Purification and some properties of L-tyrosine carboxy-lyase from barley roots

L-Tyrosine carboxy-lyase (E. C. 4. 1. 1. 25) was extracted fromthe roots of barley seedlings and purified approximately 25fold. Optimum pH for the enzyme activity was found to be 7.3.The Km value for L-tyrosine was calulated as 4.5?10^–4M.D-Isomer did not react with the enzyme. L-DOPA, m-tyrosine ando-tyrosine were decarboxylated to some extent. Pyridoxal phosphateactivated the enzyme 4 fold. Caffeic acid and p-coumaric acidare competitive inhibitors. Ki values were 4.5?10^–5M forcaffeic acid and 1.6?10^–4M for p-coumaric acid. L-DOPAand m-tyrosine had an inhibitory effect on the decarboxylationof L-tyrosine. Hydroxylamine, semicarbazide, p-CMB, Fe⁺⁺, Cu⁺⁺,and Hg⁺⁺ inhibited the decarboxylation of tyrosine. Enzyme activitywas also found in extracts from Triticum aestivum, Zea maysand Cytisus scoparius. (Received November 30, 1973; )     

Characterization of membrane-bound Mg++-activated ATPase isolated from the lower epidermis of tobacco leaves

Membrane-bound Mg⁺⁺-activated ATPase was separated from thelower epidermis of tobacco leaves (Nicotiand tabacum L. SamsunNN) on stepwise sucrose density gradient centrifugation. Membrane-bound epidermal ATPase was localized in the interfaceof densities in sucrose of 1.12 to 1.16 in the sedimentary fractionbetween 1,500?g to 10,000?g from the homogenate of the lowerepidermis. The epidermal ATPase activity was activated by divalentcations (Mg⁺⁺>Mn⁺⁺Co⁺⁺>Fe⁺⁺>Zn⁺⁺>Ca⁺⁺) and furtherstimulated by KCl by ca. 20%. The pH optimum for Mg⁺⁺-activationof the epidermal ATPase was ca. 6.0. The enzyme hydrolyzed ATPmore rapidly than other nucleoside triphosphates. The optimumtemperature for activation of the epidermal ATPase activitywas ca. 40?C. 50% of the epidermal ATPase activity was lostin 18 min at 55?C and in 2.5 days at 2.5?C. The apparent Kmvalue of the epidermal ATPase was 4.7?10^–4 M and Vmaxwas 65.4 nmoles Pi/mg protein/min. The epidermal ATPase wasstrongly inhibited by N, N'-dicyclohexylcarbodiimide (DCCD)in vitro whereas oligomycin, carbonyl cyanide m-chlorophenylhydrazone(CCGP), indoleacetic acid (IAA) and abscisic acid (ABA) wereinsensitive to the epidermal ATPase activity. (Received May 23, 1978; )     

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     

Effect of Sudden Salt Stress on Ion Fluxes in Intact Wheat Suspension Cells

Although salinity is one of the major problems limiting agriculturalproduction around the world, the underlying mechanisms of highNaCl perception and tolerance are still poorly understood. Theeffects of different bathing solutions and fusicoccin (FC),a known activator of plasma membrane ATPase, on plasma membranepotential (E_m) and net fluxes of Na⁺, K⁺and H⁺were studied inwheat suspension cells (Triticum aestivum) in response to differentNaCl treatments. E_mof cells in Murashige and Skoog (MS) mediumwas less negative than in cells exposed to a medium containing10 mM KCl + 0.1 m M CaCl₂(KSM) and to a basic salt medium (BSM),containing 1 m M KCl and 0.1 m M CaCl₂. Multiphasic Na⁺accumulationin cells was observed, peaking at 13 min after addition of 120m M NaCl to MS medium. This time scale was in good agreementwith net Na⁺flux changes measured non-invasively by moving ion-selectivemicroelectrodes (the MIFE system). When 120 m M NaCl was addedto all media studied, a quick rise of Na⁺influx was reversedwithin the first 20 min. In both 120 and 20 m M NaCl treatmentsin MS medium, net Na⁺efflux was observed, indicating that activeNa⁺transporters function in the plant cell response to saltstress. Lower external K⁺concentrations (KSM and BSM) and FCpre-treatment caused shifts in Na⁺fluxes towards net influxat 120 m M NaCl stress. Copyright 2000 Annals of Botany Company Sodium, potassium, proton, membrane potential, fusicoccin, salt stress, wheat, Triticum aestivum     

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     

Further Studies on Signaling Pathways for Ecdysteroidogenesis in Crustacean Y-Organs

