Nutrient Uptake and Organic Acid Anion Metabolism in Lupins and Peas Supplied with Nitrate

Unlike many plants reported in the literature, lupins do notexcrete OH^- in amounts equivalent to the net excess of inorganicanion uptake over inorganic cation uptake. To investigate themechanisms involved in the maintenance of charge balance, nutrientuptake and organic anion accumulation of lupins and peas suppliedwith a range of NO^-₃ concentrations, were compared. Lupins absorbed less NO^-₃ than peas on a dry weight basis, whichlargely accounted for the smaller excess of anion uptake overcation uptake in lupins than in peas at the same NO^-₃ supply.When anion uptake exceeded cation uptake, peas excreted an equivalentcharge of OH^-, whereas lupins excreted much smaller amountsof OH^- than the excess of anion over cation uptake. It was calculatedthat lupins excreted significant amounts of organic anions whenanion uptake exceeded cation uptake, whereas organic anion excretionfrom peas was negligible, regardless of their NO^-₃ supply andcation-anion balance. In this study, organic anion excretion was measured from lupinroots grown in near-sterile conditions while supplied with NO^-₃at 0, 500 and 2000 µM. Although complete sterility wasnot achieved, there was close agreement between the organicanion excreted and the excess anion over cation uptake.Copyright1994, 1999 Academic Press Lupinus angustifolius L., Pisum sativum L., organic acid, nutrient uptake     

H+/OH- Excretion and Nutrient Uptake in Upper and Lower Parts of Lupin (Lupinus angustifolius L.) Root Systems

The cultivation of narrow-leafed lupins (Lupinus angustifoliusL.) increase rates of subsoil acidification, and this is thoughtto be partly related to their pattern of nutrient uptake andH⁺/OH^- excretion. The main hypothesis of this study was thatH⁺ and OH^- excretion is not distributed evenly over the entirelength of the root system but is limited to zones where excesscation or anion uptake occur. Seedlings of nodulated lupinswere grown in solution culture using vertically split pots thatallowed the upper and lower zones of the root system to be suppliedwith varying concentrations of K⁺ and NO^-₃. Net H⁺/OH^- excretionwas equated to the addition of NaOH/HCl required to maintaina constant pH in the nutrient solution during a 4-d treatmentperiod and nutrient uptake was measured by depletion from solutionin each zone of the split pots. The excess of cation over anion uptake was positively correlatedwith H⁺ excretion in each rooting zone. In zones where K⁺ wassupplied at 1200 µM, cation uptake was dominated by K⁺and up to twice as much H⁺ was excreted than in zones whereK⁺ was absent. In zones where NO^-₃ was supplied at 750 µM,the anion/cation uptake was balanced, however H⁺ excretion continuedto occur in the zone. When NO^-₃ was supplied at 5000 µM,anion uptake exceeded cation uptake but there was no OH^- excretion.Organic acid anions may be excreted by lupins to maintain theirinternal electroneutrality when anion uptake exceeds cationuptake. Rhizosphere pH would not increase unless the pK_a ofthe excreted organic anions was greater than the external pH.Copyright1993, 1999 Academic Press Lupinus angustifolius L., H⁺/OH^- excretion, nutrient uptake, cation-anion balance, vertical split root     

The Uptake of Phosphate by Plants from Flowing Nutrient Solution: II. GROWTH OF LOLIUMPERENNEL. AT CONSTANT PHOSPHATE CONCENTRATIONS

