New expression system tightly controlled by zinc availability in Lactococcus lactis

Here we developed the new expression system P(Zn) zitR, based on the regulatory signals (P(Zn) promoter and zitR repressor) of the Lactococcus lactis zit operon, involved in Zn(2+) high-affinity uptake and regulation. A P(Zn) zitR-controlled expression vector was constructed, and expression regulation was studied with two reporter genes, uspnuc and lacLM; these genes encode, respectively, a protein derived from Staphylococcus aureus secreted nuclease and Leuconostoc mesenteroides cytoplasmic beta-galactosidase. Nuclease and beta-galactosidase activities of L. lactis MG1363 cells expressing either uspnuc or lacLM under the control of P(Zn) zitR were evaluated on plates and quantified from liquid cultures as a function of divalent metal ion, particularly Zn(2+), availability in the environment. Our results demonstrate that P(Zn) zitR is highly inducible upon divalent cation starvation, obtained either through EDTA addition or during growth in chemically defined medium, and is strongly repressed in the presence of excess Zn(2+). The efficiency of the P(Zn) zitR expression system was compared to that of the well-known nisin-controlled expression (NICE) system with the same reporter genes cloned under either P(Zn) zitR or P(nisA) nisRK control. lacLM induction levels reached with both systems were on the same order of magnitude, even though the NICE system is fivefold more efficient than the P(Zn) zitR system. An even smaller difference or no difference was observed after 3 h of induction when nuclease was used as a reporter for Western blotting detection. P(Zn) zitR proved to be a powerful expression system for L. lactis, as it is tightly controlled by the zinc concentration in the medium.     

Response of Fresh-Water Protozoan Artificial Communities to Metals*

An artificial fresh-water protozoan community was subjected to different concentrations of Zn and Cu in a test system consisting of Plexiglas troughs through which pond water flowed continuously. Although the % survival of colonizing species exposed to Cu or Zn fluctuated greatly at each concentration, the range of toxicity for each compound allowed comparison of protozoa with other organisms with respect to resistance to heavy metal toxicity. Individual protozoan species also were exposed for 3 hr to Zn, Cu, Cr, phenol, Pb, Mn, Co, HNO₃, acetic acid, Al, Sn, and HCl to derive time to death curves. Protozoa tested appeared to be more resistant than Daphnia to phenol, K₂C₂O₇, and Cu; however, some species were more sensitive than Daphnia to Zn, nitric acid, and HCl. This suggests that the sensitivity of protozoa to toxicants may be either more or less than that of macroinvertebrates and that information does not suffice to predict sensitivity. Moreover, the relative sensitivity of protozoa to various toxicants will not always be the same, i.e. species X may be twice as tolerant to a toxicant as species Y but its relative sensitivity may be quite different for another toxicant.     

Evaluation of elemental allelopathy in Acroptilon repens (L.) DC. (Russian Knapweed)

Although Acroptilon repens (L.) DC. (Russian knapweed) is known to concentrate zinc (Zn) in upper soil layers, the question of whether the elevated Zn has an allelopathic effect on restoration species has not been addressed. Experiments were conducted to investigate whether soils collected from within infestations of A. repens (high-Zn) inhibit the germination or growth and development of desirable restoration species, compared to soils collected adjacent to an A. repens infestation (low-Zn). Four bioassay species [Sporobolus airoides (Torrey) Torrey (alkali sacaton), Pseudoroegneria spicata (Pursh) A. Love (bluebunch wheatgrass), Psathyrostachys juncea (Fischer) Nevski (Russian wildrye) and A. repens] were germinated in a growth chamber and grown in a greenhouse in both soils and received treatments for the alleviation of Zn toxicity (P, Fe, Fe-oxide, and soil mixing) to isolate the effects of elevated soil Zn on plant performance. Percent germination, total plant biomass, tiller and stem number, inflorescence number, and tissue metal levels were compared among soil types and treatments for each species. There was no evidence from any of the indicators measured that high-Zn soils reduced plant performance, compared to low-Zn soils. Tissue Zn levels barely approached the lower range of phytotoxic levels established for native grasses. Older plants with longer exposure times may accumulate higher Zn concentrations. S. airoides and A. repens both had higher biomass in the high-Zn soil, most likely due to increased macronutrient (N and P) availability. As the Zn levels in the soils used in this study were much higher than any levels previously reported in soils associated with A. repens, it is unlikely that the elevation of soil Zn by A. repens will hinder germination or growth and development of desirable grasses during establishment.     

