The Distribution of Zinc-65 in Agrostis tenuis sibth. and A. stolonifera L. Tissues4

The distribution of ⁶⁵Zn in zinc-tolerant and copper-tolerantplants of Agrostis spp. from toxic mine-tailings in Enflandand Wales was compared with zinc distribution in non-tolerantplants. Isotope was applied in culture solution in which theplants were growing. No differences could be demonstrated betweenthe plants were growing. No differences could be demonstratedbetween the plants by whole-plant radioautography, or by zincanalyses of the tops. Root/shoot ratios calculated from specificactivity values varied with population, the non-tolerant plantshaving the lowest and the zinc-tolerant plants the highest ratio.After solvent (80 per cent ethanol and water) extractions, theroot residue of zinc-tolerant plants contained a higher percentageof ⁶⁵Zn than that of non-tolerant plants. Chemical fractionationof the roots revealed that the main high difference was thatthe amount of ⁶⁵Zn in the pectate extract of the cell wall washigh in zinc-tolerant plants and low in non-tolerant plants.The ⁶⁵Zn distribution in the copper-tolerant plants was similarto that in the non-tolerant plants, indicating that the tolerancemechanisms for the elements are different. Soluble protein andRNA preparations were made but they contained low levels of⁶⁵Zn. An exception was the relatively high value for RNA fromzinc-tolerant A. stolonifera shoots. An anionic complex of ⁶⁵Znin the soluble fraction was investigated. This complex accountedfor most of the radioactivity in A. tennis extracts of shootsbut the concentration of the complex was low in A. stoloniferashoots, and in root extracts of all plants examined.     

The Water Relations of Young Olive Trees in a Mediterranean Winter: Measurements of Evaporation from Leaves and Water Conduction in Wood

We report measurements of evaporation rate, leaf resistanceto evaporation and water conduction in the stems of young olivetrees (Olea europea L.) growing in Messina, Italy, during thewinter and early spring. We have measured what Zimmermann calls‘leaf specific conductivity’ (LSC) of stem segmentsexcised from olive trees. The LSC is a measure of the specifichydraulic conductivity of stem segments normalized per unitarea of leaves supplied by the stem segment rather than perunit area of sapwood cross-sectional area. We find that theLSC's of primary stems were the largest followed in magnitudeby the LSC's of secondary stems and tertiary stems. Under winterand early spring conditions the maximum evaporative flux fromCoratina and Nocellara varieties of olive trees is about 2.6x 10^–5 kg 8^–1 m^–2. From this and the LSC measurementswe calculate that the pressure gradients needed to maintainthis rate of evaporation in the steady state is 65 kPa m^–1in primary stems, 170 kPa m^–1 in secondary stems and 560kPa m^–1 in tertiary stems. Olive, Olea europea L, evaporation, leaf specific conductivity, hydraulic conductivity, leaf resistance     

Uptake and distribution of 65Zn and 54Mn in wheat grownat sufficient and deficient levels of Zn and Mn: I. During vegetative growth

The uptake and distribution of ⁶⁵Zn and ⁵⁴Mn by wheat (Triticumaestivum cv. Aroona) was investigated. Plantswere grown in achelate-buffered nutrient solution with either sufficient Znand Mn, low Zn or low Mn. A single representative seminal rootfrom 14-d-old and 42-d-old plants was dual-labelled with ⁶⁵Znand ⁵⁴Mn. The 14-d-old plants were harvested every 10 min from10–140 min of labelling, whilst the 42-d-old plants wereharvested after 2 h of labelling. At harvest, each plant wasseparated into leaves, main stem, unexposedroots, and tillers.In addition, the crown was separatedfrom the stem in the 14-d-oldplants In the control plants labelled at 14 d, ⁶⁵Zn was firstdetectedand accumulated in the crown of the roots after 40–60min. Labelled Zn was then detected in the stem, followed bythe leaves. The oldest and youngest leaves received less ⁶⁵Znthan the second and third oldest leaves. The plants grown underlow Zn conditions accumulated more ⁶⁵Zn in their older leavesand transferred ⁶³Zn to the unexposed roots. Distribution of⁵⁴Mn was similar in the controls to that of ⁶⁵Zn, except theolder leaves received no HMn, At the second harvest, a similardistribution pattern of ⁶⁵Zn and ⁵⁴Mn was observed with regardto leaf age. Large amounts of ⁶⁵Zn and ⁵⁴Mn were detected withinthe unexposed roots of all treatments. It is suggested thatthe distribution of root-supplied Zn and Mn may be determinedby micronutrient status and its relationship with leaf transpirationrates. Key words: Distribution, manganese, vegetative growth, wheat, zinc     