The effect of phosphate concentration in flowing solution cultureat a range between 0.04 and 32 mmol m^–3 P on the growthof perennial ryegrass was studied in two experiments, each lastingabout 45 d after sowing. Phosphorus contents of seedlings wereaffected by the concentration in solution within about 5 d fromgermination, and dry weight differences were first observedat about 6 d after this. The rate of uptake of phosphate byseedlings was affected by the concentration in solution beforethe root fresh weight or root/emdash shoot ratio had changed.Young plants (less than 4 weeks old) were more sensitive tophosphate concentration in solution than older ones. In conditionsof high rate of growth, older plants required a solution concentrationbetween 0.1 and 0.4 mmol m^–3 P to achieve maximum potentialgrowth rate, whereas for plants of similar age but less dryweight, 0.04 mmol m^–3 P was adequate. Towards the endof the experimental period, plants growing at a nominal solutionconcentration of 0.04 mmol m^–3 P were able to obtain phosphatefrom a solution of about 0.01 mmol m^–3P. Phosphate toxicity was not observed, nor were there visual symptoms(other than reduced growth) of phosphate deficiency in plantswhose growth was limited by phosphate concentration in solution. Key words: Lolium perenne, Phosphate uptake     

RESPIRATORY METABOLISM IN THE PHLOEM, XYLEM AND CAMBIUM OF CARROT ROOT

Comparisons of respiratory metabolism among the phloem, cambiumand xylem of mature carrot root were carried out and the followingresults were obtained. 1) The activity of O₂ uptake on a dry weight basis in the cambiumwas higher than that in the xylem. The phloem showed the lowestactivity. 2) RQ was close to unity in all the three tissues.3) The inhibition rate of O₂ uptake by malonate was considerablyhigh in the three tissues. The highest inhibition was observedin the cambium. 4) In the phloem and xylem the malonic inhibitionrate of ¹⁴CO₂ release from G-U-¹⁴C was not so high as comparedwith that of O₂ uptake. In the cambium, however, those of O₂uptake and ¹⁴CO₂ output were comparable. 5) Malonate treatmentdid not cause any significant change in RQ. 6) The C₆/C₁ ratiowas highest in the cambium and lowest in the phloem. From these it is concluded that the participation of the TCAcycle in respiration is great in all the three tissues of carrotroot, and among the three the cambium has the highest activityof the cycle. As for the PP pathway, the xylem and phloem havea higher activity than the cambium. (Received April 11, 1967; )     

青海省三江源区人工草地生态系统CO2通量

 了解三江源人工草地净生态系统CO₂交换(Net ecosystem CO₂ exchange, NEE)的季节变化规律和主要生物因子及环境因子对这些过程的影响将有助于认识青藏高原人工草地生态系统碳循环、生态价值、功能,以及对三江源区的生态安全的重要意义。该研究利用涡度相关技术,于2005年9月1日至2006年8月31日对位于青海腹地的垂穗披碱草（Elymus nutans）人工草地的NEE及生物和环境因子进行观测, 阐明NEE及其组分的动态变化特征和影响因子。三江源区人工草地生态系统的日最大吸收量为2.38 g C·m^－2·d^－1,出 现在7月30日。日间最大吸收率和最大排放率都出现在8月,分别为-6.82和2.95μmol CO₂·m^－2·s^－1。在生长季, 白天的NEE主要受光合有效辐射(Photosynthe tically active rad iation, PAR)变化控制,同时又与叶面积指数和群落多样性交互作用,共同调节光合速率和光合效率的强度。最大光合同化速率为2.46～10.39μmol CO₂·m^－2·s^－1,表观初始光能利用率为0.013～0.070μmol CO₂·μmol^－1 PAR。 在碳交换日过程中,NEE并不完全随着 PAR的增加而增大,当PAR超过某一值（>1 200μmol ·m^－2·s^－1）时,NEE随PAR的增加而降低。受温度的影响,生长季的生态系统的呼吸商Q₁₀（1.8）小于非生长季节的 2.6）。 生态系统呼吸主要受温度的控制,同时也受到叶面积指数的显著影响。生长季昼夜温差大并不利于生态系统的碳获取。 三江源区人工草地生态系统是一个较强的碳汇,为-49.35 g C·m^－2·a^－1。     

