Responses of nitrogen-fixing and nitrate-supplied alfalfa (Medicago sativa L.) to iron chelates in an alkaline hydroponic medium

Ferric ethylenediamine di-(o-hydroxyphenylacetate) (FeEDDHA) and ferric hydroxyethylethylenediaminetriacetic acid (FeHEDTA) were evaluated as Fe sources for hydroponic growth of alfalfa (Medicago sativa L., cv. Mesilla), either dependent on N₂ fixation or supplied with NO₃. The hydroponic medium was maintained at pH 7.5 by addition of CaCO₃. Nitrogen-fixing cultures were inoculated with Rhizobium meliloti 102 F51 and grown in medium without added nitrogen. After five to seven weeks of growth under greenhouse conditions, plants were harvested. Nitrogen fixation was measured by the acetylene reduction method.When FeEDDHA was supplied, growth of alfalfa, whether dependent on N₂ fixation or supplied with NO₃, was severely limited at concentrations typically used in hydroponic medium (10 or 20 M). Maximum yield of NO₃-supplied alfalfa was obtained at 100 M while maximum yield of N₂-fixing alfalfa was obtained in the range of 33 to 200 M FeEDDHA. Nodule fresh weights and N₂ fixation rates increased with FeEDDHA concentration up to 33 M and remained essentially constant up to 200 M. With FeHEDTA, maximum yields of both NO₃-grown and N₂-fixing alfalfa were obtained at 10 M. Growth of NO₃-supplied plants was inhibited at 200 M FeHEDTA while growth of N₂-fixing plants was inhibited at 100 M FeHEDTA. The numbers of nodules per plant increased between 3.3 and 10 M FeHEDTA; however, inhibition of nodule formation occurred at a concentration of 33 M or higher. Nodule weights per plant and N₂ fixation rates were depressed at 3.3 M as well as at 100 M FeHEDTA. The results suggest that alfalfa dependent on N₂ fixation is more sensitive to limited Fe availability than alfalfa supplied with NO₃.     

Growth,N2 fixation and photosynthesis in a cyanobacterium,Trichodesmium sp., under Fe stress

Trichodesmium sp., isolated from the Great Barrier Reef lagoon, was cultured in artificial seawater media containing a range of Fe concentration. Fe additions stimulated growth, N₂ fixation, cellular chlorophyll a content, light-saturated chlorophyll a-specific gross photosynthetic capacity (P_m ^chla) and the dark respiration rate (R_d ^chla). Cell yields only doubled for 9 nM Fe relative to zero added Fe, whereas N₂ fixation increased 11-fold considerably for 450 nM Fe. The results suggest that N₂ fixation of Trichodesmium is more sensitive to Fe limitation than are the cell yields.     

Siderophores of Pseudomonas putida as an iron source for dicot and monocot plants

Iron uptake from ferrated (⁵⁹Fe) pseudobactin (PSB), a Pseudomonas putida siderophore, by various plant species was studied in nutrient solution culture under short term (10 h) and long term (3 weeks) conditions. In the short term experiments, ⁵⁹Fe uptake rate from ⁵⁹FePSB by dicots (peanuts, cotton and sunflower) was relatively low when compared with ⁵⁹Fe uptake rate from ⁵⁹FeEDDHA. Iron uptake rate from ⁵⁹FePSB was pH and concentration dependent, as was the Fe uptake rate from ⁵⁹FeEDDHA. The rate was about 10 times lower than that of Fe uptake from the synthetic chelate. Results were similar for long term experiments.Monocots (sorghum) in short term experiments exhibited significantly higher uptake rate of Fe from FePSB than from FeEDDHA. In long term experiments, FePSB was less efficient than FeEDDHA as an Fe source for sorghum at pH 6, but the same levels of leaf chlorophyll concentration were obtained at pH 7.3.Fe uptake rates by dicots from the siderophore and FeEDDHA were found to correlate with Fe reduction rates and reduction potentials (E₀) of both chelates. Therefore, it is suggested that the reduction mechanism governs the Fe uptake process from PSB by dicots. Further studies will be conducted to determine the role of pH in Fe aquisition from PSB by monocots.     