Mercury(II) binding to metallothioneins. Variables governing the formation and structural features of the mammalian Hg-MT species.

With the aim of extending our knowledge on the reaction pathways of Zn-metallothionein (MT) and apo-MT species in the presence of Hg(II), we monitored the titration of Zn7-MT, Zn4-alphaMT and Zn3-betaMT proteins, at pH 7 and 3, with either HgCl2 or Hg(ClO4)2 by CD and UV-vis spectroscopy. Detailed analysis of the optical data revealed that standard variables, such as the pH of the solution, the binding ability of the counter-ion (chloride or perchlorate), and the time elapsed between subsequent additions of Hg(II) to the protein, play a determinant role in the stoichiometry, stereochemistry and degree of folding of the Hg-MT species. Despite the fact that the effect of these variables is unquestionable, it is difficult to generalize. Overall, it can be concluded that the reaction conditions [pH, time elapsed between subsequent additions of Hg(II) to the protein] affect the structural properties more substantially than the stoichiometry of the Hg-MT species, and that the role of the counter-ion becomes particularly apparent on the structure of overloaded Hg-MT.     

Arbuscular mycorrhizas are beneficial under both deficient and toxic soil zinc conditions

Background and aims

Arbuscular mycorrhizas (AM) play different roles in plant Zn nutrition depending on whether the soil is Zn-deficient (AM enhancement of plant Zn uptake) or Zn-toxic (AM protection of plant from excessive Zn uptake). In addition, soil P concentration modifies the response of AM to soil Zn conditions. We undertook a glasshouse experiment to study the interactive effects of P and Zn on AM colonisation, plant growth and nutrition, focusing on the two extremes of soil Zn concentration—deficient and toxic.

Methods

We used a mycorrhiza-defective tomato (Solanum lycopersicum) genotype (rmc) and compared it to its wild-type counterpart (76R). Plants were grown in pots amended with five soil P addition treatments, and two soil Zn addition treatments.

Results

The mycorrhizal genotype generally thrived better than the non-mycorrhizal genotype, in terms of biomass and tissue P and Zn concentrations. This was especially true under low soil Zn and P conditions, however there was evidence of the ‘protective effect’ of mycorrhizas when soil was Zn-contaminated. Above- and below-ground allocation of biomass, P and Zn were significantly affected by AM colonisation, and toxic soil Zn conditions.

Conclusions

The relationship between soil Zn and soil P was highly interactive, and heavily influenced AM colonisation, plant growth, and plant nutrition.     

Comparison of the metal-ion-promoted dephosphorylation of the 5'-triphosphates of adenosine, inosine, guanosine and cytidine by Mn2+, Ni2+ and Zn2+ in binary and ternary complexes.

The dependence of the rate of dephosphorylation of ATP, ITP, GTP and CTP (= NTP), expressed as first-order rate constants (50 degrees C; I = 0.1 M, NaClO4), on pH (2 to 10), in the absence and presence of Mn2+, Ni2+, and Zn2+, was investigated. The reaction is accelerated by Zn2+ and passes through a pH optimum at about 8 for the system Zn2+-ATP or 9 for Zn2+-ITP and Zn2+-GTP; this is analogous to observations made earlier with the corresponding Cu2+ systems. By computing the pH dependence of the distribution of the several species present in these systems it is shown that the highest rates are observed in the pH regions where the concentration of Zn(ATP)2-, Zn(ITP-H)3-, or Zn(GTP-H)3- dominates. By evaluating the pH dependence evidence is given that the attacking nucleophile is OH- or H2O for Zn (ATP)2- and H2O for Zn (ITP-H)3- or Zn(GTP-H)3-. For all these complexes metal-ion/nucleic-base interactions are known, leading to the formation of macrochelates. These metal-ion/nucleic-base interactions are crucial for the observation of a metal-ion-promoted dephosphorylation; in agreement with this, and the small tendency of the cytosine moiety to coordinate, the CTP systems are rather stable towards dephosphorylation. It should be noted that these experimental results do not necessarily mean that the macrochelates usually described are the reactive complexes, but only that the active complex must be closely related to them (e.g. isomers, etc). Although for the Ni2+ systems with ATP, ITP, and GTP, and for the Mn2+-ATP system a metal-ion/nucleic-base interaction is also known, these systems are not very sensitive to hydrolytic cleavage of the terminal P-O-P bond. The only known significant structural difference between the Ni2+-NTP or the Mn2+-ATP complexes and those of Cu2+ or Zn2+ is that Ni2+ Mn2+ coordinate to all three phsophate groups, whereas Cu2+ and Zn2+ involve only the beta and gamma ones. This structure-reactivity relationship is rationalized by the suggestion that in the active species the metal ion should be coordinated to the alpha,beta-phosphate groups leaving the gamma-group open to nucleophilic attack. Obviously, an initial beta,gamma-coordination is suitable for a shift of the metal ion along the phosphate back-bone into the reactive alpha-beta-position, while for an alpha,beta,gamma-coordination only the less favorable removal of the coordinated gamma-group remains. The metal-ion/nucleic-base interaction is considered as being important for achieving this reactive structure. The connection between trans-phosphorylation in vitro and in vivo is discussed. It is also shown that the formation of mixed-ligand or ternary complexes inhibits the dephosphorylation process. This is on the one hand of interest with regard to the transport of hydrolysis-sensitive phosphates in nature, while on the other it casts doubts on conclusions based on experiments carried out in the presence of buffers, because these contain weak bases and hence potential ligands.     

