Modifying Effects of Non-toxic Levels of Aluminium on the Uptake and Transport of Phosphate in Ryegrass

Apparent uptake and transport of H₂³²PO₄^– from nutrientsolutions containing 100 mmol m^–3 phosphate were characterizedasfunctions of time, concentration and pH in ryegrass seedlings.On a log/log plot, concentration versus uptake to the root resolvedintotwo linear phases, suggesting a change in uptake mechanism orefflux at the break. These results were compared with thosefor ³²P uptake and transport in solutions containing Al rangingfrom 0–185 mmol m^–3. Al addition depressed pH, butbecauseuptake of P was unaffected by pH below 5–0, noadjustments were attempted. Uptake time-courses revealed clearlythe usualinitial adsorption shoulder in the uptake curve, increasingwith Al concentration up to 37 mmol m^–3. Beyond about2 h, P uptaketo the root became linear, at rates increasingwith external Al concentration up to 37 mmol m^–3. Concentrationsof Al muchabove 100 mmol m^–3 were toxic. Al treatmentsdid not affect P transport to the shoot and absorbed Al wasconfined to the root.The quantities of P and Al taken up intothe root indicated storage in cortex cell vacuoles, lockingup significant amounts of P.Experiments with tillering plantsshowed similar characteristics to those with seedlings. Sequesteringof P with Al within the rootcortex cells was evident, particularlyin plants which had been grown in nutrient containing Al fromsoon after germination. Aland P solution chemistry is discussedin the context of this work and the consequences of effectson P uptake for the economy ofphosphate poor upland soils wereconsidered. Key words: Phosphate, aluminium, adsorption, uptake, Lolium perenne L     

A carrier-mediated mechanism for pyridoxine uptake by human intestinal epithelial Caco-2 cells: regulation by a PKA-mediated pathway

Vitamin B₆ is essential for cellular functions and growth due to its involvement in important metabolic reactions. Humans and other mammals cannot synthesize vitamin B₆ and thus must obtain this micronutrient from exogenous sources via intestinal absorption. The intestine, therefore, plays a central role in maintaining and regulating normal vitamin B₆ homeostasis. Due to the water-soluble nature of vitamin B₆ and the demonstration that transport of other water-soluble vitamins in intestinal epithelial cells involves specialized carrier-mediated mechanisms, we hypothesized that transport of vitamin B₆ in these cells is also carrier mediated in nature. To test this hypothesis, we examined pyridoxine transport in a model system for human enterocytes, the human-derived intestinal epithelial Caco-2 cells. The results showed pyridoxine uptake to be 1) linear with time for up to 10 min of incubation and to occur with minimal metabolic alteration in the transported substrate, 2) temperature and energy dependent but Na⁺ independent, 3) pH dependent with higher uptake at acidic compared with alkaline pHs, 4) saturable as a function of concentration (at buffer pH 5.5 but not 7.4) with an apparent Michaelis-Menten constant (K_m) of 11.99 ± 1.41 µM and a maximal velocity (V_max) of 67.63 ± 3.87 pmol · mg protein^-1 · 3 min^-1, 5) inhibited by pyridoxine structural analogs (at buffer pH 5.5 but not 7.4) but not by unrelated compounds, and 6) inhibited in a competitive manner by amiloride with an apparent inhibitor constant (K_i) of 0.39 mM. We also examined the possible regulation of pyridoxine uptake by specific intracellular regulatory pathways. The results showed that whereas modulators of PKC, Ca⁺²/calmodulin (CaM), and nitric oxide (NO)-mediated pathways had no effect on pyridoxine uptake, modulators of PKA-mediated pathway were found to cause significant reduction in pyridoxine uptake. This reduction was mediated via a significant inhibition in the V_max, but not the apparent K_m, of the pyridoxine uptake process. These results demonstrate, for the first time, the involvement of a specialized carrier-mediated mechanism for pyridoxine uptake by intestinal epithelial cells. This system is pH dependent and amiloride sensitive and appears to be under the regulation of an intracellular PKA-mediated pathway. vitamin B₆; intestinal transport; transport regulation; Caco-2 cell     

Oxidative Damages and Repair in Euglena gracilis Exposed to Ozone II. Membrane Permeability and Uptake of Metabolites