The Uptake of Growth Substances: I. FACTORS CONTROLLING THE UPTAKE OF PHENOXYACETIC ACIEDS BY LEMNA MINOR

An analysis has been made of the factors controlling the uptakeof 2:4-dichloro-phenoxyacetic acid (2:4-D) and related compoundsby Lemna minor, grown under controlled conditions. All compoundswere labelled with ¹⁴C in the carboxyl group and the ¹⁴C inthe plants, after liquid combustion, assayed as barium carbonate. With 2:4-D the rate of entry from concentrations of 8 to 32mg./l. is maximal in the first 20 minutes, then falls progressivelyuntil the rate is zero between 1 and 2 hours and in the thirdphase (up to 24 hours) there is a net loss from the plants dueto an outward movement of the compound back into the externalsolution. When the chlorine substitutions are in the 2:6-positionthe course of uptake is similar, save that loss to the solutiontakes place later. For the 2-chloro-substitution, after theinitial phase of uptake there is some loss but this is followedby a period of recovery and uptake again proceeds, but slowly.In contrast, the course of uptake over 24 hours of phenoxyaceticacid is normal, i.e. there is a steady accumulation. When plants are pretreated with labelled 2:4-D for 30–60minutes and transferred to water or culture solution then over90 per cent. of the ¹⁴C is found in the external medium after4:5 hours and this release cannot be accounted for in termsof exchange processes since the addition of unlabelled compoundto the medium retards the rate of loss. This loss is slowedbut not stopped at 1.25°C. and up to 22.5°C. the rangeof the Q₁₀ is 1.6–1.9. Uptake in the first 30 minutes is temperature sensitive between7.5 and 30°C. (Q₁₀ 2.3–2.6) and in general is positivelyand curvilinearly related to the external concentration. Pretreatmentwith unlabelled compound up to 2 hours progressively depressesthe subsequent initial uptake of labeled growth regulator. Itis concluded that initially the rate of entry greatly exceedsthe rate of loss but that with time the ratio steadily diminishesto less than unity. Initial uptake of 2:4-D is markedly dependent on the pH of thesolution and closely but not completely correlated with theexternal concentration of undissociated molecules. On the otherhand, the outward movement is relatively unaffected by the externalpH. Combinations of concentration and time of exposure which bringabout loss to the solution need not cause any permanent retardationof growth. In fact, exposure for 1 hour to 8 mg./l. significantlyaccelerated growth in the next 8 days. These results are discussed in relation to previous findingsand it is concluded that the pattern of uptake is determinedon the one hand by the chemical structure of the growth substanceand on the other by specific physiological differences at celllevel.     

Intercellular Transport in Plants: I. THE RATE OF TRANSPORT OF CHLORIDE AND THE ELECTRIC RESISTANCE

A quantitative study has been made of the intercellular movementof chloride in Chara corallina, using pairs of joined internodalcells. One cell of the pair (cell 1) was exposed to a solutioncontaining ³⁶Cl; the distribution of this tracer between thecells was determined at the end of the uptake period. Of thechloride taken up, 0.29 was transported out of cell 1 for alluptake times from 1.8 to 22 ks and 0.57 was transported to thevacuole of cell 1, in experimental series I. In series II, thefraction transported out of cell 1 was 0.43 at 22 and 43 ks,but 0.17 at 600 s. These results represent a rate of transport of 4 to 60 pmols^–1, across an intercellular wall of area 1.5 x 10^–6m²; the wall has 0.03 to 0.04 of its area occupied by plasmodesmata.The estimate of transport rate is based on an attempt to determinethe specific activity of the cytoplasm of cell 1. The electricresistance of the node was found to be 47 m m². The observed transport rate can be explained by diffusion inthe plasmodesmata, without the need to postulate active processes.Diffusion in the plasmodesmata is slower than in free solutionby an ‘impediment factor’ of 7 to 700, dependingon the assumed chloride concentration of the ground-plasm. Ifthe plasmodesmata offer the major conducting path for electriccurrent, the electric impediment factor is 390. Chloride entersthe plasmodesmata from the same small kinetic compartment whichsupplies the flux to the vacuole, or from a smallintermediatecompartment.     

