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Movement to the xylem exudate of rubidium accumulated in the apex of corn roots.

Apical segments of roots of corn (Zea mays, L.) were excised and mounted in experimental salt solutions containing (86)RbCl and CaCl(2). Xylem exudates were collected hourly beginning at the 21st hour. At the 24th hour, experimental solutions were replaced with solutions of other chlorides (nonlabeled) of Ca(2+) +/- some monovalent cation.Following replacement of the labeled solutions, the rate of (86)Rb(+) release to the exudates from root segments declined by about the same proportional amount regardless of the initial steady-state output rate or the kinds of salts used to replace the RbCl. The content of (86)Rb(+) in the roots also declined with time, but to a relatively less degree. Loss of accumulated (86)Rb(+) to the various external solutions following replacement was less than 6% in 6 hours. It is concluded that movement of accumulated Rb(+) to the exudate is dependent on the concentration of Rb(+) in the root but is largely independent of exchange between accumulated ions and other ions in transit across the root.

Journal Article↗

Transport of divalent cations: cation exchange capacity of intact xylem vessels.

The cation exchange capacity of the intact xylem vessels in cut shoots of papyrus (Cyperus papyrus spec.) has been determined. The cation exchange capacity is independent of the cation concentration in the transpiration stream, and is equal for Ca and Co. The high value of the cation exchange capacity (0.6 to 1 x 10(-7) equivalents per square centimeter vessel wall surface) leads to the hypothesis that the porous structure of the vessel wall, and not only the inner vessel wall surface, acts as a cation exchanger.Differences between anion ([(32)P]phosphate, [(45)Ca]EDTA(2-), [(115)Cd(m)]-EDTA(2-)), and cation ([(45)Ca](2+), [(115)Cd(m)](2+)) movement are explained in terms of transport with the transpiration flux or by exchange reactions. The competition between exchange sites and natural or synthetic ligands for the divalent cations is discussed.

Journal Article↗

Units of freezing of deep supercooled water in woody xylem.

The low temperature exotherms (LTE) of 1-year-old twigs of Haralson apple (Malus pumila Mill.), shagbark hickory (Carya ovata [Mill.] K. Koch), green ash (Fraxinus pennsylvanica Marsh), honey locust (Gleditsia triacanthos L.), American chestnut (Castanea dentata [Marsh] Borkh.), and red oak (Quercus rubra L.) were determined by differential thermal analysis (DTA). In one type of experiment freezing during a DTA experiment was halted for up to 2.5 hours after part of the supercooled water had frozen at temperatures between -25 and -42 C. Upon resumption of cooling the freezing started within 2 C of the stopping temperature. In a second type of experiment living and dead cells were microscopically observed in the same ray after partial freezing in the DTA apparatus. In another experiment, the LTE persisted even after tangential and radial sectioning of the twig to 0.13 millimeters. In a final experiment the LTE of a single multiseriate ray of red oak had the same shape as the LTE of wood with many uniseriate rays.These experiments confirm that the deep supercooled water in woody xylem or pith freezes in numerous independent events over a span of as much as 20 C. The units which freeze in an event are single cells or small groups of cells. Ice grows very slowly if at all from these units, and water moves very slowly from unfrozen cells to frozen ones. Deep supercooling of ray parenchyma does not require an intact ray.

Journal Article↗

Metal Complexation in Xylem Fluid : II. THEORETICAL EQUILIBRIUM MODEL AND COMPUTATIONAL COMPUTER PROGRAM.

Theoretical considerations of metal complex formation in aqueous solutions were used to develop a computer program (CHELATE) to calculate all equilibrium species (free metal ions, metal complexes, etc.) in any user-defined system, such as xylem fluid. Mass-balance equations were established to describe each free metal ion and each free ligand concentration as a function of solution pH, total metal or total ligand, hydrogen-association constants, and the stability constants of known metal complexes. A default data base can be altered by the user to define any desired system covered by the stored equilibrium data. The program can currently handle nine metal ions, 35 ligands, and 500 complex species. The validity of the program was confirmed by using experimental test systems in which free-metal ion activity measurements were made with ion-selective electrodes.Program CHELATE was used to calculate the distribution of six metals in 0- to 1-hour exudate from soybean (Glycine max L. Merr.) and tomato (Lycopersicon esculentum Mill.) plants grown in normal and Zn-phytotoxic nutrient solutions. The results indicated that Fe is bound by citric acid, and Cu is bound by several amino acids in the normal-Zn exudate. Most of the Cu in soybean exudate is bound to asparagine and histidine. In tomato, Cu is bound to histidine, glutamine, and asparagine. Zinc, Mn, Ca, and Mg are bound primarily by citric acid and malic acid in both species; the per cent bound for these metals is lower than that for Fe and Cu. Zinc phytotoxicity caused equilibrium concentration shifts and resulted in the formation of several additional metal complexes not found in the normal-Zn exudate.