Iron-efficient and iron-inefficient oats and corn respond differently to iron-deficiency stress

Iron-efficient (WF9 corn and Coker 227 oat) and Fe-inefficient (ys₁ corn and TAM 0–312 oat) cultivars were comparatively tested for their response to Fe-deficiency stress induced by the use of either ferrous or ferric chelators. Corn and oats were grown in 20 M Fe with 0, 60, and 120 M BPDS and 40 M Fe with 0, 120, and 240 M BPDS and 20 M Fe with 0 and 40 M EDDHA. All four cultivars tested, both Fe-efficient and Fe-inefficient, continuously reduced Fe³⁺ to Fe²⁺ at a low level as evidenced by the production of Fe²⁺ (BPDS)₃ in test nutrient solutions over time. Severity of chlorosis increased as more BPDS was added to the nutrient solutions for both WF9 and ys₁ corn, but unlike corn, Coker 227 and TAM 0-312 oats were both able to obtain Fe from the Fe²⁺ (BPDS)₃ complex and were less chlorotic as a result. In short-term (4-hour) in vivo measurements, iron-stressed WF9 (Fe-efficient) corn reduced more Fe³⁺ to Fe²⁺ than similarly stressed ys₁ corn, Coker 227 oat or TAM 0-312 oat. Thus, at the same time that Fe-efficient WF9 corn reduces more Fe than the other cultivars, it is also unable to compete with BPDS for that Fe in the nutrient solution. These differences coupled with the observation that only Coker 227 oat produced measureable iron solubilizing substances (phytosiderophores) suggest that these two species differ in their mechanisms for obtaining Fe during Fe-deficiency stress.     

Isoelectric Focusing of Plant Plasma Membrane Proteins : Further Evidence that a 55 Kilodalton Polypeptide Is Associated with beta-1,3-Glucan Synthase Activity from Pea

The role of the root apoplasm for iron acquisition was studied in wheat (Triticum aestivum L. cv Ares) grown in nutrient solution under controlled environmental conditions. To obtain different levels of Fe in the root apoplasm, plants were supplied in the dark for 5 hours (preloading period) with various ⁵⁹Fe-labeled Fe compounds [Fe(III) hydroxide; microbial siderophores: Fe rhodotorulic acid (FeRDA) and ferrioxamin (FeDesferal³), and synthetic Fe chelate (FeEDDHA)], each at a concentration of 5 micromolar. Large pools of apoplasmic Fe were formed after supplying Fe(III) hydroxide or FeRDA, but no such pools were observed after supplying FeDesferal or FeEDDHA. Depending on plant Fe nutritional status (preculture ± 0.1 millimolar FeEDTA), apoplasmic Fe was used to different extent for translocation to the shoot. Under Fe deficiency, a much greater fraction of the apoplasmic Fe was utilized than in Fe-sufficient plants, as a result of the different rates of phytosiderophore release. Because of the diurnal rhythm in release of phytosiderophores in Fe-deficient plants, the utilization of the apoplasmic Fe for translocation into the shoot started 2 hours after onset of the light period and was dependent on the concentration of Fe in the apoplasm, which followed the order: Fe(III) hydroxide FeRDA FeDesferal = FeEDDHA. From these results, it can be concluded that in soil-grown plants the apoplasmic Fe pool loaded by various indigenous Fe compounds such as siderophores in the soil solution can be an important Fe source in graminaceous species, particularly during periods of limited Fe supply from the soil.     