EFFECTS OF CHLORAMPHENICOL AND KINETIN ON UPTAKE AND INCORPORATION OF AMINO ACIDS BY TOBACCO LEAF DISKS

The effects of chloramphenicol and kinetin on uptake and incorporationof ³⁵S-methionine and some ₁₄C-amino acids have been investigatedin leaf-disks of Nicotiana rustica in light and dark. Chloramphenicolin a concentration of 1 mg per ml inhibits the uptake of aminoacids from 30 to 60 per cent compared with the water control.The incorporation of amino acids into bulk protein is stronglyinhibited in light (40 to 70 per cent), but only to a smalldegree in dark (10 to 20 per cent), as revealed also by ¹⁴CO₂-photosynthesisof the disks and following treatment with chloramphenicol indark. The stimulating effect of kinetin on uptake and incorporationof amino acids is dependent upon its concentration (10^–5to 10^–6 M ; but 10^–4 M solution inhibits stronglyboth uptake and incorporation). The stimulation seems to influencemore incorporation than uptake processes. Possible interactionsof chloramphenicol and kinetin in the protein metabolism oftobacco leaves have been discussed. (Received April 27, 1964; )     

ON THE NATURE OF MOSS OXALIC ACID OXIDASE

Moss oxalic acid oxidase freed from catalase by boiling is stronglyinhibited by the "metal-complexing" compounds such as thiocyanate,azide, diethyldithiocarbamate, and hydrosulfite. Inactivatedby dialysis against thiocyanate or azide, the enzyme can bereactivated to a considerable extent by the addition of ferricsalt, cytochrome-c or hemoglobin, not by other metal ions, suchas Cu²⁺, Zn²⁺ , Mn²⁺, and Fe²⁺. Nitrate, chlorate, monoiodoacetate,and iodide also act as strong inhibitors towards moss oxalicacid oxidase. Some enzyme fractions which were obtained by the sodium sulfateprecipitation method were stimulated by Fe³⁺, but not by cytochrome-cor by other metallic ions. This stimulation was inhibited bythiocyanate, azide and monoiodoacetate. ¹ Present address: Biological Institute, University of Toyama,Toyama     

Conductive pathways for chloride and oxalate in rabbit ileal brush-border membrane vesicles

To evaluate thepossibility that an apical membrane conductive pathway for oxalate ispresent in the rabbit distal ileum, we studied oxalate([¹⁴C]oxalate) andchloride (³⁶Cl) uptake intobrush-border membrane vesicles enriched 15- to 18-fold in sucraseactivity. Voltage-sensitive pathways for oxalate and chloride wereidentified by the stimulation of uptake provided by an inwardlydirected potassium diffusion potential in the presence of valinomycin.Additionally, outwardly directed oxalate (or chloride) gradientsstimulated[¹⁴C]oxalate (or³⁶Cl) uptake to a greater degreein the absence of valinomycin (when intracellular and extracellularpotassium are equal) than in the presence of valinomycin.Voltage-dependent anion uptake was poorly saturable: apparent affinityconstants were 141 ± 17 and 126 ± 8 mM for chlorideand oxalate, respectively. Activation energies for thevoltage-dependent uptake processes were low: 4.7 and 6.3 kcal/mol forchloride and oxalate, respectively. Sensitivity profiles ofvoltage-dependent chloride and oxalate uptake to anion transport inhibitors were similar. We conclude that an anion conductance ispresent in the apical membranes of ileal enterocytes and that thisconductance is a candidate pathway for oxalate efflux from theenterocyte during transepithelial oxalate secretion.

