Isolation of MTX-resistant Cell Line NP-19 of Nicotiana plumbaginifolia: Phenotypic, Genetic and Biochemical Study

Several cell lines of Nicotiana plumbaginifolia resistant tomethotrexate (MTX) were isolated upon gradual elevation of theMTX concentration in the growth medium. One of the MTX-resistantcell lines, NP-19, acquired resistance to 10 mmol m^–3MTX, that is, at least 50-fold higher than the lethal dose forthe wild-type cell suspension. Its resistance was stably maintainedupon prolonged withdrawal of the drug. The acquisition of resistancewas accompanied by severe deterioration of regeneration potential.Regenerated shoots still manifested resistance to 50 mmol m^–3.MTX-resistant colonies and tiny shoots were recovered followingasymmetric fusion between -irradiated NP-19 protoplasts andN. plumbaginifolia leaf mesophyll-derived protoplasts on a mediumcontaining 10 mmol m^–3 MTX. The development of the shootsderived both from NP-19 calli and following somatic fusion wasarrested at the stage of 6–8 leaves. No difference wasfound in uptake of ³H-MTX between cell suspension of NP-19 andthe parental cell line. A 30-fold increase in binding of ³H-MTXto protein extract was found in cell line NP-19, suggestingdifferential capacity of MTX binding as a mechanism involvedin the MTX resistance of this cell line. Since differentiatedorgans seem more sensitive to MTX than undifferentiated tissues,this cell line is a promising source for a gene(s) conferringenhanced MTX-tolerance both in non-differentiated and differentiatedtissues. Key words: Methotrexate resistance, MTX-binding capacity, MTX uptake, tobacco cell culture     

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     

Effects of Aluminium on Honeylocust (Gleditsia triacanthos L.) Seedlings in Solution Culture

Honeylocust (Gleditsia triacanthos L.) seedlings were grownin solution culture at pH 4.0) with 50, 150, 600 and 1500 mmolm^–3 aluminium. All levels of aiuminium reduced the sizeand weight of roots, shoots and leaves with the exception ofroot elongation at 50 mmol m^–3 Al. Aluminium content ofroots was 50 to 100 times that of shoots. With increasing concentrationof aluminium, aluminium content of leaves and roots increasedexponentially while a linear increase was observed for stems.The nutrient content of seedlings was improved in 50 mmol m^–3where increases in shoot calcium, magnesium, and phosphorusconcentrations were observed. Aluminium concentrations greaterthan 50 mmol m^–3 reduced shoot nutrient content. Presenceof aluminium increased the root phosphorus and calcium levelsbut had no effect on potassium and magnesium concentrations.Results show that honeylocust is an aluminium sensitive treespecies whose growth may be reduced by high soil Al levels. Key words: Aluminium toxicity, Gleditsia triacanthos, nutrient solution     

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     

Phosphate fluxes, compartmentation and vacuolar speciation in root cortex cells of intact Agrostis capillaris seedlings: effect of non-toxic levels of aluminium

From compartmental analysis of ³²P elution measurements, concentrationsand fluxes of orthophosphate were estimated for root corticalcells of intact seedlings of the indigenous grass Agrostis capillarisL. cv. Highland when in complete nutrient solution containing10 mmol m^–3 or 100 mmol m^–3 phosphate with or without3.7 and 37 mmol m^–3 Al, respectively, during loading andelution. When plotted as counts min^–1 remaining in thetissue as a function of time, the data failed to meet the criteriafor first order kinetics. Transformation of the data to meetthe kinetic criteria gave corrected values for compartmentalconcentrations and fluxes of phosphate, and estimates of theotherwise unresolved slowly exchanging compartment within thevacuole considered to be the cause of the discrepancy in fluxanalysis. In the control, the discrepancy was considered dueto sequestration of phosphate with Ca in the vacuole and a small,but not significant, increase in sequestered P occurred in thepresence of Al, the presence of which was confirmed by X-raymicroanalysis. A chemical speciation model was used to demonstrate,for various values of pH and carboxylic acid concentration,the possibility that phosphate was precipitated in root cellvacuoles as Ca and Al phosphates (hydroxy-apatite and variscite).The ecological significance of the ability of A. capillarisplants to make efficient use of scarce P resources by minimizingthe sequestration of P by Al in vacuoles, compared with Loliumperenne, was recognized. Key words: Phosphate sequestration, phosphate use efficiency, X-ray microanalysis, super-saturation     

The Role of Glucose on Flower Bud Formation in Thin-Layer Tissue 12Nicotiana tabacum L.

