Inhibition of ethylene production by cobaltous ion

The effect of Co²⁺ on ethylene production by mung bean (Phaseolus aureus Roxb.) and by apple tissues was studied. Co²⁺, depending on concentrations applied, effectively inhibited ethylene production by both tissues. It also strongly inhibited the ethylene production induced by IAA, kinetin, IAA plus kinetin, Ca²⁺, kinetin plus Ca²⁺, or Cu²⁺ treatments in mung bean hypocotyl segments. While Co²⁺ greatly inhibited ethylene production, it had little effect on the respiration of apple tissue, indicating that Co²⁺ does not exert its inhibitory effect as a general metabolic inhibitor. Ni²⁺, which belongs to the same group as Co²⁺ in the periodic table, also markedly curtailed both the basal and the induced ethylene production by apple and mung bean hypocotyl tissues.     

Action of ferric and aluminium ions on the dormancy breakage of <Emphasis Type="Italic">Stylosanthes humilis</Emphasis> seeds

Dormancy of scarified seeds of Stylosanthes humilis was broken by acidic Al³⁺ and Fe³⁺ solutions. Fe⁺³-stimulated seeds exhibited a high activity of 1-aminocyclopropane-1-carboxylate (ACC) oxidase and produced great amounts of ethylene, which showed correlated with the germination process. In addition, specific inhibitors of ethylene biosynthesis and action largely depressed the Fe³⁺-stimulated germination, leading to the conclusion that the ion broke dormancy by triggering ethylene production by the seeds. By contrast, inhibitors of ethylene biosynthesis and action did not impair germination of Al³⁺-stimulated dormant seeds. Moreover, ethylene production and activity of ACC oxidase of Al³⁺-treated seeds was substantially decreased by inhibitors of ethylene biosynthesis, but germination kept large. Together these data suggest that ethylene biosynthesis was not required in the chain of events triggered by Al³⁺ leading to dormancy breakage. Methyl viologen (MV), a reactive oxygen species-generating compound, broke dormancy of seeds to the same extent as Al³⁺ did. Germination of both Al³⁺- and MV-stimulated dormant seeds was inhibited by sodium selenate, an antioxidant compound; selenate, however had no effect on germination of Fe³⁺-stimulated seeds. Together these data indicate that the mechanisms underlying the germination of Al³⁺- and Fe³⁺-treated seeds are not the same.     

The relationship between pathogenic fungal metabolities (fusaric and picolinic acid), endogenous ethylene evolution and the development of ethylene-like symptoms

Summary The effects of fusaric acid (5-n-butylpicolinic acid), picolinic acid (2-pyridine carboxylic acid), and picloram (4-amino-3, 5, 6-trichloropicolinic acid) on endogenous ethylene production by tomato cuttings and elongation growth of oat coleoptile sections were measured. Ethylene production by tomato cuttings was substantially stimulated by treatment with 1×10⁻³ and 1×10⁻⁵ M picoloram and to a lesser extent by 1×10⁻³ M fusaric acid; picolinic acid had little effect. The ethylene levels produced in response to fusaric acid are not high enough to account for the ethylene injury observed in Fusarium wilt. Fusaric acid inhibited oat coleoptile extension, picolinic acid had little effect, and picloram promoted growth.     

Cu(II): a “signaling molecule” of the mangrove endophyte <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> ZZF51?

We previously reported the isolation of Cu–fusaric acid (Cu–FA) complex from the mangrove endophyte Fusarium oxysporum ZZF51. In this study, we explored the mechanism of Cu–FA production in the strain ZZF51 by comparing with that of another endophyte Fusarium sp. B2, which produced FA but not Cu–FA in the same culture condition. The results allowed us to hypothesize that Cu²⁺ may act as a “signaling molecule” to awaken the silent FA biosynthetic genes in ZZF51, inducing intracellular production of FA followed by chelation with Cu²⁺. This signaling network was triggered specifically by Cu²⁺ and may be interfered by other metal ions.     