Journal Article↗

Rapid increase in deep supercooling of xylem parenchyma.

Malus pumila Mill. twigs were collected from September through December and stored at 5 degrees C until the low temperature exotherms of the xylem were determined by differential thermal analysis. During the differential thermal analysis, cooling was interrupted, and temperatures of 5 to -18 degrees C were held for 0.4 to 10 hours before cooling to -50 degrees C was resumed. Control twigs were cooled to -50 degrees C without interruption. Holding the twigs at 1.3 to -5 degrees C shifted the start of the low temperature exotherm from about -20 to -30 degrees C. Slightly higher (2.6 degrees C) and lower (-10 degrees C) temperatures were occasionally effective. The shift began within 20 to 30 minutes and increased progressively to 150 minutes. The acclimation was reversibly inhibited by N(2) atmosphere.

Journal Article↗

Transmission of the Monocarpic Senescence Signal via the Xylem in Soybean.

During monocarpic senescence in soybean (Glycine max [L.] Merrill cv. Anoka) there is a remobilization of nitrogen from the leaves to the seeds, and it has been hypothesized that this loss of nitrogen from the leaves induces foliar yellowing. The phloem in a small segment of the petiole between the pods and the target leaf can be inactivated with a jet of steam. When a plant is depodded except for a single pod cluster in the center of the plant, the pod cluster induces yellowing of the nearest leaf even if the petiole contains a zone of dead phloem, whereas most of the rest of the plant remains green. The nitrogen content of these leaves with a dead phloem zone in their petioles does not decrease greatly, even though the leaves turn yellow. A similar treatment of a single leaf on a fully depodded plant (leaves stay green) does not cause that leaf to turn yellow. Since nutrients would have to be withdrawn from the leaves via the phloem, the pods do not induce yellowing by pulling nutrients out of the leaf and must be able to exert their influence via the xylem.

Journal Article↗

Ethephon-Induced Gummosis in Sour Cherry (Prunus cerasus L.) : I. Effect on Xylem Function and Shoot Water Status.

Ethephon, (2-chloroethyl)phosphonic acid, was sprayed at concentrations up to 69.2 millimolar to enhance gum formation in 1-year-old shoots of mature Prunus cerasus L. cv Montmorency trees. Gum accumulation caused rupturing of the shoot periderm, followed by gum extrusion. Lower ethephon concentrations were required to induce gum formation in spring and early summer (1.7-3.5 millimolar) then in late summer and fall (13.8-69.2 millimolar). The number of functional vessels, shoot hydraulic conductance, and water potential of both leaf and internode tissue decreased as gum content of shoots increased. Nontreated control shoots also contained small quantities of gum. There was no difference in neutral sugar composition of gum exuded by the tree, obtained from aqueous shoot extracts, or flushed from the vessels of shoots, whether induced by ethephon or not. Severe decrease in shoot and leaf water potential was associated with shoot die-back. Recovery of xylem function may occur where gummosis is less severe. Discrepancy between measured and predicted hydraulic conductance increased as shoot gum content increased, suggesting that decrease in number of functional vessels alone was not sufficient to explain the effects of gum on loss of shoot hydraulic conductance. Increased gum content in those vessels remaining functional would increase vessel sap viscosity and further reduce hydraulic conductance. The viscosities necessary to account for discrepancy between measured and predicted hydraulic conductance were calculated. Gum concentration less than 1.0% (w/v) would produce these viscosities.

Journal Article↗

Role of cytokinin in differentiation of secondary xylem fibers.

