Water Pumping Activity of the Root System in the Process of Cross-Adaptation of Sunflower Plants to Hyperthermia and Water Deficiency

To elucidate the mechanisms of cross-adaptation, we investigated the effect of heat shock (HS, for 2 h at 45°C) on leaf tolerance to overheating and exudation by roots detached from 25–30-day old sunflower (Helianthus annuus L.) plants. It was demonstrated that preheating enhanced considerably leaf tolerance and activated root exudation, especially under water deficiency produced by plant transfer to the hypertonic NaCl solution (17 mM). Under water deficiency conditions, the roots of HS-treated plants pumped water against the osmotic pressure (OP) gradient between the exudate and the external solution. Therefore, we concluded that this pumping was realized due to a metabolic (non-osmotic) constituent of root pressure. In the roots of plants that were not treated with HS, the OP gradient became positive. This fact implies that the HS-pretreatment of plants retarded the penetration of sodium and chlorine ions into roots. The data obtained demonstrate that HS induced a cross-adaptation of plants to high temperature and water deficiency. Such cross-adaptation involves, as an important component, an acceleration of water metabolism, including an enhanced water pumping activity of root system.     

Effects of Coupled Solute and Water Flow in Plant Roots with Special Reference to Brouwer's Experiment

This paper presents a further examination of the effects ofcoupled solute and water flow as it applies to plant roots.A cylindrical two-compartment membrane root model can accountfor many diverse root phenomena. Among these are (1) non-linearpressure flow, (2) interval osmotic dilution, (3) negative resistanceeffects, (4) non-osmotic water flux, and (5) the internal osmoticcompensation effect. In addition, we have now demonstrated thatthe same model is capable of reproducing the complex patternsof root resistance profile change noted by some workers. Innone of these instances is there need to invoke any change inthe hydraulic conductivity of the system to explain the observedeffects. The only features necessary to explain these phenomenaare a membrane-like structure and a mechanism for actively accumulatingsolutes.     

Onion root water transport sensitive to water channel and K+ channel inhibitors

Transroot osmotic water flux (Jos) and radial hydraulic conductivity (Lpr) in onion roots were greatly increased by three means; infiltration of roots by pressurization, repetition of osmosis and chilling at 5 degrees C. Jos was strongly reduced by the water channel inhibitor HgCl2 (91%) and the K+ channel inhibitor nonyltriethylammonium (C9, 75%), which actually made the membrane potential of root cells less sensitive to K+. C9 decreased the rate of turgor reduction induced by sorbitol solution to the same extent as HgCl2. Thus, C9 is assumed to decrease the hydraulic conductivity (Lp) of the plasma membrane by blocking water channels, although possible inhibition of the plasmodesmata of the root symplast by C9 cannot be excluded. Onion roots transported water from the tip to the base in the absence of the osmotic gradient. This non-osmotic water flux (Jnos) was equivalent to Jos induced by 0.029 M sorbitol. Jnos increased when Jos was increased by repetition of osmosis and decreased when Jos was decreased by either HgCl2 or by C9. The correlation between Jnos and Jos suggests that non-osmotic water transport occurs via the same pathways as those for osmotic water transport.     

The Nature of Water Transport across Frog Skin

A method has been developed for determining simultaneously shortcircuit currents and net water fluxes across frog skin. The basis of the water flux measurement is the determination of changes in weight of a plastic chamber containing the skin and external solution. The accuracy of this method permits net water flows larger than 0.5 mg cm^-2hr.^-1 to be detected, and the apparatus has been used to investigate the relationship between active Na transport and non-osmotic water flow across the skin. Measurement of Na transport and net water influx across completely short-circuited skins provides no good correlation between the two flows. However, skins exhibiting no net water movement in sulfate Ringer displayed an apparent electroosmotic flow of about 40 water molecules per Na ion when depolarizing current densities of 50 and 100 μA cm^-2 are used. It is concluded from this and other evidence that the net water influx across frog skin may be partially electroosmotic in character and that there remains another component of water flow unrelated to active Na transport. A theoretical model, based on irreversible thermodynamics, has been developed to explain the non-osmotic water flow across frog skin.     

