Effect of EDTA on transport forms of manganese in maize xylem exudate

Xylem exudates were collected from 20-day-old maize seedlings grown for 24 h in labelled manganese chloride solution (10 ppm) with or without EDTA (18 mgl^-1). The distribution of radioactivity on paper ohromatograms of exudates did not show the existence of free Mn. Fractionation of exudate constituents by gel filtration on Sephadex G-26 showed that⁵⁴Mn is associated with three UV (254 nm) -absorbing fractions. Two fractions contain amino acids (M. wt.:≧ 10 000 and ∼ 900). The third fraction (M. wt. ∼ 600) however, appears to be a carrier of a different type, which is practically amino acid free, although it contains 10.23% N (by elementary analysis). The properties of this carrier are discussed. Application of EDTA has favoured the; transport of Mn in xylem sap with fractions of relatively lower molecular weights.     

Cation and anion balance in the xylem exudate of tobacco roots

Summary The sum of Na, K, Ca, Mg in the exudate of tobacco generally exceeded the sum of mineral anions. Insufficient organic acids were present to account for the differences and bicarbonate appeared to be the other anion involved. Amino acids were present in very low concentrations relative to mineral cations. When nitrate salts only were in the external solutions, the anions were mostly, but not entirely, nitrate. When chloride salts only were in the external solutions, the cations far exceeded the level of mineral anions in the exudate. It is postulated that nitrate is actively transported when nitrate salts are in the external solution regardless of the cation, but when anions other than nitrate are in the external solution, the cations are actively transported with the anions passively following. Nitrate transport was via a symplasm, but that of the other anions seemed to be different. When bicarbonate is the only anion in the external solution and when present at relatively high concentrations (5 × 10⁻³ M or higher), the volume of exudate is decreased. It appears that the organic acids which were synthesized as a result of the bicarbonate absorption were not transferred to the xylem vessels.     

Transport of copper and manganese to the xylem exudate of sunflower

Abstract Absorption of copper and manganese by sunflower roots from solution cultures of varying composition was followed by measuring the concentrations of the metals appearing in whole roots, root cell sap and xylem exudate. Total copper in the fibrous roots was linearly related to the concentration of copper in the external solution but the concentration of copper released to the xylem exudate was buffered somewhat against the changes made externally. No such buffering was observed for managenese. A copper-sensitive electrode, responsive only to free cupric ions was used in conjunction with total copper analysis by atomic absorption spectrophotometry to show that little of the copper (usually < 1%) existed as a free ion in any phase of the system. Copper in the xylem exudate may be strongly complexed. An electron paramagnetic resonance spectrum of the xylem exudate indicated that manganese probably was a free divalent ion. Calculation of the electrochemical potential gradient for free cupric ions showed that no special metabolically-linked mechanism need be postulated to account for absorption of copper (or manganese) other than that necessary to maintain the transmembrane potential.     

The effect of cropping and fertiliser nitrogen rates on nitrogen balance in soil

Summary Fertilizer/soil N balance of cropped and fallow soil has been studied in a pot experiment carried out with grey forest soil (southern part of Moscow region) at increasing rates of¹⁵N labelled ammonium sulfate (0; 8; 16; 32 mg N/100 g of soil). The fertilizer¹⁵N balance has been shown to depend upon its application rate and the presence of growing plants. Fertilizer N uptake efficiency was maximum (72.5%) and gaseous losses-minimum (12.5%) at the application rate of 16 mg N/100 g of soil. Fertilizer N losses from the fallow soil were 130–220% versus those from the cropped soil. At the application of fertilizer N the plant uptake of soil N was 170–240% and the amount of soil N as N–NH₄ exchangeable + N–NO₃ in fallow was 350–440% as compared to the control treatment without nitrogen (PK).After cropping without or with N fertilizer application at the rates of 8 and 32 mg N/100 g of soil, a positive nitrogen balance has been found which is likely due to nonsymbiotic (associative) N-fixation. It has been shown that biologically fixed nitrogen contributes to plant nutrition.     