Significant injuries to the plasma membrane were detected inEuglena gracilis cells during ozone exposure (240 µ1.liter¹,delivery rate of l µmol.min^–1), as assessed by measuringthe alterations of vitamin B₁₂ and acetate uptakes and the leakageof intracellular K⁺ (Rb⁺). A rapid decrease in the uptake ofvitamin B₁₂ and acetate was observed within 15 min of treatment,indicating that both transport systems are very sensitive toO₃. On the other hand, the leakage of intracellular K⁺ ions,as measured by the efflux of ⁸⁶Rb⁺ from prelabelled cells, couldonly be detected after 30 min of O₃ exposure. These resultssuggest that the initial metabolic symptoms of injury is atthe level of the two transport systems examined and that thealteration of the membrane permeability to K⁺ ions appears asa second step in the cascade of oxidative events at the plasmamembrane level. When Euglena cells were allowed to recover underautotrophic growth conditions following O₃ treatment, vitaminB₁₂ and ⁸⁶Rb⁺ (K⁺) ions uptakes returned gradually to controllevel within 5 h of the recovery period. Acetate uptake returnedto control level at a slower rate and needed 20 h for completerecovery. These results indicate that the cells were able toactively repair most of the initial oxidative damages inducedby O₃. The metabolic significance of the repair mechanism(s)is discussed. (Received December 25, 1989; Accepted July 23, 1990)     

Role of Coccoliths in the Utilization of Inorganic Carbon by a Marine Unicellular Coccolithophorid, Emiliania huxleyi: a Survey Using Intact Cells and Protoplasts

The utilization of inorganic carbon and role of the coccolithswere investigated in intact cells and protoplasts of a marineunicellular calcareous alga, Emiliania huxleyi. Protoplastswith high photosynthetic activity were obtained by artificialdecalcification with 50 mM MES-NaOH (pH5.5). (1) The kineticsof the photosynthetic evolution of O₂ at various concentrationsof externally added NaHCO₃ were the same for intact cells andprotoplasts, indicating that the kinetic properties with respectto dissolved inorganic carbon (DIC) were not affected by thepresence or absence of the coccoliths on the cell surface. Double-reciprocalplots and plots of the concentration of substrate divided byvelocity (s/v) against the concentration of substrate (s) werebiphasic in the case of both intact cells and protoplasts. TheCO₂-utilization reaction was, therefore, considered to involvetwo processes with different values of K_m and V_max. From thekinetic analyses, K_m and V_max [µmoles O₂ (ml PCV)^–1h^–1] were deduced to be 92 µM and 76.3 for a "low-K_m"reaction and 4.1 mM and 252 for a "high-K_m" reaction, respectively.(2) In short-term (40-min) experiments, time courses of thetotal uptake of ¹⁴C-DIC and the incorporation of ¹⁴C into acid-stableproducts of photosynthesis and the internal pool of DIC, determinedas acid-labile compounds, under CO₂-limiting conditions (80µM) were very similar for intact cells and protoplasts.However, incorporation of ¹⁴C into CaCO₃ apparently occurredmore slowly in protoplasts than in intact cells. (3) In longterm (24-h) experiments, patterns of incorporation of ¹⁴C werealmost same for intact cells and protoplasts, with the exceptionthat the amount of ¹⁴C incorporated into CaCO₃ was much smallerin the former than the latter. The production of Ca¹⁴CO₃ increasedduring the course of 10 h after a 4-h lag. However, after 10h the level of Ca¹⁴CCO₃ started to decrease. The decrease wasaccompanied by an increase in ¹⁴C in the products of photosynthesis,suggesting that CaCO₃ was reutilized for the photosyntheticfixation of CO₂ and, therefore, that the coccoliths functionas sites of storage of DIC. However, the internal level of DICremained at the same level even after the supply of externalDIC has been almost completely depleted. (Received July 25, 1995; Accepted December 11, 1995)     

Phosphate uptake and transport in Agrostis capillaris L Effects of non-toxic levels of aluminium and the significance of P and Al speciation