The Y-organs of crustaceans secrete ecdysteroids (molting hormones)and are regulated (negatively) by a neurosecretory peptide,molt-inhibiting hormone (MIH). Signaling path(s) in Y-organswere explored that connect MIH receptors ultimately with suppressionof receptor number for the uptake of cholesterol (ecdysteroidprecursor) and of gene expression of steroidogenic enzymes.Experiments were conducted in vitro with Y-organs of crabs (Cancerantennarius, Menippe mercenaria) and crayfishes (Orconectessp.). It was confirmed in all species that steroidogenesis occursin the absence of external calcium (Ca⁺⁺), but increases toa maximum as Ca⁺⁺ is increased to 1 to 10 mM and is substantiallyinhibited at higher Ca⁺⁺ concentrations. MIH does not requireexternal Ca⁺⁺ for inhibitory action, but inhibition is eliminatedby high Ca⁺⁺concentrations. Several experimental approachesfailed to find evidence of phospholipase C activation, turnoverof inositol triphosphate or diacylglycerol generation connectedwith steroidogenesis. Unbinding or chelation of intracellularCa⁺⁺ with thapsigargin or TMB-8, respectively both caused dose-dependentinhibition of ecdysteroid output. Blockade of Ca⁺⁺ channelswith verapamil, nifedipine or nicardipine also inhibited steroidogenesis;highest doses inhibited profoundly to below Ca⁺⁺-free basallevels. Inhibition also was obtained with all doses of the Ca⁺⁺channel agonist/antagonist (–) BAY K 8644 in crabs, butin crayfishes lower doses were stimulatory. However, if thecrayfish cells were depolarized, allowing greater Ca⁺⁺ influx,the previously stimulatory doses of BAY K 8644 became inhibitory.Y-organ protein kinase C (PKC) is Ca⁺⁺-sensitive. Activationof PKC was uniformly stimulatory in crabs, but inhibitory incrayfishes. Cytochalasin D, which disrupts the actin cytoskeleton,and which causes moderate Ca⁺⁺ influx, stimulated hormone formation.These results are interpreted to indicate a regulatory rolefor Ca⁺⁺ in ecdysteroidogenesis, involving a local, submembranecirculation of Ca⁺⁺ through ion channels and Ca⁺⁺ pumps andinteraction with PKC in phosphorylating key proteins. An optimallocal Ca⁺⁺ environment fostering hormone synthesis is evidentsince too little or too much Ca⁺⁺ is inhibitory. Methyl farnesoate (MF) had no effect on ecdysone productionin crab or crayfish Y-organs in 24-hr incubations with MF at100 pM to 10 µM.     

Studies on NADP-isocitrate dehydrogenase from maturing castor bean seeds

NADP-specific isocitrate dehydrogenase from the soluble fractionof maturing castor bean endosperm was partially purified (approximately180-fold) and some of its enzymatic properties were studied.Mg⁺⁺, Mn⁺⁺, Cd⁺⁺, Ba⁺⁺, Co⁺⁺, Zn⁺⁺, and Sr⁺⁺ were activatorsof the enzyme reaction at a concentration of 6.7x10 M. The optimumpH of this enzyme was about 8.5. The enzyme was stable in thenarrow range from pH 7.0 to pH 8.0. Km values for isocitrateand NADP at pH 8.5 were 3.5x10^–6 M and 3.6x10^–6M, respectively. Enzyme stability was not affected by NaCl concentrationand enzyme reaction was inhibited at 5x10^–6 M PCMB (80%inhibition). It is suggested that the condensation product ofglyoxylate and oxalacetate also inhibits the reaction. NADP-IDHin the crude extract from maturing castor bean endosperm washeat-stable but the dialyzed enzyme preparation and the partiallypurified enzyme were labile against heat treatment at 57°C.When Mg⁺⁺ was added to the partially purified enzyme in thepresence of isocitrate or NADP, the enzyme was stabilized againstheat treatment. Mn⁺⁺, Ca⁺⁺, Co⁺⁺, Sr⁺⁺ or Ba⁺⁺ could be substitutedfor Mg⁺⁺. Addition of only one of the factors, Mg⁺⁺, isocitrateor NADP, had no effect on the heat stability. Moreover, a combinationof isocitrate and NADP did not establish stabilization. A divalentcation plays a central role, while adenine nucleotide, especiallyATP, may have an important part in stabilization. (Received August 14, 1972; )     

