A trans-zeatin riboside in root xylem sap negatively regulates adventitious root formation on cucumber hypocotyls

Shoot cultures of cucumber were used to analyse the roles of root-derived substances in adventitious root formation on hypocotyl tissues. Xylem sap collected from the roots of squash had a strong inhibitory effect on the formation of hypocotyl adventitious roots. Double-solvent extraction followed by fractionation with both normal and reverse phase column chromatographies and analysis by liquid chromatography/tandem mass spectrometry identified trans-zeatin riboside (ZR) as the primary suppressor of adventitious root formation. ZR was the predominant cytokinin present in the xylem sap, occurring at a concentration of 2x10(-8 )M. Application of ZR at concentrations from 3.16x10(-9) M effected inhibition of adventitious root formation. These results suggest that ZR transported from roots via xylem sap may act as an endogenous suppressor of hypocotyl adventitious root formation in planta.     

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.     

Perturbation of polyamine catabolism can strongly affect root development and xylem differentiation

Spermidine (Spd) treatment inhibited root cell elongation, promoted deposition of phenolics in cell walls of rhizodermis, xylem elements, and vascular parenchyma, and resulted in a higher number of cells resting in G(1) and G(2) phases in the maize (Zea mays) primary root apex. Furthermore, Spd treatment induced nuclear condensation and DNA fragmentation as well as precocious differentiation and cell death in both early metaxylem and late metaxylem precursors. Treatment with either N-prenylagmatine, a selective inhibitor of polyamine oxidase (PAO) enzyme activity, or N,N(1)-dimethylthiourea, a hydrogen peroxide (H(2)O(2)) scavenger, reverted Spd-induced autofluorescence intensification, DNA fragmentation, inhibition of root cell elongation, as well as reduction of percentage of nuclei in S phase. Transmission electron microscopy showed that N-prenylagmatine inhibited the differentiation of the secondary wall of early and late metaxylem elements, and xylem parenchymal cells. Moreover, although root growth and xylem differentiation in antisense PAO tobacco (Nicotiana tabacum) plants were unaltered, overexpression of maize PAO (S-ZmPAO) as well as down-regulation of the gene encoding S-adenosyl-l-methionine decarboxylase via RNAi in tobacco plants promoted vascular cell differentiation and induced programmed cell death in root cap cells. Furthermore, following Spd treatment in maize and ZmPAO overexpression in tobacco, the in vivo H(2)O(2) production was enhanced in xylem tissues. Overall, our results suggest that, after Spd supply or PAO overexpression, H(2)O(2) derived from polyamine catabolism behaves as a signal for secondary wall deposition and for induction of developmental programmed cell death.     

Flowering as a condition for xylem expansion in Arabidopsis hypocotyl and root

In dicotyledons, biomass predominantly represents cell-wall material of xylem, which is formed during the genetically poorly characterized secondary growth of the vasculature. In Arabidopsis hypocotyls, initially proportional secondary growth of all tissues is followed by a phase of xylem expansion and fiber differentiation. The factors that control this transition are unknown. We observed natural variation in Arabidopsis hypocotyl secondary growth and its coordination with root secondary growth. Quantitative trait loci (QTL) analyses of a recombinant inbred line (RIL) population demonstrated separate genetic control of developmentally synchronized secondary-growth parameters. However, major QTL for xylem expansion and fiber differentiation correlated tightly and coincided with major flowering time QTL. Correlation between xylem expansion and flowering was confirmed in another RIL population and also found across Arabidopsis accessions. Gene-expression analyses suggest that xylem expansion is initiated after flowering induction but before inflorescence emergence. Consistent with this idea, transient activation of an inducer of flowering at the rosette stage promoted xylem expansion. Although the shoot was needed to trigger xylem expansion and can control it in a graft-transmissible fashion, the inflorescence stem was not required to sustain it. Collectively, our results suggest that flowering induction is the condition for xylem expansion in hypocotyl and root secondary growth.     