The effect of glucose on flower bud formation was studied inthin-layer tissue cultures of epidermal strips from flower stalksof Nicotiana tabacum L. cv. Samsun. A minimum concentration of 30 mol m^–3 glucose in the MS-mediumcontaining 1.0 mmol m^–3 of both NAA and BA was necessaryfor flower bud formation. With 150 mol m^–3 glucose a minimumstay of 10 d was required for optimal flower bud formation. Withholding glucose for a limited period at different time intervalsafter the onset of culture caused a delay in flower bud formationand did not affect previous development on glucose. The resultsindicated that competence for flower bud initiation is not restrictedto the early stage of culture. The process may start at anytime later at the appropriate glucose concentration. However,for both optimal initiation and further development of flowerbuds the presence of a metabolizable sugar is required. Incubationof the tissue on glucose is associated with higher respirationrate. Key words: Flower formation, Glucose, mannitol, Nicotiana tabacum, Respiration, tissue culture     

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     

The Effect of External pH on the Apparent CO2 Affinity of Chlorella saccharophila

The carbon dioxide compensation point of the unicellular greenalga, Chloretla saccharophila, was determined in aqueous mediumby a gas chromatographic method. Compensation points decreasedmarkedly from 63 cm³ m^–3 at an external pH of 4.0 to 3.2cm³ m^–3 at pH 8.0 and were not affected by the O₂ concentrationof the medium. The calculated CO² concentration required tosupport the half-maximum photosynthetic rate of the algal cellsranged from 6.0 mmol m_–3 at an external pH of 60 to 1.5mmol m^–3 at pH 8.0 and these values were not affectedby O₂ concentration. The K_m(CO₂) of nbulose-l,5-bisphosphatecarboxylase isolated from cells grown either at pH 4.0 or pH8.0 was determined to be 64 mmol m^–3. These results indicatethat loss of CO₂ by photorespiration does not occur in C. saccharophilacells at acid pH and the disparity between the apparent affinityfor CO₂ of the intact cells and that of the carboxylase indicatesthe operation of a ‘CO₂ concentrating mechanism’in this alga at acid pH. Key words: Acidophilic alga, bicarbonate transport, Chlorella saccharophila, compensation point, CO₂ affinity, PH, RuBP carboxylase     

Modifying Effects of a Non-Toxic Level of Aluminium on Phosphate Fluxes and Compartmentation in Root Cortex Cells of Intact Ryegrass Seedlings

From compartmental analysis of ³²P elution measurements, concentrationsand fluxes of phosphate were estimated for root cortical cellsof intact Lolium perenne L. plants, when in complete nutrientsolution containing 0.1 mol m^–3 phosphate with and without37 mmol m^–3 Al during loading and elution. Failure ofthe data, when plotted as ct. min^–1 remaining in the tissueas a function of time, to meet the criteria for first orderkinetics led to a discussion of the relative importance of transportto the shoot, assimilation, and complexation in the vacuole,in causing this discrepancy. It was concluded that complexationwas the most important factor. Transformation of the data tomeet the criteria for first order kinetics gave corrected valuesfor compartmental concentrations and fluxes of phosphate, andestimates of the size of the otherwise unresolved slowly exchangingcompartment within the vacuole equated with condensed phosphate.In the control this was 2.5 mol m^–3 P but in Al treatmentsa much larger amount of phosphate was complexed with Al (6.5mol m^–3 P). Phosphate transport to the shoot was unaffectedby Al. Instead, levels of Al, common in solution in upland soils,sequestered in the root vacuoles quantities of phosphate significantfor stressing the phosphate economy of nutrient-poor grassland. Key words: Lolium perenne L. phosphate, aluminium, compartmentation, complexation     

Flexible Coupling of Phosphate Uptake in Dunaliella acidophila at Extremely Low pH Values

