Probing short-term root exudation in Zea mays

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

A model for water transport in the stele of plant roots

Summary The mechanism of water movement across roots is, as yet, not well understood. Some workable black box theories have already been proposed. They, however, assumed unrealistic cell membranes with low values of , or were based on a poor anatomical knowledge of roots. The role of root stele in solute and water transport seems to be especially uncertain. An attempted explanation of the nature of root exudation and root pressure by applying the apoplast canal theory (Katou andFurumoto 1986 a, b) to transport in the root stele is given. The canal equations are solved for boundary conditions based on anatomical and physiological knowledge of the root stele. It is found that the symplast cell membrane, cell wall and net solute transport into the wall apoplast are the essential constituents of the canal system. Numerical analysis shows that the canal system enables the coupled transport of solutes and water into a xylem vessel, and the development of root pressure beyond the level predicted by the osmotic potential difference between the ambient medium and the exudate. Observations on root exudation and root pressure previously reported seem to be explained quite well. It is concluded that the movement of water in the root stele although apparently active is essentially osmotic.Abbreviations J _v ^ex volume exudation per root surface - J₀ non-osmotic exudation - L^r overall radial hydraulic conductivity of an excised root - reflection coefficient - C_s difference in the osmotic concentration between the bathing medium and the exudate - R gas constant - T absolute temperature - C_K molar concentration of K⁺ - C_Cl molar concentration of Cl^– - C_j molar concentration of ion species j - P_j membrane permeability of ion j - z_j valence of ion j - F Faraday constant - V_ix intracellular electric potential with reference to the canal     

Short-term responses of Brassica rapa plants to low root temperature: Effects on nitrate uptake and its translocation to the shoot

Brassica rapa L. plants were grown hydroponically for 5 or 6 weeks at 20°C and then half batches of plants were transferred to tanks in which the root temperature was lowered decrementally over 1 h to 7°C. Changes in nitrate uptake rate (NUR) and nitrate transfer from roots were studied in relation to transpiration and root pressure xylem exudation flow rates over a 48- or 72-h period. The response of plants following the root temperature decrease was biphasic. During phase 1, NUR and water and solute flow rates through the root decreased sharply. Coping mechanisms came into operation during phase 2, and tended to offset the effects of low temperature. The 3-h cold-treated roots exhibited a very low NUR but 48-h cold-treated roots partly recovered their ability to absorb nitrate. Transpiration rate decreased more slowly (during 24 h) than both root xylem exudation and parameters of root conductivity (during 6 h). Beyond these respective times, transpiration rate was balanced while root xylem exudation clearly increased, but without returning to the level of control plants. Nitrate transfer to the root xylem was strongly and rapidly affected by low root temperature, but the subsequent readjustment was such that no or little difference compared with the control was apparent after 48 h. Water and solute flows were strongly decreased when nitrate was replaced by chloride in the culture solution during exudation sampling. The major role of nitrate in root hydraulic conductivity and root xylem exudation is discussed.     

Observations on the Exchange of Salt Between the Xylem and Neighbouring Cells in Zea mays Primary Roots

Comparison of two preparations of Zea mays primary roots hasshown that the salt concentration in the xylem fluid is alteredas it passes through a region of the root where no absorptionof salts or water from the external bathing medium can occur.Salts are secreted to the xylem by such a region, presumablyfrom the vacuoles of the cortical cells. It has also been demonstratedthat longitudinal water movement through the root cortex isnegligibly small during root-pressure exudation.     

A Correlation between Structure and Function in the Root of Zea mays

The exudation rates of fluid and potassium ions from isolatedmaize roots were determined before and after excision of certainlengths of root tip. The results of this study suggest thatexcised maize roots possess the ability to absorb potassium(and presumably chloride) ions and concomitant amounts of waterover a considerable distance (10 cm) from the tip. Moreover,the observed power of absorption of ions and water into thetranslocatory pathway decreases in passing from the tip towardsthe base of the root. Both light and electron microscope techniques were used to examinethe anatomy of primary roots similar to those used in the physiologicalexperiments. The principal observation was that the xylem vesselsnear the root tip contain membrane-bounded cytoplasm with organelles.The number of mature xylem vessels, i.e. without cytoplasm,progressively increased in transverse sections cut from 1 to10 cm from the root tip; above 10 cm from the root tip all ofthe xylem vessels were found to be completely mature. It isevident that prima facie a connexion exists between this singleaspect of root anatomy and fluid exudation from excised roots. The uptake of tritiated water by roots and its transport intoexudates was examined. These data were analysed on the assumptionthat the exchange of external labelled water with the exudatewas achieved by the fluid exudation itself; this analysis indicatedthat an operational volume, similar to that of the total xylemvolume within the root, must become labelled during the formationof the exudate.     