Effects of methyl jasmonate and excess copper on root and leaf growth

A short time effects of 25 and 150 μM Cu²⁺ or 50 μM methyl jasmonate (MJ) on growth of roots and leaves of Phaseolus coccineus, Allium cepa and Zea mays were investigated. Both Cu²⁺ and MJ inhibited root growth. Jasmonate synthesis inhibitors (ibuprofen, IB, salicylhydroxamic acid, SHAM, and propylgallate, PG) partially reversed the inhibitory effect of Cu²⁺ in P. coccineus, but in A. cepa this effect was not clear. Pretreatment with NADPH oxidase inhibitor (20 mM imidazole, IM), and especially ethylene inhibitor (silver thiosulphate, STS) mostly weakened Cu²⁺ effect on root growth in P. coccineus and A. cepa. The growth of P. coccineus leaves also slowed down by Cu²⁺ and this effect was partially ameliorated by IB, PG and IM, and completely by SHAM and STS. In Z. mays the effect of STS was considerably lower than that of PG and SHAM which reversed the effect of Cu²⁺. These results indicate that jasmonate, ethylene and NADPH oxidase activity may be involved in Cu²⁺ inhibitory action on the roots of dicotyledon plants, but in A. cepa only ethylene and NADPH oxidase are involved. However, leaf growth inhibition induced by excess Cu²⁺ is connected in Z. mays especially with jasmonate, and in P. coccineus with ethylene, NADPH oxidase and, to a minor degree, with jasmonate.     

Rapid Ethylene Production in Soybean in Response to the Cupric Ion

Very rapid accumulation of free 1-aminocyclopropane-1-carboxylicacid (ACC), followed by stimulation of ethylene production wereinduced by a Cu²⁺ in soybean cuttings. The accumulation mustbe attributed to an increase in ACC synthesis, because: (1)it was completely inhibited by aminoethoxyvinylglycine (AVG);and (2) the ethylene stimulation was inhibited by AVG, indicatingthat free ACC cannot be released from its conjugated form. Theactivity of the ethylene-forming enzyme slightly decreased followingthe Cu²⁺ pulse, and this event was accompanied by a slight increasein electrolyte leakage from the treated soybean tissues. Glycine max L., soybean, ethylene, cupric ion     

Enhanced heavy metal immobilization by a bacterial-chitosan complex in soil

Soil contaminated with heavy metals (e.g., Pb²⁺, Cu²⁺, and Zn²⁺) was treated with aminopoly-saccharide chitosan alone or a strain of the bacterium Bacillus subtilis to determine their effects on metal ion accumulations. Phosphatase and Chitosanase production were also assayed. The combined bacterial-chitosan treatment showed the greater accumulation of Zn²⁺ followed by Cu²⁺ as compared to single treatments, while Pb²⁺ did not show a marked accumulation by either single or combined treatments.     

Cellular responses of developing Fucus serratus embryos exposed to elevated concentrations of Cu2+

Elevated concentrations of Cu²⁺ can have inhibitory effects on early development in plants and algae by targeting specific cellular processes. In the present study the effects of elevated Cu²⁺ on developmental processes in embryos of the brown algae Fucus serratus (Phaeophyceae) were investigated. Elevated Cu²⁺ was shown to inhibit fixation of the zygotic polar axis but not its formation. Actin localization was unaffected by elevated Cu²⁺ but polarized secretion, which occurs downstream, was inhibited. Significant differences in tolerance to Cu²⁺ were observed for polarization and rhizoid elongation of embryos derived from adults from Cu²⁺‐contaminated and uncontaminated locations. Moderate Cu²⁺ exposure inhibited the generation of cytosolic Ca²⁺ signals in response to hypo‐osmotic shocks. In contrast, cytosolic Ca²⁺ was elevated by treatments with high [Cu²⁺] and this coincided with production of reactive oxygen species. The results indicate that direct effects on signalling processes involved in polarization and growth may in part explain complex, concentration‐dependent effects of Cu²⁺ on early development.     