The acidophilic alga Dunaliella acidophila exhibits optimalgrowth at pH 1. We have investigated the regulation of phosphateuptake by this alga using tracer techniques and by performingintracellular phosphate measurements under different growthconditions including phosphate limitation. In batch culturewith 2·2 mol m^–3 phosphate in the medium the uptakeof phosphate at micromolar phosphate concentrations followeda linear time dependence in the range of minutes and rates werein the range of 1 µmol phosphate mg^–1 chl h^–1,only. However, under discontinuous phosphate-limited growthconditions, tracer influx revealed a biphasic pattern at micromolarphosphate concentrations: An initial burst phase resulted ina 10⁴-fold internal phosphate accumulation and levelled offafter about 10 s. A double reciprocal plot of the initial influxrates obtained for phosphate-limited and unlimited algae exhibitedMichaelis-Menten kinetics. Phosphate limitation caused a significantactivation of the maximum velocity of uptake, yielding V_maxup to 1 mmol mg^–1 chl h^–1 as compared to valuesin the order of 50 µmol phosphate mg^–1 chl h^–1for the second phase (this magnitude is also representativefor non-limited batch cultures). Concomitantly the Michaelisconstant was altered from 4 mmol m^–3 to 0·7 mmolm^–3. The rapid uptake of phosphate was inhibited by arsenateand FCCP and was not stimulated by Na⁺. The pH dependence oftracer accumulation and measurements of the intracellular phosphatepool under different growth conditions indicate that at lowpH and low external phosphate concentrations the high protongradient present under these conditions is utilized for a H₃PO₄uptake or a H⁺/H₂PO^–4 cotransport. However, when the externalphosphate concentration was increased to levels sufficientlyhigh for transport to be driven by the positive membrane potential(10 mol m^–3 phosphate), the pH dependence of phosphateuptake was more complex, but could be explained by the uptakeof H₃PO₄ or a H⁺/H₂PO^–₄-cotransport at low pH and a differenttype H₂PO^–4-transport (with unknown type of ion coupling)at high pH-values. It is suggested that this flexible couplingof phosphate transport is of essential importance for the acidresistance of Dunaliella acidophila. Key words: Acid resistance, Dunaliella acidophila, phosphate cotransport, phosphate limitation, plasma membrane, sodium     

The Uptake of Phosphate by Plants from Flowing Nutrient Solution: III. EFFECT OF CHANGED PHOSPHATE CONCENTRATIONS ON THE GROWTH AND DISTRIBUTION OF PHOSPHATE WITHIN PLANTS OF LOLIUM PERENNE L.

Perennial ryegrass (Lolium perenne L.) was grown from seed for29 d in flowing solution culture containing 0.1, 0.4 or 6.4mmol m^–3 P before the concentrations were changed (0.1and 0.4 raised to 6.4; 6.4 lowered to 0.4; controls unchanged)for an experimental period of two weeks to test the hypothesisthat after the seedling stage, the maximum rate of plant growthcould be sustained by a lower concentration of phosphate atthe root/solution interface than was necessary for the maximumrate of seedling growth. During the 29 d seedling period growthwas greatest on 6.4 mmol m^–3 P achieving 179 mg per plantdry weight compared with 122 and 26 mg on 0.4 and 0.1 mmol m^–3P respectively. During the experimental period growth on thetreatment 6.4 lowered to 0.4 mmol m^–3 P continued at thesame rate as the 6.4 control achieving 981 and 983 mg per plantdry weight respectively. Similarly growth of the treatment 0.4raised to 6.4 mol m^–3 P was unaffected by the change inconcentration and was comparable with the 0.4 control. Bothresults support the hypothesis for seedlings exceeding about100 mg per plant dry weight. In contrast the small plants ofthe treatment 0.1 raised to 6.4 mmol m^–3 P behaved similarlyto seedlings and responded rapidly to the increased concentrationof phosphate in solution, achieving high rates of phosphateuptake and increasing the growth of shoot more than the growthof root so that the ratio of root: shoot declined from 065 to0.34, a value similar to that for the seedlings grown on 6.4mmol m^–3 P. Key words: Lolium perenne L, Phosphate concentration, Seedling growth     