Effect of abscisic acid on membrane-bound epidermal ATPase from tobacco leaves

Membrane-bound Mg⁺⁺-activated ATPase activity in epidermal stripsfrom tobacco leaves (Nicotiana tabacum L. Samsun NN) was stimulatedby abscisic acid (ABA) when the strips were floated on ABA solutionin light or in darkness. The optimum ABA concentrations in lightand in darkness were 10^–5 M and 10^–6 M, respectively.Carbonyl cyanide m-chlorophenylhydrazone (CCCP) and N, N'-dicyclohexylcarbodiimide(DCCD) completely blocked the basal level membrane-bound epidermalATPase activity. ABAinduced membrane-bound epidermal ATPaseactivity was completely inhibited by CCCP, but only partly byDCCD. H⁺-influx into epidermal strips on a solution in light was lowerthan that in darkness. ABA stimulated H⁺-influx into epidermalstrips in light and in darkness. CCCP suppressed basal levelH⁺-influx, whereas DCCD did not. CCCP also suppressed ABA-inducedH⁺-influx, whereas DCCD did not. Interaction between H⁺-influxand membranebound epidermal ATPase activity is discussed. (Received May 23, 1978; )     

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     

Enzymes involved in the last steps of the biosynthesis of mannitol in brown algae

Mannitol-1-phosphate dehydrogenase (EC 1.1.1.17 [EC] ) and mannitol-1-phosphatase(EC number yet unassigned) were detected in the brown algae,Spatoglossum pacificum and Dictyota dichotoma. The enzymes wereextracted from algal fronds and their properties were investigatedusing partially purified preparations. Mannitol-1-phosphatase shows maximum activity at pH 7. The enzymehad a narrow substrate specificity. The Km value for mannitol-1-phosphateis 8.3x10^–4 M (30°C, pH 7.0). The enzyme is activatedby Mg⁺⁺ and Mn⁺⁺and is strongly inhibited by PCMB, Hg⁺⁺and NaF. Mannitol-1-phosphate dehydrogenase showed maximum activitiesat pH values 6.5 and 10.2 in reductive and oxidative reactions,respectively. The dehydrogenase also showed narrow substratespecificity; mannitol-1-phosphate and NAD or fructose-6-phosphateand NADH₂ are utilized, respectively, in oxidative and reductivereactions by the enzyme. Km values for these substrates andthe coenzymes are 2.5x10^–4 M and 7.1x10^–5 M forthe first pair and 2.8x10^–4 M and 1.3x10^–5 M forthe latter pair. This enzyme was strongly inhibited by PCMBand Hg⁺⁺, but was only slightly affected by adenosine phosphates. Possible roles of these enzymes in the biosynthesis of mannitolin brown algae are discussed. ¹ Contributions from the Shimoda Marine Biological Station ofTokyo Kyoiku University, No. 233. This work was supported inpart by a Grant-in-Aid for Co-operative Research from the Ministryof Education, Japan and in part by a grant to one of us (T.Ikawa) from the Matsunaga Science Foundation. ² Present address: Chemical and Physical Laboratory, HoechstJapan Research Laboratory, Minamidai, Kawagoe, Japan. (Received February 22, 1972; )     

The ionic requirements for the initiation of action potentials in insect muscle fibers

Electrical properties of locust leg muscle fibers were studied by means of intracellular electrodes. In most fibers, a depolarizing current pulse initiated a local response. A delayed decrease in membrane resistance appeared with more than about 10 mv depolarization. In some fibers a regenerative response also was found. Membrane constants were measured, applying the short cable model. The value of the space constant λ was 1.6 mm and the calculated value of R_m was about 1750 ohm cm². Action potentials could be elicited when the bathing fluid contained more than 2–5 mM Ba or Sr. Similar responses were seen with 2 mM Ca in the presence of tetraethylammonium (TEA). The overshoot of these action potentials increased with increasing [Ca⁺⁺]_o, [Sr⁺⁺]_o, or [Ba⁺⁺]_o, the increment for a 10-fold increase being about 29 mv for Ca and Sr and between 40 and 50 mv for Ba. These action potentials were inhibited by Mn ions but were not affected by tetrodotoxin or procaine. In solutions containing Ba or Sr, action potentials generated were suppressed by addition of Ca. The removal of Na ions did not change the configuration of the action potential. The results suggest that an increase in permeability to Ca, Ba, or Sr ions makes a major contribution to the initiation of action potentials in this tissue.     