The Effects of Temperature on the Exudation Process of Excised Primary Roots of Zea mays

The fluid exudation rates and the ionic compositions of theexudates of excised maize roots have been determined in bathingmedia of 0.1 mM, 1.0 mM, 10.0 mM, and 50.0 mM KCl, each containing0.1 mM CaCl₂, at temperature intervals between 10 °C and30 °C.Analysis of these data in terms of an osmotic modelfor excised root exudation shows that the observed temperaturevariation in fluid exudation rate is accounted for by the observedtemperature variation in the osmotic driving force, the saltconcentration difference from xylem fluid to bathing medium.Temperature variation in the osmotic permeability of the rootand of the non-osmotic water flow are not significantly differentfrom zero.     

Seasonal variations in water relations of roses (Rosa hybrida cv. Sonia)

Parameters of water relations were measured throughout the season in phenologically identical leaves of rose plants grown in a heated greenhouse. The increase in osmotic potential observed from spring to early autumn seems to result from the decrease in the non-osmotic water fraction, and, conversely, its decrease from early autumn to winter seems to be due to an increase in the non-osmotic water fraction. These observations support the hypothesis that the non-osmotic water fraction may be correlated to starch concentration in rose leaves.     

Hydraulic Resistance of Rough Lemon Roots

A pressure chamber technique was used to estimate hydraulic root resistance in rough lemon (Citrus jambhiri Lush.) seedlings. The effect of previous water stress on hydraulic root resistance was evaluated. A factorial 3 × 3 design with four replications was established with potted rough lemon seedlings in a growth chamber. Three water-stress treatments were applied by watering at intervals of 1, 2 and 3 days, and root resistance measurements were made after 6, 12 and 18 days of treatment. Plants that had experienced mild and severe water stress (irrigation interval of 2 and 3 days, respectively) had higher hydraulic root resistances after several drying cycles than those plants irrigated daily. Additional cycles had no significant effect. The increase in root resistance was not due to decreased root growth but apparently to changes in the permeability of the root cell membranes or to increased suberin deposition in the cell walls of the cortical cells. In a short-term experiment (1 h), temperature strongly affected water flow through rough lemon roots in the range 5 to 35°C. Temperature influenced root membrane permeability, since reduced blow could not be explained by changes in water viscosity.     

Reverse flow in roots of Sesbania rostrata measured using the constant power heat balance method

This investigation was performed to examine qualitatively and quantitatively the reverse flow in partially dried roots of Sesbania rostrata using the constant power heat balance method. First, a semi-empirical technique for estimating sheath conductance of sap-flow sensors without assuming that sap flow is zero at night was proposed. Sap flow measured with the heat balance method was compared with water uptake as measured by a potometric method. Sap flow was overestimated by 56·1% for a 3·3-mm-diameter root, and by 40·0% for 6·1 mm and 33·3% for 8·8 mm roots. However, high correlation coefficients between the rates of water uptake and sap flow demonstrated that calibration would provide reliable values for root sap flow. To detect reverse flow, a split root experiment was conducted using a S. rostrata plant with its root system divided between dry and wet compartments. Daily sap flow of the drying compartment declined whereas that in ‘wet’ root increased, suggesting that the decrease in water uptake by ‘dry’ roots was offset by the ‘wet’ roots. Reverse flow was observed at night in the root on the dry side of the container when the soil water potential was less than –0·30 MPa. The total amount of water released into the soil during the night period was estimated to be 22·5 g.     

Cycloheximide inhibits root water flow and stomatal conductance in aspen (Populus tremuloides) seedlings

Aspen (Populus tremuloides Michx.) roots were treated with cycloheximide, a protein synthesis inhibitor, to examine the role of protein synthesis in root water transport and plant water relations. Within less than 30 min following root application, cycloheximide inhibited steady‐state root water flow rates and 1 h after the application of 1 mm cycloheximide, root hydraulic conductivity had decreased by 85% compared with control roots. However, stomatal conductance showed a significant inhibition only after 2 h following cycloheximide treatment. The reduction in root hydraulic conductivity was accompanied by an almost three‐fold increase in the apoplastic water flow ratio as determined by the trisodium 3‐hydroxy‐5,8,10‐pyrenesulphonate tracer dye. Cycloheximide‐treated roots showed a decrease in the immunostaining intensity of a 32 kDa microsomal protein band that immunoreacted with the AnthPIP1; 1 antibody suggesting a decrease in the membrane aquaporin expression. These changes occurred without severe metabolic disruptions as measured by root respiration. The results point to the importance of protein‐mediated transport in roots and the rapidity of response suggests that protein synthesis may be used as a principal regulatory mechanism in root water transport in aspen.     