Investigations on the effects of low levels of Al on P adsorption,uptake and translocation in seedlings of the indigenous grassAgrostis capillaris were undertaken. Apparent uptake and transportof H₂ ³²PO₄ from nutrient solutions containing 10 or 100mmolm^–3 phosphate were characterized as functions of timeand concentration. Experiments on ³²P uptake and transport insolutions containing no Al (control) or Al ranging from 3.7to 185 mmol m^–3 at pH ranging from 4.3 to 4.6, showedthat in 10 mmol m^–3 P, effects of Al at 3.7 and 37 mmolm^–3 on the size of the initial uptake shoulder were small,but some increase in subsequent P uptake to the roots was observed,though transport to the shoots was suppressed. With 37 mmolm^–3 Al in nutrient solution containing 100 mmol m^–3P, the uptake shoulder was much increased above the control.Subsequent root uptake was stimulated but transport was unaffected.Lack of toxicity of the Al concentrations used was indicatedby a lack of significant effect on plant fresh weight. AbsorbedAl was almost totally retained in the root in all treatments.Speciation calculations showed that the major species in Alamended nutrient solution at pH 4.4 were H₂PO₄^–, AI³⁺and AIHPO₄⁺, together with substantial amounts of AISO₄⁺ andsoluble aluminium hydroxy complexes (AIOH²⁺, AI(OH)₂⁺), dependingon the relative concentrations of P and Al. The effects of Al,with 10 mmol m^–3 P, on adsorption of complexed P werepartly accounted for in terms of preferential cell wall adsorptionof Al complexes not containing P. Conclusions were drawn aboutthe P-economy of A. capillaris plants growing on soils withlow levels of P and Al. Key words: Phosphorus, aluminium, speciation, Agrostis capillahs L     

The Interaction Between Silicon and Aluminium in Sorghum bicolor (L.) Moench: Growth Analysis and X-ray Microanalysis

Seeds of Sorghum bicolor (L.) Moench. were germinated on moistfilter paper for 6 d, before the seedlings were transferredto pots containing 500 µmol l^-1 Ca(NO₃)₂ for 2 d. Theseedlings were then treated with 0 or 100 µmol l^-1 Alin factorial combination with 0, 1400 or 2800 µmol l^-1Si for 8 d. The background solution used throughout was 500µmol l^-1 Ca(NO₃)₂. Aluminium treatment reduced root growthand caused a significant increase in shoot/root ratio. The presenceof silica in the solution significantly ameliorated the effectsof aluminium on root growth. Three treatment were selected for a microanalytical investigationof the basal region of the root: 2800 µmol l^-1 Si only;100 µmol l^-1 Al only; and a combination of the two. Inthe 2800 µmol l^-1 treatment silica was deposited in theendodermis, with the greatest accumulation being in the innertangential wall (ITW). When plants were treated with 100 µmoll^-1 Al only, aluminium concentration was highest in the outertangential wall (OTW) of the epidermis. The element was presentin the hypodermal walls and OTW of the endodermis and was notdetectable in the stele. With both 2800 µmol l^-1 Si and100 µmol l^-1 Al in the nutrient solution the two biomineralizationsites were the ITW of the endodermis, where silicon was themajor element deposited, and atypically in the OTW of the epidermiswhere both aluminium and silicon were present. The sequestrationof aluminium in the Al-Si deposit in the OTW of the epidermismay represent the mechanism that allows greater root growthin this treatment.Copyright 1993, 1999 Academic Press Sorghum bicolor (L.) Moench., aluminium, silicon, calcium, root, toxicity, biomineralization, X-ray microanalysis, freeze substitution     

The Transport and Metabolism of Urea in Chara australis: II. UREASE AND THE DISTRIBUTION OF TRANSPORTED UREA

The ureolytic enzyme in Chara was investigated. This enzymewas shown to be a urease with an unusually high affinity forurea(K_m = 158 mmol m^-3). Little inhibition of urease activitywas found when intact Chara cells were exposed to the ureaseinhibitors hydroxyurea, acetohydroxamic acid and N-ethylmaleimide,although there was some inhibition of urea uptake. The distribution of radioactivity amongst the amino acid, organicacid and sugar/neutral fractions, determined by ion-exchangechromatography, was very similar whether the Chara internodeswere exposed to ¹⁴C-urea or to H¹⁴CO₃. This suggests that thefraction of the urea-carbon liberated by the urease as CO₂ andretained by the cell is used in photosynthetic carbon-fixation.During the initial 15 min of ¹⁴C-urea uptake, label appearsin the vacuole only in the form of unmetabolized urea. Afterthis time a variety of labelled compounds appear in the vacuole,presumably reflecting the gradual movement of carbon-fixationproducts from the chloroplasts to the cytoplasm and thence intothe vacuole. Key words: Urea transport, metabolism, Chara, urease     

Utilization Modes of Inorganic Carbon for Photosynthesis in Various Species of Chlorella

