Beryllium uptake by excised barley roots

The uptake of beryllium by excised barley roots from a solutioncontaining 111 µM beryllium and 500 µM calcium wasstudied. The exchange-adsorbed portion was 15% of the totalBe uptake after 30 min of exposure. The total Be content couldbe reduced to 55% of the control in 1 hr with demineralizedwater. At pH levels above 6, Be uptake was increased dramatically.A Q₁₀ of 1.2 was found to exist between 1 and 23°C. Mg (111µM) did not influence Be absorption while Zn and Al atthe same concentration reduced it, possibly competitively, 27and 62% of the control, respectively. Dinitro-phenol (500 µM),cyanide (500µM), azide (500 µM) and m-chlorocarbonylcyanidephenylhydrazone (100 µM) at least doubled it indicatingthe existence of a possible metabolic barrier to beryllium entrywhich was disrupted by these compounds. Absorption kineticsover an external beryllium concentration range of 11 to 444µM suggested a number of binding sites. Above 444 µMa scattering effect was noted due possibly to its toxic effect. ¹Pesticide Programs, Environmental Protection Agency (P.S. -769),Washington, D.C. 20460, U.S.A. (Received August 13, 1979; )     

Effects of temperature on orthophosphate absorption by excised corn roots

The uptake of orthophosphate (³²P) by excised corn roots, Zea mays L. was studied using roots grown on 0.2 mm CaSO₄. Nine concentrations of KH₂PO₄ from 1 to 256 μm were used at temperatures of 20°, 30°, and 40°. Enzyme kinetic analysis was applied to the data obtained. Two apparent mechanisms (sites) of phosphate uptake were observed, 1 dominating at high P concentrations and 1 at low P concentrations. A Km of 1.36 × 10⁻⁴ and a Vmax of 177 × 10⁻⁹ moles per gram of roots per hour at 30° was calculated for the mechanism dominating at high P concentrations. Similar calculations gave a Km of 6.09 × 10⁻⁶ and a Vmax of 162 × 10⁻⁹ moles per gram of roots per hour at 30° for the mechanism dominating at low P concentrations. The Q₁₀ for both mechanisms was approximately 2. Calculation of thermodynamic values from the data gave ΔF of − 5200 cal, ΔH of − 950 to − 1400 cal, and a enthalpy of activation (A) of 10,300 to 13,800 cal per mole for the mechanism dominating at high P concentrations. Similar calculations from the data for the mechanism dominating at low P concentrations gave a ΔF of − 7300 cal, ΔH of − 10,700 to − 8200 cal, and a A of 9300 to 18,900 cal per mole. If the dual mechanism interpretation of this kind of data adequately describes this system, then both mechanisms of P absorption by corn roots involve chemical reactions.     

Effects of ouabain and low temperature on the sodium efflux pump in excised corn roots

Ouabain (0.05 millimolar) and low temperature (4 C) both caused the tissue Na⁺ content of excised 5-day-old corn roots to increase, indicating that there is an inhibition of the Na⁺ efflux pump. Na⁺ efflux was measured utilizing three different methods. Each method gave similar results in terms of rate and ouabain sensitivity. With one of these methods, the compartmental efflux method, it was demonstrated that rates for Na⁺ efflux increase as the external Na⁺ concentration is increased; e.g. the efflux rates are 0.529, 1.78, and 3.64 microequivalents per gram fresh weight per hour for external NaCl concentrations of 1, 10, and 30 millimolar, respectively. The data indicate that the Na⁺ efflux pump is located in the plasmalemma of root cells.     

Cation uptake from bentonite suspensions by excised barley roots

Summary The net uptake of Na, K, or Ca by excised barley roots as influenced by the amount and kind of other cation was studied from bi-ionic and tri-ionic bentonite suspensions. The net uptake of Na or K from the Na-Ca or K-Ca system followed the concentration of soluble Na or K in the system. Calcium in both systems was taken up by the excised roots only at the 80 and 100 per cent Ca levels. At lower levels of Ca, the roots lost some of their initial calcium contents to the suspensions. The net uptake of Na or K from the Na-K system was in agreement with the concentrations of soluble Na or K in the system. At the various levels of Na or K, some of the initial calcium of the roots was lost to the suspensions. In the Na-K-Ca system, Na uptake gradually decreased with the increase of Ca level. Calcium was not absorbed by the roots at the various Ca levels. However, calcium greatly enhanced K uptake from this system. Part of a dissertation submitted by the senior author in partial fulfillment of requirements for the Ph. D. degree.     

Further studies on cation uptake from bentonite suspensions by excised barley roots

Summary The net uptake of Na, K, Li, and Ca or Mg by excised barley roots was studied from bi-ionic and tri-ionic bentonite suspensions. The net uptake of Li from Li-Ca system progressed linearly with progressive Li levels and was related to the concentration of soluble lithium. Calcium in this system was taken up only at the 100 per cent Ca level. At lower Ca levels calcium was lost from the roots to the suspensions. In K-Mg and Na-Mg systems the net uptake of Na or K by the excised roots was related to the concentration of the cation in the solution phase. Magnesium uptake took place at 80 and 100 per cent Mg levels. It was much less than that of K or Na at similar levels. At lower levels of Mg the roots lost some of their initial Mg contents to the suspensions. In the Na-K-Mg system magnesium was not taken up by the excised roots. Sodium uptake was not practically affected by the Mg level, but K uptake was greatly enhanced by magnesium.     