     

THE KINETICS OF PENETRATION : XIV. THE PENETRATION OF IODIDE INTO VALONIA

When 0.1 M NaI is added to the sea water surrounding Valonia iodide appears in the sap, presumably entering as NaI, KI, and HI. As the rate of entrance is not affected by changes in the external pH we conclude that the rate of entrance of HI is negligible in comparison with that of NaI, whose concentration is about 10⁷ times that of HI (the entrance of KI may be neglected for reasons stated). This is in marked contrast with the behavior of sulfide which enters chiefly as H₂S. It would seem that permeability to H₂S is enormously greater than to Na₂S. Similar considerations apply to CO₂. In this respect the situation differs greatly from that found with iodide. NaI enters because its activity is greater outside than inside so that no energy need be supplied by the cell. The rate of entrance (i.e. the amount of iodide entering the sap in a given time) is proportional to the external concentration of iodide, or to the external product [N⁺]_o [I^-l_o, after a certain external concentration of iodide has been reached. At lower concentrations the rate is relatively rapid. The reasons for this are discussed. The rate of passage of NaI through protoplasm is about a million times slower than through water. As the protoplasm is mostly water we may suppose that the delay is due chiefly to the non-aqueous protoplasmic surface layers. It would seem that these must be more than one molecule thick to bring this about. There is no great difference between the rate of entrance in the dark and in the light.     

Regulation of Sulphate Transport in a Tropical Legume, Macroptilium atropurpureum, cv. Siratro

When young plants of Macroptilium atropurpureum, cv. Siratrowere deprived of external sulphate (-S plants) growth of shootsand roots continued at rates comparable to those in plants wellsupplied with sulphate (control) for 3 d and 5 d respectively.Dilution of internal sulphur therefore took place and redistributionof sulphur occurred between inorganic and organic forms andbetween roots and younger leaves. Even when S-deficiency limitedgrowth, plants contained 16% of their total sulphur as sulphate,but most of this was retained in old leaves and redistributedslowly to growing zones. The capacity for sulphate uptake increased in roots of –Splants very soon after they were deprived of external sulphate;within 24 h the absorption from 0.25 mol m^–3 SO₄^2–was more than five times that of control roots. Maximum increasedcapacity was reached after 2–3 d stress when the V_maxof system 1 was 1948 nmol h^–1g^–1root fr. wt. in–S plants and 337 nmol h^–1g^–1root fr. wt.in controls. The K^mfor system 1 did not change significantlywith S-stress being between 5–8 µM in both setsof plants. Absorption of L-cysteine was not stimulated by S-stress. There was a close, positive relationship between plant growthrate and the rate at which sulphate uptake capacity was enhancedby withholding sulphate from culture solutions. When –S plants were replaced in sulphate-containing solutiontheir capacity for SO₄^2– declined to the control levelwithin 24 h. Very marked repression of capacity was also foundwhen –S plants were treated with L-cysteine, but therewas no immediate effect with methionine. Roots of this species appear to have a very active system fordegrading L-cysteine to sulphate, 30% of the label in ³⁵S-cysteineabsorbed by roots was recovered in ³⁵SO₄^2– after 20 minor 2 h incubation. By contrast, roots had a very weak abilityto reduce sulphate. When part of the root system was in solution lacking sulphatethere was enhanced uptake of sulphate by other parts which themselveswere amply supplied with sulphate. This is seen as an exampleof compensatory absorption. The response to S-stress is specific and there were no positiveinteractions between S-stress and the absorption of phosphate,or P-stress and the uptake of sulphate. The results are discussed in relation to the close control ofsulphate uptake by internal sulphate concentration, redistributionof forms of sulphur during stress and mobility of sulphate inthe phloem. Key words: Kinetics, Amino-S, Sulpholipid, Repression;, Deficiency     

Whole Plant Studies using Radioactive 13-Nitrogen: II. A COMPARTMENTAL MODEL FOR THE UPTAKE AND TRANSPORT OF NITRATE IONS BY Zea mays

