Behaviour of iron chelates in calcareous soils

Summary An examination was made of the changes with time in the composition of aqueous extracts of a highly calcareous clay that had been treated with one of the four chelates Fe-DTPA, Fe-CDTA, Fe-HEEDDA and Fe-EDHPA (Fe-Chel 138).It was found that a rapid fall in the recovery of soluble iron took place with both Fe-DTPA and Fe-HEEDDA due to sorption by the clay and also to replacement of iron in the chelate by calcium from the soil. By using Fe⁵⁹-labelled compounds, it was found that for both these chelates considerable isotopic exchange occurred between the Fe⁵⁹ and the natural soil iron, and that this exchange increased with decreasing rate of application of chelated iron.Fe-CDTA and Fe EDHPA were found to be comparatively unaffected by contact with the soil; over 80% recovery was obtained after 15 days, with treatments ranging from 2.5 to 100 ppm chelated iron. Negligible isotopic exchange took place with these two chelates.These results are discussed in relation to previously published results of laboratory experiments with iron chelates and to the treatment of soils for the control of iron chlorosis.     

Behaviour of iron chelates in calcareous soils

Summary Fe-EDTA and Fe-HEEDTA, labelled with Fe⁵⁹, were applied at rates of 10 and 100 ppm iron to 50-g samples of dry soil, which were then stored for periods ranging from one to fifteen days before extracting with water.The analyses of the extracts lead to the following general conclusions.1. The decrease in soluble iron could be attributed to both sorption of chelate anions by the clay and to replacement of iron by calcium in the chelate molecule.2. The amount of each chelate sorbed changed little with time and was proportional to the quantity applied. More HEEDTA was sorbed than EDTA.3. The initial decrease in the concentration of soluble iron was rapid for both chelates, but was the greater for Fe-HEEDTA.4. The precipitation of iron from the soluble Fe-HEEDTA was slower than from Fe-EDTA, so that fifteen days after treatment more soluble iron was obtained from the Fe-HEEDTA treatments.5. Marked increases in the recoveries of Fe-EDTA were made when the treated soils were sealed within small containers, compared with those allowed free access to the air. Comparable treatments using Fe-HEEDTA had a much smaller effect.6. Isotopic exchange of Fe⁵⁹ with natural soil iron was greater in the treatments with 10 ppm chelated iron than the 100 ppm level. Fe-HEEDTA was subject to more exchange than Fe-EDTA at both levels.These results are discussed in relation to the treatment of soils with iron chelates for the control of lime-induced chlorosis, and to the importance of isotopic exchange when using chelates labelled with radioactive iron in soil.     

Reactivity of synthetic Fe chelates with soils and soil components

The most effective and common Fe fertilisers in general are EDDHA and EDDHMA Fe chelates because they are highly stable ferric complexes in neutral and alkaline solutions. EDDHSA and EDDCHA iron chelates were introduced in the market recently. Commercial Fe chelates have two Fe fractions, chelated Fe and non-chelated Fe. The latter is bonded to by-products produced during the synthesis of the chelating agent. The effectiveness of Fe chelates depends on their ability to maintain Fe in the soil solution despite simultaneous equilibrium of Fe chelate with many cations, such as Ca²⁺. The main aim of this work was to test the possible agricultural use of EDDHSA and EDDCHA Fe chelates. The pH-Ca²⁺ effect on soluble and chelated Fe (pH ranging from 2 to 12) and the interaction of Fe chelates with soils and soil phases (ferrihydrite, acid peat, calcium carbonate and Ca montmorillonite) are presented. The results demonstrated that EDDHA, EDDHMA, EDDHSA and EDDCHA in solution remain fully associated with Fe from pH 4 to 9 despite competition with Ca. Among soil materials, ferrihydrite and acid peat retain both chelated and non-chelated Fe to the greatest extent. The type of chelating agent is a factor that affects chelated Fe availability in soil. FeEDDHA and FeEDDHMA were retained more by soil surfaces than FeEDDHSA and FeEDDCHA. Commercial Fe chelates present a large amount of soluble, non-chelated Fe and make Cu soluble in soils, which may be due to non-chelated Fe being displaced by Cu.     