Cation binding sites on synaptic vesicles

The protein-sensitized fluorescence of Tb³⁺ was used as a probe for cation binding sites on synaptic vesicles. Competition studies show that the order of affinity for the sites is Cu²⁺ > Mn²⁺ > Ca²⁺ > Mg²⁺ and Zn²⁺ is inactive. Fluorescence quenching studies indicate that the site is superficial and the effect of pH suggests that histidine is involved in the binding. Measurements of enzyme activities in the presence of lanthanides reveal that the metal binding site identified by Tb³⁺ fluorescence is not the Cu²⁺ site associated with dopamine-β-hydroxylase. Terbium inhibits Ca²⁺-stimulated ATPase but not Mg²⁺-stimulated ATPase activities of the synaptic vesicle fraction. A kinetic analysis indicates that the site monitored by Tb³⁺ fluorescence may be a component of the Ca²⁺-stimulated ATPase. It is also suggested that Mg²⁺ and especially Cu²⁺ may bind to the sites in vivo, serving as a bridge between vesicles and other synaptic components such as the presynaptic plasma membrane.     

Temporal and spatial patterns of soil water following wildfire-induced changes in plant communities in the Great Basin in Nevada,USA

Infection of tall fescue (Festuca arundinacea Schreb.) with its endemicNeotyphodium coenophialum-endophyte (Morgan-Jones and Gams) Glenn, Bacon and Hanlin appears to reduce copper (Cu) concentrations in forage and serum of grazing animals, contributing to a range of immune-related disorders. A greenhouse experiment was conducted to identify effects of novel endophyte strains on Cu acquisition by tall fescue (Festuca arundinacea Schreb.) varieties Grasslands Flecha and Jesup infected with a novel, non ergot producing endophyte strain AR542, and two perennial ryegrass (Lolium perenne L.) varieties Aries and Quartet infected with a novel, non lolitrem B producing strain AR1, and their noninfected (E−) forms. Individual endophyte/grass associations were cultivated in nutrient solutions at 1.0 (P+) and 0.0 mM (P−) phosphorus concentrations. The Cu²⁺-binding activity of extracellular root exudates, and concentrations of Cu and other heavy metals in roots and shoots were measured. Extracellular root exudates of AR542-infected vs. E− tall fescue had higher Cu²⁺-binding activity only in P− nutrient solution as shown by lower concentration of free Cu²⁺ (0.096 vs. 0.188 mmol Cu²⁺ g⁻¹ root DM, respectively). The Cu²⁺-binding activity by root exudates of perennial ryegrass was not affected by endophyte infection, but was higher (i.e., lower concentration of free Cu²⁺) in P− vs. P+ nutrient solution (0.068 vs. 0.114 mmol Cu²⁺ g⁻¹ root DM). In this hydroponic experiment, Cu concentrations in shoots of both grasses were not a function of Cu²⁺-binding activity and endophyte effects on heavy metal concentrations in shoots and roots were specific for each variety. The Cu²⁺-binding activity of extracellular root exudates may affect Cu accumulation by field-grown, endophyte-infected tall fescue under P-limiting growth conditions and warrants verification by more specific methods.     

Effect of Cu Toxicity on Growth of Cowpea (Vigna unguiculata)

Accurate determination of the rhizotoxicity of Cu in dilute nutrient solutions is hindered by the difficulty of maintaining constant, pre-determined concentrations of Cu (micromolar) in solution. The critical Cu²⁺ activity associated with a reduction in the growth of solution-grown cowpea (Vigna unguiculata (L.) Walp. cv. Caloona) was determined in a system in which Cu was maintained constant through the use of a cation exchange resin. The growth of roots and shoots was found to be reduced at solution Cu²⁺ activities ≥1.7 μM (corresponding to 90% maximum growth). Although root growth was most likely reduced due to a direct Cu²⁺ toxicity, it is considered that the shoot growth reduction is attributable to a decrease in tissue concentrations of K, Ca, Mg, and Fe and the formation of interveinal chlorosis. At high Cu²⁺ activities, roots were brown in color, short and thick, had bent root tips with cracking of the epidermis and outer cortex, and had local swellings behind the roots tips due to a reduction in cell elongation. Root hair growth was reduced at concentrations lower than that which caused a significant reduction in overall root fresh weight.     