Nitrate ion uptake by the roots of hydroponically grown maizeseedlings was measured using the short-lived isotope ¹³N. Itis shown to be described by a four compartment model, recognizablynitrogen in the root bathing solution, nitrogen which is readilyexchangeable from the root, nitrogen bound in the root, andnitrogen transported from the root. Some of the absorbed activity leaks back into the root bathingsolution with the efflux from the root, as a fraction of theinflux, increasing with concentration to be greater than 0–8at external nitrate ion concentrations above about 1.0 mol m^–3.The capacity of the exchangeable root pool increases with externalnitrate ion concentration, approaching the expected cytoplasmicnitrate ion content at the highest external nitrate ion concentrationsstudied (70 mol m^–3). The investigation has highlighted the problems of interpretinguptake profiles in experiments for which the 10 min half-lifeof ¹³N dictates experimental times that are comparable withthe times for saturation of root pools. Key words: Zea mays, ¹³N, Compartmental model, Nitrate uptake     

THE INCORPORATION OF 32P ORTHOPHOSPHATE INTO THE PHOSPHOLIPIDS OF ELONGATING AVENA COLEOPTILES

Four phospholipids of Avena coleoptile tissue were identifiedas phosphatidyl inositol, phosphatidyl glycerol, phosphatidylethanolamine and phosphatidyl choline. IAA caused an increase in total uptake of ³²P and incorporationof ³²P in phospholipids. IAA also caused a shift in the proportionsof identified ³²P phospholipids. Incorporation of ³²P into phosphatidylcholine was greater while incorporation into phosphatidyl glyceroland phosphatidyl ethanolamine was less in IAA-treated tissuecompared with untreated control tissue. ¹Contribution No. 338 from the Department of Botany, PennsylvaniaState University and 3001 from the Pennsylvania AgriculturalExperiment Station. ²Present address: Juniata College, Huntingdon, Pennsylvania,U.S.A.     

Microbiological Synthesis of Fat: THE EFFECTS OF NITROGEN LEVEL ON THE METABOLISM OF PENICILLIUM LILACINUM THOM IN A SUCROSE MEDIUM

1. A study has been made of the relationships between the synthesesof carbohydrate, protein, and fat by Penicillium lilacinum Thomin presence of different amounts of sodium nitrate us a definedsucrose salts medium.
2. Under the defined experimental conditionsincreases in the concentrationof NO^–₂ in the medium werefollowed by increases in therates at which nitrogen and sugarwere taken up by the fungus,in the quantities assimilated,and in total and protein nitrogenin the felt. These conditionsprevailed so long as unassimilatedsugar was available.
3. Mediaof lower NO^–₃ concentration (for example, 0·32or 0·64 per cent. (w/v) NaNO^–₂;) yielded feltsricher in carbohydrate than were those grown in media of higherNO^–₂; content (0·96 or 1·28 per cent. (w/v)NaNO₃ The carbohydrate content of the felts increased graduallyuntil the sugar in the medium was exhausted; carbohydrate contentthen decreased.
4. Media of lower NO^–₃; concentration weremore conduciveto fat synthesis than those of higher NO^–₃;content.

     

Light-dependent potassium uptake by Pisum sativum leaf fragments

A net K⁺ influx into chopped pea leaves bathed in 5 mM KCl,0.26 M sucrose and illuminated with 4000 lux amounted to about7.5 µmoles/g fresh weight-hr, while essentially no netflux occurred in the dark. This light-dependent K⁺ uptake waslinear with time for nearly 2 hr and continuously increasedas the light intensity was raised to 9000 lux. Over half ofthe K⁺ uptake was balanced by H⁺ release into the bathing solution,possibly by a mechanism in which bicarbonate was the anion accompanyingK⁺. The replacement of Cl^– by HCO₃^– increased thelight-dependent K⁺ uptake to 56 µmoles/g fresh weight-hr.About 23% of the light-dependent K⁺ uptake in 5 mM KCl was accompaniedby a Cl^– uptake. This net Cl^– influx was less sensitiveto the uncoupler tri-Fl-CCP and more sensitive to DCMU in thebathing solution than was the K⁺ uptake. The remaining net K⁺influx into pea leaf fragments was balanced by effluxes of sodium(accounting for 5%), magnesium (8%) and calcium (1%). (Received March 31, 1969; )     