Der Einfluss verschiedener Salze und verschiedener Inhibitoren auf den Wurzeldruck von Zea mays

The effect of different salts and inhibitors on the root pressure of Zea mays.— The influence of various salt solutions and inhibitors on the exudation rate has been investigated with young excised primary roots of Zea mays. The following results were obtained. — The effect of chlorides on the exudation rate was higher than the effect of sulphates K⁺ and Na⁺ effected higher flux rates than Ca²⁺ and Mg²⁺ The highest exudation rate was obtained with KCl. — In comparison to an isotonic Lutrol-solution (a liquid condensed polyethylenoxid) a 0.5525 molar KCl-solution, applicated on the root stump, increased the exudation rate considerably. — Metabolic inhibitors and anaerobic conditions decreased the exudation rate. — Experiments, concerning the influence of metabolic inhibitors on the exudation and on the Rb-uptake showed a highly significant positive correlation (r =+0.72***) between the exudation rates and the Rb-concentrations in the exudates. The Rb-accumulation in the root tissue was not correlated to the exudation rate. — The experimental data agree with the concept of a transversal water transport in the root tissue, effected by osmotic forces. The root pressure is based on the osmotic gradient between the xylem sap and the outer solution. This gradient is built up by the metabolic secretion of ions into the xylem sap. It is supposed that the transversal water transport in the roots mainly goes through the free space of the cortex.     

A model for radial water transport across plant roots

Summary A model based on the canal theory (Katou andFurumoto 1986 a, b) is proposed for the absorption of solute and water at the root periphery. The present canal model in the periphery and the model which was previously proposed for the exudation in the stele (Katou et al. 1987), are organized into a model for radial transport across excised plant roots, in the light of anatomical and physiological knowledge of maize roots. The canal equations for both canals are numerically solved to give quite a good explanation for the observed exudation of maize roots. It is found that the regulation of solute transport has a primary importance in the regulation of water transport across excised roots. The internal cell pressure of the symplast adjusts the water absorption at the root periphery to the water secretion into the vessels. There seems no need for this explanation of the radial water transport across roots to assume cell membranes with low reflection coefficient or variable water permeability. It would seem that the apoplast wall layers play a crucial role in metabolic control of water transport in roots as well as in hypocotyls.Abbreviations J _s ^ex* the theoretically estimated rate of solute exudation per unit surface area of model maize roots - J that of volume exudation per unit surface area of model maize roots - the reflection coefficient of the cell membrane against solutes     

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.     

Abscisic Acid promotes both volume flow and ion release to the xylem in sunflower roots

The effect of abscisic acid (ABA) on the exudation process in excised sunflower root was investigated. A promotion of both ion flux and volume exudation rate was observed. Cutting off the external supply of ions to the roots markedly increased the magnitude of the ABA effect. The promotive effect of ABA on exudation rate was extremely rapid (less than 6 minutes) and seemed to be biphasic. Reasons are given for relating the ABA effect to two separate actions, namely: (a) on water permeability; and (b) on the release of ions both from vacuoles to the cytoplasm and from symplasm to the xylem. Kinetin inhibited movement of ions to the xylem and its effect did not appear to be related to that of the ABA.     

Effects of synthetic plant growth retardants and abscisic acid on root functions of Brassica rapa plants exposed to low root-zone temperature

Possible interactions of two synthetic plant-growth retardants during the short-term response of Brassica rapa L. ssp. oleifera (DC.) Metzger plants to low root-zone temperature were investigated by pretreating with mefluidide or paclobutrazol. Water and solute transfers were studied by measuring xylem sap volume flow (under root pressure exudation) and ion flow from the roots. Relations with nitrate uptake rate were also considered. Root pretreatment with paclobutrazol strongly restricted the cold-inducible processes which normally restore water and solute flow from the root xylem. Paclobutrazol decreased the rates of nitrate uptake and exudation flow from the root xylem (principally by reducing root hydraulic conductivity) with dramatic consequences for ion flow, especially that of nitrate.
The effects of root ABA pretreatment on plant response to root cooling were then studied separately or in association with a pretreatment with paclobutrazol. Despite a slight decrease in nitrate uptake rate, ABA pretreatment of the roots enabled the plant to develop rapid mechanisms for adaptation to cold constraint at the root level. Moreover, this action of exogenous ABA greatly reduced the effect of a simultaneous paclobutrazol pretreatment and partly restored water and solute flows.
Thus, the improvement of plant resistance to cold conditions brought about by treatments with mefluidide and paclobutrazol (previously shown in long-term experiments) cannot simply be explained by their short-term effects.     