Characterization of Cu and Zn Resistant Nicotiana plumbaginifolia Suspension Cultures

The mechanisms of Cu and Zn resistance in cultured Nicotianaplumbaginifolia cells were investigated. When crude extractsof both the Cu and Zn resistant cells were passed through SephadexG-75 gel filtration columns, two metal containing peaks wereidentified. In each case the Cu or Zn was associated with ahigh molecular weight fraction (ca. 70,000) and a low molecularweight fraction (less than 3,000) with the majority being associatedwith the low molecular weight fraction. The low molecular weightfraction in particular increased upon exposure of the cellsto the metals. Both the Cu and Zn resistant cells accumulated higher levelsof citrate and malate than the wild type cells in the presenceof the metals. The addition of citrate or malate, but not fumarateor succinate to the culture medium prevented Cu or Zn toxicityto wild type cells. These results suggest that the N. plumbaginifoliacells selected as resistant to Cu or Zn are resistant, at leastin part, because of their ability to accumulate the chelators,citrate and malate, in the presence of the metals. ¹Present address: Plantech Research Institute, MCI ResearchCenter, 1000 Kamoshida-cho, Midoriku, Yokohama 227 Japan (Received June 30, 1986; Accepted December 1, 1986)     

An Unusual Pattern of 65Zn Distribution in the Leaves of Some Grasses

Autoradiographs of leaves of ryegrass plants (Lolium sp.) takingup radioactive zinc (⁶⁵Zn) from soil showed an unusual patternof label distribution. Tracer accumulated in a regular seriesof pulses along leaf veins. Pulses in adjacent veins occurredin juxtaposition forming the pattern of a band across the leaf. The possibilities of artifacts resulting from plant-culturetechnique or methods of preparing leaves for autoradiographywere eliminated. The band effect in leaves of Loliumand other grass species occurredwith ⁶⁵Zn but not ¹⁹Fe or ⁵⁴Mn. Leaves of Hordeum vulgare, Zeamays, Bromus rigidus, and two non-gramineous species did notshow the band phenomenon with ⁶⁵Zn. There was a close relationshipbetween the frequency of bands and the rate of growth of a leaf.The band pattern was shown to be responsive to changes in daylengthand temperature during growth. Possible origins of the band phenomenon are discussed.     

The Potential of Two Perennial C4 Grasses and a Perennial C4 Sedge as Ligno-cellulosic Fuel Crops in N.W. Europe. Crop Establishment and Yields in E. England

Three perennial C₄ rhizomatous species, Cyperus longus L., Spartinacynosuroides and Spartina pectinata Link, were examined as potentialrenewable energy crops. These species are unusual among C₄ plantsin showing natural distributions which extend into cool temperateregions. This study examined whether these species could beestablished in the cool temperate climate of eastern Englandand whether they could consistently attain the relatively highdry matter yields associated with C₄ plants of warmer regions.Clonally produced material was planted in 1986, on two siteswith contrasting soil types in Essex, eastern England. Plantingwas within a randomized-block design incorporating replicatedplots of each species, both with and without fertilizer. Survivorshipand stem demography were monitored at monthly intervals from1986 to Jun. 1989 for stem recruitment and to Dec. 1991 forstem density. Yields were determined from 1987 (the year followingestablishment) to 1993. Survivorship of the planted propagules over the first 12 monthswas 92% for S. pectinata , 96% for S. cynosuroides and 100%for C. longus. Recruitment of new stems peaked in Apr. of mostyears, although a significant number of new stems appeared asearly as Feb. Stem death peaked in Sep. or early Oct. and allabove-ground stems had died by mid-Nov. Stem density trendsindicated that 2-4 years were required to reach a steady-statedensity, depending upon species. The stem density of the twoSpartina species had reached more than 1000 m^-2 in 1989 althoughthat of S. pectinata fluctuated considerably in the subsequentyears. C. longus stem density rose to approx. 600 m^-2 by 1988and did not change significantly in the subsequent years. In the 6 years following establishment, annual yields averagedacross all fertilizer treatments and both sites were 1·0,1·1 and 1·3 kg m^-2 for C. longus, S. cynosuroidesand S. pectinata, respectively. The average annual yield ofall three species at the site with the heavier soil was 1·3kg m^-2. This was significantly greater than the 1·0 kgm^-2 on the lighter soil. Nitrogen addition did not significantlyincrease yield. Even in the absence of any nitrogen addition,the annual yield of S. pectinata averaged 1·2 kg m^-2over the 6 years, with no evidence of any decline with the increasingage of the stands.Copyright 1995, 1999 Academic Press Cyperus longus, Spartina cynosuroides, Spartina pectinata, energy crop, dry matter yield     

The Effects of Light Intensity and External Potassium Level on Root/Shoot Ratio and Rates of Potassium Uptake in Perennial Ryegrass (Lolium perenne L.)