Influence of xylem development on axial hydraulic conductance within Prunus root systems

The effect of secondary growth on the distribution of the axial hydraulic conductance within the Prunus root system was investigated. Secondary growth resulted in a large increase in both the number (from about 10 to several thousand) and diameter of xylem vessels (from a few micrometres to nearly 150 µm). For fine roots (<3 mm), an increase in root diameter was correlated with a slight increase in the number of xylem vessels and a large increase in their diameter. Conversely, for woody roots, an increase in root diameter was associated with a dramatic increase in the number of xylem vessels, but little or no change in vessel diameter. The theoretical axial conductivity (Kh, m⁴.s^-1.MPa^-1) of root segments was calculated with the Poiseuille-Hagen equation from measurements of vessel diameter. Kh measured using the tension-induced technique varies over several orders of magnitude (7.4᎒^-11 to 5.7᎒^-7 m⁴.s^-1.MPa^-1) and shows large discrepancies with theoretical calculated Kh. We concluded that root diameter is a pertinent and useful parameter to predict the axial conductance of a given root, provided the root type is known. Indeed, the relationship between measured Kh and root diameter varies according to the root type (fine or woody), due to differences in the xylem produced by secondary growth. Finally, we show how the combination of branching pattern and axial conductance may limit water flow through root systems. For Prunus, the main roots do not appear to limit water transfer; the axial conductance of the main axes is at least 10% higher than the sum of the axial conductance of the branches.     

Perfusion of onion root xylem vessels: a method and some evidence of control of the pH of the xylem sap

We describe a method for perfusing the xylem in the stele of excised onion roots with solutions of known composition under a pressure gradient. Tracer studies using [¹⁴C] polyethylene glycol 4000 and the fluorescent dye, Tinopal CBSX, indicated that perfusing solutions passed exclusively through the xylem vessels. The conductance of the xylem was small over the apical 100 mm of the root axis but increased markedly between 100 and 200 mm. Unbuffered perfusion solutions supplied in the range pH 3.7–7.8 emerged after passage through the xylem adjusted to pH 5.2–6.0, indicating the presence of mechanisms for absorbing or releasing protons. This adjustment continued over many hours with net proton fluxes apparently determined by the disparity between the pH of the perfusion solution and the usual xylem sap pH of about 5.5. Mild acidification of the xylem sap by buffered perfusion solutions increased the release of ⁸⁶Rb (K⁺) and ³⁵SO₄ ^2- from the stelar tissue into the xylem stream. The ion-transporting properties of onion roots seemed little changed by excision from the bulbs, or by removal of the apical zones of the root axis. The pH of sap produced by root pressure resembles that found in the outflow solutions of perfused root segments.     

The haustorium of the root parasite Triphysaria (Scrophulariaceae), with special reference to xylem bridge ultrastructure

Haustoria of Triphysaria pusilla and T. versicolor subsp. faucibarbata from a natural habitat were analyzed by light and electron microscopy. Secretory trichomes (root hairs) participate in securing the haustorium to the surface of the host root. The keel-shaped intrusive part of the secondary haustorium penetrates to the depth of the vascular tissue of the host. Some of the epidermal interface cells differentiate into xylem elements. A significant number of haustoria do not differentiate further, but in most haustoria one to five of the epidermal xylem elements terminate a similar number of xylem strands. The strands mostly consist of vessel members and they connect host xylem or occasionally host parenchyma to the plate xylem adjacent to the stele of the parasite root. Each strand of this xylem bridge is accompanied by highly protoplasmic parenchyma cells with supposed transfer cell function. Increased surface area of the plasmalemma occurs in these cells as it does in interface parenchyma cells. Graniferous tracheary elements are restricted to the haustorium and occur most frequently in the plate xylem. The plate xylem is also accompanied by highly protoplasmic parenchyma cells. Hyphae of mycorrhizal fungi of the host root occasionally penetrate into the distal part of the xylem bridge. We combine structural observations and physiological facts into a hypothesis for translocation of water and nutrients between host and parasite. Some evolutionary aspects related to endogeny/exogeny of haustoria are discussed, and it is argued that the Triphysaria haustorium represents a greatly advanced and/or reduced condition within Scrophulariaceae.     