Bioaccumulation and Physiological Response of Five Willows to Toxic Levels of Cadmium and Zinc

We evaluated the phytoremediation potential of Salix spp. exposed to high cadmium (Cd) and zinc (Zn) concentrations to select feasible plant materials for restoration and revegetation of mining soil contaminated by heavy metals on the basis of their Cd and Zn accumulation, Cd-Zn interaction on bioaccumulation, and the changes of photosynthetic parameters. The Cd and Zn concentrations were in the order of root > leaf > stem, regardless of the species. In the combined Cd and Zn treatment, the leaf and stem Cd concentration in all species were higher relative to Cd-alone treatment. In contrast, the Zn concentration in plant tissues when exposed to the combined Cd + Zn treatment decreased relative to the Zn-alone treatment. The translocation factor (TF) of Cd and Zn from root to leaf was generally higher compared to TF from root to stem than those in the single treatment. The Cd + Zn treatments resulted in enhanced translocation of Cd from root to aboveground tissue (synergistic), while the same treatment suppressed the Zn translocation from root to leaf and stem (antagonistic). The reduction of photosynthetic parameters in Zn alone and Cd + Zn treatments was generally higher than that of Cd-alone treatment. Among the different species, S. caprea and P. alba×glandulosa have the lowest photosynthetic reduction relative to the control. Overall, S. caprea could be a potential candidate for phytoremediation of Cd- and Zn-contaminated sites.     

Molecular design of an acid–base cooperative catalyst for RNA cleavage based on a dizinc complex

The effects of donor groups of dizinc complexes, formed from a 2:1 mixture of Zn(II) and a dinucleating ligand, on adenylyl(3'-5')adenosine (ApA) cleavage have been studied. Two dinucleating ligands were used: one had two 2-pyridylmethyl and two 2-hydroxyethyl moieties on the 1,3-diamino-2-propanol linker moiety (2), and the other had two 2-pyridylmethyl and two carboxymethyl moieties on the 1,3-diamino-2-propanol linker moiety (3(2-)). The dizinc complex with2 [(Zn(2+))(2)-2] showed higher activities toward ApA cleavage than the dizinc complex using an analogous dinucleating ligand having four 2-pyridylmethyl donor moieties [(Zn(2+))(2)-1] at pH 5-8. The former showed a bell-shaped pH-rate constant profile, whereas the latter showed a sigmoidal pattern. The differences in the pH-rate constant profile are attributable to the various distributions of the monohydroxo-dizinc species, i.e. dideprotonated species, which are responsible for ApA cleavage. The monohydroxo species of (Zn(2+))(2)-2 has two acidic protons, which are not present in the corresponding monohydroxo species of (Zn(2+))(2)-1. The existence of both intracomplex acid (ROH or H(2)O) and base catalysts (RO(-) or OH(-)) in (Zn(2+))(2)-2 can explain its higher activity toward ApA cleavage than that of (Zn(2+))(2)-1. In contrast, (Zn(2+))(2)-3(2-) showed lower activity toward ApA cleavage at pH 7.0, which can be ascribed to the absence of the monohydroxo-dizinc species under these conditions.     