Reactions of iron chelates in calcareous soil and their relative efficiency in iron nutrition of corn

Summary Laboratory incubation studies on the reactions of Fe-DTPA, Fe-EDTA, Fe-citrate and Fe-fulvate with a calcareous soil indicated that Fe³⁺ was very rapidly displaced by Ca²⁺, Mg²⁺, Zn²⁺ and Cu²⁺ ions. The displacement of iron was in the reverse order of the stability of the Fe-chelates. The activity of Fe³⁺, Ca²⁺, Mg²⁺, Zn²⁺ and Cu²⁺ tended to attain a constant value with time. Application of chelating agents to a calcareous soil mobilized different amounts of iron as defined by their relative stability and cation competition. The degree of mobilization increased with increasing levels of applied chelating agents. A significant negative correlation (r = –0.77)^* was observed between pH and DTPA-extractable iron. Results of greenhouse experiment showed significant increase in the dry matter yield and iron uptake by corn plants upon application of iron-chelates. The chelates enhance the uptake of both native and applied sources. The effectiveness of the chelates used was in the order of their capacity to maintain iron in soluble form in the soil solution. These results suggest that iron nutrition of plants in calcareous soils can be effectively regulated by the application of iron chelated by natural or synthetic water-soluble chelating agents.     

Electron transfer in ruthenium-modified proteins

Photochemical techniques have been used to measure the kinetics of intramolecular electron transfer in Ru(bpy)₂(im)(His)²⁺-modified (bpy = 2,2-bipyridine; im = imidazole) cytochromec and azurin. A driving-force study with the His33 derivatives of cytochromec indicates that the reorganization energy () for Fe²⁺Ru³⁺ ET reactions is 0.8 eV. Reductions of the ferriheme by either an excited complex,^*Ru²⁺, or a reduced complex, Ru⁺, are anomalously fast and may involve formation of an electronically excited ferroheme. The distance dependence of Fe²⁺Ru³⁺ and Cu⁺Ru³⁺ electron transfer in 12 different Ru-modified cytochromes and azurins has been analyzed using a tunneling-pathway model. The ET rates in 10 of the 12 systems exhibit an exponential dependence on metal-metal separation (decay constant of 1.06 å^–1) that is consistent with predictions of the pathway model.     

Production of citric acid with immobilized Yarrowia lipolytica

Summary Citric acid was produced with immobilized Yarrowia lipolytica yeast in repeated batch-shake-flask and air-lift fermentations. In active and passive immobilization methods calcium alginate, -carrageenan, polyurethane gel, nylon web and polyurethane foams were tested as carriers in repeated-batch fermentations. The highest citric acid productivity of 155 mg l^–1 h^–1 was reached with alginate-bead-immobilized cells in the first batch. A decrease in bead diameter from 5–6 mm to 2–3 mm increased the volumetric citric acid productivity threefold. In an air-lift bioreactor the highest citric acid productivity of 120 mg l^–1 h^–1 with a product concentration of 16.4 g l^–1 was obtained with cells immobilized in -carrageenan beads. Offprint requests to: H. Kautola     

Nitrogen and phosphorus requirements for raising mycorrhizal seedlings of Leucaena leucocephala in containers

A greenhouse study was undertaken to determine the nitrogen and phosphorus fertilization requirements for raising mycorrhizal seedlings in soil in containers. Seedlings of Leucaena leucocephala were grown for 40 days in dibble tubes containing fumigated or nonfumigated soil uninoculated or inoculated with Glomus aggregatum. The soil was fertilized with NH₄NO₃ solution to obtain 25–200 mg N kg^-1 soil, and with a KH₂PO₄ solution to establish target soil solution P concentrations of 0.015–0.08 mg P l^-1. At the end of 40 days, seedlings were transplanted into pots containing 5-kg portions of fumigated soil. Posttransplant vesicular arbuscular mycorrhizal fungal (VAMF) effectiveness, measured as pinnule P content, plant height, shoot dry weight and tissue N and P concentrations, was significantly increased by pretransplant VAMF colonization in both soils. The best posttransplant mycorrhizal colonization and mycorrhizal growth responses were observed if the nonfumigated pretransplant soil was amended with 50 mg N kg^-1 soil and 0.04 mg P l^-1 or if the fumigated pretransplant soil was amended with 100 mg N kg^-1 soil and 0.04 mg P 1^-1. There was no relationship between NP ratios of nutrients added to the pretransplant soil medium and shoot NP ratios observed after transplanting. Shoot NP ratio was also not correlated with root colonization level.Contribution from the Hawaii Institute of Tropical Agriculture and Human Resources Journal Series No. 4025     