Sequential depolarization of root cortical and stelar cells induced by an acute salt shock - implications for Na(+) and K(+) transport into xylem vessels

Early events in NaCl-induced root ion and water transport were investigated in maize (Zea mays L) roots using a range of microelectrode and imaging techniques. Addition of 100 mm NaCl to the bath resulted in an exponential drop in root xylem pressure, rapid depolarization of trans-root potential and a transient drop in xylem K(+) activity (A(K+) ) within ～1 min after stress onset. At this time, no detectable amounts of Na(+) were released into the xylem vessels. The observed drop in A(K+) was unexpected, given the fact that application of the physiologically relevant concentrations of Na(+) to isolated stele has caused rapid plasma membrane depolarization and a subsequent K(+) efflux from the stelar tissues. This controversy was explained by the difference in kinetics of NaCl-induced depolarization between cortical and stelar cells. As root cortical cells are first to be depolarized and lose K(+) to the environment, this is associated with some K(+) shift from the stelar symplast to the cortex, resulting in K(+) being transiently removed from the xylem. Once Na(+) is loaded into the xylem (between 1 and 5 min of root exposure to NaCl), stelar cells become more depolarized, and a gradual recovery in A(K+) occurs.     

A method for measuring xylem hydraulic conductance and embolism in entire root and shoot systems

Current methods for determining the influence of xylem cavitationon hydraulic conductance are limited to unbranched stem or rootsegments with hydraulic conductances above c. 2 mmol s^–1MPa^–1. Lower conductances and/or highly branched systemsare encountered in seedlings, arid-land shrubs, herbs, and distalportions of shoot and root systems of trees. In order to quantifythe hydraulic impact of cavitation in such systems, existingtechniques have been modified. Branched shoot or root systemswere prepared for measurement by removal of leaves, or roottips, respectively. The shoot or root system was enclosed ina vacuum chamber with the proximal end protruding and suppliedwith perfusing solution. Flow through the xylem was inducedby chamber vacuum. Hydraulic conductance was determined fromthe slope of the flow rate versus pressure relationship. Xylemembolism was quantified from the increase in hydraulic conductancefollowing high pressure (100 kPa) perfusion of solution throughthe plant. Examples are provided of the application of the methodto cavitation studies in the cold desert shrub Artemisia tridentata. Key words: Hydraulic conductance, xylem cavitation, embolism, whole root/shoot system     

Xylem perfusion of tap root segments of Plantago maritima: the physiological significance of electrogenic xylem pumps

Abstract A method is described for perfusing xylem vessels in tap root segments of the halophyte P. maritima. Use of excised segments allowed recording of the trans-root potential (TRP) at both ends of a segment. It was shown that there can be a spatial variation of electrogenic ion pump activity along the xylem in one root segment. The pH of perfusion solutions, differing in buffering capacity, was adjusted by the root segment to pH 5.1–5.6 during How through the xylem. This pH range was similar to that of sap produced by root pressure. The K⁺ activity in the outflow solution (K⁺_out) was rather constant at 12–13 mol m^?l3 despite input K⁺ activities ranging from 8 to 20 mol m^?l3. Addition of fusicoccin (10^?l2 mol m^?l3) to the perfusion solution induced a strong acidification of the xylem sap, a decrease in K⁺_out and an increase in Na⁺_out. Inhibition of aerobic respiration through anoxia inhibited electrogenic proton pumping into the xylem and led to an increase in K⁺_out and a decrease in Na⁺_out. It is suggested that transport of K⁺ and Na⁺ to the shoot of the halophyte P. maritima is regulated in the tap root by means of ion exchange between xylem vessels and xylem parenchyma and that this exchange is energized by proton translocating ATPases.     