Membrane fluxes and comparative toxicities of aluminium, scandium and gallium

Measurements were made of the membrane fluxes and toxicitiesof three cations with trivalent forms, Al, Ga and Sc, in internodalcells of the giant alga Chara corallina. With this species itwas possible to separate the cell wall from the cell contentsto obtain membrane fluxes which were not complicated by adsorptionof cations to the cell wall. Net uptake of Al was low, approximately1.5 pmol m^–2 s^–1, compared to the influxes of thedivalent cation ⁴⁵Ca of 82 pmol m^–2 s^–1 and themonovalent cation ²²Na of 1100 pmol m^–2 s^–1 at thesame external concentration. Traditional desorption methodsfor removing cell wall cations were found to be relatively ineffectivein the case of trivalent cations and, consequently, influx measuredwithout separating the cell wall component would greatly overestimatethe true membrane flux, possibly by several orders of magnitude.Al, Ga and Sc all inhibited growth at 20 mmol m^–3 at pH4.4. Toxicity decreased in the order Sc>Al>Ga. Sc andAl were also toxic to mature non-growing cells. Influx of ⁴⁶Scincreased with increasing pH, consistent with membrane permeationby hydroxy Sc rather than Sc³⁺. However, Sc was more toxic atlow pH where Sc³⁺ was the dominant species and where influxwas low and binding to cell walls was high. These results argueagainst Sc acting intracellularly and favour a toxicity mechanismwhich is initiated extracellularly. Key words: Aluminium toxicity, trivalent cations, Chara corallina, scandium influx, gallium     

Aluminium and the Hydraulic Conductivity of Quercus rubra L. Root Systems

Two hydroponic experiments were conducted to determine the effectsof brief and prolonged AI³⁺ exposures on the hydraulic conductivity(Lp) of northern red oak (Quercus rubra L.) root systems. RootLp was determined using the pressure chamber method of Fiscus(1977). In the first experiment, 28- to 40-d-old seedlings weretreated for 4 d with complete nutrient solutions containingone of three Al concentrations (0.04, 1.85 or 3.71 mol m^–3)and either 0 or 50 mmol m^–3 P. Neither Lp nor daily transpirationwas affected by treatment. In Experiment II, seedlings were grown for 48–63 d incomplete solutions containing one of three Al concentrations(0, 0.75 or 2.00 mol m^–3) and either 10 or 250 mmol m^–3Ca. Lp and leaf area to root length ratio (LA/RL) were reducedwhen (AI³⁺/ Ca²⁺), the solution activity ratio, was 2.9 andhigher. Lp and LA/RL were also negatively correlated with Alconcentration and Al/Ca concentration ratio in the roots. Lpwas positively correlated with LA/RL in both experiments. Itis unclear whether Lp in the second experiment was reduced directlyby solution and root chemistry or whether Lp changed in responseto altered leaf/root balance. Key words: Al phytotoxicity, Al x Ca interaction, Quercus rubra, root hydraulic conductivity     

Isolation, Culture, and Plant Regeneration of Protoplasts of Two Shrubby Oxalis Species from South America

Protoplasts were successfully isolated from internodal callustissues of both Oxalis glaucifolia and O. rhombeo-ovata whenthey were digested in a solution containing 0.1% (w/v) MacerozymeR-10, 0.5% (w/v) cellulase Onozuka R-10 and 0.3 mmol m^–3sucrose. Protoplasts proliferated to give cell colonies on Gamborget al.'s B5 medium supplemented with 0.3 mmol m^–3 mannitol,0.5 mg dm^–32, 4-D, and 2.0 mg dm^–3 kinetin. Calluswas produced upon transfer of cell colonies to Murashige andSkoog medium containing 2.0 mg dm^–3 l-naphthaleneaceticacid (NAA) and 0.1 mg dm^–3 kinetin for O. glaucifolia,or with 5.0 mg dm^–3 NAA and 0.5 mg dm^–3 6-benzylaminopurine,for O. rhombeo-ovata. Plants were regenerated from O. glaucifoliaprotoplasts on a medium containing 0.1 mg dm–3 NAA, 1.0mg dm^–3 kinetin and 1.0 mg dm^–3 gibberellic acid,but only vascular nodules were differentiated by O. rhombeo-ovataprotoplast-derived calli. Key words: Tissue culture, protoplasts, plant regeneration, Oxalis spp     

Effects of Two Plasmalemma ATPase Inhibitors on H+ Extrusion and Intracellular pH in Elodea densa Leaves