OsPIN2, which encodes a member of the auxin efflux carrier proteins,is involved in root elongation growth and lateral root formation patterns via the regulation of auxin distribution in rice

Auxin flow is important for different root developmental processes such as root formation, emergence, elongation and gravitropism. However, the detailed information about the mechanisms regulating the auxin flow is less well understood in rice. We characterized the auxin transport‐related mutants, Ospin‐formed2‐1 (Ospin2‐1) and Ospin2‐2, which exhibited curly root phenotypes and altered lateral root formation patterns in rice. The OsPIN2 gene encodes a member of the auxin efflux carrier proteins that possibly regulates the basipetal auxin flow from the root tip toward the root elongation zone. According to DR5‐driven GUS expression, there is an asymmetric auxin distribution in the mutants that corresponded with the asymmetric cell elongation pattern in the mutant root tip. Auxin transport inhibitor, N‐1‐naphthylphthalamic acid and Ospin2‐1 Osiaa13 double mutant rescued the curly root phenotype indicating that this phenotype results from a defect in proper auxin distribution. The typical curly root phenotype was not observed when Ospin2‐1 was grown in distilled water as an alternative to tap water, although higher auxin levels were found at the root tip region of the mutant than that of the wild‐type. Therefore, the lateral root formation zone in the mutant was shifted basipetally compared with the wild‐type. These results reflect that an altered auxin flow in the root tip region is responsible for root elongation growth and lateral root formation patterns in rice.     

Effect of Hydrotropism on Root System Development in Soybean (Glycine max): Growth Experiments and a Model Simulation

To observe root system development, soybean plants (Glycine max) were grown in root boxes that were set horizontally to reduce the effect of gravity. Along with the root system development, the two-dimensional distribution of soil water content in the root boxes was measured continuously by the time domain reflectometry (TDR) method. Root system development and its morphological architecture were strongly affected by the positions of the water supply. It is suggested that root hydrotropism plays the dominant role in root system development. In addition to root hydrotropism, the importance of root compensatory growth is suggested. A combined model of root system development and soil water flow considering root hydrotropism and compensatory growth was used to simulate root system development and soil water flow. The morphological architecture of root systems and the distribution of soil water content obtained in the experiment were successfully explained by the model simulation. These results confirmed that root hydrotropism and compensatory growth are dominant factors in root system development under a reduced effect of gravity. The validity of the model was confirmed, and its applications for various purposes were suggested.     

Effect of Hydrotropism on Root System Development in Soybean ( Glycine max): Growth Experiments and a Model Simulation

To observe root system development, soybean plants (Glycine max) were grown in root boxes that were set horizontally to reduce the effect of gravity. Along with the root system development, the two-dimensional distribution of soil water content in the root boxes was measured continuously by the time domain reflectometry (TDR) method. Root system development and its morphological architecture were strongly affected by the positions of the water supply. It is suggested that root hydrotropism plays the dominant role in root system development. In addition to root hydrotropism, the importance of root compensatory growth is suggested. A combined model of root system development and soil water flow considering root hydrotropism and compensatory growth was used to simulate root system development and soil water flow. The morphological architecture of root systems and the distribution of soil water content obtained in the experiment were successfully explained by the model simulation. These results confirmed that root hydrotropism and compensatory growth are dominant factors in root system development under a reduced effect of gravity. The validity of the model was confirmed, and its applications for various purposes were suggested.     

Translocation of nitrogen in osmotically stressed wheat seedlings

Wheat (Triticum aestivum L., cv. Drabant) seedlings were grown in a ‘split root’ system where either the whole root system or one root half was subjected to osmotic stress for 24 h, using 200 g polyethylene glycol (PEG, molecular weight 4000) dm^?3 nutrient solution. ¹⁵N-Labelled nitrate was fed to one of the root compartments and total N and ¹⁵N-labelling were measured in plant material and xylem sap. Untreated plants translocated 87% of the N taken up to the shoot, and 10% of this was then retranslocated back to the root. Recalculated on a root nitrogen basis, 36% of the label recovered in the root after 24 h had passed through the shoot. Significant labelling of xylem sap collected from non-labelled roots indicated cycling of organic N through the roots. PEG-treatment of the whole root system caused significant water loss in both roots and shoots. Uptake of nitrate and retranslocation of N to roots were inhibited, whereas cycling of organic nitrogen through the root was still measurable. Treatment of half the root system with PEG had minor effects on shoot water content, but reduced the water content of the treated root part. The total uptake of nitrate by the root system was unaffected, and the effect on the treated root half was comparatively small. Nitrate reductase activity (NRA) declined in PEG-treated roots even if high nitrate uptake rates were maintained. Shoot NRA was unaffected by osmotic stress. The data indicate that the reduction in water content of the root per se has only small effects on nitrate uptake. Major inhibition of nitrate uptake was observed only after treatment of a sufficiently large portion of the root system to given an effect on shoot water content.     