The differentiation of secondary xylem fibers was studied in cultured hypocotyl segments of Helianthus annuus L. It is shown that cytokinin is both a limiting and controlling factor in the early stages of fiber differentiation. In the absence of kinetin or zeatin, there was no fiber differentiation. However, cytokinin could induce fiber differentiation only in the presence of indoleacetic and gibberellic acids. First fibers were observed in the tissue after 12 days in culture, and their number increased linearly during the following 2 weeks. At low cytokinin levels, there was a positive correlation between cytokinin concentration in the medium and the number of fibers formed in the explants. A similar correlation was also found at low gibberellic acid concentrations. At high concentration, zeatin was more effective than kinetin. It seems that later stages of fiber differentiation can occur in the absence of cytokinin. It is proposed that the mechanism which controls and determines the early stages of fiber differentiation is based on an interaction of three major hormonal signals: indoleacetic acid plus gibberellic acid from the leaves with zeatin from the root apices.

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Identification of Cytokinins from Xylem Exudate of Phaseolus vulgaris L.

The principal biologically active cytokinins in xylem exudate of young Phaseolus vulgaris L. plants were identified by bioassay, high-performance liquid chromatography, enzymic degradation and combined gas chromatography-mass spectrometry (selected ion monitoring) a zeatin riboside, zeatin nucleotide, dihydrozeatin riboside, dihydrozeatin nucleotide, O-glucosyl zeatin, O-glycosyl dihydrozeatin, O-glucosyl dihydrozeatin riboside, and O-glucosyl dihydrozeatin nucleotide. Trace amounts of O-glucosyl zeatin riboside and O-glucosyl zeatin nucleotide were also detected.

Journal Article↗

The influence of dwarfing interstocks on the distribution and metabolism of xylem-applied [h]gibberellin a(4) in apple.

The influence of an interstock of the dwarfing cultivar M9 and the nondwarfing cultivar MM115 on the distribution and metabolism of labeled gibberellic acid A(4) ([(3)H]GA(4)) of high specific radioactivity (5.18 x 10(10) becquerel per millimole) applied to the xylem of the rootstock in grafted apple (Malus x domestica Borkh.) trees was compared. Free [(3)H] GA-like metabolites of [(3)H]GA(4), including putative GA(1), GA(2), GA(3), and GA(34), as well as various (3)H-putative GA glucosyl conjugates were detected in stem segments from both cultivars. M9 interstocks reduced the total uptake of [(3)H]GA(4) and decreased the proportion of (3)H metabolites transported to the shoots and leaves of scions. The M9 interstock tissue and adjacent rootstock and scion tissue retained a much greater amount and a higher proportion of the label than did comparable tissue of the nondwarfing MM115 interstock. In addition, the amount and proportion of free [(3)H]GAs was higher, and the proportion of putative [(3)H]GA glucosyl conjugates lower, in M9 interstocks compared to MM115. These effects of the dwarfing interstock on GA distribution and metabolism indicate a significant role for GAs in any satisfactory explanation of the dwarfing mechanism in apple.

Journal Article↗

Evidence for the involvement of a specific cell wall layer in regulation of deep supercooling of xylem parenchyma.

Current theory indicates that the structure of the cell wall is integral to the ability of a tissue to exhibit deep supercooling. Our previous work has indicated that the structure of the pit membrane and/or amorphous layer (protective layer), rather than the entire cell wall, may play a major role in deep supercooling (21, 22). The present study indicates a shift in the low-temperature-exotherm of current year shoots of peach can be induced by soaking twigs in water over 3 to 10 days. Alternatively, these shifts can be inhibited by exposing tissues to 10(-4) molar cycloheximide. Ultrastructural observations indicated a marked alteration of the amorphous layer in xylem parenchyma of water-soaked tissue. Alterations consisted of an apparent loosening or partial dissolution of portions of the amorphous layer. Changes in the density or uniformity of the amorphous layer in cycloheximide-treated tissues were not as readily apparent. The appearance of the protoplast in tissue soaked in water for up to 10 days was characteristic of deacclimated cells. However, in tissue soaked in cycloheximide for the same period these changes were not evident. These observations further support our contention that the structure of the amorphous layer may play a key role in establishing and regulating the ability of a cell to exhibit deep supercooling.

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Gradients of turgor, osmotic pressure, and water potential in the cortex of the hypocotyl of growing ricinus seedlings : effects of the supply of water from the xylem and of solutes from the Phloem.

