Plasma membrane anion channels in higher plants and their putative functions in roots

Recent years have seen considerable progress in identifying anion channel activities in higher plant cells. This review outlines the functional properties of plasma membrane anion channels in plant cells and discusses their likely roles in root function. Plant anion channels can be grouped according to their voltage dependence and kinetics: (1) depolarization-activated anion channels which mediate either anion efflux (R and S types) or anion influx (outwardly rectifying type); (2) hyperpolarization-activated anion channels which mediate anion efflux, and (3) anion channels activated by light or membrane stretch. These types of anion channel are apparent in root cells where they may function in anion homeostasis, membrane stabilization, osmoregulation, boron tolerance and regulation of passive salt loading into the xylem vessels. In addition, roots possess anion channels exhibiting unique properties which are consistent with them having specialized functions in root physiology. Most notable are the organic anion selective channels, which are regulated by extracellular Al3+ or the phosphate status of the plant. Finally, although the molecular identities of plant anion channels remain elusive, the diverse electrophysiological properties of plant anion channels suggest that large and diverse multigene families probably encode these channels.     

酸雨胁迫下樟树林降水中无机阴离子变化特征

对城市大气降水及城市樟树林穿透水、树干茎流、地表径流中的SO2-4、NO-3、Cl-、NO-2、F-、Br-、PO3-47种水溶性无机阴离子进行了定位测定,Br-、PO3-4未检出;主要的阴离子为SO2-4、NO-3,二者占总阴离子含量比例最大,占到75.6%—89.0%,且在水文学各分量中变异系数均值都很稳定,变动幅度在0.40—0.47;在各分量中SO2-4、NO-3、Cl-、NO-2、F-含量变化较大:大气降水、树干茎流中均为SO2-4NO-3Cl-F-NO-2,穿透水为SO2-4NO-3Cl-NO-2F-,地表径流为NO-3SO2-4Cl-F-NO-2;穿透水SO2-4、NO-3、Cl-、F-、NO-2淋溶系数分别为3.83、2.61、4.18、4.32、16.06,树干茎流SO2-4、NO-3、Cl-、F-、NO-2淋溶系数分别为7.52、3.55、4.29、2.76、3.10,地表径流SO2-4、NO-3、Cl-、F-、NO-2淋溶系数分别为2.60、4.74、5.00、3.01、3.56,表明阴离子在樟树林水文学过程中均表现出不同程度的富集效应;大气降水中PH均值为4.77,SO2-4/NO-3当量比值为1.1,属于硫酸-硝酸混合型酸雨;大气降水SO2-4、NO-3、Cl-、F-与穿透水中SO2-4、NO-3、Cl-、F-对应呈显著性正相关(P0.01),大气降水NO-3与树干茎流NO-3呈显著性正相关(P0.01),地表径流中SO2-4、Cl-与穿透水、树干茎流中SO2-4、Cl-对应呈显著性正相关(P0.01),说明樟树林能够吸附大气中的酸性物,让酸性养分于自身系统内部吸收、迁移、络合,从而起到净化大气的效用;穿透水p H与地表径流NO-3、NO-2呈显著性负相关(P0.01),与地表径流SO2-4呈显著性正相关(P0.05),表明NO-3、NO-2有助于促进地表水的酸化,而SO2-4有利于缓解地表水的酸化;穿透水p H均值5.62,对酸雨起到中和作用,树干茎流p H均值4.61,有被酸化趋势,地表径流p H均值7.19,接近中性。     

Ionic content in plant extracts determined by ion chromatography with conductivity detection