Root water uptake by kiwifruit vines following partial wetting of the root zone

Rates of sap flow and root-water uptake by two 7-year old kiwifruit vines (Acinidia deliciosa) were studied in an orchard with the aim of determining the ability of the vines to alter their spatial pattern of root-water uptake following differential wetting of the root zone. Time-domain reflectometry (TDR) was used to monitor changes in the soil's volumetric water content, . The heat-pulse technique was used to monitor sap flow not only in the stem but also in several large roots to see how root flow responded with local changes in soil water availability. Prior to irrigation there was a broad correspondence between the pattern of water uptake and the distribution of root-length density. However, following irrigation, we observed a preferential uptake of water from the wetter parts of the soil and a corresponding decline in water uptake from the drier parts of the soil. Observations of root uptake by TDR following irrigation also revealed the inordinate activity of near-surface roots. The vine would preferentially draw upon near-surface water if it were available. Kiwifruit vines are able to shift rapidly their pattern of uptake, in a matter of days, away from drier parts of the root zone and begin to extract water preferentially from those regions where it is more freely available. Upon full wetting of the root zone, previously inactive roots in the dry soil of the root zone were quickly able to recover their activity. Indeed their activity following rewatering was found to be greater than it had been prior to the period of soil dryness. A rapid flush of new root growth is considered to be the mechanism that leads to this enhanced activity.     

Estimation of hydraulic conductance within field-grown apricot using sap flow measurements

Using the heat pulse and other techniques, the hydraulic architecture of apricot trees was mapped out. The flows (overall flow, flow across the four main branches) and forces (water potential differences between xylem and leaves) measured allowed us to quantify hydraulic conductance of branches and of the root/soil resistance. The experiment was carried out in a commercial orchard of 11-year-old apricot trees (Prunus armeniaca L., cv. Búlida, on Real Fino apricot rootstock) during 1 week (October 27–November 3, 1998). Three representative trees with a cylindrical trunk divided into four main branches of different sizes, orientation and local microclimate were chosen for the experiment. Sap flow was measured throughout the experimental period. Twelve sets of heat-pulse probes were used, one for each main branch. The diurnal course of the environmental conditions, the fraction of the area irradiated and leaf water relations were also considered in each main branch. The relationships between leaf water potential, xylem water potential and transpiration were established for different branches and also for the total plant. Using the slopes of these regressions, total plant conductance, the hydraulic conductance of the stem and root pathway, the hydraulic conductance of the canopy and the hydraulic conductance of each branch were estimated. Our findings show that the root conductance and the canopy hydraulic conductance are similar in magnitude. Leaf hydraulic conductance per leaf area unit was similar for each of the four branch orientations, indicating that, while the light microclimate has a dominant influence on transpiration, in this case it had little effect on the hydraulic properties of the canopy.     

Stomatal closure with soil water depletion not associated with changes in Bulk leaf water status

Summary It has previously been reported that canopy water loss by cowpea (Vigna unguiculata) decreases with small depletions in soil water. In these studies, under field conditions, it was demonstrated that with small changes in soil water status leaf conductance of cowpea decreases in a manner which is consistent with the sensitive regulation of canopy water loss.However, treatments which differed in leaf conductance, and presumably stomatal aperture, had similar leaf water potentials. It is hypothesized that the stomatal closure which results from soil water depletion is mediated by changes in root water status through effects on the flow of information from root to shoot. An efficient mechanism of this type could be partially responsible for the extreme drought avoidance exhibited by this plant.Dedicated to Dr. K. Springer     

MAIN ROOT-LATERAL ROOT JUNCTIONS OF TWO DESERT SUCCULENTS: CHANGES IN AXIAL AND RADIAL COMPONENTS OF HYDRAULIC CONDUCTIVITY DURING DRYING