To evaluate the possible role of solute transport during extension growth, water and solute relations of cortex cells of the growing hypocotyl of 5-day-old castor bean seedlings (Ricinus communis L.) were determined using the cell pressure probe. Because the osmotic pressure of individual cells (pi(i)) was also determined, the water potential (psi) could be evaluated as well at the cell level. In the rapidly growing part of the hypocotyl of well-watered plants, turgor increased from 0.37 megapascal in the outer to 1.04 megapascal in the inner cortex. Thus, there were steep gradients of turgor of up to 0.7 megapascal (7 bar) over a distance of only 470 micrometer. In the more basal and rather mature region, gradients were less pronounced. Because cell turgor approximately pi(i) and psi approximately 0 across the cortex, there were also no gradients of psi across the tissue. Gradients of cell turgor and pi(i) increased when the endosperm was removed from the cotyledons, allowing for a better water supply. They were reduced by increasing the osmotic pressure of the root medium or by cutting off the cotyledons or the entire hook. If the root was excised to interrupt the main source for water, effects became more pronounced. Gradients completely disappeared and turgor fell to 0.3 megapascal in all layers within 1.5 hours. When excised hypocotyls were infiltrated with 0.5 millimolar CaCl(2) solution under pressure via the cut surface, gradients in turgor could be restored or even increased. When turgor was measured in individual cortical cells while pressurizing the xylem, rapid responses were recorded and changes of turgor exceeded that of applied pressure. Gradients could also be reestablished in excised hypocotyls by abrading the cuticle, allowing for a water supply from the wet environment. The steep gradients of turgor and osmotic pressure suggest a considerable supply of osmotic solutes from the phloem to the growing tissue. On the basis of a new theoretical approach, the data are discussed in terms of a coupling between water and solute flows and of a compartmentation of water and solutes, both of which affect water status and extension growth.

Journal Article↗

Identification and characterization of the Arabidopsis PHO1 gene involved in phosphate loading to the xylem.

The Arabidopsis mutant pho1 is deficient in the transfer of Pi from root epidermal and cortical cells to the xylem. The PHO1 gene was identified by a map-based cloning strategy. The N-terminal half of PHO1 is mainly hydrophilic, whereas the C-terminal half has six potential membrane-spanning domains. PHO1 shows no homology with any characterized solute transporter, including the family of H(+)-Pi cotransporters identified in plants and fungi. PHO1 shows highest homology with the Rcm1 mammalian receptor for xenotropic murine leukemia retroviruses and with the Saccharomyces cerevisiae Syg1 protein involved in the mating pheromone signal transduction pathway. PHO1 is expressed predominantly in the roots and is upregulated weakly under Pi stress. Studies with PHO1 promoter-beta-glucuronidase constructs reveal predominant expression of the PHO1 promoter in the stelar cells of the root and the lower part of the hypocotyl. There also is beta-glucuronidase staining of endodermal cells that are adjacent to the protoxylem vessels. The Arabidopsis genome contains 10 additional genes showing homology with PHO1. Thus, PHO1 defines a novel class of proteins involved in ion transport in plants.

Alleles↗

Control of cellulose synthase complex localization in developing xylem.

Cellulose synthesis in the developing xylem vessels of Arabidopsis requires three members of the cellulose synthase (CesA) gene family. In young vessels, these three proteins localize within the cell, whereas in older vessels, all three CesA proteins colocalize with bands of cortical microtubules that mark the sites of secondary cell wall deposition. In the absence of one subunit, however, the remaining two subunits are retained in the cell, demonstrating that all three CesA proteins are required to assemble a functional complex. CesA proteins with altered catalytic activity localize normally, suggesting that cellulose synthase activity is not required for this localization. Cortical microtubule arrays are required continually to maintain normal CesA protein localization. By contrast, actin microfilaments do not colocalize with the CesA proteins and are unlikely to play a direct role in their localization. Green fluorescent protein-tagged CesA reveals a novel process in which the structure and/or local environment of the cellulose synthase complex is altered rapidly.

Arabidopsis↗

EgMYB2, a new transcriptional activator from Eucalyptus xylem, regulates secondary cell wall formation and lignin biosynthesis.