Nitrate reductase activity and nitrogen compounds in xylem exudate of Juglans nigra seedlings: relation to nitrogen source and supply

Nitrogen (N) limits plant productivity and its uptake and assimilation may be regulated by N source, N availability, and nitrate reductase activity (NRA). Knowledge of how these factors interact to affect N uptake and assimilation processes in woody angiosperms is limited. We fertilized 1-year-old, half-sib black walnut (Juglans nigra L.) seedlings with ammonium (NH₄ ⁺) [as (NH₄)₂SO₄], nitrate (NO₃ ⁻) (as NaNO₃), or a mixed N source (NH₄NO₃) at 0, 800, or 1,600 mg N plant⁻¹ season⁻¹. Two months following final fertilization, growth, in vivo NRA, plant N status, and xylem exudate N composition were assessed. Specific leaf NRA was higher in NO₃ ⁻-fed and NH₄NO₃-fed plants compared to observed responses in NH₄ ⁺-fed seedlings. Regardless of N source, N addition increased the proportion of amino acids (AA) in xylem exudate, inferring greater NRA in roots, which suggests higher energy cost to plants. Root total NRA was 37% higher in NO₃ ⁻-fed than in NH₄ ⁺-fed plants. Exogenous NO₃ ⁻ was assimilated in roots or stored, so no difference was observed in NO₃ ⁻ levels transported in xylem. Black walnut seedling growth and physiology were generally favored by the mixed N source over NO₃ ⁻ or NH₄ ⁺ alone, suggesting NH₄NO₃ is required to maximize productivity in black walnut. Our findings indicate that black walnut seedling responses to N source and level contrast markedly with results noted for woody gymnosperms or herbaceous angiosperms.     

Changes in the xylem exudate composition of poplar Populus tremula x P. alba)-dependent on the nitrogen

The capacity of poplar (P. tremula x P. alba, clone INRA 717 1/B4) to respond to changes in the nutrient supply with modifications in the xylem exudate collected after decapitation was investigated with special respect to N-compounds. The composition (inorganic ions and amino-N) was analysed with respect to (a) the time after decapitation at different times of day, (b) a change in the nitrogen concentration from NO^-3 to N-free medium, a change in the nitrogen source from NO^-3 to NH⁺4, (d) an increase in NO^-3 supply from 1 mM to 8 mM, and (e) the withdrawal of K⁺ supply. (a) The ion concentration in the xylem exudate was not affected up to 15 min after decapitation of the plants. Later a continuous increase in the concentration was observed. This increase was large if decapitation was performed in the middle of the light period and small at night-time. In both types of experiments (b,c) the NO^-3 concentration dropped immediately after the transfer, indicating the close connection between NO^-3 uptake and xylem loading. (b) After transfer to N-free medium poplar did not balance the charge in the xylem by increasing the concentration of other inorganic anions or decreased xylem loading of cations within 3 d of treatment. The N-status of the xylem exudate was reduced within 15 min. After transfer of the NO^-3-grown plants to NH⁺4, as the sole N-source, the charge compensation in the xylem exudate was maintained by reducing the loading of cations, and 3 d later by an enhanced xylem loading of mainly SO²4-. The N-status in the xylem exudate was maintained by an immediate increase in glutamine concentration (2-fold in 15 min). (d) Increasing the NO^-3 supply to 8 mM had no effect on the ionic composition or the N-status of the xylem exudate. (e) The withdrawal of K⁺ from the medium for 11 d resulted in a limitation of the S- and N-supply of the plant, causing a decrease in the N-status of the xylem sap. The data are discussed with respect to charge compensation after changes in the nutrient supply and to the maintenance of the nitrogen status in the xylem sap.     

Translocation of nickel in xylem exudate of plants

Topped plants of tomato (Lycopersicon esculentum), cucumber (Cucumis sativus), corn (Zea mays), carrot (Daucus carota), and peanut (Arachis hypogaea) were treated with 0.5 to 50 micromolar Ni (containing ⁶³Ni) in nutrient solutions. Xylem exudate was collected for 10 hours or, in the case of corn, for 20 hours at 5-hour intervals. Electrophoresis of nutrient solution distributed all Ni cathodically as inorganic Ni²⁺. Low concentrations of Ni in tomato exudate migrated anodically, presumably bound to organic anion (carrier). However, this carrier became saturated at about 2 micromolar Ni in exudate, and excess Ni ran cathodically. Most of the Ni in cucumber, corn, carrot, and peanut exudate ran anodically, and its migration rate was identical for all exudates. Peanut root sap contained 14 to 735 micromolar Ni. The anodic Ni carriers in root sap and exudate appear identical. The carrier in root sap became saturated near 100 micromolar Ni, as shown by cathodic streaking of Ni exceeding that concentration. It appears that all five species translocate low concentrations of Ni in the same anionic form.     