Effects of low water potential on ion uptake and loss for excised roots

Summary In excised roots of barley and tomato plants, lowering the water potential of nutrient solutions to-10.4 and-20.4 atm decreased the uptake of bromide and phosphorus while increasing the loss of these ions to the external solutions.Lowering the water potential greatly increased the rate of loss of potassium and bromide from the cytoplasm, but the increases in loss from the vacuoles were much smaller. The results suggest that the mechanisms of ion uptake are not affected by low water potential and that the decrease in ion accumulation is caused by the increased leakage from the cells.     

Manganese absorption by excised barley roots

Short-term absorption studies with 5-day-old excised barley roots revealed that the basic aspects of Mn absorption were similar to those of other metabolically absorbed cations. Following an initial non-metabolic equilibration with the root, Mn was absorbed for several hours at a slower steady-state rate comparable to that of other inorganic cations. Complete or nearly complete inhibition of the steady-state phase by low temperature, dinitrophenol, and azide provides strong evidence that Mn transport into this tissue was metabolically mediated. Within limits, the rate of transport was strongly dependent upon the concentrations of Mn and the hydrogen ions in the ambient solution. Absorption increased rapidly with increasing concentrations of Mn up to 1 meq per liter. Above this concentration, the rate leveled off, apparently due to a saturation of the transport mechanism. Within the physiological pH range in which Mn is soluble (below pH 7), absorption increased greatly with decreasing hydrogen-ion concentration.     

Absorption of cobalt by excised barley roots

Co absorption by excised roots of Hordeum vulgare L. appearsto be an active process exhibiting a Q₁₀ of 2.2. The effectsof poisons also indicate active absorption. After 1 hr difluorodinitrobenzeneat a concentration of 5000 µM inhibited it to 20% of thecontrol, while dinitrophenol (500 µM) and carbonyl cyanidephenylhydrazone (100 µM) inhibited it to 43% and 38% respectively.A study of absorption kinetics over a concentration range of1 through 100 µM CoCl₂ indicated that a number of carriersites were available for absorption depending upon the concentration.Results which were consistent with this interpretation wereobtained when the concentration of a competing ion, Ni, wasvaried. An exchangeable, absorbed fraction amounting to about25% of the total cobalt present in the tissue after 1 hr ofabsorption was found. (Received September 28, 1973; )     

Regulation of sulfate uptake by excised barley roots in the presence of selenate

Active transport of SO₄²⁻ and SeO₄²⁻ has been evaluated during 60-hour contact of barley roots with nutrient solutions containing either ³⁵SO₄²⁻ or ⁷⁵SeO₄²⁻, or both ions, at 0.1 milli-equivalent per liter. In the SO₄²⁻ solution the time course of active transport follows a straight line; if SeO₄²⁻ is also present transport is strongly inhibited after 20 to 30 hours for both ions. The S-Se uptake ratio remains 1.4 during the 60 hours; S-Se ratio shifts from 3.0 to 3.3 in proteins and falls to 0.6 in free amino acids. S-Se discrimination is mainly operating at the level of amino acid incorporation into proteins. The presence of Se-amino acids blocks this incorporation and brings about an accumulation of free amino acids; at the same time carrier activity is inhibited. The addition of methionine or Se-methionine causes a 60 to 80% inhibition of the active transport.     

Boron uptake by excised barley roots: I. Uptake into the free space

At 2 C, all boron accumulated by excised barley roots (Hordeum vulgare L. cv. Herta) remains in the free space; i.e. active uptake is nil at this temperature. Three component fractions of free space B were apparent: (a) a surface contaminant film of B on blotted roots, (b) water free space B, and (c) B reversibly bound in the cell walls. A stoichiometric release of H⁺ from the roots in the presence of B indicated that B was bound by borate complexes with polysaccharides in the cell walls. Polysaccharide-borate complexes are much less stable than those of monosaccharides, and the bound B fraction could be readily removed by rinsing the roots in the presence of a monomeric polyol possessing the necessary cis-diol configuration. Cell wall material separated from excised barley roots had a B binding capacity 66% greater than that of intact roots.     

