Acetate as the main energy substrate for the sulfate-reducing bacteria in Lake Eliza (South Australia) hypersaline sediments

Abstract Simultaneous measurements of sulfate reduction and acetate oxidation using ³⁵S and ¹⁴C tracers showed that acetate was the main energy substrate for the sulfate-reducing bacteria in Lake Eliza sediments. Sulfate reduction rates calculated from acid-volatile sulfide data only, correlated with acetate oxidation at around 0.5:1. However, the rates calculated from acid-volatile plus pyrite sulfur data correlated with acetate oxidation at a ratio of around 1:1. Molybdate completely inhibited sulfate reduction but acetate oxidation was not totally inhibited. From 10 to 15% of acetate oxidation was not attributable to the sulfate-reducing bacteria. There was rapid accumulation of acetate, within the first 12 h of incubation. Acetate, propionate and butyrate accumulated in the presence of molybdate.     

The link between the electrogenic and redox functions of the plasmalemma and the energy metabolism of the cell

A dependence of the plasmalemma redox activity, determined by the reduction of external electron acceptors (ferricyanide, nitro-blue tetrazolium), on the energy state of the cell, which was modified by light conditions or introduction of glucose into the media, was shown on leaves of Elodea canadensis Rich. Glucose (10 m M ) and light (40 W m^-2) caused hyperpolarization of the membrane potential and stimulated the redox activity of the plasmalemma. 3-(3,4-Dichlorophenyl)-1,1-dimethylurea (DCMU) completely inhibited the light activation of electrogenic and redox functions of the plasmalemma. The light saturation intensity for membrane potential and ferricyanide reductase activity was 10–30% of the light saturation of photosynthesis. Membrane potential, K⁺ transport and plasmalemma redox activity changed in parallel in response to light and darkness and when DCMU was added. Ferricyanide reductase activity is suggested to be a simple parameter for characterizing the energy state of the cell. The functional significance of the light-induced hyperpolarization of the membrane potential is discussed.     

A rapid non-destructive assay to quantify soybean nodule gas permeability

The low gas permeability of a diffusion barrier in the cortex of soybean nodules plays a significant role in the protection of nitrogenase from oxygen inactivation. It may also set an upper limit on nodule respiration and nitrogen fixation rates. Two methods which have been used to quantify the gas permeability of leguminous nodules are reviewed and found to be unreliable. A new assay technique for determining both the nodule activity and gas permeability is developed and tested. This ‘lag-phase’ assay is based on the time nodules require to reach steady-state ethylene production after being exposed to acetylene. The technique is rapid, insensitive to errors in biochemical parameters associated with nitrogenase, and is non-destructive. The method was tested with intact aeroponically grown soybean plants for which the mean nodule gas permeability was found to be 13.3×10⁻³ mms⁻¹. This corresponds to a layer of cells approximately 35 um thick and is consistent with previously reported values.     

Soybean nodule gas permeability,nitrogen fixation and dirunal cycles in soil temperature

While diurnal cycles in nitrogen fixation rates are sometimes assumed to result from diurnal variation in photosynthetically active radiation, contradicting evidence exists that indicate soil temperature is the primary environmental influence. These studies assessed the significance of temperature on soybean nitrogen fixation under field conditions. Two groups of intact field-grown soybean plants, one at ambient and the other exposed to a 10°C diurnal variation in soil temperature, were nondestructively assayed for acetylene reduction rates. Activity was closely associated with soil temperature (R²=0.85), even when temperature was 12 h out of phase with ambient. Data were also obtained to determine if the effects of rhizosphere temperature on nitrogen fixation are mediated through an effect on the nodule oxygen permeability. Nodule oxygen permeability of intact, aeroponically grown soybean was closely correlated with the diurnal changes in temperature (R²=0.90).     

Asymmetric Reduction of 4-Oxoisophorone Using Immobilized Thermophilic Bacteria Under Conditions of Controlled Cell Growth

Asymmetric reduction of 2,6,6,-trimethyl-2-cyclohexene-l,4-dione (4-oxoisophrone) to (6R)-2,2,6-trimethyl-1,4-cyclohexane-dione((3R)-dihydro-4-oxoisophorone) was catalysed by immobilized thermophilic bacteria, Thermomonospora curvata JTS 321. Because of leakage of entrapped cells from gel beads during reactions using culture medium, we optimized the medium to allow the microbial conversion under conditions of controlled cell growth. Of the media screened, liver infusion medium was found to be the most suitable and microbial conversion was achieved without cell leakage from the immobilized gels. Immobilized T. curvata cells were repeatedly used for the asymmetric reduction of 4-oxoisophorone, more than 15 times, with an extent of conversion of 50%.     