Reduction and transport of Fe from siderophores

Soils contain siderophores produced by bacteria and fungi; however, the role of siderophores in Fe nutrition of plants is uncertain. The Strategy I plant cucumber (Cucumis sativus L.) was used in an investigation of ferric chelate reduction activity and uptake and transport of Fe from ferric hydroxyethylethylenetriacetic acid (FeHEDTA) and ferric N,N–di–(2–hydroxybenzoyl)–ethylenediamine– N,N-diacetic acid (FeHBED) and the hydroxamate siderophores, ferric rhodotorulic acid (FeRA) and ferric ferrioxime B (FeFOB). Cucumber seedlings were grown in a hydroponic medium without Fe or supplied with 10 M FeHEDTA. Iron-deficient cucumber roots readily reduced FeHEDTA, while Fe-sufficient roots had low levels of ferric chelate reduction activity. The siderophore FeRA was reduced by Fe-deficient roots at 8% of the rate of FeHEDTA, while FeFOB was not reduced. The highly stable synthetic chelate FeHBED was reduced at 16% the rate of FeHEDTA. Fe transport to shoots by Fe-deficient seedlings from the slowly reducible complexes ⁵⁹FeRA and ⁵⁹FeHBED was, respectively, 74% and 73% of that transported from ⁵⁹FeHEDTA. The ferrous complexing agent, bathophenanthrolinedisulfonic acid (BPDS), had a strong inhibitory effect on uptake and transport of Fe from ⁵⁹FeHEDTA or ⁵⁹FeRA into shoots. An average of 11% as much Fe was transported to shoots of Fe-deficient seedlings from ⁵⁹FeFOB as from ⁵⁹FeHEDTA. Neither the Fe nutritional status of the seedlings nor the presence of BPDS influenced the uptake and transport of Fe from ⁵⁹FeFOB. It is concluded that cucumber roots may take up substantial amounts of Fe from FeRA and FeHBED following reduction, while small amounts of Fe may be taken up from FeFOB by a mechanism not involving reduction of the ferric siderophore at the root surface.     

Ferric chelate reduction by suspension culture cells and roots of soybean: A kinetic comparison

The abilities of suspension cultures and intact roots of soybean (Glycine max L. cv. Hawkeye) to reduce ferric chelate were compared. Ferric chelate was supplied as ferric hydroxyethylethylenediaminetriacetic acid (FeHEDTA) and reduction was measured spectrophotometrically using bathophenan-throlinedisulfonic acid (BPDS) as the ferrous scavenger. Ferric chelate reduction by cell suspension cultures showed typical saturation kinetics; however, no difference was observed between cells that had been continuously grown with Fe (+Fe) and those that had been grown for four days without added Fe (–Fe). Values for K_m and V_max, determined from a Lineweaver-Burk plot, were 57 M and nmoles mg^-1 dry weight for the +Fe cells and 50 M and 22 nmoles mg^-1 dry weight for the -Fe cells, respectively. Ferric chelate reduction by Fe-deficient roots also exhibited saturation kinetics, while roots grown with adequate Fe did not reduce ferric chelate. The K_m and V_max values for Fe-deficient roots were 45 M and 20 nmoles mg^-1 dry weight, respectively, and did not differ from values obtained for cells in culture. This study offers strong evidence that the mechanism responsible for the reduction of ferric chelate is the same for cultured cells and roots and that the process is controlled at the cellular level. We propose that suspension cultures can be used as an alternative to intact roots in the study of ferric chelate reduction.     

Effects of phosphorus and iron fertilizers on the growth of two soybean varieties at two soil temperatures

The influence of FeEDDHA (0, 0.2 and 2 μg Fe g⁻¹ soil) and NaH₂PO₄·H₂O (0 and 120 μg Pg⁻¹ soil) on the growth of two Fe-ineffective soybean (Glycine max L. Merr.) varieties (anoka and T203) on a calcareous soil at two soil temperatures (16 and 24°C) was compared under greenhouse conditions. The two soybean varieties differed in the following respects: (a) T203 accumulated smaller concentrations of Fe in washed tops than Anoka under comparable conditions; (b) T203 was more susceptible to Fe deficiency and its accentuation by high levels of fertilizer P than Anoka; (c) T203 accumulated lower quantities of Mn in tops than Anoka under comparable conditions; (d) T203, but not Anoka, developed Mn deficiency symptoms when treated with P and 2 μg Fe g⁻¹ at 16°C. Fe deficiency was more severe in both varieties at the higher soil temperature due apparently to: (a) greater plant concentration of P in tops at 24°C; and/or (b) an increased rate of plant growth and greater dilution of Fe in young tissue at 24°C. Foliar P concentration was increased much more than foliar Fe concentration by an increase in soil temperature. Severely Fe deficient T203 plants grown without FeEDDHA at 24°C accumulated less foliar Mn than their FeEDDHA counterparts. Comparisons of Fe effectiveness of various soybean cultivars based on relative responses to FeEDDHA can be influenced by differential effects on Mn nutrition.     