Potassium transport in salt-stressed barley roots

Salinization of the medium inhibits both K⁺ uptake by excised barley (Hordeum vulgare L.) roots and K⁺ release from their stele, as measured by short-term ⁸⁶Rb uptake and xylem exudation, respectively. Although inhibition was not specific to chloride, mannitol caused a different response from that of inorganic sodium salts, indicating that inhibition was at least partly the result of an ion effect. In roots previously exposed to low levels of NaCl, NaCl stress directly affected stelar K⁺ release, whereas in low-sodium roots stelar K⁺ release was much less salt-sensitive than K⁺ uptake.Abbreviation chCl choline chloride     

Are developing xylem vessels the site of ion exudation from root to shoot?

Abstract This paper describes experiments to test the suggestion that developing xylem vessels are the site of exudation of ions from the root to the shoot. Electron microscopy is used to define the stage of development of xylem vessels in young barley roots along the length of the root. The amino acid analogue p-fluorophenyl-alanine (FPA) is used to inhibit ion transport from the stele to the xylem vessels at varied distances from the apex. In the presence of FPA protein synthesis is not inhibited but ineffective proteins are formed. It is shown that exudation of Cl^? from the root can be inhibited in this way in parts of the root where all the xylem vessels are mature. This is in contradiction to the suggestion that root exudation is due to the activity of developing vessels. The hypothesis is thus strengthened that ion transport proceeds into the xylem vessels, which are fully mature and devoid of cytoplasm, and is due to release from the xylem parenchyma cells.     

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.     

The influence of temperature on the exudation of xylem sap from detached root systems of rye (Secale cereale) and barley (Hordeum vulgare)

Summary Roots of intact plants of rye and barley which had been growing at 20° were cooled for 12–72 h at 8–14° C while the shoots were kept at 20°. The roots were then excised and placed in solutions at temperatures ranging from 2.5–22.5° C. The rate of exudation of xylem sap and the chemical composition and osmotic potential of the sap were measured and compared with controls which had been kept at 20° C during the pretreatment period. Pre-cooling increased the fluxes of K⁺, Ca²⁺ and H₂PO ₄ ^- into the xylem sap of both species by factors of two to three; the total volume of exudate rose by larger factors. Thus the concentrations of these ions were lower in the sap exuding from cooled roots than in that from controls. Measurements of the osmotic potential of the sap from barley roots indicated that the osmotic driving force in cooled and control roots was similar even though flow in the former was much greater.The enhancement of exudation was shown to be dependent on the duration and the temperature experienced by the roots during pretreatment, and was lost rapidly when roots of intact plants were returned to 20°.Analysis of the temperature coefficients for exudation and Arrhenius plots revealed very distinct changes in the activation energy for exudation above and below a transition temperature. In control plants of barley and rye this temperature was around 10° C, but in cooled roots of rye there was a significant shift in the transition temperature to 5° C. Activation energies for exudation of control and cooled roots above or below the transition temperature were broadly similar, thus pre-cooling roots did not alter the temperature sensitivity of exudation but merely its rate at a given temperature.The results are discussed in relation to active ion transport, membrane fluidity and the resistance of the root to water flow.Abbreviation ABA abscisic acid     

Does hydraulic lift exist in shallow-rooted species? A quantitative examination with a half-shrub Gutierrezia sarothrae

Hydraulic lift occurs in some deep-rooted shrub and herbaceous species. In this process, water taken up by deep roots from the moist subsoil is delivered to the drier topsoil where it is later reabsorbed by shallow roots. However, little is known about the existence of hydraulic lift in shallow-rooted xeric species. The objectives of this study were 1) to ascertain whether hydraulic lift exists in Gutierrezia sarothrae (broom snakeweed), a widespread North American desert species with a shallow root system, grown in pot and field conditions and 2) if it does, how much water can be transferred from the subsoil to the 30 cm topsoil during the night. Snakeweed seedlings were transplanted in buried pots allowing the deeper roots to grow into the subsoil 30 cm below the surface. Soil water content inside and outside of the pot was measured seasonally and diurnally with time domain reflectometry technique (TDR). An increase in water content was detected in the pot after the plant was covered for 3 h by an opaque plastic bag during the day, suggesting hydraulic lift from deeper depths and exudation of water into the drier topsoil. Root exudation was also observed on native range sites dominated by snakeweed. Water efflux in the pot was 271 g per plant per night. which was equivalent to 15.3% of the extrapolated, porometer-derived whole-plant daily transpiration. Hydraulic lift observed in Gutierrezia improved water uptake during the day when evaporative demand is high and less water is available in the topsoil. We concluded that hydraulic lift might help snakeweed to alleviate the effect of water stress.     