Loliun perenne L. (cv.S. 23) was grown on vermiculite in winterin a heated greenhouse for 8 weeks under factorial combinationsof two potassium regimes (nominally 6 parts/10⁶ and 156 parts/10⁶in Hewitt's solution) and three densities of artificially supplementedvisible radiation flux (36.1, 7.3, and 2.2 W m^–2). Growthand potassium uptake were studied through the calculation ofvarious growth functions from fitted curves. There was little effect of potassium treatment but the experimentalmaterial responded markedly to light. Leaf-area ratio in thethree treatments showed extreme plasticity in increasing from2–3 x 10^–2 through 6 x 10^–2 to 8–9 x10^–2 m² g^–1 as light intensity decreased. Correspondingdecreases in unit leaf rate, however, caused over-all reductionsin relative growth rate. Specific absorption rates for potassium (A_K, dry-weight basis)were strongly reduced at the lower light intensities but alsodisplayed complex ontogenetic drifts. Values of the allometricconstant, k (the ratio of root and shoot relative growth rates),decreased from c. 0.7 at 36.1 W m^–2 through c. 0.3 at7.3 W m^–2 to a value not significantly different fromzero (P < 0.05) at 2.2 W m^–2. In material grown under the two higher light intensities a constantinverse relationship was found between the mass ratio of rootand shoot and the corresponding activity ratio. The resultsconform to this model: Mass ratio = –0.001+45.0 (1/activityratio) where activity ratio is expressed as specific absorptionrate for potassium (in µg g root^–1 h^–1)/unitshoot rate (rate of increase of whole-plant dry weight per unitshoot dry weight, in mg g shoot^–1 h^–1). The implicationsof this relationship are discussed.     

Inhibition of Pinus radiata Primary Needle Epicuticular Wax Biosynthesis by Trichloroacetate

High concentrations (1000 parts 10^–6) of trichloroacetate(TCA) inhibit germination of Pinus radiata seed. Seedlings growingin the presence of lower concentrations (100 parts 10^–6)take up TCA where it becomes concentrated in cotyledons anddeveloping primary needles. TCA inhibits biosynthesis of nonacosan-10-oland long chain diol constituents of the primary needle epicuticularwax concomitant with a reduction in appearance of tubular waxexcept around stomatal pores. Epicuticular wax from TCA-treatedplants consists mainly of alkyl esters, and is amorphous. Itis suggested that P. radiata stomatal subsidiary cells possesstubular epicuticular wax chemically distinct from that of epidermalcells.     

Short-term Products of 14C-acetate Assimilation by Chlorella pyrenoidosa in the Light

The kinetics of ¹⁴C-2-acetate assimilation by Chlorella pyrenoidosain the light were examined. Under aerobic conditions the primaryproduct of acetate assimilation was succinic acid which, afterten seconds, contained over 60 per cent of the ¹⁴C incorporatedby the cells. The percentage of the total ¹⁴C in succinate fellwith time, while that in citrate and glutamate increased. After1800 sec over 60 per cent of ¹⁴C was present in two compounds,glutamic acid and an unknown compound (X). Glucose-6-phosphate,fructose-6-phosphate, phosphoglyceric acid and phosphoenolpyruvicacid became labelled after 60 sec but together never containedmore than one per cent of the total ¹⁴C incorporated. Underanaerobic conditions succinate was still the primary productof acetate assimilation, and the absence of carbon dioxide resultedin a decrease in ¹⁴C incorporation into compound X. The patternof acetate assimilation in acetate grown and acetate adaptedChlorella was very similar to that in photo-autotrophicallygrown Chlorella. In the presence of 10^–6M DCMU, succinicacid was the primary product of acetate assimilation, but therewas an early Incorporation of ¹⁴C into glutamate, aspartate,and malate. 4 x10^–3M MFA did not effect the early incorporationof ¹⁴C into succinic acid, but resulted in accumulation of ¹⁴Cin citrate and a decreased amount in glutamate and in compound X.     