The regulation of anion loading to the maize root xylem

The regulation of anion loading to the shoot in maize (Zea mays) was investigated via an electrophysiological characterization of ion conductances in protoplasts isolated from the root stele. Two distinct anion conductances were identified. In protoplasts from well-watered plants, Z. mays xylem-parenchyma quickly-activating anion conductance (Zm-X-QUAC) was the most prevalent conductance and is likely to load the majority of NO(3)(-) and Cl(-) ions to the xylem in nonstressed conditions. Z. mays xylem-parenchyma inwardly-rectifying anion conductance was found at a lower frequency in protoplasts from well-watered plants than Zm-X-QUAC, was much smaller in magnitude in all observed conditions, and is unlikely to be such a major pathway for anion loading into the xylem. Activity of Z. mays xylem-parenchyma inwardly-rectifying anion conductance increased following a water stress prior to protoplast isolation, but the activity of the putative major anion-loading pathway, Zm-X-QUAC, decreased. Addition of abscisic acid (ABA) to protoplasts from well-watered plants also inhibited Zm-X-QUAC activity within minutes, as did a high free Ca(2+)concentration in the pipette. ABA was also seen to activate a Ca(2+)-permeable conductance (Z. mays xylem-parenchyma hyperpolarization activated cation conductance) in protoplasts from well-watered plants. It is postulated that the inhibition of anion loading into the xylem (an important response to a water stress) due to down-regulation of Zm-X-QUAC activity is mediated by an ABA-mediated rise in free cytosolic Ca(2+).     

Root xylem CO<Subscript>2</Subscript> flux: an important but unaccounted-for component of root respiration

Key message

In tree roots, a large fraction of root-respired CO ₂ remains within the root system rather than diffusing into the soil. This CO ₂ is transported in xylem sap into the shoot, and because respiration is almost always measured as the flux of CO ₂ into the atmosphere from plant tissues, it represents an unaccounted-for component of tree root metabolism.

Abstract

Root respiration has been considered a large component of forest soil CO₂ efflux, but recent findings indicate that it may be even more important than previous measurements have shown because a substantial fraction of root-respired CO₂ remains within the tree root system and moves internally with the transpiration stream. The high concentration of CO₂ in roots appears to originate mainly within the root. It has been suggested that plants can take up dissolved inorganic carbon (DIC) from soil, but under most conditions uptake from soil is minimal due to the root-to-soil diffusion gradient, which suggests that most of the CO₂ in root xylem is derived from root respiration. Estimates of the internal flux of CO₂ through root xylem are based on combined measurements of sap flow and internal [CO₂]. Results quantifying root xylem CO₂ flux, obtained for a limited number of species, have raised important concerns regarding our understanding of tree respiration. Taken together, the results of these studies call into question the partitioning of ecosystem respiration into its above- and belowground components, and redefine the energetic costs of tree root metabolism and hence estimates of belowground carbon allocation. Expanding our observations of root xylem CO₂ flux to more species and at longer time scales, as well as improving the techniques used to study this process, could be fruitful avenues for future research, with the potential to substantially revise our understanding of root respiration and forest carbon cycles.

     

Bicarbonate-induced alkalinization of the xylem sap in intact maize seedlings as measured in situ with a novel xylem pH probe

In higher plants the pH of the xylem sap plays an important role in drought signaling, growth regulation, and plant nutrition. However, the interpretation of the data is very controversial. The main reason for this is that the xylem pH in intact plants was not directly accessible hitherto. We present here a novel, minimally-invasive probe based on the xylem pressure-potential probe (used for measuring directly xylem pressure and the electrical potential between root xylem sap and medium). Single-tipped, double-barreled capillaries were used, one barrel served as H(+)-selective electrode, whereas pressure and electrical potential were recorded by the other one. Upon insertion of the probe into the root xylem of maize (Zea mays) seedlings, pH values ranging between about 4.2 and 4.9 were monitored when the roots were immersed in standard nutrient solution. The pH did not respond to changes in light irradiation (up to 300 micromol m(-2) s(-1)), but increased upon exposure of the root to 5 or 20 mm bicarbonate in the bath solution. Changes in pH could also be recorded in transpiring plants when the root was cut below the insertion point of the probe and placed in media with different pH. The data support the hypothesis of Mengel ([1994] Plant Soil 165: 275-283) that upon external supply with bicarbonate Fe is immobilized in the leaf apoplast via changes in xylem pH.     