Aluminium effects on spruce and beech seedlings

Summary Natural spruce seedlings sampled on acid brown soil of the Belgian Ardennes, either on mineral soil, or on the litter layer, show higher Al content in the latter case, where substrate Al content is lower. Absorbing power of the root for Al organic chelates is probably higher than for Al⁺³ ions.Most soluble Al is chelated, especially in the holorganic and hemiorganic layers, under spruce and beech. In the mineral layers pH is above 3.8, and total Al in soil solutions is always found to be less than 2 eq/ml.     

Soil composition affects the nesting behavior of blue-tailed bee-eaters (Merops philippinus) on Kinmen Island

The blue-tailed bee-eater (Merops philippinus) is a summer migrant that breeds on Kinmen Island, located off the west coast of Taiwan, about 5 km from the southern coast of mainland China. The aim of this study was to investigate why blue-tailed bee-eaters build their nests in sandy loam and sandy clay loam, but not in clay loam. Soil chemical and physical properties, and mineralogical composition were measured for the different soil types. Clay loam had a significantly lower pH, Na, and base saturation than did sandy loam or sandy clay loam. Clay loam had a significantly higher N, cation-exchange capacity (CEC), K, and free iron (Fe_d) and aluminum oxide (Al_d) contents than the other soil types. Clay loam had significantly lower sand and higher clay content, and higher bulk density and penetration resistance than the other soil types. The correlation coefficients (r ²) between penetration resistance and Fe_d, Al_d, and clay contents were 0.997, 0.848, and 0.779, respectively. Soil strength and compaction are important criteria for bee-eaters nesting-site selection. The lower pH of clay loam would enhance the exchangeable Al and acidity, further increasing the soil aggregation. Thus, it might prevent the bee-eaters from excavating nesting burrows.     

Antioxidant effect of bovine serum albumin on membrane lipid peroxidation induced by iron chelate and superoxide

Albumin is supposed to be the major antioxidant circulating in blood. This study examined the prevention of membrane lipid peroxidation by bovine serum albumin (BSA). Lipid peroxidation was induced by the exposing of enzymatically generated superoxide radicals to egg yolk phosphatidylcholine liposomes incorporating lipids with different charges in the presence of chelated iron catalysts. We used three kinds of Fe³⁺-chelates, which initiated reactions that were dependent on membrane charge: Fe³⁺-EDTA and Fe³⁺-EGTA catalyzed peroxidation in positively and negatively charged liposomes, respectively, and Fe³⁺-NTA, a renal carcinogen, catalyzed the reaction in liposomes of either charge. Fe³⁺-chelates initiated more lipid peroxidation in liposomes with increased zeta potentials, followed by an increase of their availability for the initiation of the reaction at the membrane surface. BSA inhibits lipid peroxidation by preventing the interaction of iron chelate with membranes, followed by a decrease of its availability in a charge-dependent manner depending on the iron-chelate concentration: one is accompanied and the other is unaccompanied by a change in the membrane charge. The inhibitory effect of BSA in the former at high concentrations of iron chelate would be attributed to its electrostatic binding with oppositely charged membranes. The inhibitory effect in the latter at low concentrations of iron chelate would be caused by BSA binding with iron chelates and keeping them away from membrane surface where lipid peroxidation is initiated. Although these results warrant further in vivo investigation, it was concluded that BSA inhibits membrane lipid peroxidation by decreasing the availability of iron for the initiation of membrane lipid peroxidation, in addition to trapping active oxygens and free radicals.     

A novel eye morphology induced by a P element in somatic tissue of Drosophila melanogaster.

Summary We found a specific eye morphology designated as Square, which is induced when some Drosophila melanogaster strains harboring P elements are crossed with the 2–3 strain carrying a modified P element, P[ry ⁺, 2–3], which produces transposase in somatic tissue. This phenotype was dominant and also induced in the reciprocal crosses. Square was induced when the 2–3 strain was crossed with Q and M strains such as the sn^w (M) strain carrying three small P elements but not with P strains. Inheritance of Square was also tested and its phenotype was not transmitted to the next generation. These results suggest that Square is caused by the transposition of P elements in somatic cells.     