Dependency of H+ efflux on ATP in cells of Chara australis

The internodal cell of Chara australis was made tonoplast-freeby internal perfusion with a medium containing a Ca²⁺-chelatorEGTA and the net H⁺ efflux across the plasma membrane was estimatedeither by titration of the external medium or by measuring thechange in pH in the external medium. The amount of H⁺ effluxwas high in the presence of internal ATP and low in its absence.The ATP-dependent net H⁺ efflux was about 40 nmoles/m²/sec.This amount is smaller than that estimated for the pump currenton the basis of electrical data obtained earlier (3). This discrepancyis discussed. (Received June 18, 1980; )     

Some properties of shikimate: NADP oxidoreductase produced in sweet potato root tissue after slicing

The activity of shikimate: NADP oxidoreductase [EC 1. 1. 1.25] in sweet potato root tissue increased soon after slicing.Enzyme preparations obtained from both sliced tissue and fromfresh tissue probably contained a single enzyme component, andthey showed identical chromatographical behaviour. K_m values of the enzyme for NADP and shikimate were 1.0x10^–4Mand 1.3 x 10^–3M, respectively. Enzyme activity was potentlyinhibited by SH-inhibitors such as p-chloromercuribenzoate andoxidized glutathione. Enzyme activity was affected neither by mononucleotides suchas ATP, ADP and AMP, divalent cations, Mg⁺⁺, Ca⁺⁺ and Mn⁺⁺,nor by metabolites such as tryptophan, phenylalanine, tyrosineand t-cinnamic acid which are involved in aromatic compoundsyntheses. The enzyme rapidly lost its activity. This inactivation reactionshowed a time course consisting of two steps of the first-orderreaction. The inactivated enzyme preparation was not reactivatedby thiol compounds such as cysteine, 2-mercaptoethanol and glutathione,although these reagents, to a certain extent, protected theenzyme from inactivation. The results suggest that denaturationof the enzyme protein was involved in inactivation of the enzyme. ¹Part 74 of the phytopathological chemistry of sweet potatowith black rot and injury. ²Present address: Department of Biology, Faculty of Science,Tokyo Metropolitan University, Setagaya-ku, Tokyo. (Received August 5, 1968; )     

A Preliminary Study on the Effects of Nitrogen Supply on the Growth In Vitro of Nine Potato Genotypes (Solanum spp)

Cultures of nine potato genotypes (seven Solanum tuberosum oneS. sparsipilum and one S. oplocense genotypes) were examinedfor their response to growing on medium containing either 60mol m^–3, 40 mol m^–3 or 20 mol m^–3 nitrogen.Genotypes differed in their response to nitrogen. Reducing thenitrogen regime tended to produce taller plants with longerinternodes, shoots had larger leaves but contained less chlorophyll.No change in fresh weight or number of nodes was observed. Genotypex nitrogen interactions were significant for chlorophyll content,shoot length and internode length. Results suggest that thechanges observed were as a result of changes in the total nitrogenlevel rather than changes in the ammonium : nitrate ratio. Thisstudy suggests that for certain potato genotypes, nitrogen levelsin MS medium are too high for producing desirable microplantsin terms of leaf area and shoot length Key words: Solanum tuberosum, S. sparsipilum, S. oplocense, micropropagation, morphogenesis     

Effects of Cytokinin and Several Inorganic Cations on the Polyamine Content of Lettuce Cotyledons

Kinetin (4.7 x 10^–5 M) and 6-benzyladenine (2.22 x 10^–5M) were found to increase ca. 2-fold the putrescine contentin cotyledons of lettuce (Lactuca sativa L.) seedlings grownfor 3 days under fluorescent light. On the other hand, severalinorganic ions (K⁺, Na⁺, Ga⁺⁺, Mg⁺⁺) at a concentration of 3x 10^–2 M reduced the putrescine content. The combinationof kinetin with one of several inorganic ions at the same levelmarkedly increased the spermine content, but the putrescinecontent decreased; calcium and magnesium ions were less effective.The physiological significance of these findings is discussed. (Received July 4, 1982; Accepted November 6, 1982)     

Effect of the Osmotic Environment on K+ and Mg2+ Release from the Seed Coat and Cotyledons of Developing Seeds of Vicia faba and Pisum sativum. Evidence for a Stimulation of Efflux from the Vacuole at High Cell Turgor

After removal of the embryo from developing seeds of Vicia fabaL. and Pisum sativum L., the ‘empty’ ovules werefilled with a substitute medium (pH 5.5) and the effect of theosmolality of the medium on K⁺ and Mg²⁺ release from the seedcoat was examined. In long-term experiments (12 h or longer),with both attached and detached seed coats, the rate of K⁺ andMg²⁺ release from seed coats filled with a solution withoutosmoticum was enhanced, in comparison with release from seedcoats filled with a solution containing 400 mol m     

Spatial and Temporal Aspects of Growth in the Primary Root of Cotton Seedlings: Effects of NaCl and CaCl2