Rates of CO₂ and HCC₃^– fixation in cells of various Chlorellaspecies in suspension were compared from the amounts of ¹⁴Cfixed during the 5 s after the injection of a solution containingonly ¹⁴CO₂ or H¹⁴CO₃^–. Results indicated that irrespectiveof the CO₂ concentration during growth, Chlorella vulgaris 11h and C. miniata mainly utilized CO₂, whereas C. vulgaris C-3,C. sp. K. and C. ellipsoidea took up HCO₃^– in additionto CO₂. Cells of C. pyrenoidosa that had been grown with 1.5%CO₂ (high-CO₂ cells) mainly utilized CO₂, whereas those grownwith air (low-CO₂ cells) utilized HCO₃^– in addition toCO₂. Cells that utilized HCO₃^– had carbonic anhydrase(CA) on their surfaces. The effects of Diamox and CA on the rates of CO₂ and HCO₃^–fixation are in accord with the inference that HCO₃^– wasutilized after conversion to CO₂ via the CA located on the cellsurface. CA was found in both the soluble and insoluble fractions;the CA on the cell surface was insoluble. Independent of the modes of utilization, the apparent K_m (NaHCO₃)for photosynthesis was much lower in low-CO₂ cells than in high-CO₂ones. The fact that the CA in the soluble fraction in C. vulgarisC-3 was closely correlated with the K_m(NaHCO₃) indicates thatsoluble CA lowers the K_m. ¹ Dedicated to the late Professor Joji Ashida, one of the foundersand first president of the Japanese Society of Plant Physiologists. ⁴ On leave from Research and Production Laboratory of Algology,Bulgarian Academy of Sciences, Sofia. (Received September 14, 1982; Accepted March 1, 1983)     

The Conductance of the Plasmalemma for CO2

Permeability coefficients (P_S values) for CO₂ of the plasmamembrane (PM) of the unicellular green algae Eremosphaera viridis,Dunaliella parva, and Dunaliella acidophila, and of mesophyllprotoplasts isolated from Valerianella locusta were determinedfrom ¹⁴CO₂ uptake experiments using the rapid separation ofcells by the silicone oil layer centrifugation technique. Theexperimental P_S values were compared with calculated numbersobtained by interpolation of Collander plots, which are basedon lipid solubility and molecular size, for D. parva cells,mesophyll protoplasts isolated from Spinacia oleracea, mesophyllcells and guard cells of Valerianella, and guard cell protoplastsisolated from Vicia faba. The conductivity of algal plasma membranes for CO₂ varies between0.1 and 9 ? 10^–6 m s^–1, whereas for the plasmalemmaof cells and protoplasts isolated from leaves of higher plantsvalues between 0.3 and 11 ? 10^–6 m s^–1 were measured.By assuming that these measurements are representative for plantsand algae in general, it is concluded that the CO₂ conductivityof algal PM is of the same order of magnitude as that of thehigher plant cell PM. P_s values of plasma membranes for CO₂are lower than those for SO₂, but are in the same order of magnitudeas those measured for H₂O. On the basis of these results itis concluded that theoretical values of about 3000 ? 10^–6m s^–1 believed to be representative for higher plant cells(Nobel, 1983) and which are frequently used for computer-basedmodels of photosynthesis, lack experimental confirmation andrepresent considerable overestimations. However, with severalsystems, including higher plant cells, the conductance of thePM for CO₂ was significantly higher in light than in darkness.This suggests that in light, additional mechanisms for CO₂ uptakesuch as facilitated diffusion or active uptake may operate inparallel with diffusional uptake. Key words: Conductivity, CO₂, permeability coefficient, photosynthesis, plasmalemma     

