Preferential inhibition by ethidium bromide of photosynthetic apparatus formation in Rhodopseudomonas spheroides cells

Ethidium bromide (EB) inhibited growth of cells of the non-sulfurpurple photosynthetic bacterium Rhodopseudomonas spheroides.The inhibitory action of EB on the light-anaerobic culturedcells was stronger than on dark-aerobic cultured ones. EB alsosuppressed the induced synthesis of bacteriochlorophyll (Bchl)and carotenoids in the dark-aerobically grown cells incubatedwith gentle aeration under no-growth conditions, suggestingthat the target of the inhibitory action of EB on the photosyntheticgrowth of R. spheroides cells is chromatophore formation. EBdepressed the incorporation of ³H- or ¹⁴C-uracil into both RNAand DNA fractions from cells incubated with gentle aeration.In contrast, inhibition by EB of ³H-uracil incorporation intothe DNA fraction was not observed under vigorous aeration. Ourfindings seemed to favor the hypothesis described previously(10) that lowering the intracellular oxidation-reduction potentialmight bring about a unique synthesis or turnover of DNA responsiblefor chromatophore formation. (Received December 22, 1977; )     

Heterogeneity of the DNA preparation from light-anaerobically grown Rhodopseudomonas spheroides cells as revealed by methylated albumin kieselguhr column chromatography

Biochemical properties of DNA preparations obtained from light-anaerobicallygrowth R. spheroides cells and from dark-aerobically grown cellswere examined comparatively using methylated albumin kieselguhrcolumn chromatography. The DNA preparation from light-growncells, which formed chromatophores, exhibited considerable heterogeneityin its size or conformation. ¹ Present address: The Third Department of Medicine, TohokuUniversity School of Medicine, Sendai, Japan. (Received August 2, 1974; )     

Further studies on ribulose-l,5-diphosphate carboxylase from Rhodopseudomonas spheroides and Rhodospirillum rubrum

Some enzymic properties of RuDP carboxylase isolated from Rhodopseudomonasspheroides and Rhodospirillum rubrum, grown under autotrophic,semi-autotrophic and heterotrophic conditions were studied.Regardless of the growing condition, the nature of the enzymefrom each respective bacterium remained unchanged. The molecularweight of R. spheroides and R. rubrum RuDP carboxylase was estimatedto be approximately 2.4x10⁵ and 8.3x10⁴, respectively. Bothenzymes require Mg⁺⁺. Their kinetic properties were also examined. ¹Part XII. Structure and Function of Chloroplast Proteins ²Supported in part by research grants from the Ministry of Educationof Japan (No. 8719), USPHS (AM. 10792-03) and the Asahi Press(Tokyo) (Received March 18, 1970; )     

DNA Levels in Dividing and Developing Plastids in Expanding Primary Leaves of Avena sativa

The amounts of plastid DNA in the primary leaves of 4-d-oldlight- and dark-grown seedlings of Avena sativa were measuredby microspectrofluorometry using the DNA-fluorochrome DAPI (4',6-diamidino-2-phenylindole). In the light-grown primary leaves (40–45 mm long) therewas a marked increase in DNA level per plastid from 10.2 to18.5 ? 10^–15 g between 2.0 mm and 10 mm from the leafbase, resulting from the rate of plastid DNA synthesis beinghigher than the rate of plastid division. Beyond 30 mm the plastidDNA level was reduced to 14 ? 10^–15g due to chloroplastdivision rates being higher than the rate of plastid DNA synthesis,while from 20 mm plastid DNA levels were constant at 2.2 ? 10^–12g per cell, which corresponds to 16000 plastome copies per cell. Observations of dark-grown leaves establish that, in Avena,light is not necessary for plastid division and the dark-grownleaf cells accumulate higher amounts of plastid DNA than light-grownleaf cells. Plastid nucleoids showed a change of distribution after completionof plastid DNA synthesis in light-grown leaves. A change inthe distribution of plastid nucleoids was also observed duringthe greening of etioplasts of dark-grown leaves while plastidDNA level remained constant. Such changes in plastid nucleoiddistribution appear to be independent of plastid DNA synthesisand correlate with the formation of grana stacks. Key words: Avena sativa, microspectrofluorometry, plastid DNA     