The influence of junctions between main roots and lateral roots on water flow was investigated for the desert succulents Agave deserti and Ferocactus acanthodes during 21 d of drying in soil. Under wet conditions, the junctions did not restrict xylem water flow from lateral roots to main roots, consistent with predictions of axial conductance based on vessel diameters. Embolism caused by drying reduced such axial conductance more at the junctions than in adjoining root regions. Connective tracheary elements at the junctions were abundantly pitted and had large areas of unlignified primary wall, apparently making them more susceptible to embolism than vessels or tracheids elsewhere in the roots. Unlike the decrease in axial conductance, the overall hydraulic conductivity of the junction increased during drying because of an increase in the conductivity of the radial pathway. Despite such increases, main roots may not lose substantial amounts of water to a dry soil during drought, initially because embolism at the junctions can limit xylem flow and later because soil hydraulic conductivity decreases. Moreover, the increased root hydraulic conductivity and a potentially rapid recovery from embolism by connective tracheary elements may favor water uptake near the junctions upon soil rewetting.     

Flow sorting of wheat chromosome arms from the ditelosomic line 7BL

Flow cytometric analysis confirmed that root tip cells can be synchronized with 1.25 mM hydroxyurea (DNA synthesis inhibitor) for 12 h and 1 μM trifluralin (metaphase blocking reagent) treatment for 5 h. Chromosome suspensions prepared from homogenized tissue were suitable for chromosome sorting. A flow karyotypic histogram showed that the genome of common wheat (Triticum aestivum L.) ‘Chinese Spring’ was divided into 4 chromosome peaks, but the 7BL ditelosomic line had an additional chromosome peak. PCR amplification of sorted chromosome arms indicated that the extra chromosome peak consisted of 7BL telosomics. Some technical details of sample preparation and parameter setting for flow cytometric analysis are described.     

Comparative assessment of the sensitivity of oilseed rape and wheat to limited water supply

The drought‐sensitivity of oilseed rape (OSR, Brassica napus cv. SW Landmark) was investigated, using the more widely studied crop species wheat (Triticum aestivum cv. Tybalt) as a benchmark. The water relations of OSR and wheat were compared in lysimeter and controlled environment experiments to test the hypothesis that the growth of OSR is restricted to a greater extent by soil drying than wheat and to determine whether the greater sensitivity results from differences in root or shoot traits. Plants were grown, with or without irrigation, in 1.2 m tall lysimeters packed with a sandy clay loam soil. The experiment was conducted in an open‐sided glasshouse to encourage air flow and to resemble a field environment as far as possible; plant population densities were equivalent to commercial crops. Irrigated OSR (evapo)transpired more water than wheat (498 vs. 355 mm), but had a comparable water use efficiency (WUE; 4.1 vs. 4.4 g DW mm^?1 H₂O). Oilseed rape showed a greater reduction in above‐ground growth (52% vs. 21%) and a smaller increase in WUE (27% vs. 45%) when water was withheld. Oilseed rape also responded to soil drying at a lower soil moisture deficit than wheat; transpiration rates fell below the potential of irrigated plants when plant available water remaining in the soil profile declined from 54 to 23% compared to 38 to 9% for wheat. The root hydraulic conductivity of young OSR plants, measured on root surface area basis, was about twice that of wheat, and was comparable on a root length basis. The results show that OSR was more sensitive to a restricted water supply than the benchmark species wheat and that the greater sensitivity resulted from differences in shoot, rather than root, characteristics. The root system of OSR was at least as efficient as wheat at extracting water from soil.     

Evidence supporting a non-phloem source of water for export of solutes in the xylem of soybean root nodules

The vascular anatomy of soybean nodules [Glycine max (L.) Merr.] suggests that export of solutes in the xylem should be dependent on influx of water in the phloem. However, after severing of stem xylem and phloem by shoot decapitation, export of ureides from nodules continued at an approximately linear rate for 5h. This result was obtained with decapitated roots remaining in the sand medium, but when roots were disturbed by removal from the rooting medium prior to shoot decapitation, export of ureides from nodules was greatly reduced. Stem exudate could not be collected from disturbed roots, indicating that flow in the root xylem had ceased. Thus, ureide export from nodules appeared to be dependent on a continuation of flow in the root xylem. When seedlings were fed a mixture of ³H₂O and ¹⁴C-inulin for periods of 14–21 min, nodules had higher ³H/¹⁴C ratios than roots from which they were detached. The combined results are not consistent with the proposal that export of nitrogenous compounds from nodules is dependent on import of water via the phloem. The results do support the view that a portion of the water required for xylem export from soybean nodules is supplied via a symplastic route from root cortex to nodule cortex to the nodule vascular apoplast.