不同生态型东南景天对土壤中Cd的生长反应及吸收积累的差异性

 采用盆栽方法研究了两种生态型东南景天(Sedum alfredii)对土壤中不同含量Cd(即对照, 12.5, 25, 50, 100, 200, 300, 400 mg&;#8226;kg^－1)的生 长反应、吸收和积累Cd的差异性。结果表明,土壤添加重金属Cd后,矿山生态型东南景天生长正常,地上部和根系Cd含量随着土壤中Cd含量的 增加而增加,在400 mg&;#8226;kg^－1 Cd处理下含量分别高达2 900和500 mg&;#8226;kg^－1,其地上部显著大于根部;然而,土壤添加Cd后,非矿山生态型东 南景天的生长受到抑制,地上部和根部的生物量显著降低。当土壤Cd含量为50～100 mg&;#8226;kg^－1 时,非矿山生态型东南景天的地上部和根系Cd含 量随着土壤中Cd含量的增加而增加,而且根系Cd含量则大于地上部。当土壤Cd≤50 mg&;#8226;kg^－1时,矿山生态型东南景天根系Cd含量比非矿山生态 型高 ,但当土壤Cd≥100 mg&;#8226;kg^－1,两者之间无显著差异;然而,但在同一Cd处理水平下,矿山生态型东南景天地上部Cd含量总是高于非矿山 生态型。这些结果表明,矿山生态型东南景天有很强的忍耐和吸收土壤Cd的能力,再次证明其为一种Cd超积累植物。     

Response of Wheat Seedlings to Low O2 Concentrations in Nutrient Solution: II. K +/Na+ SELECTIVITY OF ROOT TISSUES9

This paper reports the effects of low O₂ concentration (0–01,0–055, and 0.115mol m^–3) in nutrient solutions onK⁺/Na⁺ selectivity of growing and mature root tissues of 6-to 8-d-old, intact, wheat (Triticum aestivum cv. Gamenya) seedlings. Increases in anaerobic catabolism and decreases in O₂ uptake,K⁺ uptake and K⁺/Na⁺ selectivity were all more pronounced and/oroccurred at higher external O₂ concentrations in the apex (0–2mm) than in the expanding tissues (2–4 mm); these growingtissues were, in turn, more affected than the expanded tissuesof the roots (4–12 mm). Selectivity for K⁺ over Na⁺ in roots and shoots was particularlysensitive to O₂ deficiency. For example, in apical tissues (0–2mm) K ⁺ /Na⁺ selectivity was already reduced at 0.115 mol m^–3O₂, yet at this O₂ concentration there was no effect on eithergrowth or (K⁺/Na⁺) uptake. Upon transfer from 0.01 to 0.26 mol m^–3 O₂, a detailedstudy of the 12 mm root tips showed that 70% of these tips regainedhigh (K⁺ + Na⁺) concentrations and K⁺/^Na+ ratios. In contrast,there was no recovery in the remaining 30% of the 12 mm roottips. Net K⁺ transport to the shoots during the period afterre-aeration was negative for the population as a whole. Theseverity of these effects supports the view that the root tipsand the stele were more susceptible to O2 deficiency than wasthe cortex of the fully-developed root tissues. Key words: Hypoxia, K⁺/Na⁺ selectivity, expanded and expanding tissues     

The Uptake of Phosphate by Plants from Flowing Nutrient Solution: IV. EFFECT OF PHOSPHATE CONCENTRATION ON THE GROWTH OF TRIFOLIUM REPENS L. SUPPLIED WITH NITRATE, OR DEPENDENT UPON SYMBIOTICALLY FIXED NITROGEN