Mechanism of phosphorus-induced zinc deficiency in cotton. I. Zinc deficiency-enhanced uptake rate of phosphorus

The effect of withholding Zn on the uptake, translocation and accumulation of P was studied in cotton plants ( Gossypium hirsutum L. cv. Deltapine 15/21) grown in nutrient solutions under controlled environmental conditions. The influence of P on the uptake rate, translocation and distribution of ⁶⁵Zn in the plants was also examined. Increasing the P supply resulted in severe Zn deficiency symptoms (interveinal chlorosis) as well as P toxicity symptoms, which were characterized by leaf puckering and grayish-brown marginal necrosis. Zinc deficiency markedly increased the uptake and translocation rates of P over the whole concentration range tested (5x10^-5 to 1.25x10^-3 M ). Uptake and translocation rates of P increased with both level of P and severity of Zn deficiency. This often caused P toxicity symptoms on Zn-deficient leaves. In contrast to P, the concentrations of K and Mg in the leaves were not affected by Zn deficiency. Similar results were obtained for sunflower ( Helianthus annuus L.) and buckwheat ( Fagopyrum esculentum Moench) plants. Higher P concentrations in Zn-deficient leaves or shoots could not be attributed wholly to reduced shoot growth. This was also evident when Zn deficiency was compared with other micronutrient (Fe, Mn, and Cu) deficiencies. Only Zn-deficient plants showed enhanced uptake and translocation of P. In experiments with ⁶⁵Zn, a high P supply did not depress uptake and translocation of Zn. From the results obtained it is concluded that the P-induced Zn deficiency in cotton, as well as in other species, is primarily caused by enhanced P uptake and translocation and not by inhibition of Zn uptake.     

Comparative study of metal resistance and accumulation of lead and zinc in two poplars

In our study, we tested two poplars, Populus beijingensis and Populus cathayana, as model species for their potential for phytoremediation by measuring changes in biomass, pigments, superoxide radicals (O₂^?), cellular ultrastructure and their ability for O₂^? quenching and heavy metal accumulation when exposed to Pb, Zn and their interaction in a hydroponic system. Exposure to Pb did not cause a significant decrease in biomass in either P. beijingensis or P. cathayana. Correspondingly, no obvious impairment in cellular organelles was observed in either species, although the former species translocated a higher fraction of Pb to its shoots than the latter. In contrast, there were significant decreases in biomass and pigment content, and serious impairments in ultrastructure in both species when exposed to either Zn alone or to a combined treatment. Under such conditions, P. beijingensis showed smaller losses of biomass and pigments but a greater ability to quench O₂^? and maintained relatively intact cellular organelles compared with P. cathayana. Under the combined stress, there were no obvious additive effects on biomass, pigments or cellular impairment, whereas synergistic effects on metal absorption and accumulation in both species were observed when compared with exposure to either alone. Thus, the attribute of synergistic uptake and translocation in both species validates their potential to remediate soil contaminated by multiple metals. Moreover, our results indicated that P. beijingensis is a better potential candidate for phytoremediation than P. cathayana, due to its greater phytoremediation efficiency as well as its higher tolerance capacity.     

Compartmentation of Zinc in Roots and Leaves of the Zinc Hyperaccumulator Thlaspi caerulescens J & C Presl

Thlaspi caerulescens is a metallophyte that is able to hyperaccumulate Zn. In the present study the subcellular compartmentation of Zn was investigated in roots and leaves of this species by means of X-ray microanalysis. Leaves accumulated higher average Zn concentrations than roots. In roots of plants exposed to 10 μM Zn, Zn concentrations in the apoplast were similar to those in vacuoles, while in plants treated with 100 μM Zn considerably higher Zn concentrations were detected in vacuoles than in the apoplast. In epidermal and sub-epidermal cells of leaves of plants from both treatments, Zn mainly accumulated in vacuoles and, to a lesser extent, in the apoplast. In vacuoles from plants exposed to 100 μM Zn, high Zn concentrations were associated with variable amounts of P, Ca and K. In leaves, the highest Zn concentrations (13,600 μg g^?1 d.m.) were found in globular crystals present in many vacuoles of epidermal and subepidermal cells. Smaller deposits with a variable Zn concentration between 1,000 and 18,300 μg g^?1 d.m. were observed in the epidermal and subepidermal cells of roots. Both the high Zn/P element ratios found in the crystals and the absence of Mg indicate that, in contrast to other plant species, myo-inositol hexaphosphate (phytate) is not the main storage form for Zn in Thlaspi caerulescens.     