Effects of root temperature on uptake of nitrate and ammonium ions by barley grown in flowing-solution culture

Summary Barley plants (Hordeum vulgare L.) grown from seed for 28 days in flowing solution culture were subjected to different root temperatures (3, 5, 7, 9, 11, 13, 17, 25°C) for 14 days with a common air temperature of 25/15°C (day/night). Uptake of NH₄ and NO₃ ions was monitored separately and continuously from solutions maintained at 10 M NH₄NO₃ and pH 6.0. Effects of root temperature on unit absorption rate , flux and inflow were compared. After 5 days , and increased with temperature over the range 3–11°C for NH₄ ions and over the range 3–13°C for NO₃ ions, with little change for either ion above these temperatures. Q₁₀ temperature coefficients for NH₄ ions (3–13°C) were 1.9, 1.7 and 1.6 for , and respectively, the corresponding values for NO₃ ions being 5.0, 4.5 and 4.6. For both ions, , and changed with time as did their temperature dependence over the range 3–25°C, suggesting that rates of ontogenetic development and the extent of adaptation to temperature may have varied among treatments.     

Mechanism of sulfate uptake by excised maize roots

The mechanism of uptake of sulfate by excised maize roots (Zea mays L. cv. Ganga-5) was investigated. It was found that in the concentration range l × 10^?9 M ? 5 × 10^?2 M, the sulfate uptake could be represented by a single multiphasic isotherm having four phases. Each phase covered a limited concentration range and obeyed Michaelis-Menten kinetics, which indicates mediation in sulfate uptake by a structure which changes characteristics (kinetic constants) at certain discrete salts concentrations (transition points).     

Effect of adsorbed cations on phosphorus uptake by excised roots

Pretreatment of excised roots of Hordeum vulgare, Zea mays, and Glycine max with various salt solutions affected their subsequent rate of phosphorus absorption from 2 × 10⁻⁵m KH₂PO₄. The rate of absorption was greatest for roots pretreated with trivalent cations, intermediate with divalent cations and lowest with monovalent cations. It appeared that the pretreatment involved a rapid exchange reaction at the root surface which was reversible. A 1 min pretreatment was effective for more than 20 min. The acceleration of phosphorus uptake by roots produced by polyvalent cations may be due partly or entirely to a greater reduction in the electrical potential at the root surface or within the pores of the negatively charged cell wall by polyvalent cations than by monovalent cations.     

Role of the accompanying anion in the effect of calcium salts on potassium uptake by excised barley roots

The effect of addition of Ca salts on accumulation of K from 5 mN KCl or K₂SO₄ solutions was found to depend on whether Ca was added as Cl or SO₄ salt. Chloride as well as K uptake was increased when Ca and Cl concentrations in culture solutions were increased. Pre-treatment of roots with CaCl₂ stimulated subsequent K uptake from K₂SO₄ solutions as compared to pre-treatment with distilled water but pre-treatment with CaSO₄ did not. The results indicate that addition of Ca salts to KCl or K₂SO₄ solutions increased anion uptake and the effect of the addition of the Ca salts on K uptake was in part the result of increased anion uptake and not entirely a direct effect of Ca.     

Multiphasic uptake of phosphate by corn roots

Abstract The concentration dependence of phosphate uptake was studied using root sections of corn (Zea mays L. cv. Ganga 5). Detailed and wide-range (57 concentrations in the range 1 μmol m^?3-75 mol m^?3), precise (average SEM < 2.5%, n= 6) and reproducible (similar patterns in three independent experiments and for 5, 10, 15, 20, 25 and 30°C) data revealed six (or seven) concentration-dependent phases separated by ‘jumps’ or sharp breaks. These transitions were independent of temperature and occurred over relatively narrow concentration ranges (0.0001–0.0004, 0.08–0.31, 1.0–3.5, (7.5–10), 18–20 and 57–59 mol m^?3). The intermediate phases obeyed Michaelis-Menten kinetics, whereas sigmoidal kinetics were observed at lower concentrations. Uptake within each of the two highest phases increased more rapidly with increasing external phosphate concentration than predicted from Michaelis-Menten kinetics but also saturated more rapidly. The latter finding is not consistent with free diffusion across the plasmalemma at high external phosphate concentrations. Kinetic models yielding continuous isotherms, e.g. the sum of one or two Michaelis-Menten terms and a diffusion term, cannot account for the data.     

Effect of calcium on sulfate uptake by barley roots

Abstract Sulfate uptake by excised roots of barley (Hordeum vulgare L.) was maximal in the presence of about 3x10^-3M CaCl₂. Kinetic studies contraindicate a stoichiometric binding of calcium to the carrier for sulfate, in contrast to findings of Cuppoletti and Segel (Biochemistry 14: 471–4718, 1975) for the filamentous fungus Penicillium notatum. In barley, calcium affects the K_m but not the V_max for sulfate uptake, presumably by altering the conformation and, thereby, the affinity of the carrier. Calcium also affects the transition site for sulfate uptake.     

Loss of rubidium and potassium from barley roots on sudden chilling

Sudden chilling of roots of barley pretreated in a labeled Rb⁺ solution at 25 C causes a transient loss of label from the nonfree space. The half-time of the release is about 10 minutes. Within the first few hours of uptake the loss increases proportionally to the root Rb⁺ content but varies between experiments as a fraction of this content: values from 4 to 20% were observed. The same phenomenon is found for K⁺ labeled with ⁸⁶Rb⁺