Breeze, V. G. and Hopper, M. J. 1987. The uptake of phosphateby plants from flowing nutrient solution. IV. Effect of phosphateconcentration on the growth of Trifolium repens L. suppliedwith nitrate, or dependent upon symbiotically fixed nitrogen.—J.exp. Bot. 38: 618–630. Nodulated white clover plants were subjected to a range of phosphateconcentrations in flowing solution culture (0.32 to 8.0 mmolm^–3 P) at 41 d from sowing, either supplied with nitrateor dependent on symbiotically-fixed nitrogen. No effect of phosphateconcentration in solution on dry matter production, relativegrowth rate, root/shoot ratio, or water soluble carbohydrateconcentration of the plant tissue was observed after 24 d fromthe start of the experiment, although the plants supplied withnitrate yielded more than the others. Phosphate uptake throughoutthe experimental period was related to the solution concentration,but the source of nitrogen did not affect the phosphorus concentrationsof the shoots. However, the roots of the plants dependent onsymbiotically-fixed nitrogen had higher concentrations of phosphorusthan those supplied with nitrate, but this did not appear tobe due to an increased phosphorus requirement for nitrogen fixation,because the amount fixed was unaffected by the phosphate concentrationin solution. The cation-anion balance showed that plants dependenton nitrogen fixation had no larger requirement for calcium thanplants supplied with nitrate, but a requirement for hydroxylions equivalent to over 130 kg lime per tonne of dry shoot.It is suggested that the enhanced phosphate uptake by plantsdependent on nitrogen fixation is due to this need for a cation-chargebalancing anion. Key words: Phosphate uptake, nitrogen fixation, Trifolium repens L., repens L., cation-anion balance, flowing solution culture     

Palytoxin-induced cell death cascade in bovine aortic endothelial cells

The plasmalemmal Na⁺-K⁺-ATPase (NKA) pump is the receptor for the potent marine toxin palytoxin (PTX). PTX binds to the NKA and converts the pump into a monovalent cation channel that exhibits a slight permeability to Ca²⁺. However, the ability of PTX to directly increase cytosolic free Ca²⁺ concentration ([Ca²⁺]_i) via Na⁺ pump channels and to initiate Ca²⁺ overload-induced oncotic cell death has not been examined. Thus the purpose of this study was to determine the effect of PTX on [Ca²⁺]_i and the downstream events associated with cell death in bovine aortic endothelial cells. PTX (3–100 nM) produced a graded increase in [Ca²⁺]_i that was dependent on extracellular Ca²⁺. The increase in [Ca²⁺]_i initiated by 100 nM PTX was blocked by pretreatment with ouabain with an IC₅₀ < 1 µM. The elevation in [Ca²⁺]_i could be reversed by addition of ouabain at various times after PTX, but this required much higher concentrations of ouabain (0.5 mM). These results suggest that the PTX-induced rise in [Ca²⁺]_i occurs via the Na⁺ pump. Subsequent to the rise in [Ca²⁺]_i, PTX also caused a concentration-dependent increase in uptake of the vital dye ethidium bromide (EB) but not YO-PRO-1. EB uptake was also blocked by ouabain added either before or after PTX. Time-lapse video microscopy showed that PTX ultimately caused cell lysis as indicated by release of transiently expressed green fluorescent protein (molecular mass 27 kDa) and rapid uptake of propidium iodide. Cell lysis was 1) greatly delayed by removing extracellular Ca²⁺ or by adding ouabain after PTX, 2) blocked by the cytoprotective amino acid glycine, and 3) accompanied by dramatic membrane blebbing. These results demonstrate that PTX initiates a cell death cascade characteristic of Ca²⁺ overload. necrosis; vital dyes; membrane blebs; time-lapse video microscopy; fura-2