A simple method is described for the determination of the ionic content of vegetable samples by ion chromatography with suppressed conductivity detection. Extracts of leaves of cucumber (Cucumis sativus), leaves and cotyledons of watermelon (Citrullus lanantus), cotyledons of zucchini (Cucurbitapepo), and leaves and roots of olive (Olea europaea) obtained at room temperature yielded chromatographic profiles with substantial differences in the relative contents of Cl-, NO3-, HPO4(2-) and SO4(2-) as well as of Na+, NH4+, K+, Mg2+ and Ca2+. Although NO3-, Cl- and K+ were common to each extracted sample and accounted for most of the ions present, two additional anion peaks (i.e. malate and oxalate) were detected. Among the vegetable tissues investigated, olive roots contained a considerable amount of oxalate (37 mg/g dry weight), while Na+, which is present in very low amount in extracted samples of leaves and cotyledons, represented ca. 30% of the cationic content of olive roots. In all the examined tissue extracts, K+ was the main cation (16-55 mg/g dry weight) and NO3-, Cl- and HPO4(2-) were the main inorganic anions.     

Elemental composition of the cyanobacterium Anabaena flos-aquae collected from different depths within a stratified lake

The elemental composition of cells of Anabaena flos-aquae was investigated by energy-dispersive electron probe X-ray microanalysis (10?kV) of mixed phytoplankton samples obtained from different depths within a stratified eutrophic freshwater lake. Preliminary studies had indicated that at 10?kV, X-ray information was derived mainly from the peripheral region (up to 1–2?μm) of cells. Routinely detectable elements (present in at least 50% of cells) included Mg (overall mean 112?mmol?kg^?1 dry mass), Si (1850), P(313), S (113), Cl (92), K (300) and Ca (56). Peaks of Na and Al were also frequently present. Individual elements showed a wide range of intracellular concentrations at each depth within the lake. Si had a clear bimodal distribution at depths of 1–8?m, indicating both low-Si (mean concentrations 54–290–3800?mmol?Si?kg^?1) and high-Si (2770–3800?mmol?Si?kg^?1) cell subpopulations. Low-Si cells had significantly higher concentrations of other detectable elements compared with high-Si cells, but could not be distinguished from the latter in terms of morphology or stage of cell cycle (comparison of dividing and non-dividing cells). High-Si cells were intermixed with low-Si cells in individual Anabaena colonies and decreased proportionally with depth, ranging from approximately 75% in the surface sample to 10% at 8?m. With the exception of Si, mean elemental concentrations of Anabaena populations were closely similar throughout the epilimnion. These differed from the metalimnion sample, which had significantly higher concentrations of Mg, P, S and K, and a lower concentration of Si. Mean ratios of certain elements (Mg, P, S and K) and ion groups (monovalent/divalent cations) were highly constant throughout the sampled water column. At each depth, most of the cell elements were significantly and positively inter-correlated (Pearson analysis), with Mg, P, S and K typically constituting the major group (factor analysis). Correlations with Si were invariably negative, suggesting a separate (possibly cell wall) location of this element in the cell.     

In vitro and in vivo hepatic transport of the magnetic resonance imaging contrast agent B22956/1: role of MRP proteins

The molecular mechanisms of the hepatic transport of B22956/1, a new gadolinium complex from the class of intravascular contrast agents for MRI, which undergoes extensive biliary elimination, were studied. Biliary and urinary elimination of B22956/1 were measured in normal and in mutant MRP2 lacking rats (TR(-)); cellular trafficking of the compound was assessed in wild and MRP1 or MRP2 transfected MDCKII cells. Eight hours after IV injection of B22956/1, 90+/-8% of the dose was recovered in the bile of normal rats. By contrast, in TR(-) rats, the biliary excretion was significantly lower (14+/-3%) while 55+/-9% of the compound was found in urine. In vitro, the cellular accumulation of B22956/1 was significantly lower in both MRP1 and MRP2 transfected cells as compared to wild type MDCKII cells, and the cellular efflux was prevented by the MRP inhibitor MK571, indicating the involvement of both MRP2 and MRP1 in the transport of B22956/1. Due to the distinct cellular localization of the proteins, MRP2 accounts for the biliary and urinary excretion of the compound, while MRP1 prevents cellular accumulation of the MRI agent. B22956/1 may be useful in clinical conditions where a defective biliary transport is present.     