Transport of sodium into the xylem exudate of tobacco

When tobacco (Nicotiana tabacum L. var. Virginia Gold) plants were pretreated with Na (²²Na) several days before detopping, from 2.3 to 4.9% of Na previously accumulated in roots appeared in the xylem exudate in 7 days after detopping. Na from the external medium, however, was readily transported to the exudate. Moreover, the amount of the pretreatment Na that was transported to the exudate was not influenced by the presence of Na in the external medium. When Na was present in the external medium after detopping, about 4% (with an NaNO₃ post treatment) to 10% (with an NaCl post treatment) of the Na transported to the xylem in the 7 days following detopping originated in the vacuoles. Nitrate salts of K or Na in the external medium after detopping resulted in transport of large quantities of the respective cation to the exudate, but not in increased transport of the pretreatment Na. A much larger percentage of the K that was accumulated after detopping than of the Na similarly accumulated was transferred to the xylem exudate.     

The effect of local application of fertilizer on the content of cytokinins in the xylem sap of maize

Cytokinin content in xylem sap was higher in plants grown under local supply of fertilizers as compared to those grown under homogenous distribution of nutrients in soil. The separate assay of cytokinins in xylem exudate from split root system showed that roots, which were in contact with fertilizer mainly contributed to cytokinins transported from roots to shoots. This revised version was published online in July 2006 with corrections to the Cover Date.     

Translocation of iron citrate and phosphorus in xylem exudate of soybean

Soybean plants, Glycine max (L.) Merrill, in standard solution received 2.5 μm ferric ethylenediamine di(o-hydroxyphenylacetate (FeEDDHA) and 0 to 128 μm phosphorus. Their stem exudates contained: 32 to 52 μm Fe, 120 to 5000 μm P, and 120 to 165 μm citrate. Electrophoresis of exudates with high P caused Fe trailing that precluded identification of any major form of Fe. Exudate with low P gave an anodic band of Fe citrate as the major Fe compound. Phosphate added to exudate in vitro depressed the Fe citrate peak and cause Fe trailing. EDDHA added to exudate in vitro pulled Fe from Fe citrate; citrate then migrated as a slower form and Fe migrated as FeEDDHA. A modified preculture system, involving 2-day renewals of 0.2 μm FeEDDHA with 3.2, 9.6, or 16 μm P and low levels of other ions, controlled pH depression and produced considerable change in citrate and P levels. The exudates contained: 45 to 57 μm Fe, 200 to 925 μm P, and 340 to 1025 μm citrate. The high citrate was from plants grown with low P. The major form of Fe in the exudates was Fe citrate. This is probably the form translocated in the plants.     

Prediction of long-term fertilizer nitrogen requirements of maize in the tropics using a nitrogen balance model

A simple N balance model was used to calculate fertilizer requirement for a target N uptake by maize. Nitrogen uptake from soil sources and target uptake of N with fertilizer N additions were obtained from fertilizer trials in Africa and Latin America. Most experiments had data for only one cropping period, although some from Latin America had data for four to six crops. The transfer coefficient of fertilizer N to the crop was adjusted to realize maximum recovery of fertilizer N under best methods of fertilizer application. The time constants of transfer of soil N to the crop were allowed to vary and were affected mainly by soil texture. Where 4 to 6 cropping periods were available good agreement between actual and predicted fertilizer N requirements was obtained. With this approach long-term fertilizer N requirements for 14 sites were predicted using first cropping period N uptake. This study showed that pools of organic N in more coarse-textured soils were usually smaller and declined more rapidly than in fine-textured soils. Labile organic N pools declined with time under all simulations, but approached equilibrium within 10 croppings seasons. Equilibrium N uptake from the soil organic N pool was predicted to be 31 kg ha^–1 for the more coarse-textured soils and 36 kg ha^–1 for the fine-textured soils. Long-term projections of fertilizer requirements using input data of the field experiments were reasonable, and effects of legume green manures and other amendments could be clearly evaluated.     