On the Prebiotic Role of Iron and Sulfur

Thermodynamic data (Sillén, L. G.: 1966, Arkiv Kemi, 25, 159) indicate that there is little support for the idea that metabolic sulfur cycles were involved in prebiotic and early life processes, since under reducing conditions the equilibrium concentration of sulfur in the primordial seas should have been very low, < 10^–8 M. However, it is suggested that metabolic sulfur cycles may have become important when oxygen evolved, when iron(II) ions disappeared from the seas, and when large amounts of sulfur were released from their iron sulfide sources.     

Smectite clays in Mars soil: Evidence for their presence and role in Viking biology experimental results

Summary Various chemical, physical and geological observations indicate that smectite clays are probably the major components of the Martian soil. Satisfactory ground-based chemical simulation of the Viking biology experimental results was obtained with the smectite clays nontronite and montmorillonite when they contained iron and hydrogen as adsorbed ions. Radioactive gas was released from the medium solution used in the Viking Labeled Release (LR) experiment when interacted with the clays, at rates and quantities similar to those measured by Viking on Mars. Heating of the active clay (mixed with soluble salts) to 160°C in CO₂ atmosphere reduced the decomposition activity considerably, again, as was observed on Mars. The decomposition reaction in LR experiment is postulated to be iron-catalyzed formate decomposition on the clay surface. The main features of the Viking Pyrolytic Release (PR) experiment were also simulated recently (Hubbard, 1979) which the iron clays, including a relatively low 1st peak and significant 2nd peak.The accumulated observations on various Martian soil properties and the results of simulation experiments, thus indicate that smectite clays are major and active components of the Martian soil. It now appears that many of the results of the Viking biology experiments can be explained on the basis of their surface activity in catalysis and adsorption.     

Effects of mutations of conserved Lys-155 and Thr-156 residues in the phosphate-binding glycine-rich sequence of the F1-ATPase beta subunit of Escherichia coli.

beta Lys-155 in the glycine-rich sequence of the beta subunit of Escherichia coli F1-ATPase has been shown to be near the gamma-phosphate moiety of ATP by affinity labeling (Ida, K., Noumi, T., Maeda, M., Fukui, T., and Futai, M. (1991) J. Biol. Chem. 266, 5424-5429). For examination of the roles of beta Lys-155 and beta Thr-156, mutants (beta Lys-155-->Ala, Ser, or Thr; beta Thr-156-->Ala, Cys, Asp, or Ser; beta Lys-155/beta Thr-156-->beta Thr-155/beta Lys-156; and beta Thr-156/beta Val-157-->beta Ala-156/beta Thr-157) were constructed, and their properties were studied extensively. The beta Ser-156 mutant was active in ATP synthesis and had approximately 1.5-fold higher membrane ATPase activity than the wild type. Other mutants were defective in ATP synthesis, had < 0.1% of the membrane ATPase activity of the wild type, and showed no ATP-dependent formation of an electrochemical proton gradient. The mutants had essentially the same amounts of F1 in their membranes as the wild type. Purified mutant enzymes (beta Ala-155, beta Ser-155, beta Ala-156, and beta Cys-156) showed low rates of multisite (< 0.02% of the wild type) and unisite (< 1.5% of the wild type) catalyses. The k1 values of the mutant enzymes for unisite catalysis were lower than that of the wild type: not detectable with the beta Ala-156 and beta Cys-156 enzymes and 10(2)-fold lower with the beta Ala-155 and beta Ser-155 enzymes. The beta Thr-156-->Ala or Cys enzyme showed an altered response to Mg2+, suggesting that beta Thr-156 may be closely related to Mg2+ binding. These results suggest that beta Lys-155 and beta Thr-156 are essential for catalysis and are possibly located in the catalytic site, although beta Thr-156 could be replaced by a serine residue.     