A spectroelectrochemical and chemical study on oxidation of 7,8-dihydroxy-4-methylcoumarin (DHMC) and some related compounds in aprotic medium

Electrochemical and chemical oxidation of 7,8-hydroxy-4-methylcoumarin (DHMC 1) and 7,8-diacetoxy-4-methylcoumarin (DAMC 4) were studied to investigate the mechanisms occurring in their antioxidant activities in acetonitrile, under electron transfer and H-atom transfer conditions. Electrolysis and chemical reactions were followed on-line by monitoring the UV spectral changes with time.     

Acidification of Soil in a Dry Land Winter Wheat-sorghum/corn-fallow Rotation in the Semiarid U.S. Great Plains

Soil pH is decreasing in many soils in the semiarid Great Plains of the United States under dry land no-till (NT) cropping systems. This study was conducted to determine the rate of acidification and the causes of the acidification of a soil cropped to a winter wheat (Triticum aestivum L.)-grain sorghum [Sorghum bicolor (L.) Moench]/corn (Zea mays L.)-fallow rotation (W-S/C-F) under NT. The study was conducted from 1989 to 2003 on soil with a long-term history of either continuous NT management [NT(LT)] (1962–2003) or conventional tillage (CT) (1962–1988) then converted to NT [NT(C)] (1989–2003). Nitrogen was applied as ammonium nitrate (AN) at a rate of 23 kg N ha⁻¹ in 1989 and as urea ammonium nitrate (UAN) at an average annual rate of 50 kg N ha⁻¹ from 1990 to 2003 for both NT treatments. Soil samples were collected at depth increments of 0–5, 5–10, 10–15, and 15–30 cm in the spring of 1989 and 2003. Acidification rates for the NT(LT) and NT(C) treatments were 1.13 and 1.48 kmol H⁺ ha⁻¹ yr⁻¹ in the 0–30 cm depth, respectively. The amount of CaCO₃ needed to neutralize the acidification is 57 and 74 kg ha⁻¹ yr⁻¹ for the NT(LT) and NT(C) treatments, respectively. A proton budget estimated by the Helyar and Porter [1989, Soil Acidity and Plant Growth, Academic Press] method indicated that NO₃⁻ leaching from the 30 cm depth was a primary cause of long-term acidification in this soil. Nitrate leaching accounted for 59 and 66% of the H⁺ from the acid causing factors for NT(LT) and NT(C) treatments, respectively. The addition of crop residues to the soil neutralized 62 and 47% of the acidity produced from the leaching of NO₃⁻, and 37 and 31% of the acid resulting from NO₃⁻ leaching and the other acid-causing constituents for the NT(LT) and NT(C) treatments, respectively. These results document that surface soils in dry land W-S/C-F rotations under NT are acidifying under current management practices. Improved management to increase nitrogen uptake efficiency from applied fertilizer would help reduce the rate of acidification. The addition of lime materials to prevent negative impacts on grain yields may be necessary in the future under current management practices. A contribution of the university of Nebraska Agricultural Research Division, Lincoln, NE 68583. Journal series No. 15120     

Cluster-root production and organic anion exudation in a group of old-world lupins and a new-world lupin