DYNAMIC RESPONSE OF ROOT BORDER CELLS AND THEIR ASSOCIATED MUCILAGE EXUDATION IN SOYBEAN TO AI STRESS AND RECOVERY

 以耐铝性明显差异的两个大豆(Glycine max)基因型‘浙秋2号’(耐性)和‘浙春3号’(敏感)为材料, 研究根尖边缘细胞比活度、粘液分泌和根长对铝胁迫和解除胁迫的反应, 明确边缘细胞的粘液分泌对策在铝毒环境中的生态学意义。结果表明, ‘浙秋2号’在100~400 µmol&;#8226;L^–1 Al³⁺处理的3~12 h, 边缘细胞比活率呈递减趋势, 12 h后比活率又略有上升。‘浙春3号’在300和400 µmol&;#8226;L^–1 Al³⁺处理的变化与前者一致。两个大豆基因型的粘液层随着Al³⁺浓度增加和时间延长而增厚, 并于400 µmol&;#8226;L^–1 Al³⁺处理24 h时达到最大(>17 µm)。‘浙秋2号’在低浓度Al3+ (100和200 µmol&;#8226;L^–1)处理3~6 h后就会分泌大量粘液, ‘浙春3号’则在300 µmol&;#8226;L^–1 Al³⁺处理12 h后才有类似的变化。‘浙秋2号’在400 µmol&;#8226;L^–1 Al³⁺处理下的根相对伸长率均高于100~300 µmol&;#8226;L^–1 Al³⁺处理, ‘浙春3号’则表现为Al³⁺浓度越高, 根伸长受抑越明显。Al³⁺胁迫解除后, ‘浙秋2号’的粘液分泌速度和分泌量急剧下降, ‘浙春3号’在胁迫解除后的24 h, 仍会持续、大量地分泌粘液(>19 µm)。可见, 耐性大豆通过在铝胁迫初期快速、大量地分泌粘液以维持较高的边缘细胞活性和解除胁迫后迅速降低粘液的分泌速度及分泌量来适应铝毒害环境。     

Aluminate-Induced Changes in Morphology and Ultrastructure of Thinopyrum Roots

The effects of aluminate [Al(OH)_4$$$] on the morphology andultrastructure of root cells were studied in the salt-tolerantgrasses Thinopyrum bessarabicum (Savul. and Rayss) A. Lve (2) and Thinopyrum junceum (L.) A. Lve (6) by light and transmissionelectron microscopy. Seedlings were grown in nutrient solutioncontaining 1 mol m^–3 [Al] and 5 mol m^–3 Na₂CO₃ atpH 100. Light microscopy revealed that root tips of [Al]-treated plantsdisplayed bending. Many cells of the cortex in the elongatingregion contained a fibrillar/granular material which renderedthem densely staining. Radial (anticlinal) walls of the epidermalcells were either cleft apart of unusually thickened. Amyloplastsof the central root cap cells contained fewer starch grains,while their distribution was disturbed. Electron microscopy showed that the most serious effects of[Al] toxicity occurred at the cell walls of the epidermal androot cap cells, as they lost their fibrillar fine structureand contained an amorphous electron-dense material distributedall over the wall section. Electron-opaque droplets were encounteredat the plasma membrane region of epidermal cells, while theelectron-dense material observed in the vacuoles of cortex cellscould be aluminate which had accumulated there. Thus, despitethe presence of a barrier to aluminate uptake, some [Al] doesenter the symplast. However, the cytoplasm of many epidermalcells displayed a normal fine structure and contained the usualsubcellular components. Dictyosomes, in particular, were abundantand surrounded by many vesicles denoting an active state. Theseobservations stress the role of cell walls as the major [Al]pool and of the plasma membrane as the ultimate barrier thatprotects the cytoplasm. Results are further discussed in relation to the findings inother plant species and it is concluded that, although aluminateis less toxic than Al³⁺, it causes morphological, structuraland, presumably, functional damage-to the roots of the speciesinvestigated. Key words: Thinopyrum, aluminate toxicity, cell walls, root bending     

Uptake and Utilization of Fructose by Anabaena variabilis ATCC 29413. Effect on Respiration and Photosynthesis

Anabaena variabilis ATCC 29413 showed a constitutive mechanismfor fructose uptake which was further enhanced by growing thecells with fructose. The uptake process was energydependentas indicated the inhibitory effect of the uncoupler carbonylcyanidem-chlorophenylhydrazone (CCCP) and the reduction induced byincubating the cells in the dark or in the light with DCMU.Cells adapted to growth on fructose showed increased rates ofrespiration both in the dark and in the light. The rate of ¹⁴CO₂evolved from radiolabelled fructose was lower in the light thanin the dark or in the presence of DCMU. This fact can be partiallyexplained by the photosynthetic reutilization of respiratory¹⁴CO₂. Modifications in photosynthesis were observed in fructose-growncells. PS I and PS II activity measured in spheroplasts obtainedby lysozyme treatment were enhanced by fructose probably asa way to compensate the lower concentration of chorophyll showedby fructose-grown cells. The photosynthetic affinity for externalCO₂ and the rate of photosynthesis dependent on external inorganiccarbon were reduced by fructose. (Received September 24, 1991; Accepted February 14, 1992)     