Seedlings of cotton (Gossypium hirsutum L. cv. Acala SJ-2) weregrown in modified Hoagland nutrient solution with various combinationsof NaCl and CaCl₂. Marking experiments and numerical analysiswere conducted to characterize the spatial and temporal patternsof cotton root growth at varied Na/Ca ratios. At 1 mol m^–3Ca, 150 mol m^–3 NaCl reduced overall root elongation rateto 60% of the control, while increasing Ca to 10 mol m^–3at the same NaCl concentration restored the elongation rateto 80% of the control. Analysis of the spatial distributionof elongation revealed that the presence of 150 mol m^–3NaCl in the medium shortened the growth zone by about 2 mm fromthe approximate 10 mm in the control and also reduced the relativeelemental elongation rate (i.e. the longitudinal strain rate,defined as the derivatives of displacement velocity of a cellularparticle with respect to position on root axis). Supply of 10mol m^–3 Ca at the high salt condition restored partiallythe relative elemental elongation rate, but not the length ofthe growth zone. Compared to the control, the growth trajectoriesshowed that at 1 mol m^–3 CaCl2 it took more time for acellular particle to move through the growth zone at 150 molm^–3 NaCl, while at 10 mol m^–3 CaCl it took lesstime and there was no difference between the NaCl treatments Key words: Gossypium hirsutum, salinity stress, root growth kinematics     

Effect of Root Zone Temperature on the Mineral Composition of Xylem Sap and Plasma Membrane K+-Mg++-ATPase Activity of Grafted-Cucumber and -Figleaf Gourd Root Systems

Cucumber (Cucumis sativus L.) seedlings were grafted onto cucumber-(CG) or figleaf gourd- (FG, Cucurbita ficifolia Bouché)seedlings in order to determine the effect of solution temperature(12, 22, and 32°C) on the mineral composition of xylem sapand the plasma membrane K⁺-Mg⁺⁺-ATPase activities of the roots.Low solution temperature (12°C) lowered the concentrationof NO^–₃ and H₂PO^–₄ in xylem sap of CG plants butnot of FG plants. Concentrations of K⁺, Ca⁺⁺ and Mg⁺⁺ in xylemsap were less affected than anions by solution temperature.The plasma membrane of FG plants grown in 12°C solutiontemperature showed the highest K⁺- Mg⁺⁺-ATPase activity at allATP concentrations up to 3 mM and at low reaction temperatureup to 12°C, indicating resistance of figleaf gourd to lowroot temperature. (Received December 27, 1994; Accepted March 10, 1995)     

Induction of Solute Release from Nicotiana tabacum Tissue 10

For determination of the effects of polymyxin B, polymyxin E,or ethylenediamine tetra-acetic acid (EDTA) on plant cell membranes,the rates at which three solutes, K⁺, P₁, and sugar, leakedfrom treated tissue culture cell suspensions of Nicotiana tabacumwere measured. The kinetics of leakage from cells treated witheither of the polymyxins was biphasic, whereas kinetics forcells treated with EDTA was monophasic. Only K⁺ leaked frompolymyxin-treated cells during the first phase, and all threesolutes leaked during the second phase. The slower first phaseis interpreted as leakage of K⁺ from the Donnan free space andcytoplasm, and the faster second phase as the leakage of solutesfrom the vacuole. The monophasic kinetics of EDTA treatmentindicated that solutes were leaking simultaneously from cytoplasmand vacuole. Of the divalent cations tested, only Ca⁺⁺ and Mn⁺⁺counteracted the effects of polymyxin and EDTA. Ca⁺⁺ even restoredP¹ and sugar uptake. Addition of Mg⁺⁺ or Sr⁺⁺ to polymyxin-treatedcells did not stop solute leakage but actually enhanced theleakage rates. A model is presented that suggests that polymyxinor EDTA induces solute leakage by forming pores in plant cellmembranes. The effects of divalent cations on membranes oncethe pores are formed are also discussed. Key words: Polymyxin, EDTA, Nicotiana tabacum, Solute leakage     

Light-stimulated incorporation of L-leucine into the gibberellin of Gibberella fujikuroi

Cultures of Gibberella fujikuroi grown under continuous illuminationof 1800 lux incorporated over 60% more leucine into the gibberellinsthan dark controls. In the dark at a C/N ratio of 37.6 Ca⁺⁺increased the incorporations of leucine into A₃ to essentiallythe same level as the light-stimulated incorporation. The failureof Ca⁺⁺ to increase gibberellin synthesis in the dark at a C/Nratio of 9.4 suggested that light and Ca⁺⁺ were exerting theirregulatory roles at different sites. (Received October 15, 1969; )