Rate of soil acidification under wheat in a semi-arid environment

The rate of acidification under wheat in south-eastern Australia was examined by measuring the fluxes of protons entering and leaving the soil, using the theoretical framework of Helyar and Porter (1989). Monthly proton budgets were estimated for the root zone (0–90 cm layer) and for the 0–25 and 25–90 cm layers. After an annual cycle, the root zone was alkalinized by 0.5 to 3.1 kmol OH^- ha^-1. The alkalinity originated from the mineralization of the organic anions contained in the organic matter. The budget was near neutrality in the 0–25 cm layer (range: –1.0 to 1.4 kmol H⁺ ha^-1), whereas there was net alkalinization in the 25–90 cm layer (1.7 to 2.3 kmol OH^- ha^-1). In the 0–25 cm layer, the acidity produced in autumn by mineralization of organic nitrogen was counterbalanced by the alkalinity released from crop residues. The main acidifying factor in this layer was leaching of NO₃ ^- during early winter (2.4 kmol H⁺ ha^-1). Nitrate added through leaching was the main alkalinizing factor in the 25–90 cm layer, as added NO₃ ^- was taken up by the roots or denitrified in this layer. Urea fertilization had almost no effect on the rate of acidification, as little NO₃ ^- was leached out of the root zone. The factors acidifying the soil under wheat were limited in this environment because of the small amout of NO₃ ^- leached and the retention of the crop residues.     

Effects of W-7, W-5, Verapamil and Diltiazem on vacuolar proton transport. Comparison of vacuolar H+-ATPase and H+-PPase from roots of Zea mays

The vacuolar membrane of plant cells is characterized by two proton pumps: the vacuolar H⁺-ATPase (V-ATPase; EC 3.6.1.3) and the vacuolar H⁺-PPase (V-PPase; EC 3.6.1.1). Recently, Du Pont and Morrissey reported that Ca²⁺ stimulates hydrolytic activity of purified V-ATPase (Arch. Biochim. Biophys., 1992. 294: 341–346). Since this effect may be due to degradation during purification further investigation of Ca²⁺ regulation of native V-ATPase was done. However, native tonoplast membranes contain a Ca²⁺/H⁺ antiport activity, which interferes with effects of calcium ions on proton transport activity of vacuolar ATPase. Therefore, the effects of anti-calmodulin drugs (W-7, W-5, calmidazolium), and calcium channel antagonists (Verapamil, Diltiazem) on proton transport activities of the vacuolar-type H⁺-ATPase and H⁺-PPase in tonoplast enriched membrane vesicle preparations from roots of Zea mays L. were studied. The concentrations for half maximal inhibition of vacuolar H⁺-ATPase (H⁺-PPase) were: 71 (191) μM W-7, 470 (> 800) μM W-5, 26 (24) μM calmidazolium (= compound R 24571). 398 (700) μM Verapamil, and 500 (1 330) μM Diltiazem. Estimation of Hill coefficients (n_H) for the inhibition by Verapamil showed a further difference between the two vacuolar proton pumps (H⁺-ATPase, n_H= 2.02; H⁺-PPase, n_n= 0.96). The data indicate that the vacuolar H⁺-ATPase itself is affected by these chemicals. It is suggested that some biological activities of W-7, W-5, Verapamil, and Diltiazem are due to their effects on proton translocation by the vacuolar-type H⁺-ATPase.     

Direct HPLC separation of enantiomers of pantoprazole and other benzimidazole sulfoxides using cellulose-based chiral stationary phases in reversed-phase mode

A direct, isocratic, and simple reversed-phase HPLC method was described for the separation of enantiomers of the proton pump inhibitor, rac-pantoprazole (PAN) using cellulose-based chiral stationary phases (Chiralcel OD-R and Chiralcel OJ-R). Some structurally related chiral benzimidazole sulfoxides, rac-omeprazole (OME) and raclansoprazole (LAN), were also studied. Chiralcel OJ-R was successful in the resolution of enantiomers of rac-PAN and rac-OME, while Chiralcel OD-R was most suitable for resolving the enantiomers of rac-LAN. Highest enantioselectivity to rac-PAN and rac-OME was achieved on Chiralcel OJ-R by using acetonitrile as an organic modifier, whereas methanol afforded better resolution of rac-LAN on Chiralcel OD-R than acetonitrile. Increases in buffer concentration and column temperature decreased retention and did not improve the resolution of the enantiomers on both columns. Using a mixture of 50 mM sodium perchlorate solution and acetonitrile as a mobile phase at a flow rate of 0.5 ml/min, maximum separation factors of 1.26 and 1.13 were obtained for the enantiomers of rac-PAN and rac-OME using a Chiralcel OJ-R column, while maximum separation factor of 1.16 was obtained for the enantiomers of rac-LAN using a Chiralcel OD-R column. © 1995 Wiley-Liss, Inc.     