The Cellular Pathway of Radial Transfer of Photosynthates in Stems of Phaseolus vulgaris L.: Effects of Cellular Plasmolysis and p-Chloromercuribenzene Sulphonic Acid

Cellular plasmolysis with l M solutions of mannitol appearedto sever plasmodesmatal interconnections between all cells ofthe stems of Phaseolus vulgaris plants except the sieve element-companioncell (se—cc) complexes. Phloem loading and uptake of [¹⁴C]sucroseby the storage cells of the stems was unimpaired by cellularplasmolysis followed by rehydration of the stem tissues. Accumulationof phloem-transported ¹⁴C-photosynthates of the treated stemswas inhibited in summer-grown plants and unaffected in winter-grownplants indicating that phloem unloading follows a symplasticand a free-space route respectively depending on growth season.At a concentration that did not interfere with cellular metabolism,p-chloromercuribenzene sulphonic acid (PCMBS) applied to thestems blocked [¹⁴C]sucrose loading into the phloem and storagecells of the stem, but had no effect on the pool size of free-spacesugars. This latter response is consistent with a facilitatedmechanism of sugar unloading to the stem free-space. Accumulationof phloem-transported ¹⁴C-photosynthates was stimulated by PCMBSand this effect was most pronounced in winter-grown plants.Cellular plasmolysis followed by rehydration abolished the PCMBSaction on ¹⁴C-photosynthate accumulation. This effect is consistentwith a PCMBS induction of phloem unloading through the stemsymplast. It is proposed that phloem unloading in bean stemsmay follow either a free-space or symplastic route and thatthe latter route is entrained under sink-limited conditions. Phaseolus vulgaris, french bean, stem, phioem unloading, free-space, symplast     

Effect of Nitrogen on Nitrate Reductase Activity in the Nodules and Leaves of Summer Moong (Vigna radiata)

The relation of the in vivo nitrate reductase (NR) activityto growth period was studied in the nodules and the leaves ofthe summer moong (Vigna radiata). The maximum NR activity wasobserved 31 days after sowing (DAS) in the leaves and 28 DASin the case of the nodules. In a pot experiment, the effectof the various nitrogen concentrations, namely 0, 3, 6, 9 and12 mg kg^–1 was studied on NR activity at three growthstages. The maximum NR activity was observed at 6 mg kg^–1N during the pre-flowering stage (26 DAS). Though the noduleshave higher NR activity, its expression was limited by substrateavailability. The NR activity in the leaf could be used as anindex of NR activity in the nodules. Nitrate reductase, nitrogen, nitrate, moong, Vigna radiata     

Zinc Mobility in Wheat: Uptake and Distribution of Zinc Applied to Leaves or Roots

Little is known about transport of Zn from leaves to other plantorgans. The present study tested a range of Zn forms appliedfoliarly for their suitability to provide adequate Zn nutritionto wheat (Triticum aestivum L.). Transport of⁶⁵Zn applied eitherto leaves or to one side of the root system was also studied.Inorganic (ZnO, ZnSO₄) and chelated sources of Zn (ZnEDTA, glycine-chelatedBiomin Zn) applied foliarly provided sufficient Zn for vigorousgrowth. Zinc concentrations in roots and shoots were in thesufficiency range, except in the -Zn control. Foliar treatmentswith ZnSO₄and chelated Zn forms resulted in shoot Zn concentrationsin 7-week-old plants being about two-fold greater than thosein plants supplied with Zn in the root environment or via foliarspray of ZnO. Adding surfactant to foliar sprays containingchelated forms of Zn did not cause negative growth effects,but surfactant added to ZnO or ZnSO₄foliar sprays decreasedshoot growth. Adding urea to the ZnO foliar spray had no effecton shoot growth. Foliarly-applied⁶⁵Zn was translocated to leavesabove and below the treated leaf as well as to the root tips.Stem girdling confirmed that⁶⁵Zn transport toward lower leavesand roots was via the phloem. Split-root experiments showedintensive accumulation of⁶⁵Zn in the stem and transport to allleaves as well as to the root tips in the non-labelled sideof the root system. Foliar application of Zn in inorganic ororganic form is equally suitable for providing adequate Zn nutritionto wheat. Phloem transport of Zn from leaves to roots was demonstrated.Copyright 2001 Annals of Botany Company Foliar spraying, phloem, surfactant, urea, xylem, wheat, zinc     