Studies on structure activity relationship of some dihydroxy-4-methylcoumarin antioxidants based on their interaction with Fe(III) and ADP

Three dihydroxy-4-methylcoumarin (DHMC) derivatives, namely 7,8-DHMC, 6,7-DHMC and 5,7-DHMC alone and complexed with Fe (III) and ADP have been tested for their antioxidative potential. Chemical speciation studies and formation constants reveal the formation of strong DHMC–Fe–ADP (1:1:1) ternary complex. In vitro studies were done for their antioxidative property by scavenging the superoxide radicals (O ₂ ^– ) generated by xanthine + xanthine oxidase (XO) reaction. The IC₅₀ values for 7,8-DHMC, 6,7-DHMC and 5,7-DHMC and their ternary complexes with Fe (III)–ADP worked out to be 34.0 M, 62.0 M, 8.80 mM and 10.5, 11.5 and 148.5 M, respectively. The results indicate that O ₂ ^– scavenging potential of all the three DHMCs increased significantly after forming the ternary complex with Fe(III) and ADP. The structure activity relationship studies suggest that the introduction of hydroxyl group at 7th and 8th positions in the coumarins, irrespective of Fe(III)–ADP complexation, increases the antioxidative efficacy. No change in uric acid production in the reactions done for all studies further reveals that the coumarin derivatives and their complexes were the only causative factors for O ₂ ^– scavenging and not the suppression of the enzyme, xanthine oxidase.Published online: March 2005     

Low-spin, mononuclear, Fe(III) complexes with P,N donor hemilabile ligands: A combined experimental and theoretical study

Two new mononuclear Fe(III) complexes, [FeCl₃{PPh₂(p-C₆H₄NMe₂)-P}₃](1) (PPh₂(p-C₆H₄NMe₂): 4-(dimethylamino)phenyldiphenylphosphine) and [FeCl₃(PPh₂py-P)(PPh₂py-P,N)] (2) (PPh₂py: diphenyl(2-pyridyl)phosphine) were synthesized by reacting anhydrous FeCl₃ with respective ligand in acetonitrile solution under refluxing condition. Both the complexes were characterized by elemental analysis, FAB-Mass, FTIR, UV-Vis, ESR, Cyclic Voltammetry and magnetic measurement. The FAB mass spectra of complexes 1 and 2 show molecular ion peak at m/z 1078 [M]⁺ and m/z 687 [M−1]⁺, respectively, indicating mononuclear nature of the complexes. UV-Vis spectra of the complexes were consistent with low-spin, octahedral geometry. The variable temperature magnetic susceptibility measurement (73-323 K) of these complexes is also consistent with the paramagnetic nature of the complexes with a ground state spin S = ½. The Fe(III) centers of these two complexes remain low-spin, both at room temperature and liquid nitrogen temperature, was also indicated by the ESR analysis. Cyclic Voltammetry of both the complexes show an irreversible oxidation wave attributed to Fe³⁺ → Fe⁴⁺ + e⁻ along with the peak for ligand oxidation. Theoretical calculations (B3LYP) of the complexes show that for complex 1, a trans geometry of the two phosphorous atoms and for complex 2, a mer,cis structures are the most favored geometrical isomer. TDDFT calculations were performed to interpret the observed bands in the UV-Visible spectra.     