Pluripotency of Arabidopsis xylem pericycle underlies shoot regeneration from root and hypocotyl explants grown in vitro

We have established a detailed framework for the process of shoot regeneration from Arabidopsis root and hypocotyl explants grown in vitro . Using transgenic plant lines in which the GUS or GFP genes were fused to promoters of developmental genes ( WUS , CLV1 , CLV3 , STM , CUC1 , PLT1 , RCH1 , QC25 ), or to promoters of genes encoding indicators of the auxin response ( DR5 ) or transport ( PIN1 ), cytokinin (CK) response ( ARR5 ) or synthesis ( IPT5 ), or mitotic activity ( CYCB1 ), we showed that regenerated shoots originated directly or indirectly from the pericycle cells adjacent to xylem poles. In addition, shoot regeneration appeared to be partly similar to the formation of lateral root meristems (LRMs). During pre-culture on a 2, 4-dichlorophenoxyacetic acid (2, 4-D)-rich callus-inducing medium (CIM), xylem pericycle reactivation established outgrowths that were not true calli but had many characteristics of LRMs. Transfer to a CK-rich shoot-inducing medium (SIM) resulted in early LRM-like primordia changing to shoot meristems. Direct origin of shoots from the xylem pericycle occurred upon direct culture on CK-containing media without prior growth on CIM. Thus, it appeared that the xylem pericycle is more pluripotent than previously thought. This pluripotency was accompanied by the ability of pericycle derivatives to retain diploidy, even after several rounds of cell division. In contrast, the phloem pericycle did not display such developmental plasticity, and responded to CKs with only periclinal divisions. Such observations reinforce the view that the pericycle is an 'extended meristem' that comprises two types of cell populations. They also suggest that the founder cells for LRM initiation are not initially fully specified for this developmental pathway.     

Radial transport of sodium and chloride into tomato root xylem

Transport of Na and Cl across exuding tomato (Lycopersicon esculentum Mill.) roots was determined as a function of ambient NaCl concentrations in the ranges of both systems 1 and 2. Kinetics of radial transport under steady-state conditions and the effect of dinitrophenol indicate that Na and Cl were transported by two different mechanisms. Sodium was neither accumulated against a concentration gradient nor directly inhibited by dinitrophenol from diffusing into the xylem. Chloride was accumulated in the xylem and its transport was nearly completely blocked by dinitrophenol. A comparison of the radial transport isotherms for Na and Cl for intact and decapitated plants indicates that the separate mechanisms were not unique to excised roots. It is concluded that radial Na transport in tomatoes was facilitated by a passive convective type process with the rate-limiting barrier located at the outer cortical plasmalemma. Chloride transport in both concentration ranges involved, either directly or indirectly, a metabolic mechanism. Absorption and retention of Na in the root tissue was negligible. Chloride was accumulated by the tissue but was unaffected by dinitrophenol.     

Ultrastructure of xylem parenchyma cells of barley roots in relation to ion transport to the xylem

Summary The structure of xylem parenchyma cells is examined in relation to transport of ions through the root. Measurement of uptake of ⁸⁶Rb⁺ and its transport through the root at different distances from the apex show that this is a general activity along the length of the root and not confined to a limited region. Thus transport through the root is not stopped by removal of that part of the root tip containing metaxylem vessels with living contents. The structure of xylem parenchyma appears to be suitable for involvement in ion transport from the stele to the xylem. At 1 cm behind the tip, where metaxylem vessels have no living contents but ion uptake and transport are going on at high rates, xylem parenchyma cells are rich in cytoplasm with extensive rough endoplasmic reticulum and well-developed mitochondria. Their cell walls contain numerous plasmodesmata, establishing the possibility of a symplastic pathway across the stele up to the vessels. The results are discussed in relation to regulation of ion transport to the xylem vessels in roots.Dedicated to Professor O. Stocker on the occasion of his 85th birthday.     

Vascular tissue-specific gene expression of xylem sap glycine-rich proteins in root and their localization in the walls of metaxylem vessels in cucumber