Changes in the Organic Acid Content of some Cultivated Plants Induced by Mineral Ion Deficiency

Four plant species (Zea mays, Lolium italicum, Phaseolus vulgaris,and Pisum sativum) were grown either with full nutrient supplyor with calcium, potassium, or nitrogen deficiency. After 4–6weeks the shoots were analysed for their content of K⁺, Ca²⁺,Mg²⁺, and Na⁺, of total anions, and of organic acids. The qualitativepattern of organic acid components, characteristic of each species,remained, with few exceptions, unchanged by deficiency treatments.Striking differences were detected in the total acid contentand in the quantities of the respective main organic acid components,especially in potassium-deficient plants. These changes showedstrong correlations with alterations of cation excess and demonstratethat aconitate in Zea mays, malonate in Phaseolus vulgaris,and citrate in Pisum sativum contribute to charge balance.     

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     

Haustoria of Cuscuta japonica, a Holoparasitic Flowering Plant, Are Induced by the Cooperative Effects of Far-Red Light and Tactile Stimuli

When seedlings of Cuscuta japonica were grown with Vigna radiata(the host plant) in a flower pot for 6 d under white light andthen irradiated with far-red or blue light (ca. 6 µmolphotons m^–2 s^–1), the seedlings parasitized V. radiata.However, no parasitism of the seedlings was observed under redor white light or in darkness. The parasitic behavior of seedlingsof C. japonica was observed even if an acrylic rod was usedas a substitute for the host plant. Upon incubation under far-redlight, the seedling twined tightly around the rod and developedhaustoria towards it. Haustoria also developed when apical andsubapical regions of seedlings were held between two glass platesthat were about 0.7 mm apart and were irradiated with far-redlight. However, no haustoria were induced by either the holdor irradiation alone. These results indicate that parasitismof Cuscuta japonica is controlled by the cooperative effectsof two physical signals, far-red light and appropriate tactilepressure. Our findings suggest that parasitism by the genusCuscuta involves a novel strategy. (Received April 10, 1996; Accepted August 21, 1996)     

Salt Tolerance in the Halophyte Suaeda maritima L. Dum.: Intracellular Compartmentation of Ions

The time-course of exchange of sodium and potassium ions fromroot and leaf material of the halophyte Suaeda maritima hasbeen followed and the data analysed according to the phenomenologyof efflux, or compartmental, analysis. Sodium ions were exchangedmuch more slowly (c. 4 times) from the vacuoles of leaf cellsof plants grown in sodium chloride than were potassium ionsfrom the vacuoles of leaf cells of plants grown either in similarconcentrations of potassium chloride or in low concentrationsof potassium. In plants grown in sodium chloride, sodium ionswere exchanged 9 times more slowly from the vacuoles of leafcells than from the vacuoles of root cells. The concentration of sodium ions in the cytoplasm of leaf cellsof plants growing in 340 mol m^–3 sodium chloride was estimatedto be 165 mol m^–3 when the average concentration in theleaf tissue was about 600 mol m^–3. As measured by movement from mature to developing leaves inintact plants; there was less in vivo retranslocation of ²²Naand ³⁶CI in plants growing in sodium chloride than there wasof ⁸⁶Rb in plants growing either in potassium chloride or innon-saline conditions. The results are discussed in terms of the concept and energeticsof compartmentation of ions in the cells of halophytes.     

Effect of Applied Nitrogen on N2 Fixation, Assimilation of Nitrate and Ammonia in Nodules of Field-grown Moong (Vigna radiata)

A field experiment was conducted to study the effect of nitrogenapplication at 15, 30 and 45 kg ha^–1 of urea at pre-flowering(PF) and pod initiation (PI) stages on the activity of nitrogenase(N₂ase), nitrate reductase (NR) and other related parametersin the nodules of moong (Vigna radiata). Nitrogen applied atPF or PI stage was found to be inhibitory to N₂ase and glutaminesynthetase (GS) activities except at 15 kg N ha^–1 whenapplied at PF in the case of N₂ase. At both the stages therewas increase in NR and glutamate dehydrogenase (GDH) activitieswith the application of nitrogen. Seed yield increased by 18per cent with the application of 15 kg N ha^–1 at PI stagewhereas nitrogen application at PF stage only increased strawyield significantly. Nitrate reductase, nitrogenase, nitrogen application, ammonia assimilation, Vigna radiata     