Protective mechanisms of Pycnogenol® in ethanol‐insulted cerebellar granule cells

Pycnogenol® (PYC), a patented combination of bioflavonoids extracted from the bark of French maritime pine (Pinus maritima), inhibits apoptosis and necrosis of developing neurons exposed acutely to ethanol (EtOH). The present study shows that the protective mechanisms of PYC in EtOH‐exposed postnatal day 9 cerebellar granule cells (P9 CGCs) include (1) reduction of reactive oxygen species (ROS) production; (2) counteraction of suppressed copper/zinc superoxide dismutase (Cu/Zn SOD) and glutathione peroxidase/reductase (GSH‐Px/GSSG‐R) system activities; (3) upregulation of Cu/Zn SOD protein expression; (4) mitigation of the EtOH‐mediated exacerbation of catalase (CAT) activity; and, (5) specific binding and inhibition of active caspase‐3. These results indicate that the mechanisms by which PYC antagonizes EtOH‐induced oxidative stress include oxidant scavenging and modulation of endogenous, cellular proteins. Using findings from the present and previous studies, a model delineating the mechanisms of EtOH effects on the system of antioxidant enzymes in developing CGCs is presented. © 2004 Wiley Periodicals, Inc. J Neurobiol, 2004     

Bark beetle-mediated fungal infections of susceptible trees induce resistance to subsequent infections in a dose dependent manner

1 Experiments were conducted to determine whether propagule loads on the twig beetles Pityophthorus setosus and Pityophthorus carmeli (Coleoptera: Scolytidae) influence the pathogen infection of the host tree in the Monterey pine- Fusarium circinatum system.
2 On an average, F. circinatum was isolated from 2.6% and 3.3% of trapped P. setosus and P. carmeli , respectively, although the isolation percentages varied over the season, being highest in the spring and lowest in late summer and fall for both species. Mean pathogen load was 13.4 and 22.6 propagules per beetle, on P. setosus and P. carmeli , respectively, and decreased from May to November for both species. The pathogen was also isolated from approximately 55% of both beetle species that emerged from infected branches. Mean propagule load on emerged P. setosus and P. carmeli was 39 and 66.5, respectively.
3 On the basis of these data, beetle species were treated with one of three propagule loads (low, medium, high) and caged onto live branches to determine whether they could transmit the pathogen. At all propagule loads, both species transmitted the pathogen, and transmission percentage and lesion length, a measure of tree susceptibility, were positively correlated with propagule load.
4 To investigate further whether the previous transmission by beetles could affect response of the same trees to subsequent infection with F. circinatum , different branches were inoculated on the same trees used in the transmission study, and lesion lengths were measured. Lesion lengths were lower on trees that had been previously exposed to beetles treated with high or medium propagule loads than on trees that had previously been exposed to beetles treated with low propagule loads. This suggests that the initial infection by beetles carrying high or medium propagule loads induced resistance to subsequent infections of the host, whereas infections caused by beetles with low propagule loads did not.     

Intracellular zinc release-activated ERK-dependent GSK-3β-p53 and Noxa-Mcl-1 signaling are both involved in cardiac ischemic-reperfusion injury