Beffagna, N. and Romani, G. 1988. Effects of two plasmalemmaATPase inhibitors on H⁺ extrusion and intracellular pH in Elodeadensa leaves.—J. exp. Bot. 39: 1033–1043. Elodea leaves in the dark show very little exchange of H⁺ withthe medium in the external pH range between 5.0 and 6.0. Thepresence of fusicoccin and potassium in the medium markedlystimulates H⁺ extrusion. Fusicoccin- and K⁺ -induced H⁺ extrusionis inhibited by either erythrosin B (EB) or Na-orthovanadate,two inhibitors of H⁺ transporting plasma membrane ATPase. EBcompletely inhibits it from the first 30 min of treatment, whensupplied at pH 5.5 at a concentration of 30 mmol m^–3.Vanadate also inhibits H⁺ extrusion, this effect becoming evidentonly after 45 min of treatment. After this time inhibition iscomplete with 250 mmol m^–3 vanadate but only partial forlower concentrations. In the presence of either inhibitor the intracellular pH, measuredas cell sap pH, is significantly lowered. When the intracellularpH changes are determined on vacuole and, separately, on cytoplasmby the weak acid and base distribution method, acidificationof both compartments is found to accompany the blocking of H⁺extrusion by either of the inhibitors. Key words: Intracellular pH, vanadate, erythrosin B, H⁺pumping     

Phosphate Uptake in the Cyanobacterium Synechococcus R-2 PCC 7942

Phosphate uptake rates in Synechococcus R-2 in BG-11 media (anitrate-based medium, not phosphate limited) were measured usingcells grown semi-continuously and in continuous culture. Netuptake of phosphate is proportional to external concentration.Growing cells at pH_o 10 have a net uptake rate of about 600pmol m^–2 s^–1 phosphate, but the isotopic flux for³²P phosphate was about 4 nmol m^–2 s^–1. There appearsto be a constitutive over-capacity for phosphate uptake. TheK_m and V_max, of the saturable component were not significantlydifferent at pH_o 7.5 and 10, hence the transport system probablyrecognizes both H₂PO^–₄and HPO^2–₄. The intracellularinorganic phosphate concentration is about 3 to 10 mol m^–3,but there is an intracellular polyphosphate store of about 400mol m^–3. Intracellular inorganic phosphate is 25 to 50kJ mol^–1 from electrochemical equilibrium in both thelight and dark and at pH_o 7.5 and 10. Phosphate uptake is veryslow in the dark ( 100 pmol m^–2 s^–1) and is light-activated(pH_o 7.51.3 nmol m^–2 s^–1, pH_o 10600 pmol m^–2s^–1). Uptake has an irreversible requirement for Mg²⁺in the medium. Uptake in the light is strongly Na⁺-dependent.Phosphate uptake was negatively electrogenic (net negative chargetaken up when transporting phosphate) at pH_o 7.5, but positivelyelectrogenic at pH_o 10. This seems to exclude a sodium motiveforce driven mechanism. An ATP-driven phosphate uptake mechanismneeds to have a stoichiometry of one phosphate taken up perATP (1 PO_{4 in}/ATP) to be thermodynamically possible under allthe conditions tested in the present study. (Received June 16, 1997; Accepted September 4, 1997)     

Growth and Nutrient Status of Quercus rubra L. in Response to Al and Ca

Northern red oak (Quercus rubra L.) seedlings were grown for63 d in a complete nutrient solution (pH 3.8) containing oneof three concentrations of Al (0, 0.75 or 2-0 mol m^–3)and either 10 or 250 mmol m^–3 Ca. Of all solution variables,the In of (Al³⁺)/(Ca²⁺), the solution activities ratio, wasmost closely correlated with declines in shoot and root growth.Ln (Al³⁺)/(Ca²⁺) also most closely predicted leaf and root [Mg],[Al], and [Al]/[Ca]. These three variables in turn were closelyrelated to growth. Toxic levels of (Al³⁺) and (Al³⁺)/(Ca²⁺)in solution are compared to levels in forest soils. Key words: Al phytotoxicity, Al x Ca interaction, Quercus rubra     

Nitrate Transport into Chara corallina Cells using 36ClO-3 as an Analogue for Nitrate: I. INTERACTION BETWEEN 36ClO-3 AND NO-3 AND CHARACTERIZATION OF 36CIO-3/NO-3 INFLUX