Abscisic acid and Ethrel abolish the inhibition of adventitious root formation of paclobutrazol-treated bean primary leaf cuttings

Paclobutrazol (PB), a triazole growth retardant and an inhibitor of gibberellin biosynthesis, reduced at 17 μM concentration the adventitious root formation of bean primary leaf cuttings. Treatments with 5 μM ABA or 4 μM Ethrel, an ethylenereleasing compound, restored the rooting of PB-treated cuttings. Ethylene production and the content of the precursor 1-aminocyclopropane-l-carboxylic acid (ACC) were enhanced in root-forming tissues of PB-treated petioles 48 h after ABA application. The effect of ABA could be abolished by 10 μM CoCl₂, an inhibitor of ACC oxidase. Thus, ABA might stimulate rooting through its effect on ethylene release. 2 mM silver thiosulphate, an inhibitor of ethylene action, decreased the rooting of PB-treated cuttings similarly to Co²⁺, but failed to negate the ABA effect. These data indicate that the effect of PB on rhizogenesis is not associated directly with the inhibition of the biosynthesis of gibberellins Acknowledgements: We are grateful to Gabriella Biró. This work was supported by the Hungarian National Science Research Foundation (OTKA), Project No. 462.     

Stimulation and oxidative catalytic inactivation of thermolysin by copper•Cys-Gly-His-Lys

[Cu²⁺•Cys-Gly-His-Lys] stimulates thermolysin (TLN) activity at low concentration (below 10 μM) and inhibits the enzyme at higher concentration, with binding affinities of 2.0 and 4.9 μM, respectively. The metal-free Cys-Gly-His-Lys peptide also stimulates TLN activity, with an apparent binding affinity of 2.2 μM. Coordination of copper through deprotonated imine nitrogens, the histidyl nitrogen, and the free N-terminal amino group is consistent with the characteristic absorption spectrum of a Cu²⁺–amino-terminal copper and nickel binding motif (λ _max ∼ 525 nm). The lack of thiol coordination is suggested by both the absence of a thiol to Cu²⁺ charge transfer band and electrochemical studies, since the electrode potential (vs. Ag/AgCl) 0.84 V (ΔE = 92 mV) for the Cu^3+/2+ redox couple obtained for [Cu²⁺•Cys-Gly-His-Lys] was found to be in close agreement with that of a related complex [Cu²⁺•Lys-Gly-His-Lys]⁺ (0.84 V, ΔE = 114 mV). The N-terminal cysteine appears to be available as a zinc-anchoring residue and plays a critical functional role since the [Cu²⁺•Lys-Gly-His-Lys]⁺ homologue exhibits neither stimulation nor inhibition of TLN. Under oxidizing conditions (ascorbate/O₂) the catalyst is shown to mediate the complete irreversible inactivation of TLN at concentrations where enzyme activity would otherwise be stimulated. The observed rate constant for inactivation of TLN activity was determined as k _obs = 7.7 × 10⁻² min⁻¹, yielding a second-order rate constant of (7.7 ± 0.9) × 10⁴ M⁻¹ min⁻¹. Copper peptide mediated generation of reactive oxygen species that subsequently modify active-site residues is the most likely pathway for inactivation of TLN rather than cleavage of the peptide backbone.     

Ethylene biosynthesis in Fusarium oxysporum f. sp. tulipae proceeds from glutamate/2-oxoglutarate and requires oxygen and ferrous ions in vivo