Stimulatory effects of adrenalin and noradrenalin on root water-pumping activity and the involvement of G-proteins

To evaluate the involvement of G-proteins in the signal transduction during stimulatory action of neurotransmitters, adrenalin and noradrenalin, on root exudation and the ivolvement of G-proteins in water transport in the root and creation of the root pressure, we tested the effects of guanosinethiodiphosphate, an inhibitor of GTP-binding capacity of G-proteins, and guanosinethiotriphosphate, a stimulator of this capacity. Experiments were performed with detached roots of 5–7-day-old maize (Zea mays L.) seedlings and the mittens produced from them due to the removal of the vascular cylinder. The latter are a convenient model to study the nature of the root pressure due to its strongly limited possibility to function as an osmometer during the early step of exudation. In the “mittens,” adrenalin and noradrenalin enhanced exudation, increased its temperature coefficient (Q₁₀), root pressure, and its metabolic component much stronger than in detached roots with the vascular cylinder retained (conventionally named as “intact” roots). In control treatment (with water), guanosinethiodiphosphate retarded exudation on the average by 30% in intact roots and by 50% in mittens, simultaneously reducing Q₁₀ from 3.0 to 1.7 in intact roots and from 4.0 to 1.3 in mittens. Guanosinethiotriphosphate exerted an opposite action: it stimulated exudation on the average by 30% in intact roots and by 60% in mittens; the Q₁₀ value increased from 3.0 to 3.6 in intact roots and from 4.0 to 5.8 in mittens. These data indicate that G-proteins are involved in the control of water transport and creation of the root pressure (without any other treatments). Guanosinethiodiphosphate neutralized completely adrenalin-and noradrenalin-induced stimulation of exudation, resulting in the level substantially below the control one, especially in mittens. Guanosinethiotriphosphate enhanced stimulatory effects of both neurotransmitters, mainly in mittens, whereas its effect on intact roots was relatively weak, especially in experiments with noradrenalin. It should be emphasized that the mittens responded to both neurotransmitters and the regulators of G-protein activity much stronger than intact roots. The data obtained argue for the G-protein involvement in (1) transduction of adrenalin and noradrenalin signals stimulating root water-pumping activity and (2) the control of water transport and creation of the root pressure under normal conditions. Experiments with mittens indicate that this G-protein involvement could by mainly related to the functioning of the root cortex parenchymal cells and the formation of the metabolic component of the root pressure.     

Effect of nitrate on water transfer across roots of nitrogen pre-starved maize seedlings

The addition of 10 mM KNO₃ to the solution bathing the roots of young nitrogen-starved seedlings of Zea mays L. enhanced root water transfer within 15 h, compared with 10 mM KCl addition. The free exudation flux was 2.2–3.9 times higher in excised KNO₃-treated roots than in KCl-treated ones. Cryo-osmometry data for xylem sap suggested that, compared with chloride, nitrate treatment increased the steady solute flux into the xylem, but did not modify the osmotic concentration of sap. Root growth was not significantly modified by nitrate within 15 h. Root hydraulic conductances were measured by using either hydrostatic-pressure or osmotic-gradient methods. During hydrostatic experiments, the conductance (k_p), which is thought to refer mainly to the apoplasmic pathway, was 1.6 times larger in KNO₃-than in KCl-treated plants. From experiments in which polyethylene glycol (PEG) 8000 was used as external osmolyte, osmotic conductances (k_s) were found to be smaller by 5–20 times than k_p for the two kinds of plants. The KCl-treated roots were characterized by a low k_s which was the same for influx or efflux of water. By contrast, KNO₃-treated roots exhibited two distinct conductances k_s1 and k_s2, indicating that influx of water was easier than efflux when the water flow was driven by the osmotic pressure gradient. Infiltration of roots with KNO₃ solution supported the idea that nitrate might enhance the efficiency of the cell-to-cell pathway. The low k_s value of KCl-treated roots and the existence of two contrasting k_s values (k_s1 and k_s2) for KNO₃-treated roots are discussed in terms of reversible closing of water channels.     