Oxidative stress and nitrosative stress are both suggested to be involved in cardiac ischemia-reperfusion (I/R) injury. Using time-lapse confocal microscopy of cardiomyocytes and high-affinity O(2)(-?) and Zn(2+) probes, this study is the first to show that I/R, reactive oxygen species (ROS), and reactive nitrogen species (RNS) all cause a marked increase in the [O(2)(-?)](i), resulting in cytosolic and mitochondrial Zn(2+) release. Exposure to a cell-penetrating, high-affinity Zn(2+)(i) chelator, TPEN, largely abolished the Zn(2+)(i) release and markedly protected myocytes from I/R-, ROS-, RNS-, or Zn(2+)/K(+) (Zn(2+)(i) supplementation)-induced myocyte apoptosis for at least 24?h after TPEN removal. Flavonoids and U0126 (a MEK1/2 inhibitor) largely inhibited the myocyte apoptosis and the TPEN-sensitive I/R- or Zn(2+)(i) supplement-induced persistent extracellular signal-regulated kinase 1 and 2 (ERK1/2) phosphorylation, dephosphorylation of p-Ser9 on glycogen synthase kinase 3β (GSK-3β), and the translocation into and accumulation of p-Tyr216 GSK-3β and p53 in, the nucleus. Silencing of GSK-3β or p53 expression was cardioprotective, indicating that activation of the ERK-GSK-3β-p53 signaling pathway is involved in Zn(2+)-sensitive myocyte death. Moreover, the ERK-dependent Noxa-myeloid cell leukemia-1 (Mcl-1) pathway is also involved, as silencing of Noxa expression was cardioprotective and U0126 abolished both the increase in Noxa expression and in Mcl-1 degradation. Thus, acute upstream Zn(2+)(i) chelation at the start of reperfusion and the use of natural products, that is, flavonoids, may be beneficial in the treatment of cardiac I/R injury.     

Effects of human presence on two social wasp species

Abstract.  1. The effects of human presence on the behavioural response and productivity of a native wasp species of North America ( Polistes fuscatus ) and an invasive wasp species ( Polistes dominulus ) were examined in a field experiment. Over a 3-month period, colonies of each species were exposed to human presence at either low (biweekly) or high (daily) levels. At 2-week intervals, wasp response to human presence was recorded for all colonies via an index, ranging from no response (0) to an attack (4).
2. The levels of human presence had no effect on wasp response or colony productivity but foundresses of both species increased their response level the closer a human was to the nest. In addition, foundresses of P. fuscatus became more aggressive over time, corresponding to after adult offspring emerged, whereas foundresses of P. dominulus did not change their response level after the emergence of offspring.
3. The adult offspring of P. dominulus responded to human presence at a level similar to that of the foundress, whereas adult offspring of P. fuscatus exhibited a lower response to human presence than that of the foundress. The more distinctive aposematic coloration of P. dominulus may play a role in the behavioural differences between species; it may be more effective in deterring predators, and with the result that leaving the nest to defend it is less often necessary. Accordingly, the less aggressive response to human presence by P. dominulus may contribute to its success as an introduced species in North America.     

Protective mechanisms of pycnogenol in ethanol-insulted cerebellar granule cells

Pycnogenol (PYC), a patented combination of bioflavonoids extracted from the bark of French maritime pine (Pinus maritima), inhibits apoptosis and necrosis of developing neurons exposed acutely to ethanol (EtOH). The present study shows that the protective mechanisms of PYC in EtOH-exposed postnatal day 9 cerebellar granule cells (P9 CGCs) include (1) reduction of reactive oxygen species (ROS) production; (2) counteraction of suppressed copper/zinc superoxide dismutase (Cu/Zn SOD) and glutathione peroxidase/reductase (GSH-Px/GSSG-R) system activities; (3) upregulation of Cu/Zn SOD protein expression; (4) mitigation of the EtOH-mediated exacerbation of catalase (CAT) activity; and, (5) specific binding and inhibition of active caspase-3. These results indicate that the mechanisms by which PYC antagonizes EtOH-induced oxidative stress include oxidant scavenging and modulation of endogenous, cellular proteins. Using findings from the present and previous studies, a model delineating the mechanisms of EtOH effects on the system of antioxidant enzymes in developing CGCs is presented.     

Zinc stoichiometry in Escherichia coli alkaline phosphatase. Studies by 31P NMR and ion-exchange chromatography.

31P nuclear magnetic resonance spectra and enzymatic activities are compared for alkaline phosphatase (orthophosphoric-monoester phosphohydrolase (alkaline optimum), EC 3.1.3.1) species with different zinc contents. The enzyme containing two Zn2+ per protein dimer exists in two forms; one, prepared by dialysis of native enzyme, has full enzymatic activity and a 31P magnetic resonance spectrum similar to but distinguishable from that of the native enzyme containing four or more Zn2+. The other form, prepared by restoring two Zn2+ to apoenzyme, has low enzymatic activity and a 31P magnetic resonance spectrum that indicates stoichiometric binding of phosphate, but otherwise altered properties. Reconstituted enzyme with four Zn2+ is similar to but distinguishable from native enzyme with four Zn2+. Chromatography on DEAE-cellulose can separate apoenzyme and enzyme containing two Zn2+ and suggests that the binding of a pair of Zn2+ is cooperative.     