Iron deficiency-induced changes in carbon fixation and leaf elemental composition of sugar beet (Beta vulgaris) plants

In this experiment we (i) tested the hypothesis that, besides decreasing leaf C fixation, lime induced iron (Fe) deficiency increases root C fixation via PEP carboxylase and (ii) assessed the Fe-induced modifications in the elemental composition of plant tissues. Sugar beet plants were grown in nutrient solutions with Fe (45 M Fe-EDTA; +Fe control) or in a similar nutrient solution without Fe (–Fe) and in presence of CaCO₃ (1.0 gL^–1), either labelled with ¹³C (20 at. %) or unlabelled. After 7 and 17 days from treatment imposition, plants were harvested and single organs analysed for total O, C, H, macro and micronutrients. ¹³C abundance was also assessed in control, unlabelled and labelled –Fe plants. Iron deficiency caused significant growth reductions; chlorophyll and net photosynthesis decreased markedly in Fe-deficient plants when compared to the controls, whereas leaf transpiration rates and stomatal conductance were not affected by Fe deficiency. Iron deficient plants had leaf biomass with lower C (2 to 4%) and higher O (3 to 5%) concentrations than +Fe plants. The ¹³C was higher (less negative) in +Fe than in –Fe unlabelled plants. Iron deficient plants grown in the nutrient solution enriched with labelled CaCO₃ absorbed a relatively small amount of labelled C, which was mainly recovered in the fine roots and accounted for less than 2% of total C gain in the 10 d treatment period. Evidences suggest that iron deficient sugar beets grown in the presence of CaCO₃ do not markedly shift their C fixation from leaf RuBP to root PEPC.     

Water-extractable humic substances enhance iron deficiency responses by Fe-deficient cucumber plants

The ability of Fe-deficient cucumber plants to use iron complexed to a water-extractable humic substances fraction (WEHS), was investigated. Seven-day-old Fe-deficient plants were transferred to a nutrient solution supplemented daily for 5 days with 0.2 μM Fe as Fe-WEHS (5 μg org. C mL^-1), Fe-EDTA, Fe-citrate or FeCl₃. These treatments all allowed re-greening of the leaf tissue, and partial recovery of dry matter accumulation, chlorophyll and iron contents. However, the recovery was faster in plants supplied with Fe-WEHS and was already evident 48 h after Fe supply. The addition of 0.2 μM Fe to the nutrient solution caused also a partial recovery of the dry matter and iron accumulation in roots of Fe-deficient cucumber plants, particularly in those supplied with Fe-WEHS. The addition of WEHS alone (5 μg org. C mL^-1, 0.04 μM Fe) to the nutrient solution slightly but significantly increased iron and chlorophyll contents in leaves of Fe-deficient plants; in these plants, dry matter accumulation in leaves and roots was comparable or even higher than that measured in plants treated with Fe-citrate or FeCl₃. After addition of the different iron sources for 5 days to Fe-deficient roots, morphological modifications (proliferation of lateral roots, increase in the diameter of the sub-apical zones and amplified root-hair formation) and physiological responses (enhanced Fe(III)-chelate reductase and acidification of the nutrient solution) induced by Fe deficiency, were still evident, particularly in plants treated with the humic molecules. The presence of WEHS caused also a further acidification of the nutrient medium by Fe-deficient plants. The Fe-WEHS complex (1 μM Fe) could be reduced by intact cucumber roots, at rates of reduction higher than those measured for Fe-EDTA at equimolar iron concentration. Plasma membrane vesicles, purified by two-phase partition from root microsomes of Fe-deficient plants, were also able to reduce Fe-WEHS. Results show that Fe-deficient cucumber plants can use iron complexed to water soluble humic substances, at least in part via reduction of complexed Fe(III) by the plasma membrane Fe(III)-chelate reductase of root cells. In addition, the stimulating effect of humic substances on H⁺ release might be of relevance for the overall response of the plants to iron shortage. This revised version was published online in June 2006 with corrections to the Cover Date.     