Salinity Reduces Water Use and Nitrate-N-use Efficiency of Citrus

Five-month-old Cleopatra mandarin (Citrus reticulata Blanco)(CM) and Volkamer lemon (Citrus volkameriana Ten. and Pasq.)(VL) seedlings were grown in a glasshouse in 2·3-1 potsof Candler fine sand. Plants were irrigated with either non-saline(EC_e = 0·23 dS m^-1) or saline (6·13 dS m^-1) waterusing 3:1 NaCl:CaCl₂ solution over a 4-week period. A singleapplication of K¹⁵NO₃ (19·64 atom % excess ¹⁵N) at 212mg N1^-1, was substituted for a normal weekly fertilization after3 weeks and plants were harvested 7 d later. The transpirationrate, uptake of nitrogen, growth and nitrogen-use efficiency(NUE) on a dry weight basis (mg d. wt mg^-1 N) of both specieswas reduced by salinity. Based on growth, water-use and chloride(Cl) accumulation in leaves, VL was more salt-sensitive thanCM, but ¹⁵N uptake was equally reduced by salinity in both species.Salinity reduced ¹⁵N uptake relatively more than shoot growthover the 7-d period, such that the ¹⁵NUE (mg d. wt µg^-115N) of new shoot growth of both species increased. There wasno evidence of Cl antagonism of nitrate (NO₃) uptake but totalplant ¹⁵NO₃ uptake was positively correlated with whole planttranspiration in both species. Thus, it appears that reductionsin NO₃ uptake are more strongly related to reduced water usethan to Cl antagonism from salt stress.Copyright 1993, 1999Academic Press Sodium, chloride, salinity, calcium, nitrate, ¹⁵NO₃ uptake, nitrogen allocation, nitrogen-use efficiency, water use, Citrus reticulata, Citrus volkameriana     

A comparison of nitrate transport in four different rice (Oryza sativa L.) cultivars

As rice can use both nitrate (NO₃^-) and ammonium (NH₄⁺), we have tested the hypothesis that the shift in the pattern of cultivars grown in Jiangsu Province reflects the ability of the plants to exploit NO₃^- as a nitrogen (N) source. Four rice cultivars were grown in solution culture for comparison of their growth on NO₃^- and NH₄⁺ nitrogen sources. All four types of rice, Xian You 63 (XY63), Yang Dao 6 (YD), Nong Keng 57 (NK) and Si You 917 (SY917), grew well and produced similar amounts of shoot biomass with 1 mmol/L NH₄⁺ as the only N source. However, the roots of NK were significantly smaller in comparison with the other cultivars. When supplied with 1 mmol/L NO₃^-, YD produced the greatest biomass; while NK achieved the lowest growth among the four cultivars. Electrophysiological measurements on root rhizodermal cells showed that the NO₃^--elicited changes in membrane potential (ΔE_m) of these four rice cultivars were significantly different when exposed to low external NO₃^- (<1 mmol/L); while they were very similar at high external NO₃^- (10 mmol/L). The root cell membrane potentials of YD and XY63 were more responsive to low external NO₃^- than those of NK and SY917. The ΔE_m values for YD and XY63 rhizodermal cells were almost the same at both 0.1 mmol/L and 1 mmol/L NO₃^-; while for the NK and SY917 the values became larger as the external NO₃^- increased. For YD cultivar, ΔE_m was measured over a range of NO₃^- concentrations and a Michaelis-Menten fit to the data gave a K_m value of 0.17 mmol/L. Net NO₃^- uptake depletion kinetics were also compared and for some cultivars (YD and XY63) a single-phase uptake system with first order kinetics best fitted the data; while other cultivars (ND and SY917) showed a better fit to two uptake systems. These uptake systems had two affinity ranges: one had a similar K_m in all the cultivars (0.2 mmol/L); the other much higher affinity system (0.03 mmol/L) was only present in NK and SY917. The expression pattern of twelve different NO₃^- transporter genes was tested using specific primers, but only OsNRT1.1 and OsNRT2.1 expression could be detected showing significant differences between the four rice cultivars. The results from both the physiological and molecular experiments do provide some support for the hypothesis that the more popular rice cultivars grown in Jiangsu Province may be better at using NO₃^- as an N source.     