Coupled cyclic electron transport in intact chloroplasts and leaves of C3 plants: Does it exist? if so,what is its function?

Transthylakoid proton transport based on Photosystem I-dependent cyclic electron transport has been demonstrated in isolated intact spinach chloroplasts already at very low photon flux densities when the acceptor side of Photosystem I (PS I) was largely closed. It was under strict redox control. In spinach leaves, high intensity flashes given every 50 s on top of far-red, but not on top of red background light decreased the activity of Photosystem II (PS II) in the absence of appreciable linear electron transport even when excitation of PS II by the background light was extremely weak. Downregulation of PS II was a consequence of cyclic electron transport as shown by differences in the redox state of P700 in the absence and the presence of CO₂ which drained electrons from the cyclic pathway eliminating control of PS II. In the presence of CO₂, cyclic electron transport comes into play only at higher photon flux densities. At H⁺/e=3 in linear electron transport, it does not appear to contribute much ATP for carbon reduction in C3 plants. Rather, its function is to control the activity of PS II. Control is necessary to prevent excessive reduction of the electron transport chain. This helps to protect the photosynthetic apparatus of leaves against photoinactivation under light stress.     

Stimulation of H+transport activity of vacuolar H+ATPase by activation of H+PPase in Kalanchoë blossfeldiana

In Kalanchoë blossfeldiana cv. Tom Thumb the initial rate of ATP-dependent H+-transport into tonoplast vesicles was stimulated up to three times if the H+-ATPase (EC 3.6.1.3) was energized a few minutes after pre-energization of the H+-PPase (EC 3.6.1.1). H+-PPase-activated ATP-dependent H+-transport was observed in plants of K. blossfeldiana cultivated in short day (SD) or long day (LD) conditions expressing different degrees of crassulacean acid metabolism (CAM). However, based on the higher activity and protein amount of H+-PPase and H+-ATPase present in the vacuolar membrane of SD plants the maximum H+-transport activity in the stimulated mode of the H+-ATPase was significantly higher in tonoplast vesicles of SD plants than of LD plants. Hence, a co-ordinated action of the H+-PPase and H+-ATPase at the tonoplast of Kalanchoë could allow a higher transport capacity at the vacuolar membrane when plants perform high CAM. Immunoprecipitation experiments with an antiserum raised against the A-subunit of the vacuolar H+-ATPase of Mesembryanthemum crystallinum L. showed that in SD and LD plants of K. blossfeldiana the H+-PPase was co-precipitated with the vacuolar H+-ATPase holoenzyme. The co-percipitation of the two transport proteins indicates a close structural localization of the H+-PPase and the A-subunit of the vacuolar H+-ATPase.     

Interaction of electron leak and proton leak in respiratory chain of mitochondria——Proton leak induced by superoxide from an electron leak pathway of univalent reduction of oxygen

By incubating the isolated rat myocardial mitochondria with xanthine-xanthine oxidase, anexogenous superoxide (O2) generating system, and by ischemia-reperfusion procedure of isolated rat heart as an endogenous O2 generating system, it was found that both sources of O2 showed the same injurious effects on mitochondrial function resulting in (i) increasing proton leak rate, lowering proton pumping activity and Ht/2e ratio of respiratory chain, and (ii) decreasing transmembrane potential of energized mitochondria] inner membrane by succinate oxidation. The injurious effects of O2 on these mitochondrial bioenergitical parameters mentioned above exhibited a dosage- or reaction time-dependent mode. (X has no effects on the electron transfer activity and transmembrane potential of nonenergized mitochondria. Being a superoxide scavenger, 3, 4-dihydroxylphenyl lactate showed obvious protection effects against damage of both exogenous superoxide sources from xanthine-xanthine oxidase system and endogenous Or sou     

Kinetics of nitrate and ammonium absorption and accompanying H+ fluxes in roots of Lolium perenne L. and N2-fixing Trifolium repens L.