Effect of calcium on transport of cations to the xylem exudate of tobacco

Summary Roots of detopped tobacco plants (Nicotiana tabacum var. Virginia Gold) were exposed to Na, K, and Ca salts or to water, and cation transfer to xylem vessels was measured. In some cases plants had been exposed to Na in addition to regular nutrient solutions before detopping. Calcium in the external medium greatly depressed the transport of Na from the external medium to the xylem vessels and it often stimulated the transfer of K from the external medium to the xylem vessels. The K/Na ratio in the exudate thus was dependent upon the Ca content of the external medium under these conditions. In contrast, externally applied Ca or Ca deficiency had very little effect on the transfer of preaccumulated K and Na from compartments within roots to the xylem vessels. The K/Na ratio in the exudate under these conditions was not related to Ca levels nor to mild Ca deficiency. The ratios decreased with time after detopping regardless of Ca level. Intact plants accumulated more Na than did root systems of detopped plants in a 6-day period.Riverside University of CaliforniaSoil Science and Agricultural Engineering     

Characterization of the maize xylem sap proteome

The xylem in plants has mainly been described as a conduit for water and minerals, but emerging evidence also indicates that the xylem contains protein. To study the proteins in xylem sap, we characterized the identity and composition of the maize xylem sap proteome. The composition of the xylem sap proteome in maize revealed proteins related to different phases of xylem differentiation including cell wall metabolism, secondary cell wall synthesis, and programmed cell death. Many proteins were found to be present as multiple isoforms and some of these isoforms are glycosylated. Proteins involved in defense mechanisms were also present in xylem sap and the sap proteins were shown to have antifungal activity in bioassays.     

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+).     

The effect of manganese on the production of Phanerochaete flavido-alba ligninolytic peroxidases in nitrogen limited cultures

Manganese supplementation of culture medium affected Phanerochaete flavido-alba FPL 106507 growth, glucose consumption and extracellular protein accumulation. Both the titre and time of detection of lignin peroxidase (LiP) were affected by manganese concentration in the medium, whereas with manganese peroxidase (MnP) only the titre was affected. In high Mn(II) containing cultures highest manganese peroxidase levels and a decrease in extracellular veratryl alcohol accumulation were observed. After FPLC a number of haemprotein peaks showing manganese peroxidase activity were detected in Mn(II) supplemented cultures. On the contrary, only haemprotein peaks of lignin peroxidase were detected in culture medium not supplemented with Mn(II).     

Diurnal and temporal changes in the chemical profile of xylem exudate from Vitis rotundifolia

Positive root pressure in Vitis rotundifolia Michx. cv. Noble was employed to quantify diurnal and temporal changes in the chemical profile of xylem exudate. Xylem fluid osmolarity (7.2 to 16.8 m M ), water flux (8.2 to 18.5 ml h⁻¹) and solute flux (0.7 to 2.2 mmol h⁻¹) from a cut spur exhibited a diurnal pattern with maxima during midday and minima at night. Total osmolarity was similar to the sum of all organic and inorganic entitites quantified, indicating that the major solutes have been identified. Total amino acid and organic acid concentration were about equal (2 to 7 m M ), and sugars accounted for a minor fraction of the total profile (<0.2 m M ). Glutamine represented ca 80% of the organic N and 70% of the total N transported in the xylem fluid. A circadian rhythm in water flux and net flux of most organic and inorganic entities was observed with maxima during midday and minima at night. The increase in xylem fluid osmolarity occurring during midday was primarily a consequence of increased organic acid (oxalic, citric, tartaric, malic and succinic acids) and ion (NH₄⁺, No₃⁻, P and Ca) concentration. A diurnal cycle in amino acid concentration was less clear. The concentration of individual organic and inorganic entities varied asynchronously with time. Xylem solute was comprised of 80% organic and 20% inorganic components when collected 5 min to 2 h after the commencement of bleeding, but the ratio of organic to inorganic components fell to about 50% after 7 days.