Al Binding in the Epidermis Cell Wall Inhibits Cell Elongation of Okra Hypocotyl

Al inhibits root elongation at micromolar concentrations, butthe mechanisms leading to this process are unknown. In thesestudies, Al-induced inhibition of cell elongation was examinedusing hypocotyl of okra (Abelmoschus esculentus Moench cv. ClemsonSpineless) as an experimental model. One-h exposure to Al (0.5mM A1Cl₃) in the presence of 10 µM auxin in 0.5 mM CaCl₂,pH 4.0 significantly inhibited auxin-induced cell elongationof okra hypocotyl segments. Elongation was further suppressedwith increasing Al concentrations up to 1 mM. Treatment of thehypocotyl with 1 mM citrate for 10 minutes after 2-h exposureto Al resulted in significant recovery of elongation. The amountof Al in the cell wall relative to the total in the tissue was96.0, 96.2, and 85.4%, respectively, following 1-, 2-, and 3-hexposure to the Al solution. The total and cell wall Al contentwas decreased by half after the citrate desorption treatment.Further-more, 95% of Al was found in the epidermis, and 95%of the Al in the epidermis was associated with the cell wall.Experiments using split hypocotyl segments showed that Al exposureincreased the outward bending of hypocotyl segments, suggestingthat the epidermis elongation was specifically inhibited byAl. Al inhibited the autolysis of epidermis by about 20%, buthad little effect on the autolysis of core tissue. Taken together,these results suggest that Al binding in the epidermal cellwall inhibits critical components in cell wall loosening mechanism,resulting in inhibition of cell elongation.     

Kinetic Studies on the Active Species of Inorganic Carbon Absorbed by the Cells of Dunaliella tertiolecta

Cells of Dunaliella tertiolecta which had been grown in ordinaryair (low-CO₂ cells) had high carbonic anhydrase (CA) activityon the cell surface and mainly utilized HCO₃^– for photosynthesis.When CA activity on the cell surface was inhibited by Diamoxor subtilisin, the cells utilized CO₂. When bovine CA was added,the subtilisin-treated low-CO₂ cells utilized mainly HCO₃^–.When grown in air containing 2% CO₂, the cells had low CA activityon the cell surface, and preferred CO₂ to HCO₃^–. Kineticanalysis of these results indicated that low-CO₂ cells of D.tertiolecta absorb CO₂ which was converted from HCO₃^–via the CA located on the cell surface. (Received June 29, 1985; Accepted October 9, 1985)     

Inoculation with Azospirillum lipoferum Affects Growth and Gibberellin Status of Corn Seedling Roots

Maize (Zea mays L., hybrid Cargill 147) seedlings, grown inaseptic conditions, were inoculated with three strains of Azospirillumlipoferum (Al op 33, Al iaa 320, and ATCC 29708) or culturedin different concentrations of indol-3-acetic acid (IAA) orgibberellin A₃ (GA₃). After 48 h, root length, root surfacearea, root dry weight, and shoot dry weight were measured inall treatments. Gibberellin content was evaluated in selectedroots of control and Azospirillum inoculated seedlings by gaschromatography-mass spectrometry-selected ion monitoring withthe use of deuterated gibberellins as internal standards. Thethree strains of A. lipoferum, IAA (2 ng ml^–1), and GA₃(40 to 400 pg ml^–1) significantly enhanced root growth.Improvement of root hair growth and density was obtained mainlywith A. lipoferum strain Al op 33 and GA₃ 40 pg ml^–1.GA₃ was identified by gas chromatography-mass spectrometry (byboth, full scan and selected ion monitoring) in a free acidfraction from roots of the seedlings inoculated with A. lipoferum.In the roots of the non inoculated seedlings GA₃ was found afterhydrolysis of a fraction expected to contain glucosyl conjugates. (Received April 26, 1993; Accepted September 27, 1993)     

Aluminium Effects on Pollen Germination and Tube Growth of Chamelaucium uncinatum. A Comparison with Other Ca2+Antagonists

An interaction between aluminium (Al) and calcium (Ca) may bea cause of Al toxicity in plants. The pollen tube is a suitablesystem to test the interaction between Al and Ca since Ca ionsplay a pivotal role in pollen germination and tube growth. Weinvestigated how Al and other known blockers of Ca²⁺-permeablechannels (trivalent cations, ruthenium red, verapamil and nifedipine)influence pollen of an Australian native species Geraldton waxflower(Chamelaucium uncinatum). Pollen germination was inhibited bymicromolar concentrations of trivalent cations (La³⁺>Al³⁺>Gd³⁺)and ruthenium red, but it was relatively insensitive to a micromolarconcentration of verapamil. Exposure of the growing pollen tubesto micromolar concentrations of Al³⁺and La³⁺, and a millimolarconcentration of Ca²⁺chelator ethyleneglycol-bis(ß-aminoethylether)-N,N'-tetraacetic acid (EGTA) led to rapid tip bursting.In contrast, exposure to Gd³⁺, nifedipine, ruthenium red, verapamiland the organic trivalent cation tris (ethylenediamine)cobalt(TEC³⁺) caused only inhibition of pollen tube growth. The Al³⁺-relatedpollen tube bursting was reduced significantly by increasingeither solution pH from 4.5 to 6 or activity of Ca²⁺from 0.25to 5 m M. In contrast, La³⁺-related pollen tube bursting wasinsensitive to changes in Ca²⁺activity. The results are discussedin terms of Al interactions with cell wall Ca²⁺and the plasmamembrane Ca²⁺-permeable channels. Copyright 1999 Annals of BotanyCompany Aluminium toxicity, Ca²⁺-channel blockers, cell wall, Chamelaucium uncinatum, pollen germination, pollen tube growth.     