Kinetic parameters for NH₄⁺ and NO₃^? uptake were measured in intact roots of Lolium perenne and actively N₂-fixing Trifolium repens. Simultaneously, net H⁺ fluxes between the roots and the root medium were recorded, as were the net photosynthetic rate and transpiration of the leaves. A Michaelis–Menten-type high-affinity system operated in the concentration range up to about 500 mmol m^?3 NO₃^? or NH₄⁺. In L. perenne, the V_max of this system was 9–11 and 13–14 μmol g^?1 root FW h^?1 for NO₃^? and NH₄⁺, respectively. The corresponding values in T. repens were 5–7 and 2 μmol g^?1 root FW h^?1. The K_m for NH₄⁺ uptake was much lower in L. perenne than in T. repens (c. 40 compared with 170 mmol m^?3), while K_m values for NO₃^? absorption were roughly similar (around 130 mmol m^?3) in the two species. There were no indications of a significant efflux component in the net uptake of the two ions. The translocation rate to the shoots of nitrogen derived from absorbed NO₃^?-N was higher in T. repens than in L. perenne, while the opposite was the case for nitrogen absorbed as NH₄⁺. Trifolium repens had higher rates of transpiration and net photosynthesis than L. perenne. Measurements of net H⁺ fluxes between roots and nutrient solution showed that L. perenne absorbing NO₃^? had a net uptake of H⁺, while L. perenne with access to NH₄⁺ and T. repens, with access to NO₃^? or NH₄⁺, in all cases acidified the nutrient solution. Within the individual combinations of plant species and inorganic N form, the net H⁺ fluxes varied only a little with external N concentration and, hence, with the absorption rate of inorganic N. Based on assessment of the net H⁺ fluxes in T. repens, nitrogen absorption rate via N₂ fixation was similar to that of inorganic N and was not down-regulated by exposure to inorganic N for 2 h. It is concluded that L. perenne will have a competitive advantage over T. repens with respect to inorganic N acquisition.     

Role of Intracellular pH in Proliferation, Transformation, and Apoptosis

Both cellular proliferation and apoptosis (programmed cell death) have been claimed to be modulated, perhaps even triggered by, changes in intracellular pH. In this review, we summarize the evidence that gave rise to these hypotheses. To facilitate a critical appraisal of the existing data, we briefly review the main pathways involved in cytosolic pH homeostasis and their regulation by mitogens and by apoptosis-inducing agents. The information available at present suggests that cytosolic pH plays a permissive role in cellular growth and proliferation, but is neither a trigger nor an essential step in the mitogenic signal transduction cascade. Concerning apoptosis, it is clear that lowering the pH in vitro can activate DNase II. However, the evidence linking cytosolic acidification with DNA degradation in vivois presently not convincing. We conclude that the cytosolic pH, an essential physiological parameter that is tightly controlled by multiple, complementary, or redundant systems, is unlikely to play a role in signalling either cell growth or death.     

Photosynthetic oxygen evolution: H/D isotope effects and the coupling between electron and proton transfer during the redox reactions at the oxidizing side of Photosystem II

The oxygen evolving complex (OEC) of Photosystem II (PS II) incorporates a Mn-cluster and probably a further redox cofactor, X. Four quanta of light drive the OEC through the increasingly oxidized states S0 S1 S2 S3 S4 to yield O2 during the transition S4 S0. It has been speculated that the oxidation of water might be kinetically facilitated by the abstraction of hydrogen. This implied that the respective electron acceptor is deprotonated upon oxidation. Whether YZ and X fulfill this expectation is under debate. We have previously inferred a 'chemical' deprotonation of X based on the kinetics of proton release (Haumann M, Drerenstedt W, Hundelt M and Junge W (1996) Biochim Biophys Acta 1273: 237–250. Here, we investigated the rates of electron transfer and proton release as function of the D2O/H2O ratio, the pH, and the temperature both in thylakoids and PS II core particles. The largest kinetic isotope effect on the rate of electron transfer (factor of 2.1–2.4) and the largest pH-dependence (factor of about 2 between pH 5 and 8) was found on S2 S3 where X is oxidized. During the other transitions both factors were much smaller ( 1.4). Electron transfer is probably kinetically steered by proton transfer only during S2 S3. These results corroborate the notion that X serves as a hydrogen acceptor for bound water during S4 S0. We propose a consistent scheme for the final reaction with water to yield dioxygen: two two-electron (hydrogen) transfers in series with a peroxide intermediate.