with its lower soil temperatures was approximately half as much as in the summer. For a 15-year period, predicted annual root growth of A. deserti varied more than 18-fold because of annual variations in rainfall amount and pattern as well as seasonal variation in soil temperature. Predicted annual water uptake varied 47-fold over the same period. The nurse plant, which is crucial for establishment of A. deserti seedlings, reduced seedling water uptake by 38% during an average rainfall year. Lowering the location of the root system of a medium-sized A. deserti by 0.24 m reduced its simulated annual water uptake by about 25%, reflecting the importance of shallow roots for this desert succulent. Lowering the root system of a medium-sized H. rigida by 0.28 m increased the simulated annual water uptake of an associated A. deserti seedling by 17%, further indicating the influence of root overlap on competition for water.     

Photosynthesis at low water potentials in sunflower: lack of photoinhibitory effects

The losses in chloroplast capacity to fix CO₂ when photosynthesis is reduced at low leaf water potential (ψ₁) have been proposed to result from photoinhibition. We investigated this possibility in soil-grown sunflower (Helianthus annuus L. cv IS894) using gas exchange techniques to measure directly the influence of light during dehydration on the in situ chloroplast capacity to fix CO₂. The quantum yield for CO₂ fixation as well as the rate of light- and CO₂-saturated photosynthesis were strongly inhibited at low ψ₁. The extent of inhibition was the same whether the leaves were exposed to high or to low light during dehydration. When intercellular partial pressures of CO₂ were decreased to the compensation point, which was lower than the partial pressures resulting from stomatal closure, the inhibition of the quantum yield was also unaffected. Photoinhibition could be observed only after high light exposures were imposed under nonphysiological low CO₂ and O₂ where both photosynthesis and photorespiration were suppressed. The experiments are the first to test whether gas exchange at low ψ₁ is affected by potentially photoinhibitory conditions and show that the loss in chloroplast capacity to fix CO₂ was entirely the result of a direct effect of water availability on chloroplast function and not photoinhibition.     

Nonstomatal inhibition of photosynthesis in sunflower at low leaf water potentials and high light intensities

The inhibition of photosynthesis at low leaf water potentials was studied in soil-grown sunflower to determine the degree to which photosynthesis under high light was affected by stomatal and nonstomatal factors. Below leaf water potentials of −11 to −12 bars, rates of photosynthesis at high light intensities were insensitive to external concentrations of CO₂ between 200 and 400 microliters per liter. Photosynthesis also was largely insensitive to leaf temperature between 10 and 30 C. Changes in CO₂ concentration and temperature had negligible effect on leaf diffusive resistance. The lack of CO₂ and temperature response for both photosynthesis and leaf diffuse resistance indicates that rates of photosynthesis were not limited by either CO₂ diffusion or a photosynthetic enzyme. It was concluded that photosynthesis under high light was probably limited by reduced photochemical activity of the leaves at water potentials below −11 to −12 bars.