The use of chlorate as an analogue for NO^–₃ during nitrateuptake into Chara corallina cells has been investigated. NO^–₃inhibits ³⁶C1O^–₃ influx into Chara over the concentrationrange 0–1000 mmol m^–3. Lineweaver-Burke plots ofthe data are characteristic of competitive inhibition by NO^–3in the low concentration range (0–300 mmol m^–3 ClO^–₃)and apparent K_INO3 is 140 mmol m^–3 which is of a similarorder of magnitude as apparent K_mCIO3- 180 mmol m^–3. Athigher substrate concentrations the inhibition by NO^–₃was not characteristic of competitive or uncompetitive inhibition. ³⁶C1O^–₃/NO^–₃ influx was dependent on K⁺ and Ca²⁺in the external medium and inhibited by FCCP. NO^–₃ pretreatmentor N starvation increased subsequent ³⁶C1O^–₃/NO^–₃influx into Chara. A comparison between rates of net NO^–₃uptake and ³⁶C1O^–₃/NO^–₃ influx supported the previoushypothesis that NO^–₃ efflux is an important componentin the determination of overall uptake rates. Key words: Nitrate, Chara, ³⁶CIO^–₃     

Cytoplasmic pH of Root Hair Cells of Sinapis alba Recorded by a pH-sensitive Micro-electrode. Does Fusicoccin Stimulate the Proton Pump by Cytoplasmic Acidification?

Bertl, A. and Felle, H. 1985. Cytoplasmic pH of root hair cellsof Sinapsis alba recorded by a pH-sensitive micro-electrode.Does fusicoccin stimulate the proton pump by cytoplasmic acidification?—J. exp. Bot. 36: 1142–1149. pH-sensitive micro-electrodes, filled with ion-exchanger resinhave been fabricated with a turgorinsensitive tip and have beenapplied to test the intracellular pH and changes thereof inroot hair cells of Sinapis alba. (1) The cytoplasmic pH of Sinapisroot hairs was determined to be 7.3 ±0.2 (at neutralexternal pH). (2) 10 mol m^–3 sodium azide depolarizesthe membrane potential by about 100 mV and acidifies the cytoplasmby 0.8 pH-units. (3) The change from 1.0 mol m^–3 to 10mol m^–3 external potassium causes a depolarization ofabout 45 mV, but no change in internal pH. (4) At an externalpH of 5.0, sodium acetate hyperpolarizes the plasmalemma byabout 60 mV and acidifies the cytoplasmic pH by 0.2 to 0.3 units.(5) 2.0 mmol m^–3 fusicoccin (FC) hyperpolarizes the plasmalemmaby 20–25 mV, acidifies the cytoplasm by 0.1 to 0.2 pH-units,and acidifies the external medium by about 0–3 pH-units.It is concluded that cytoplasmic acidification stimulates theelectrogenic proton pump in Sinapis root hairs, and it is suggestedthat the FC-induced effects, viz. hyperpolarization and externalacidification, can also be interpreted in this way. Key words: —Cytoplasmic pH, pH-sensitive micro-electrode, fusicoccin     

The Effects of Aluminium on the Growth of White Clover dependent upon Fixation of Atmospheric Nitrogen

Jarvis, S. C. and Hatch, D. J. 1985. The effects of aluminiumon the growth of white clover dependent upon fixation of atmosphericnitrogen.–J. exp. Bot. 36: 1075–1086. The effects of aluminium (Al^{3 +}) on the growth of white cloverdependent upon symbiotic fixation of atmospheric nitrogen wereexamined at concentrations that may be encountered in solutionsfrom soils of low pH. Well established plants were grown inflowing solution culture with carefully maintained concentrationsof Al and of P and with pH held constant at a value (4.5) atwhich insoluble precipitates are unlikely to form. After 3 weeksof treatment, there were major visual differences between treatmentsin both shoots and roots. Although added Al at 25, 50 and 100mmol m^–3 reduced dry weight, the differences between theplants were not significant. There were, however, some considerabledifferences in P and Ca contents between the treatments. Muchof the Al held by the roots was displaced when plants were transferredto solutions containing either scandium or gallium. As wellas the effects on the plant, Al had a considerable influenceon the fixation of atmospheric nitrogen. At concentrations of50 and 100 mmol m^–3 Al³⁺ nodule initiation was reducedand there was a much reduced nodule nitrogenase activity perunit of root. Even at 25 mmol m^–3 Al³⁺, when nodule numberswere not reduced, nitrogenase activity was adversely affected. Key words: —Aluminium, Trifolium repens, nitrogen fixation