Anaerobic growth on elemental sulfur using dissimilar iron reduction by autotrophic Thiobacillus ferrooxidans

Abstract Anaerobic growth on elemental sulfur using dissimilar iron reduction by Thiobacillus ferrooxidans has been demonstrated. The ferric ion reducing activity (FIR) of the anaerobic cells was double that of the aerobic cells. Significant differences in inhibition of FIR by respiratory inhibitors were observed between aerobic and anaerobic cells. A higher amount of cytochrome was detected in anaerobic cells compared to aerobic cells. Absorption minima developed with the addition of ferric sulfate in the dithionite reduced cell suspension demonstrated that the ferric ion could accept electrons from the cytochrome system of this bacterium. The possibility of two different electron transport chains in ferric ion reduction is discussed.     

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.     

A genetically related response to iron deficiency stress in muskmelon

A mutant muskmelon (Cucumis melo L.) with characteristic Fe-deficiency chlorosis symptoms was compared to related cultivars in its ability to obtain Fe via the widely known Fe-stress response mechanisms of dicotyledonous plants. The three cultivars (fefe, the Fe-inefficient mutant; Mainstream and Edisto, both Fe efficient plants) were grown in nutrient solution in either 0 or 3.5 mg L^-1 Fe as FeCl₃. None of the three cultivars released reductants or phytosiderophores, but both Edisto and Mainstream produced massive amounts of H+ ions to reduce and maintain the pH of nutrient solutions below pH 4.0. The roots of these two Fe-efficient cultivars were also capable of reducing Fe³⁺ to Fe²⁺. These responses maintained green plants, resulted in high leaf Fe in both Edisto and Mainstream, and produced Mn toxicity in Mainstream. The lack of Fe-deficiency stress response in fefe not only affected leaf Fe concentration and chlorosis, but also resulted in reduced uptake of Mn. The importance of reduced Fe (Fe²⁺) to the Fe-efficient cultivars was confirmed by growing the cultivars with BPDS (4, 7-diphenyl-1, 10-phenanthroline disulfonic acid, a ferrous chelator) and EDDHA [ethylene-diamine di (0-hydroxphenylacetic acid)] (a ferric chelator), and observing increased chlorosis and reduced Fe uptake in BPDS grown plants. The Fe-deficiency response observed in these cultivars points out the diversity of responses to Fe deficiency stress in plants. The fefe mutant has a limited ability to absorb Fe and Mn and perhaps could be used to better understand Mn uptake in plants.     

Oscillations in photosynthesis are initiated and supported by imbalances in the supply of ATP and NADPH to the Calvin cycle

Oscillations in the rate of photosynthesis of sunflower (Helianthus annuus L.) leaves were induced by subjecting leaves, whose photosynthetic apparatus had been activated, to a sudden transition from darkness or low light to high-intensity illumination, or by transfering them in the light from air to an atmosphere containing saturating CO₂. It was found that at the first maximum, light-and CO₂-saturated photosynthesis can be much faster than steady-state photosynthesis. Both Q_A in the reaction center of PS II and P₇₀₀ in the reaction center of PS I of the chloroplast electron-transport chain were more oxidized during the maxima of photosynthesis than during the minima. Maxima of P₇₀₀ oxidation slightly preceded maxima in photosynthesis. During a transition from low to high irradiance, the assimilatory force F_A, which was calculated from ratios of dihydroxyacetone phosphate to phosphoglycerate under the assumption that the reactions catalyzed by NADP-dependent glyceraldehydephosphate dehydrogenase, phosphoglycerate kinase and triosephosphate isomerase are close to equilibrium, oscillated in parallel with photosynthesis. However, only one of its components, the calculated phosphorylation potential (ATP)/(ADP)(P_i), paralleled photosynthesis, whereas calculated NADPH/NADP ratios exhibited antiparallel behaviour. When photosynthetic oscillations were initiated by a transition from low to high CO₂, the assimilatory force F_A declined, was very low at the first minimum of photosynthesis and increased as photosynthesis rose to its second maximum. The observations indicate that the minima in photosynthesis are caused by lack of ATP. This leads to overreduction of the electron-transport chain which is indicated by the reduction of P₇₀₀. During photosynthetic oscillations the chloroplast thylakoid system is unable to adjust the supply of ATP and NADPH rapidly to demand at the stoichiometric relationship required by the carbonreduction cycle.Abbreviations PGA 3-phosphoglycerate - DHAP dihydroxyacetone phosphate - P₇₀₀ electron-donor pigment in the reaction enter of PS I - Q_A quinone acceptor in the reaction center of PS II This work received support from the Estonian Academy of Sciences, the Bavarian Ministry of Science and Art and the Sonderforschungsbereich 251 of the University of Würzburg. We are grateful for criticism by D.A. Walker, Robert Hill Institute, University of Sheffield, U.K. and by Mark Stitt, Institute of Botany, University of Heidelberg, FRG.     

Weak field influence on biomolecular changes

Model equations for the kinetics of the synthesis and decay of molecular aggregates are used to show the high sensitivity of equilibrium concentrations of high-molecular aggregates to external radiation. This phenomenon is used to explain the effects of low-intensity microwave fields on the functioning of biological systems. The experimental results on the influence of SHF-radiation on ferricyanide reduction by erythrocytes are interpreted in detail.