Isolation and Characterization of Sodium Chloride-Resistant Callus Culture of Vigna radiata (L.) Wilczek var. radiata

Callus cultures were initiated from seedling root segments ofmungbean (Vigna radiata (L.) Wilczek var. radiata) cv. K 851on modified PC-L2 basal medium. Growing cells were exposed toincreasing concentrations of NaCl in the medium. A concentrationof 300 mol m^–3 NaCl proved completely inhibitory to growthof the calli. On incubation for 25 d, cells which could toleratethis concentration of NaCl grew to form cell clones. Selectedclones were characterized with regard to their growth behaviour,K⁺, Na⁺ and free proline content when grown under stress aswell as on normal media and were compared with the normal sensitivecallus. The selected callus was capable of growing on mediumcontaining NaCl at the inhibitory concentration. The K⁺ contentof the selected callus was lower in the case of the NaCl mediumthan for the normal medium. However, the selected clones maintainedhigher K⁺ and Na⁺ levels, with increased salinization comparedwith the wild-type cells. Salt-selected cells accumulated higherlevels of free proline under NaCl stress compared to wild-typecells. Under normal conditions, however, the amounts of freeproline in selected and non-selected calli were comparable. Key words: Vigna radiata, callus culture, NaCl stress     

The Pathway of Radial Transfer of Photosynthate in Decapitated Stems of Phaseolus vulgaris L.

[¹⁴C]Sucrose was found to be the predominant component of the¹⁴C-photosynthates that accumulated in the free space of decapitatedstems of P. vulgaris plants. The ¹⁴C-photosynthates appearedto occupy the entire free-space volume of the stems at totalsugar concentrations in the range of 3–12 mM. The free-spacesugar levels were found to rapidly decline once photosynthatetransfer to the stems was halted. Moreover, it was found thatestimates of the rate of in vitro sucrose uptake by the stemscould account fully for the decline in free-space sugar levels.Overall, the evidence indicated that at least part of the radialpathway of photosynthate transfer in bean stems involved thestem apoplast. It is tentatively proposed that, based on celland tissue distribution of ¹⁴C-photosynthates, the apoplasticpathway extends from the membrane boundary of the sieve element/companion-cellcomplex to all other cells of the stem. Apoplast, Phaseolus vulgaris L., bean, phloem unloading, photosynthates, symplast     

The Membrane Potential of Enzymatically Isolated Nitella expansa Protoplasts as Compared with their Intact Cells

With the enzymatically isolated Nitella protoplasts, sufficientinsertions of micro-electrodes to make a stable measurementof the membrane potential by the conventional method could notbe made because of an ‘elasticity’ of the outermembrane. We developed an effective method in which a micro-electrodecould be inserted after the outer membrane was punctured bypassing an electrical impulse through the micro-electrode. Inthis method, Ca ions play a crucial role in the ‘punching’and ‘healing’ processes of the protoplast membrane. The effects of the cations K⁺, Na⁺, Ca²⁺ and the anions Cl^–,, , on the membrane potentials of Nitella expansa protoplasts were compared with those of intactcells. The membrane potential of protoplasts was less negativethan that of intact cells when concentrations of Na or K, inthe presence of Ca, were below certain levels which increasedwith increasing Ca concentration; and it tended to become identicalto that of intact cells when Na or K concentrations were beyondthose levels. Beyond those levels for K the membrane potentialsof both protoplasts and intact cells typically seemed to bethe Nernst potentials in the presence of 0•1 to 30 molm^–3 Ca²⁺. However, for Na, the difference in potentialsbetween intact cells and protoplasts decreased at much higherconcentrations than for K. Increase of Ca always gave less negativeprotoplast potentials than those in intact cells. Replacementof Ca by Mg did not change the membrane potential of intactcells, although it was deleterious to protoplasts. The cellwall potential of intact cells was also measured by the micro-electrodetechnique and was revealed as a simple Donnan potential, assumingthe fixed negative charge density of 0•8 equivalent perdm³. The membrane potential of intact cells seems to be a significantreflection of the plasmalemma potential which is thought tobe measured directly in their protoplasts in terms of ionicselectivity and concentration dependency of the ion speciesexamined. In addition, increased sensitivity to calcium in protoplastpotentials compared to intact cells is suggested, though themembrane potential of intact cells seems to be largely preservedin their enzymatically isolated protoplasts. Key words: Membrane potential, protoplasts, Nitella expansa, cell wall potential