Liquid cultures of the deuteromycete, Fusarium oxysporum f. sp. tulipae, a tulip pathogen, produced high amounts of ethylene during stationary phase. 1-Aminocyclopropane-1-carboxylic acid, the direct precursor of ethylene in plants, was not present in the fungus. Radioactivity from [3,4-³H]glutamate as well as [U-¹⁴C]glutamate was incorporated into ethylene, indicating that it was derived from C3 and C4 of glutamate or 2-oxoglutarate. Ferrous ions markedly stimulated the rate of ethylene formation in vivo, whereas Fe³⁺, Cu²⁺ or Zn²⁺ had little or no effect. Ethylene biosynthesis was strongly inhibited by the heavy metal chelator ,-dipyridine. The effect of ,-dipyridine was fully reversed by Fe²⁺ ions and partially by Cu²⁺ and Zn²⁺ ions but not by the supply of glutamate or 2-oxoglutarate, suggesting that a step in the ethylene biosynthetic pathway downstream of 2-oxoglutarate is dependent on Fe²⁺. When stationary phase cultures were supplied with arginine, ornithine, or proline, ethylene production increased dramatically while addition of glutamate or 2-oxoglutarate had little effect. Tracer studies were performed to test the possibility that an intermediate in the catabolism of arginine to glutamate was the direct precursor of ethylene. In cultures supplied with [U-¹⁴C]arginine or [U-¹⁴C]glutamate, the specific radioactivity of ethylene was closely similar to the specific radioactivity of the endogenous glutamate pool, indicating that glutamate was on the pathway between arginine and ethylene. An enzyme system converting 2-oxoglutarate to ethylene in a reaction dependent on oxygen, ferrous ions and arginine has previously been described in extracts from Penicillium digitatum (Fukuda et al. 1986). The present results suggest that a similar enzyme system catalyzes the final step of ethylene biosynthesis in F. oxysporum.Non-standard abbreviations AdoMet S-adenosyl methionine - ACC 1-aminocyclopropane-1-carboxylic acid - EFE ethylene forming enzyme     

Study of the Action of Se and Cu on the Growth Metabolism of <Emphasis Type="Italic">Escherichia coli</Emphasis> by Microcalorimetry

The biological effect of Se and Cu²⁺ on Escherichia coli (E. coli) growth was studied by using a 3114/3236 TAM Air Isothermal Calorimeter, ampoule method, at 37°C. From the thermogenesis curves, the thermokinetic equations were established under different conditions. The kinetics showed that a low concentration of Se (1–10 μg/mL) promoted the growth of E. coli, and a high concentration of Se (>10 μg/mL) inhibited the growth, but the Cu²⁺ was always inhibiting the growth of E. coli. Moreover, there was an antagonistic or positive synergistic effect of Se and Cu²⁺ on E. coli in the different culture medium when Se was 1–10 μg/ml and Cu²⁺ was 1–20 μg/ml. There was a negative synergistic effect of Se and Cu²⁺ on E. coli when Se was higher than 10 μg/ml and Cu²⁺ was higher than 20 μg/ml. The antagonistic or synergistic effect between Se and Cu²⁺ on E. coli was related to the formation of Cu–Se complexes under the different experimental conditions chosen.     

Enhanced heavy metal immobilization by a bacterial-chitosan complex in soil

Soil contaminated with heavy metals (e.g., Pb²⁺, Cu²⁺, and Zn²⁺) was treated with aminopoly-saccharide chitosan alone or a strain of the bacterium Bacillus subtilis to determine their effects on metal ion accumulations. Phosphatase and Chitosanase production were also assayed. The combined bacterial-chitosan treatment showed the greater accumulation of Zn²⁺ followed by Cu²⁺ as compared to single treatments, while Pb²⁺ did not show a marked accumulation by either single or combined treatments.     

Biosorption of Cd, Cu, Pb, and Zn from aqueous solutions by the fruiting bodies of jelly fungi (Tremella fuciformis and Auricularia polytricha)

In this study, dried and humid fruiting bodies of Tremella fuciformis and Auricularia polytricha were examined as cost-effective biosorbents in treatment of heavy metals (Cd²⁺, Cu²⁺, Pb²⁺, and Zn²⁺) in aqueous solution. The humid T. fuciformis showed the highest capacity to adsorb the four metals in the multi-metal solutions. The Pb²⁺ adsorption rates were 85.5%, 97.8%, 84.8%, and 91.0% by dried T. fuciformis, humid T. fuciformis, dried A. polytricha, and humid A. polytricha, respectively. The adsorption amount of Pb²⁺ by dried and humid T. fuciformis in Cd²⁺ + Pb²⁺, Cu²⁺ + Pb²⁺, Pb²⁺ + Zn²⁺, Cd²⁺ + Cu²⁺ + Pb²⁺, and Cd²⁺ + Zn²⁺ + Pb²⁺ solutions were not lower than that in Pb²⁺ solutions. The results suggested that in humid T. fuciformis, Cd²⁺, Cu²⁺, and Zn²⁺ promoted the Pb²⁺ adsorption by the biomass. In the multi-metal solutions of Cd²⁺ + Cu²⁺ + Pb²⁺ + Zn²⁺, the adsorption amount and rates of the metals by all the test biosorbents were in the order of Pb²⁺ > Cu²⁺ > Zn²⁺ > Cd²⁺. Compared with the pseudo first-order model, the pseudo second-order model described the adsorption kinetics much better, indicating a two-step biosorption process. The present study confirmed that fruiting bodies of the jelly fungi should be useful for the treatment of wastewater containing Cd²⁺, Cu²⁺, Pb²⁺, and Zn²⁺.     