Favouring the bioavailability of Zn and Cu to enhance the production of lignin-modifying enzymes in Trametes versicolor cultures

Metal effect on the enzyme secretion in fungi is usually related to total concentrations but not to bioavailable metal species. In this work, we aimed at enhancing the secretion of lignin-modifying oxidoreductases in Trametes versicolor by favouring the bioavailability of essential metals. For this purpose, the fungus was exposed to Cu or Zn in liquid culture media exhibiting different complexation levels. Metal speciation was determined experimentally or theoretically to quantify free metal species, supposed to be the most bioavailable, and species complexed to ligands. Although Zn(2+) contents were high in media, Zn had no effect on the oxidoreductase production. Conversely, Cu highly induced the manganese peroxidase and laccase productions until 40 and 310 times when compared to unexposed controls. This inductive potential was highly correlated to Cu(2+) contents in media. Furthermore, in poorly complexing media, the response threshold of oxidoreductases to Cu greatly decreased and an unexpected production of lignin peroxidase occurred.     

Enhancing phosphorus and zinc acquisition efficiency in rice: a critical review of root traits and their potential utility in rice breeding

Background

Rice is the world''s most important cereal crop and phosphorus (P) and zinc (Zn) deficiency are major constraints to its production. Where fertilizer is applied to overcome these nutritional constraints it comes at substantial cost to farmers and the efficiency of fertilizer use is low. Breeding crops that are efficient at acquiring P and Zn from native soil reserves or fertilizer sources has been advocated as a cost-effective solution, but would benefit from knowledge of genes and mechanisms that confer enhanced uptake of these nutrients by roots.

Scope

This review discusses root traits that have been linked to P and Zn uptake in rice, including traits that increase mobilization of P/Zn from soils, increase the volume of soil explored by roots or root surface area to recapture solubilized nutrients, enhance the rate of P/Zn uptake across the root membrane, and whole-plant traits that affect root growth and nutrient capture. In particular, this review focuses on the potential for these traits to be exploited through breeding programmes to produce nutrient-efficient crop cultivars.

Conclusions

Few root traits have so far been used successfully in plant breeding for enhanced P and Zn uptake in rice or any other crop. Insufficient genotypic variation for traits or the failure to enhance nutrient uptake under realistic field conditions are likely reasons for the limited success. More emphasis is needed on field studies in mapping populations or association panels to identify those traits and underlying genes that are able to enhance nutrient acquisition beyond the level already present in most cultivars.     

The inhibition of mitochondrial complex I (NADH:ubiquinone oxidoreductase) by Zn2+

NADH:ubiquinone oxidoreductase (complex I) from bovine heart mitochondria is a highly complicated, membrane-bound enzyme. It is central to energy transduction, an important source of cellular reactive oxygen species, and its dysfunction is implicated in neurodegenerative and muscular diseases and in aging. Here, we describe the effects of Zn2+ on complex I to define whether complex I may contribute to mediating the pathological effects of zinc in states such as ischemia and to determine how Zn2+ can be used to probe the mechanism of complex I. Zn2+ inhibits complex I more strongly than Mg2+, Ca2+, Ba2+, and Mn2+ to Cu2+ or Cd2+. It does not inhibit NADH oxidation or intramolecular electron transfer, so it probably inhibits either proton transfer to bound quinone or proton translocation. Thus, zinc represents a new class of complex I inhibitor clearly distinct from the many ubiquinone site inhibitors. No evidence for increased superoxide production by zinc-inhibited complex I was detected. Zinc binding to complex I is mechanistically complicated. During catalysis, zinc binds slowly and progressively, but it binds rapidly and tightly to the resting state(s) of the enzyme. Reactivation of the inhibited enzyme upon the addition of EDTA is slow, and inhibition is only partially reversible. The IC50 value for the Zn2+ inhibition of complex I is high (10-50 microm, depending on the enzyme state); therefore, complex I is unlikely to be a major site for zinc inhibition of the electron transport chain. However, the slow response of complex I to a change in Zn2+ concentration may enhance any physiological consequences.