We examined the capacity of several Old-World lupin species (Lupinus luteus L., L. hispanicus Boiss. et Reuter and L. angustifolius L.) and one species of a New-World lupin (L. mutabilis Sweet) to form cluster roots under a range of conditions in solution culture. The effect of the synthetic auxin, IBA (indole-3-butyric acid), on cluster-root development in L. luteus and L. albus L. provided with an adequate phosphorus (P) supply was also investigated. In addition, the effect of a high nitrate-N (NO₃-N) supply on the efflux of citrate and malate from roots of L. angustifolius was examined to determine if specific regions of the root system exuded these organic anions. When P-deficient, L. hispanicus, L. luteus and L. mutabilis formed cluster roots that secreted organic anions. Citrate was generally the dominant organic anion exuded, although succinate was also exuded in large quantities from L. luteus. Citrate efflux by L. hispanicus and L. luteus was at least comparable to that reported for P-deficient L. albus[up to 1.092 nmol g^–1 fresh weight (FW) s^–1], but was over an order of magnitude lower in L. mutabilis (0.036 nmol g^–1 FW s^–1). Citrate and malate were not detected in significant amounts from either the lateral roots or the root tips of any species grown under P-sufficient or -deficient conditions. Citrate efflux from cluster roots of L. luteus showed a diurnal pattern, similar to that reported for L. albus, with maximum efflux during the day, and declining to a minimum before dawn. IBA added to the nutrient solution induced cluster-root formation on both L. albus and L. luteus at concentrations of P that would normally suppress the production of these roots. However, the IBA-induced cluster roots did not exude significant amounts of citrate. Although L. angustifolius did not produce cluster roots when P-deficient, it produced cluster-like root structures that exuded citrate (0.053 nmol g^–1 FW s^–1) when grown at a high nitrate-N (NO₃-N) supply. L. angustifolius did not exude significant citrate or malate from lateral roots or root tips when grown at either high or low NO₃-N supply. Our findings for L. hispanicus and L. luteus are the first reports of cluster-root formation in response to P deficiency for these Old-World species, and for L. mutabilis, it is the first report of cluster roots for a New-World lupin species. These reports indicate that evolutionary and biogeographical aspects of cluster-root formation in the genus Lupinus need to be revised. Furthermore, investigation is warranted to determine the capacity of species of the large group of New-World lupins to form cluster roots in soils of their native habitats.     

Validation of the use of the lipophilic thiocyanate anion for the determination of membrane potential in Ehrlich ascites tumor cells

Summary The utility of the lipophilic anion thiocyanate (SCN⁺) as a probe for the indirect estimation of the cell membrane potential (V _m ) in Ehrlich ascites tumor cells has been evaluated by comparison to direct electrophysiological measurements. SCN accumulation is consisten with first-order uptake into a single kinetically-identifiable cellular compartement, achieving steadystate distribution in 20–30 min at 22°C. The steady-state distribution ratio ([SCN^–] _c /[SCN^–] _e ) in physiological saline is 0.44±0.02. Treatment of the cells with proparanolol (0.13 mM), an activator of Ca²⁺ dependent K⁺ channels, reduces the steady-state distribution ratio to 0.19±0.02. Conversely, treatmetn with BACl₂ (10 mM), an antagonist of the pathway, increases the SCN^– distribution ratio to 0.62±0.01. The equilibrium potentials (V _SCN ) calculated under these conditions are virtually identical to direct electrophysiological measurements of theV _m made under the same conditions. The effect of varing extracellular [K⁺]([K⁺] _e ) in the presence of constant [Na⁺] _e =100 mM has also been tested. In control cells, elevation of [K⁺] _e from 6 to 60 mM reducesV _SCN from –20.6±1.0 to –13.2±1.2 mV. Again, microelectrode measurements give excellent quantitative agreement. Propranolol increases the sensitivity of the cells to varying [K⁺] _e , so that a 10-fold elevation reducesV _SCN by approximately 31 mV. BaCl₂ greatly reduces this reponse: a 10-fold elevation in [K⁺] _e yielding only a 4-mV rediction inV _SCN . It is concluded that the membrane potential of Ehrlich cells can be estimated accurately from SCN^– distribution measurements.     

Inhibition of mitochondrial ATPase by dicarbopolyborate,a new enzyme inhibitor

Polyborate anions were found to inhibit mitochondrial ATPase. Mercapto and chloro derivatives of dicarbononaborates showed full inhibition of the enzyme activity at 0.5–0.8 mM. The inhibitory effect of dodecaborates was lower. The inhibition was of competitive type with respect to ATP. The inhibition of soluble F₁-ATPase indicates a direct interaction of the polyborate anion with the catalytic part of the enzyme molecule.