Nitrogen Utilization in N-limited Barley during Vegetative and Generative Growth: I. GROWTH AND NITRATE UPTAKE KINETICS IN VEGETATIVE CULTURES GROWN AT DIFFERENT RELATIVE ADDITION RATES OF NITRATE-N

Growth and nitrate uptake kinetics in vegetatively growing barley(Hordeum vulgare L., cvs Laevigatum, Golf, and Mette) were investigatedin solution culture under long-term limitations of externalnitrogen availability. Nitrate was fed to the cultures at relativeaddition rates (RA) ranging from 0.02 to 0.2 d^–1. Therelative growth rate (RG, calculated for total plant dry weight)correlated well with RA in the range 0.02 to 0.07 d^–1.In the RA range from 0.07 to 0.2 d^–1 RG continued to increase,but an increasing fraction of nitrogen, added and absorbed,was apparently stored rather than used for structural growth.The RG of the roots was less affected by RA. V_max, for net nitrateuptake increased with RA up to 0.11 d^–1, but decreasedat higher RA. The decline in V_max coincided with a build-upof nitrate stores in both roots and shoots. V_max, expressedper unit nitrogen in the plants (the relative V_max, was higherthan required for maintenance of growth (up to 30-fold) at lowRA, whereas at higher RA the relative V_max decreased. Kineticpredictions of steady-state external nitrate concentrationsduring N-limited growth ranged from 0.2 to 5.0 mmol m^–3over the RG range 0.02 to 0.11 d_–1. It is suggested thatthe nitrate uptake system is not under specific regulation atlow RA, but co-ordinated with root protein synthesis and growthin general. At RA higher than 0.11 d_–1, however, specificregulation of nitrate uptake, possibly via root nitrate pools,become important. The three cultivars showed very similar growthand nitrate uptake characteristics. Key words: Barley, growth, nitrogen limitation, nitrate uptake, kinetics     

Activation of K+ channels induces apoptosis in vascular smooth muscle cells

Intracellular K⁺ playsan important role in controlling the cytoplasmic ion homeostasis formaintaining cell volume and inhibiting apoptotic enzymes in thecytosol and nucleus. Cytoplasmic K⁺ concentration is mainlyregulated by K⁺ uptake viaNa⁺-K⁺-ATPase and K⁺ efflux throughK⁺ channels in the plasma membrane. Carbonyl cyanidep-trifluoromethoxyphenylhydrazone (FCCP), a protonophorethat dissipates the H⁺ gradient across the inner membraneof mitochondria, induces apoptosis in many cell types. In ratand human pulmonary artery smooth muscle cells (PASMC), FCCP opened thelarge-conductance, voltage- and Ca²⁺-sensitiveK⁺ (maxi-K) channels, increased K⁺ currentsthrough maxi-K channels [I_K(Ca)], and inducedapoptosis. Tetraethylammonia (1 mM) and iberiotoxin (100 nM)decreased I_K(Ca) by blocking the sarcolemmalmaxi-K channels and inhibited the FCCP-induced apoptosis inPASMC cultured in media containing serum and growth factors.Furthermore, inhibition of K⁺ efflux by raisingextracellular K⁺ concentration from 5 to 40 mM alsoattenuated PASMC apoptosis induced by FCCP and theK⁺ ionophore valinomycin. These results suggest thatFCCP-mediated apoptosis in PASMC is partially due to anincrease of maxi-K channel activity. The resultant K⁺ lossthrough opened maxi-K channels may serve as a trigger for cellshrinkage and caspase activation, which are major characteristics ofapoptosis in pulmonary vascular smooth muscle cells.