Effects of Copper on T-Type Ca<Superscript>2+</Superscript> Channels in Mouse Spermatogenic Cells

Low voltage-activated, rapidly inactivating T-type Ca²⁺ channels are found in a variety of cells, where they regulate electrical activity and Ca²⁺ entry. In whole-cell patch-clamp recordings from mouse spermatogenic cells, trace element copper (Cu²⁺) inhibited T-type Ca²⁺ current (I _T-Ca) with IC₅₀ of 12.06 μM. Inhibition of I _T-Ca by Cu²⁺ was concentration-dependent and mildly voltage-dependent. When voltage stepped to −20 mV, Cu²⁺ (10 μM) inhibited I _T-Ca by 49.6 ± 4.1%. Inhibition of I _T-Ca by Cu²⁺ was accompanied by a shift of −2.23 mV in the voltage dependence of steady-state inactivation. Cu²⁺ upshifted the current–voltage (I-V) curve. To know the change of the gating kinetics of T-type Ca²⁺ channels, we analyzed the effect of Cu²⁺ on activation, inactivation, deactivation and reactivation of T-type Ca²⁺ channels. Since T-type Ca²⁺ channels are a key component in capacitation and the acrosome reaction, our data suggest that Cu²⁺ can affect male reproductive function through T-type Ca²⁺ channels as a preconception contraceptive material.     

Red cabbage anthocyanin extract alleviates copper-induced cytological disturbances in plant meristematic tissue and human lymphocytes

Red cabbage is a source of health beneficial substances with antioxidant and antigenotoxic properties. HPLC analysis specifying the content of the investigated extract indicated that mainly anthocyanins (ATH) were responsible for its abilities. Cytological research was conducted with two experimental models: plant tissues—meristematic cells of Vicia faba, and animal tissue elements—human lymphocytes. Positive influence of ATH extract on mitotic activity of Vicia cells exposed to Cu²⁺ stress, and inhibitory effect of ATH on cytotoxic actions of Cu²⁺ on lymphocytes were demonstrated. In all experimental series with ATH application in combinations with Cu²⁺, mitotic index (MI) were higher than those obtained for only Cu²⁺ stressed tissues. Preincubation in ATH before Cu²⁺ stress had the best effect. Similarly, after ATH applications in all tested series decrease in frequency of micronuclei (MN) appearance was noticed in comparison with only Cu²⁺ stressed material. In the case of Vicia cells ATH acted effectively even applied after Cu²⁺ stress. It suggests that this ATH mixture not only prevents and limits but also heals the cytological injury caused by Cu²⁺ stress.     

Effects of iron and copper ions in promotion of selective abscission and ethylene production by citrus fruit and the inactivation of indoleacetic Acid

Application of Cu²⁺ (<10^-2 M) and Fe³⁺ ions as aqueous solutions of chloride salts promoted fruit abscission, erratic rind damage, and ethylene production of various citrus species with little to no defoliation. Mixing of 10^-5 M Cu²⁺ or Fe³⁺ ions with equimolar indole-3-acetic acid resulted in a reduction of the ultraviolet absorption at 220 nanometers, and an increase at 245 nanometers. Ultraviolet irradiation accelerated the change by Fe³⁺ and Cu²⁺ ions in the absorption of indole-3-acetic acid. Pretreatment of indole-3-acetic acid with Fe³⁺ and Cu²⁺ ions for 6 hours resulted in more than 90% reduction in its growth-promoting activity in the Avena bioassay, even when cations were removed by chromatography. Acceleration of abscission by Fe³⁺ and Cu²⁺ ions could be related to both promotion of ethylene production and direct inactivation of auxin.