Enzymatic responses of cucumber roots to different levels of Fe supply

Development of the coordinated response to decreasing Fe availability was studied in cucumber plants grown in nutrient solution (NS) over a range of Fe^III-EDTA concentrations (from 0.1 to 80 M). Physiological and biochemical parameters were evaluated in intact roots, root extracts and plasma membrane (pm) vesicles. Acidification of the NS was evident in plants grown at 1 M Fe^III-EDTA and inversely related to the external Fe concentration. Fe^III-EDTA reduction by intact roots was also gradually depressed by increasing Fe supply. The rate of net nitrate uptake by the roots was directly related to the amount of Fe^III-EDTA added to the NS. Activity of pmH⁺-ATPase was significantly higher in plants grown without added Fe as compared to those grown at 80 M Fe. A lower increase, dependent on Fe concentration, was observed at 0.1, 1, 5 or 10 M Fe^III-EDTA. Activity of pmFe^III-EDTA reductase was also increased by Fe deprivation and strongly correlated with pmH⁺-ATPase activity. PEP-carboxylase activity gradually increased with decreasing Fe concentration in the NS. Changes in activity and amount of the enzyme showed a close correlation with parameters measured in intact roots (nitrate uptake, Fe^III-EDTA reduction). Results show that the development of the Fe-deficiency response in cucumber roots can be finely tuned by the level of Fe supply. Adjustments to different levels of available Fe involve a correlated modulation of pm-associated enzymes. PEP-carboxylase activity appeared to be a suitable metabolic marker of the Fe nutritional status of the plant.     

Factors affecting the growth form of Aspergillus terreus NRRL 1960 in relation to itaconic acid fermentation

The effect of medium composition on the growth form of Aspergillus terreus NRRL 1960 in relation to itaconic acid fermentation has been studied. Four types of mycelial pellets were obtained under the conditions used and may be classified as (a) frayed and loose with 0.1–0.5 mm diameter (b) compact with 0.1–0.5 mm diameter (c) loose with 0.5–2.0 mm diameter and (d) compact with 0.5–2.0 mm diameter. Their respective maximum specific rates of formation and yields of itaconic acid, based on 100 g sucrose supplied, were (a) 1.25 mol mg^–1h^–1 and 55–59 g, (b) 0.27–0.43 mol mg^–1 h^–1 and 26–38 g, (c) 0.75–0.90 mol mg^–1 h^–1 and 45–51 g and (d) 0.12 mol mg^–1 h^–1 and 10 g. The presence of Ca²⁺, Zn²⁺ and Fe²⁺ in the basal medium at concentrations of 23.3 mg/100 ml, 0.01 mg/100 ml and 0.006 mg/100 ml respectively were found to be adequate and crucial in obtaining the desired outgrowth for both high production rates and consistent yields of itaconic acid. The further addition of either commercial plaster of Paris or analytical-reagent-grade CaSO₄, especially when activated by heating to 530°C and present in excess of solubility, results in small and frayed pellets, which lead to itaconic acid yields of 55–59 g acid/100 g sugar supplied.     

Selection of soybean plant leaves which yield mesophyll cell isolates with maximal rates of CO2 and NO inf2 sup− photoassimilation

A problem often encountered when assaying mesophyll cell isolates prepared from mature soybean leaves, was that of poor reproducibility in rates of net ¹⁴CO₂ photoassimilation and NO₂ ^– photoreduction. It was known that soybean source leaves repeatedly displayed their most active net CO₂ photoassimilation in the period from attainment of maximal leaf area to approximately two to five days subsequent to that point. Advantage was taken of the fact that when soybean leaflets of each leaf reach their maximal area they also have reached their maximal leaf length from base to tip. This facilitates a more rapid determination of the point in time in which leaflet areas had reached A_max. Soybean plants (Glycine max cv. Williams) were propagated in the growth chamber with a 12 h light-12 h dark cycle, 25C, 65% RH, and 700 microeinsteins per meter squared per second. At 24 d post-emergence, the third leaf (numbered acropetally from the unifoliates) of each plant had just attained maximum leaflet areas (110 cm²) and lengths (13 cm). For this study, leaf mesophyll cells were enzymatically isolated, using commercially prepared pectinase, from leaflet sets of leaves selected from each of the second, third, and fourth leaf positions. Maximal rates of net ¹⁴CO₂ photoassimilation (with 5 mM HCO₃ ^–) for the second, third and fourth leaf (leaflet) isolates were, respectively, 27.0, 57.0, and 41.7 mol ¹⁴CO₂ assimilated per milligram chlorophyll per hour; simultaneously maximal rates of NO _inf2 ^sup– photoreduction (1 mM NO _inf2 ^sup– ) were, respectively, 4.4, 8.1, and 0.0 mol NO _inf2 ^sup– reduced per milligram chlorophyll per hour. These studies made it clear that in order repeatedly to attain reproducible maximal rates of leaf cell isolate net ¹⁴CO₂ photoassimilation and NO _inf2 ^sup– photoreduction, it always was necessary to select the newest, fully expanded leaves (e.g. leaf number 3) for cell isolation. Leaves from several plants only were pooled if they were excised from identically the same node on each of the plants.Abbreviations A_max - maximum leaflet (trifoliolate) area attained during ontogeny - CO₂ - CO₂ gas dissolved in solution - HCO _inf3 ^sup– - bicarbonate - L_max - maximum leaf blade length (midvein) attained during ontogeny - NiRase - chloroplast nitrite reductase (reduced ferredoxin) - NiPR - nitrite photoreduction - PE - post-emergence - Pn - net CO₂ photoassimilation (for leaflets and mesophyll cell isolates) - PPRC - pentose phosphate reductive cycle     