Summary EgMYB2, a member of a new subgroup of the R2R3 MYB family of transcription factors, was cloned from a library consisting of RNA from differentiating Eucalyptus xylem. EgMYB2 maps to a unique locus on the Eucalyptus grandis linkage map and co-localizes with a quantitative trait locus (QTL) for lignin content. Recombinant EgMYB2 protein was able to bind specifically the cis-regulatory regions of the promoters of two lignin biosynthetic genes, cinnamoyl-coenzyme A reductase (CCR) and cinnamyl alcohol dehydrogenase (CAD), which contain MYB consensus binding sites. EgMYB2 was also able to regulate their transcription in both transient and stable expression assays. Transgenic tobacco plants over-expressing EgMYB2 displayed phenotypic changes relative to wild-type plants, among which were a dramatic increase in secondary cell wall thickness, and an alteration of the lignin profiles. Transcript abundance of genes encoding enzymes specific to lignin biosynthesis was increased to varying extents according to the position of individual genes in the pathway, whereas core phenylpropanoid genes were not significantly affected. Together these results suggest a role for EgMYB2 in the co-ordinated control of genes belonging to the monolignol-specific pathway, and therefore in the biosynthesis of lignin and the regulation of secondary cell wall formation.

Amino Acid Sequence↗

Improvement of seed yields under boron-limiting conditions through overexpression of BOR1, a boron transporter for xylem loading, in Arabidopsis thaliana.

Soil fertilization is a common practice in modern agriculture, undertaken to prevent nutrient deficiency in crops. However, fertilization is costly and causes environmental pollution. The cultivation of plants that tolerate low nutrient supplies may circumvent this problem. Here, we report the generation of Arabidopsis thaliana plants that tolerate boron (B)-deficient conditions due to the overexpression of BOR1, an efflux B transporter that is required for efficient xylem loading of B. In several independently generated transgenic plants expressing BOR1 or BOR1-GFP under the control of the cauliflower mosaic virus 35S RNA promoter, root-to-shoot translocation of B was enhanced and shoot growth was greater under B-limiting conditions compared with wild-type plants. In addition, the transgenic plants showed increased translocation of B, especially to the shoot apex, and set seed normally under B-limiting conditions, under which wild-type plants failed to set seed. This study therefore reports plants that show improved seed yields compared with wild-type under nutrient-deficient conditions as a result of increased production of an essential mineral nutrient transporter.

Antiporters↗

Xylem transport and shoot accumulation of lumichrome, a newly recognized rhizobial signal, alters root respiration, stomatal conductance, leaf transpiration and photosynthetic rates in legumes and cereals.

* Root respiration, stomatal conductance, leaf transpiration and photosynthetic rates were measured in phytotron and field-grown plants following the application of 5 or 10 nM lumichrome, 10 nM ABA (abscisic acid) and 10 ml of 0.2 OD600 infective rhizobial cells. * Providing soybean and cowpea roots with their respective homologous rhizobia and/or purified lumichrome increased the concentration of this molecule in xylem sap and leaf extracts. Relative to control, rhizobial inoculation and lumichrome application significantly increased root respiration in maize, decreased it in lupin, but had no effect on the other test species. * Applying either lumichrome (10 nM), infective rhizobial cells or ABA to roots of plants for 44 h in growth chambers altered leaf stomatal conductance and transpiration in cowpea, lupin, soybean, Bambara groundnut and maize, but not in pea or sorghum. Where stomatal conductance was increased by lumichrome application or rhizobial inoculation, it resulted in increased leaf transpiration relative to control plants. Treating roots of field plants of cowpea with this metabolite up to 63 d after planting showed decreased stomatal conductance, which affected CO2 intake and reduction by Rubisco. * The effect of rhizobial inoculation closely mirrored that of lumichrome application to roots, indicating that rhizobial effects on these physiological activities were most likely due to lumichrome released into the rhizosphere.

Abscisic Acid↗

Rhodotorula pinicola sp. nov., a basidiomycetous yeast species isolated from xylem of pine twigs.

Three pink-colored yeast strains 3-1-3, 10-3-3 and 19-3-3 were isolated from xylem of surface-sterilized twigs of Pinus tabulaeformis collected from Dongling Mountain, Beijing, in different seasons. These strains were identified as Rhodotorula minuta (Saito) F.C. Harrison by conventional taxonomic characterization. However, molecular phylogenetic analysis based on internal transcribed spacer region (including 5.8S rDNA) and large-subunit rDNA D1/D2 domain sequences indicated that they represent a novel basidiomycetous yeast species, for which Rhodotorula pinicola is proposed (type strain: AS 2.2193(T)=CBS 9130(T)). The new species was most closely related to Rhodotorula laryngis Reiersöl in the R. minuta complex.

DNA, Ribosomal Spacer↗