Root-specific cDNAs of glycine-rich protein (cucumber root glycine rich protein-1 and -2; CRGRP-1 and CRGRP-2) were cloned previously by use of an antiserum raised against whole xylem sap of Cucumis sativus. The accumulation of the corresponding mRNA at high levels was detected in the root-hair zone of cucumber tap root [Sakuta et al. (1998) Plant Cell Physiol. 39: 1330]. The RNA gel blot analysis with the CRGRP-1- and -2-specific probes revealed that the CRGRP genes expressed only in root but not at all in aboveground organs. When the localization of these mRNAs were examined by in situ hybridization, CRGRP mRNAs were found only in the parenchyma cells in the central cylinder of young lateral roots and it was most abundant in the cells that surrounded xylem vessels in the root-hair zone of the tap root. In immunoblotting of xylem sap collected from cucumber stem with an antiserum raised against CRGRP-1 that had been produced in an E. coli expression system, the antibodies, which did not cross-react with GRP1.8 of kidney bean, reacted with two proteins, whose mobilities corresponded to those of proteins deduced from the CRGRP-1 and -2 cDNAs. Immunohistochemical staining revealed that the CRGRPs accumulated specifically in the lignified walls of metaxylem vessels in the root, stem and leaf and in the lignified cell walls of perivascular fibers in cucumber stems. Immunostaining was also detected in the walls of metaxylem vessels and in the cell walls of adjacent sclerenchyma in the hypocotyl of kidney bean. These data clearly indicate that the novel glycine-rich proteins were produced in the vascular tissue of the root, transported systemically over a long distance via the xylem sap and immobilized in the walls of metaxylem vessels and sclerechyma cells in aboveground organs.     

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.     

The concentration of xylem sap constituents in root exudate, and in sap from intact, transpiring castor bean plants (Ricinus communis L.)

Root exudates were sampled from detopped root systems of castor bean (Ricinus communis). Different volume flux rates were imposed by changing the pneumatic pressure around the root system using a Passioura-type pressure chamber. The concentrations of cations, anions, amino acids, organic acids and abscisic acid decreased hyperbolically when flux rates increased from pure root exudation up to values typical for transpiring plants. Concentrations at low and high fluxes differed by up to 40 times (phosphate) and the ratio of substances changed by factors of up to 10. During the subsequent reduction of flux produced by lowering the pneumatic pressure in the root pressure chamber, the concentrations and ratios of substances deviated (at a given flux rate) from those found when flux was increased. The flux dependence of exudate composition cannot therefore be explained by a simple dilution mechanism. Xylem sap samples from intact, transpiring plants were collected using a Passioura-type root pressure chamber. The concentrations of the xylem sap changed diurnally. Substances could be separated into three groups: (1) calcium, magnesium and amino acid concentrations correlated well with the values expected from their concentration-flux relationships, whereas (2) the concentrations of sulphate and phosphate deviated from the expected relationships during the light phase, and (3) nitrate and potassium concentrations in intact plants varied in completely the opposite manner from those in isolated root systems. Abscisic acid concentrations in the root exudate were dependent on the extent of water use and showed strong diurnal variations in the xylem sap of intact plants even in droughtstressed plants. Calculations using root exudates overestimated export from the root system in intact plants, with the largest deviation found for proton flux (a factor of 10). We conclude that root exudate studies cannot be used as the sole basis for estimating fluxes of substances in the xylem of intact plants. Consequences for studying and modelling xylem transport in whole plants are discussed.     

The pathways of calcium movement to the xylem.

Calcium is an essential plant nutrient. It is acquired from the soil solution by the root system and translocated to the shoot via the xylem. The root must balance the delivery of calcium to the xylem with the need for individual root cells to use [Ca2+]cyt for intracellular signalling. Here the evidence for the current hypothesis, that Ca2+ travels apoplastically across the root to the Casparian band which it then circumvents via the cytoplasm of the endodermal cell, is critically reviewed. It is noted that, although Ca2+ channels and Ca2+-ATPases are present and could catalyse Ca2+ influx and efflux across the plasma membrane of endodermal cells, their transport capacity is unlikely to be sufficient for xylem loading. Furthermore, there seems to be no competition, or interactions, between Ca2+, Ba2+ and Sr2+ for transport to the shoot. This seems incompatible with a symplastic pathway involving at least two protein-catalysed transport steps. Thus, a quantity of purely apoplastic Ca2+ transport to the xylem is indicated. The relative contributions of these two pathways to the delivery of Ca2+ to the xylem are unknown. However, the functional separation of symplastic Ca2+ fluxes (for root nutrition and cell signalling) and apoplastic Ca2+ fluxes (for transfer to the shoot) would enable the root to fulfil the demand of the shoot for calcium without compromising intracellular [Ca2+]cyt signals. This is also compatible with the observed correlation between transpiration rate and calcium delivery to the shoot.