Efficient aerobic oxidation of hydrocarbons promoted by high-spin nonheme Fe(II) complexes without any reductant

Fe(II)-tris(2-pyridylmethyl)amine complexes, Fe(II)-tpa, having different co-existing anions, [Fe(tpa)(MeCN)₂](ClO₄)₂ (1), [Fe(tpa)(MeCN)₂](CF₃SO₃)₂ (2) and [Fe(tpa)Cl₂] (3), were prepared. Effective magnetic moments (evaluated by the Evans method) revealed that while 1-3 in acetone and 3 in acetonitrile (MeCN) have a high-spin Fe(II) ion at 298 K, the Fe(II) ions of 1 and 2 are in the low-spin state in MeCN. The aerobic oxidation of 1-3 was monitored by UV-Vis spectral changes in acetone or MeCN under air at 298 K. Only the high-spin Fe(II)-tpa complexes were oxidized with rate constants of k_obs = 0.1-1.3 h⁻¹, while 1 and 2 were stable in MeCN. The aerobic oxidation of 1 or 2 in acetone was greatly accelerated in the presence of pure, peroxide-free cyclohexene (1000 equiv.) and yielded a large amount of oxidized products; 2-cyclohexe-1-ol (A) and 2-cyclohexene-1-one (K) (A + K: 23 940% yield based on Fe; A/K = 0.3), and cyclohexene oxide (810%). Besides cyclohexene, aerobic oxidation of norbornene, cyclooctene, ethylbenzene, and cumene proceeded in the presence of 1 in acetone at 348 K without any reductant. Essential factors in the reaction are high-spin Fe(II) ion and labile coordination sites, both of which are required to generate Fe(II)-superoxo species as active species for the H-atom abstraction of hydrocarbons.     

Effects of nitric oxide and Fe supply on recovery of Fe deficiency induced chlorosis in peanut plants

The effects of nitric oxide (NO) and/or iron (Fe) supplied to Fe deficient plants have been investigated in peanut (Arachis hypogaea L.) grown in Hoagland nutrient solution with or without Fe. Two weeks after Fe deprivation, recovery was induced by addition of 250 μM sodium nitroprusside (SNP, a NO donor) and/or 50 μM Fe (Fe-EDTA) to the Fe deprived (-Fe) nutrient solution. Activities of antioxidant enzymes, leaf chlorophyll (Chl), and active Fe content decreased, whereas activities of H⁺-ATPase, ferric-chelate reductase (FCR), nitrate reductase, and nitric oxide synthase and NO production increased in Fe deficient plants, consequently an Fe chlorosis symptom appeared obviously. In contrast, these symptoms disappeared gradually after two weeks with NO and/or Fe supply, which caused an increases in leaf Chl and active Fe content, especially following by co-treatment with NO and Fe to values found in Fe sufficient plants. Increased activities of antioxidant enzymes (superoxide dismutase, peroxidase, and catalase) and decreased accumulation of reactive oxygen species (H₂O₂, O ₂ ^?? ) and malondialdehyde enhanced the ability of resistance to oxidative stress. Supplied NO alone had the obvious effect on increased NO production and on activity of H⁺-ATPase and FCR, whereas root length and root/shoot ratio were most effectively increased by Fe supplied alone. Co-treatment with NO and Fe did the best effects on recovery peanut chlorosis symptoms by significantly increased Chl and available Fe content and adjusted distribution of Fe and other mineral elements (Ca, Mg, and Zn) in both leaves and roots.     