Culture conditions effecting plant regeneration from cotyledons of Vigna radiata (L.) Wilczek

Conditions for plant regeneration from excised cotyledons of Vigna radiata were studied. Complete plant developed from the uncallused proximal ends of cotyledons on Murashige & Skoog's (MS), Gamborg's (B₅) and C (MS salts + B₅ vitamins) basal media. The basal medium C was found to be best for plant regeneration. Regeneration frequency, however, varied with genotype, size, orientation and age of explant and the different plant growth regulators combination in the medium. Addition of cytokinins induced callusing at the proximal ends of cotyledons followed by multiple shoot formation. Out of 6-benzyl aminopurine (BAP), kinetin (KIN), N (^–2 isopentyl) adenine (2iP) and adenine sulphate (AS), only BAP and KIN were found to be more effective in enhancing the frequency of shoot regeneration. BAP at 1×10^-1M induced maximum (60%) shoot regeneration whereas maximum number of shoots (8 to 9 shoots) per explant was observed with 5×10^-6M BAP. Cotyledons excised from two-day old seedlings were most regenerative. The regenerative response of cotyledons decreased when sliced into two equal parts either longitudinally or transversely. Callusing and organogenic differentiation occured only if the petiolar end of cotyledons was in contact with medium. None of the tested treatments were effective in inducing shoot bud differentiation from subcultured callus. Well developed shoots rooted when incubated on half strength MS, MS and MS basal medium supplemented with IAA (5×10^-6M). The rooted plants were transferred to pots and later established in the field with 60% success.Abbreviations AS adenine sulphate - BAP 6-benzylaminopurine - B₅ medium after Gamborg et al. [6], - C Medium with MS salts + B₅ vitamins - 2iP N (^–2 isopentyl) adenine - IAA indole-3-acetic acid - KIN Kinetin - MS medium after Murashige & Skoog [21] - NAA 1-napthaleneacetic acid     

Mating behaviour of Ctenomyces serratus

Nineteen isolates of Ctenomyces serratus including three pairs of tester strains were crossed in all possible combinations on diluted Pablum cereal agar with added salts, oatmeal salts agar-Medium E, and soil and hair medium. Out of 190 crosses, 18 were fertile and the remaining 172 were sterile. Of the 19 isolates, seven were– mating type, 11 were + mating type, and one (UAMH 1959) was non-fertile with the other isolates. The high degree of non-fertile crosses among the 19 isolates shows that the use of mating reactions for the identification of conidial isolates of C. serratus may not be a practical tool. unless a large number of isolates belonging to both the mating types are employed. The production of exudate droplets in the fertile crosses seemed to be correlated with the degree of mating response. Ascocarp morphology corresponded closely to that of a cleistothecium, because of the completely closed nature. Ascospores were quite alike in their size, but they differed in shape, depending on their orientation.     

Free metal activity and total metal concentrations as indices of micronutrient availability to barley [Hordeum vulgare (L.) ‘Klages’]

The form in which a micronutrient is found in the rhizosphere affects its availability to plants. We compared the availability to barley of the free hydrated cation form of Fe³⁺, Cu²⁺, Zn²⁺, and Mn²⁺ versus their total metal concentrations (free ion plus complexes) in chelator-buffered solutions. Free metal ion activities were estimated using the chemical equilibrium program GEOCHEM-PC with the corrected database. In experiment 1, barley was grown in nutrient solutions with different Fe³⁺ activities using chelators to control Fe levels. Chlorosis occurred at Fe³⁺ activities of 10^–18 and 10^–19 M for barley grown in HEDTA and EDTA solutions, respectively. In experiment 2, barley was grown in nutrient solutions with the same calculated Fe³⁺ activity and the same chelator, but different total Fe concentrations. Leaf, root and shoot Fe concentrations were higher from CDTA buffered solutions which had the higher total Fe concentration indicating the importance of the total Fe concentration on Fe uptake. Results from treatments using EDTA or HEDTA, with one exception, were similar to the results from the CDTA treatment. This suggests differences in critical Fe³⁺ activities found in experiment 1 were due to differences in the total Fe concentration and not errors in chelate formation constants used to estimate the critical activities. Results for Cu, Zn, and Mn were similar to Fe; despite solutions with equal free Cu²⁺, Zn²⁺ and Mn²⁺ activities, plant concentrations of these metals were generally greater when grown in the solutions with the greater total amount of Cu, Zn, or Mn. When the free Zn²⁺ activity was kept constant while the total amount of Zn was increased from 4.4 to 49 M, leaf Zn concentration increased from 77 to 146 g g^-1. In order to predict metal availability to barley and other species in chelator-buffered nutrient solutions, both free and total metal concentrations in solution must be considered. The critical Fe³⁺ activities required by barley in this study are much higher than those from tomato and soybean, 10^-28 M, which strongly supports the Strategy 2 model of Fe uptake for Poaceae. This is related to the importance of the Fe³⁺ (barley) and the Fe²⁺ (tomato and soybean) ions in Fe uptake. Fe-stressed barley is known to release phytosiderophores which compete for Fe³⁺ in the nutrient solution, while tomato and soybean reduce Fe³⁺ to Fe²⁺ at the epidermal cell membranes to allow uptake of Fe²⁺ from Fe³⁺ chelates in solution.Abbreviations CDTA trans-1,2-diaminocyclohexane-N,N,N,N-tetracetic acid - DTPA diethylenetriaminepentacetic acid - EDTA ethylenediaminetetracetic acid - EDDHA ethylenediamine-di(o-hydroxyphenylacetic acid) - HBED-N,N di(2-hydroxybenzoyl)-ethylenediamine-N,N-diacetic acid - HEDTA-N hydroxyethylenediaminetriacetic acid - MES-2 (N-morpholino)ethanesulfonic acid - NTA nitrilotriacetic acid     