Effects of defoliation and atmospheric CO2 depletion on nitrate acquisition,and exudation of organic compounds by roots of Festuca rubra

The aim of this study was to investigate the physiological basis of increased root exudation from Festuca rubra, in response to defoliation. The hypothesis, that assimilate supply to roots is a key determinant of the response of root exudation to defoliation was tested by imposing CO₂-deplete (< 50 mol mol^–1) atmospheres to F. rubra. This was done as a non-destructive means of preventing supply of new assimilate to roots of intact and defoliated plants. F. rubra was grown in axenic sand systems, with defoliation and CO₂-depletion treatments applied to plants at 14 and 35 days after planting. Root exudation and NO₃ ^– uptake were quantified throughout, and post-treatment uptake and allocation of N were determined from the distribution of ¹⁵N label, supplied as ¹⁵NO₃ ^–. Defoliation of F. rubra resulted in significantly (P <0.01) increased root exudation, CO₂-depletion did not result in increased exudation from plants of either age. When treatments were applied to F. rubra after 14 days, defoliation and CO₂-depletion each reduced NO₃ ^– uptake significantly (P <0.05). However, in older plants, uptake of NO₃ ^– was less sensitive to defoliation and CO₂-depletion. The results indicate that increased root exudation following defoliation is not related directly to reduced assimilate supply to roots. This was evident from the lack of effect of CO₂-depletion on root exudation, and the absence of correlation between root-C efflux and the rate of NO₃ ^– uptake. The physiological basis of increased exudation following defoliation remains uncertain, but may be dependent on physical damage, either directly or as a consequence of systemic responses to wounding.     

Defoliation alters the relative contributions of recent and non-recent assimilate to root exudation from Festuca rubra

The deposition of organic compounds from plant roots is a key determinant of rhizosphere microbial activity and community structure. Consequently, C-flow from roots to soil is an important process in coupling plant and microbial productivity, via impacts on microbial nutrient cycling in soil. Experimentally, isotopic tracers (¹³C or ¹⁴C) are used to track C inputs to soil and microbial communities. However, in many such studies the relationship between labelled C-flows and total C-flows are not established, limiting the interpretative value of the results. In this study, we applied steady-state near natural abundance ¹³CO₂ labelling to determine the impact of partial defoliation of Festuca rubra on root exudation. This approach in axenic culture facilitated determination of the contribution of pre- and post-defoliation assimilates both to root C-flow and plant tissues. The results demonstrated that total root exudation was increased in the two days following defoliation. This was concurrent with reduced net CO₂ assimilation and reduced allocation of post-defoliation assimilates below-ground and to active root meristems. Through determination of the δ¹³C of root exudates, it was established that the source of the increased root exudation was pre-defoliation assimilate. However, this response was transient, with reduced deposition of pre- and post-defoliation assimilates from roots during the period 2–4 d following defoliation. The results highlight the limitations of pulse-labelling approaches as a means of quantifying impacts of treatments on root exudation, particularly where the treatment is likely to affect plant C-partitioning or the balance between deposition to, and re-mobilization from, C-storage pools.     

Na^＋ and Water Uptake in Relation to the Radial Reflection Coefficient of Root in Arrowleaf Saltbush Under Salt Stress

The response of halophyte arrowleaf saltbush (Atriplex triangularis Willd) plants to a gradient of salt stress were investigated with hydroponically cultured seedlings. Under salt stress, both the Na+ uptake into root xylem and negative pressures in xylem vessels increased with the elevation of salinity (up to 500 mol/m3) in the root environment. However, the increment in negative pressures in root xylem far from matches the decrease in the osmotic potential of the root bathing solutions, even when the osmotic potential of xylem sap is taken into consideration. The total water potential of xylem sap in arrowleaf saltbush roots was close to the osmotic potential of root bathing solutions when the salt stress was low, but a progressively increased gap between the water potential of xylem sap and the osmotic potential of root bathing solutions was observed when the salinity in the root environment was enhanced. The maximum gap was 1.4 MPa at a salinity level of 500 mol/m3 without apparent dehydration of the tested plants. This discrepancy could not be explained with the current theories in plant physiology. The radial reflection coefficient of root in arrowleaf saltbush decreased with the enhanced salt stress was and accompanied by an increase in the Na+ uptake into xylem sap. However, the relative Na+ in xylem exudates based on the corresponding NaCl concentration in the root bathing solutions showed a tendency of decrease. The results showed that the reduction in the radial reflection coefficient of roots in the arrowleaf saltbush did not lead to a mass influx of NaCl into xylem when the radial reflection coefficient of the root was considerably small; and that arrowleaf saltbush could use small xylem pressures to counterbalance the salt stresses, either with the uptake of large amounts of salt, or with the development of xylem pressures dangerously negative. This strategy could be one of the mechanisms behind the high resistance of arrowleaf saltbush plants to salt stress.