Effect of Benzylaminopurine on Rehydration of Bean Plants after Water Stress

The possibility of improving the recovery of plant photosynthesis after water stress by cytokinin-induced stimulation of stomatal opening or delay of leaf senescence was tested. The 6-benzylaminopurine (BAP) in concentrations 1 and 10 M was applied to the substrate (sand + nutrient solution) or sprayed on primary leaves of 14-d-old Phaseolus vulgaris L. plants sufficiently supplied with water or water-stressed for 4 d. The later ones having relative water content decreased to 69 % were fully rehydrated during the following three days. Parameters of photosynthesis and water relations were measured in primary leaves of 7-, 10-, 14-, and 17-d-old plants. Application of 1 M BAP slightly delayed leaf senescence: in 17-d-old control plants, net photosynthetic rate (P_N) and chlorophyll (Chl) content, and when sprayed on leaves also some of Chl a fluorescence kinetic parameters of BAP-treated leaves were slightly higher than those of untreated leaves. Both types of application of 1 M BAP slightly improved recovery of plants during rehydration after water stress in terms of increased g_ad, g_ab and P_N, i.e., parameters which were markedly decreased by mild water stress. However, contents of Chl a, Chl b and carotenoids and parameters of Chl a fluorescence kinetic were not markedly affected by mild water stress and after rehydration were not stimulated by 1 M BAP. 10 M BAP had mostly negative effects on the parameters measured.     

A method for long-term micropropagation of Phaseolus coccineus L.

A method for long-term plant regeneration of Phaseolus coccineus L, is described. Shoot-tips and cotyledonary nodes cultured on a Murashige and Skoog medium supplemented with N⁶-benzylaminopurine, 10 M, and -naphthaleneacetic acid, 1M, formed multiple bud-shoots. These shoots were transferred to medium containing BAP 1 M, NAA 0.1 M, and gibberellic acid 3 M to promote shoot growth and further shoot multiplication. Rooting was achieved in medium with 11 M indole-3-acetic acid. Rooted plants grew to maturity and were fertile. Cultures have maintained their ability to regenerate plants for more than two years. A sample of 30 regenerated plants (R₀) was tested for chromosome number, all of them being diploid; seven isozymatic systems were electrophpretically analyzed in 82 R₀ regenerated plants. No differences were observed in their electrophoretic patterns in comparison with those shown by seedlings. Histological studies revealed the origin of buds from calluses via organogenesis.Abbreviations BAP N⁶-benzylaminopurine - 2,4-D 2,4-dichlorophenoxyacetic acid - GA₃ gibberellic acid - IAA indole-3-acetic acid - MS Murashige and Skoog (1962) medium - NAA -naphthaleneacetic acid - ADH alcohol dehydrogenase - GOT glutamic-oxaloacetic transaminase - MDH malate dehydrogenase - 6PGD 6-phosphogluconate dehydrogenase - PGI Phosphoglucose isomerase - PGM phosphoglucose mutase - SK shikimate dehydrogenase     

Organic solid wastes from urban environment as iron sources for sorghum

Sorghum (Sorghum bicolor L. Moench) which is susceptible to Fe deficiency was grown in two different soils in a glasshouse with two different organic urban wastes (sewage sludge and dog manure) to ascertain their ability to supply Fe and other micronutrients to plants. One soil was calcareous with a history of Fe deficiency. Sewage sludge and dog manure at an application rate of 15,000 g/g to this soil effectively supplied Fe to plants. This effect was not present when the ash rather than the organic matter was used. Ferrous sulfate and Fe-EDDHA (Fe[ethylenediamine di-0-hydroxyphenylacetate]) likewise were not effective. Infrared spectra revealed differences in the fulvic acid for the two sources of solid wastes. The results imply that some sources of organic wastes may be useful in prevention or correction of Fe deficiency in calcareous soils.