Characterization of Iron Uptake from Ferrioxamine B by Chlorella vulgaris

Iron uptake from two Fe³⁺-hydroxamate siderophores, ferrioxamine B and Fe³⁺-rhodotorulate, by iron-stressed Chlorella vulgaris (ATCC strain 11468) was evaluated with some comparison to iron uptake from synthetic and organic acid ferric chelates. Iron-stress induced iron uptake from ferrioxamine B. Dissipation of the electrochemical gradient, via uncouplers, inhibited iron uptake. Respiratory inhibitors gave variable results, an indication that a direct link to respiration was not apparent. Vanadate inhibition of iron uptake indicated that an ATPase or phosphate intermediate could be involved in the uptake mechanism. Divalent cations manifested variable effects dependent on the cation and chelator used. These data confirm that C. vulgaris has an inducible iron-uptake system for Fe³⁺-hydroxamic acid siderophores which may involve a different mechanism than that observed for other chelates.     

Effects of β-phenylethylamine on polyphosphoinositide turnover in rat cerebral cortex

The effects of the trace amine, -phenylethylamine, on the hydrolysis of inositol phospholipids in rat cerebral cortical slices was studied using a direct assay involving prelabeling with [³H]inositol and then examining the production of [³H]inositol phosphates in the presence of lithium. Phenylethylamine exhibited two different effects. Millimolar concentrations of phenylethylamine stimulated the production of [³H]inositol phosphates to about 200% of control, while much smaller concentrations (micromolar) inhibited noradrenaline(NE)-stimulated [³H]inositol phosphate formation dose-dependently. The ₁-antagonist, prazosin, inhibited the increases in [³H]polyphosphoinositide turnover stimulated by phenylethylamine and by NE, though it inhibited phenylethylamine to a lesser extent than NE. It appears, therefore, that phenylethylamine affects [³H]inositol phosphate formation by acting as a partial ₁-agonist.     

Soil water uptake by trees using water stable isotopes (δ2H and δ18O)−a method test regarding soil moisture,texture and carbonate

Aims

Stable isotopes of oxygen and hydrogen are often used to determine plant water uptake depths. We investigated whether and to what extend soil moisture, clay content, and soil calcium carbonate influences the water isotopic composition.

Methods

In the laboratory, dried soil samples varying in clay content were rewetted with different amounts of water of known isotopic composition. Further, we removed soil carbonate from a subset of samples prior to rewetting. Water was extracted from samples via cryogenic vacuum extraction and analysed by mass spectrometry.

Results

The isotopic composition of extracted soil water was similarly depleted in both ¹⁸O and ²H with decreasing soil moisture and increasing clay and carbonate content. Soil carbonate changed the δ¹⁸O composition while δ²H was not affected.

Conclusions

Our results indicate that soil carbonate can cause artifacts for ¹⁸O isotopic composition of soil water. At low soil moisture and high carbonate content this could lead to conflicting results for δ¹⁸O and δ²H in plant water uptake studies.