Stability of rRNA genes during germination of Vicia faba seeds

Germinating Vicia embryos and the roots of 4-day-old seedlingswere assayed for the proportion of DNA that is complementaryto Vicia rRNA. Hybridization experiments which were performedusing DNA purified by ordinary procedures including RNase digestionshowed a higher level of saturation only for the DNA from theembryos. The embryo DNA showed a Tm of 74?C in 0.1?SSC, whereasthe Tm of the root DNA was 70?C. However, our final conclusionis that there is no gross amplification of rRNA genes for Viciafaba, since the saturation level for the embryo DNA did notexceed that for the root DNA when the DNAs were used after additionalpurification on CsCl gradients. ¹ Present address: Department of Botany, Faculty of Education,Utsunomiya University, Utsunomiya 320, Japan. (Received November 25, 1974; )     

Ribulose-1, 5-diphosphate carboxylase of Chromatium strain D

RuDP carboxylase isolated from autotrophically grown cells ofphotosynthetic sulfur bacterium, Chromatium strain D, was partiallypurified by (NH₄)₂SO₄ precipitation and Sephadex G-200 gel filtration.The molecular size of the bacterial RuDP carboxylase was foundto be large, analogous to that of the plant enzyme, in agreementwith results of previous workers. Sucrose density gradient centrifugationshowed the S_rel to be approximately 18; the omission of Mg⁺⁺caused no dissociation of the enzyme molecule in its subunits.Chromatium RuDP carboxylase showed similarities to the plantenzyme in some of its kinetic properties; (a) a shift of pHoptimum to the neutral side from the alkaline side on the additionof Mg⁺⁺, (b) deviation of the substrate concentration (NaHCO₃)-activityrelationship from the MICHAELIS formula and (c) a marked stimulativeeffect of Mg⁺⁺. A unique sigmoidal saturation curve of the enzymeto RuDP, which had been detected in Rhodospirillum rubrum andRhodopseudomonas spheroides RuDP carboxylase in the absenceof Mg⁺⁺, was not found. Another characteristic feature of ChromatiumRuDP carboxylase is its partial immunological response to therabbit anti-spinach RuDP carboxylase serum as detected by theinhibition of the carboxylation reaction due to the antibody-antigenreaction. ¹Part X, Structure and Function of Chloroplast Proteins. Supportedin part by research grants from the Ministry of Education ofJapan (No. 8719) and USPHS (AM-10792-03) (Received July 4, 1969; )     

Nuclear DNA Content of 26 Orchid (Orchidaceae) Genera with Emphasis onDendrobium

2C DNA content values for 70 orchid species from 26 genera,including 37Dendrobiumspecies from eight taxonomic sections,were analysed using flow cytometry. The resulting nuclear DNAcontent values for species other thanDendrobiumranged from 1.91pg 2C^-1to 15.19 pg 2C^-1nuclei forCadetia tayloriandVanilla phaeantha,respectively.Dendrobiumnuclear DNA content values ranged from1.53 pg 2C^-1to 4.23 pg 2C^-1nuclei forD. cruentumandD. spectabile,respectively. DNA content measurements varied greatly withinDendrobiumsectionsLatouria and Spatulata. Nuclear DNA content values for the sixspecies analysed within Latouria ranged from 1.88 pg 2C^-1nucleiforD. macrophyllumto 4.23 pg 2C^-1nuclei forD. spectabile. NuclearDNA content values for the 16 species analysed within Spatulataranged from 1.69 pg 2C^-1nuclei forD. discolorto 4.05 pg 2C^-1nucleiforD. samoense. The least variation in DNA content was foundwithin the section Phalaenanthe, with nuclear DNA content valuesof 1.79 pg  2C^-1, 1.86 pg 2C^-1and 1.98 pg 2C^-1forD. bigibbum,D.affineandD. phalaenopsis, respectively.Copyright 1998 Annalsof Botany Company Orchidaceae,Dendrobium, flow cytometry, propidium iodide, nuclear DNA, genome size, 2C values.     

Light-induced absorption spectrum changes of carotenoid in chromatophores of photosynthetic bacterium, Rhodopseudomonas spheroides II. Rapid and slow absorption changes of carotenoid in chromatophores

Time courses of light-induced absorption changes of carotenoid(spheroidene) in chromatophores of the photosynthetic bacterium,Rhodopseudomonas spheroides, consisting of an 8–10 nm-shiftin the carotenoid absorption spectrum towards the longer wavelength,were investigated. The whole time course of absorption changecan be expressed as the sum of two first order reactions; arapid change accomplished in several msec after the onset ofillumination with actinic light (800–900 nm) and a slowchange extending over the whole period (200 msec). The rapidchange required a high light intensity, whereas the slow changewas saturated at relatively low light intensity (ca. 5xl0³ erg/cm²sec). Electron transport inhibitors (HOQNO, piericidin A and o-phenanthroline)and Cl-CCP, at concentrations uncoupling photophosphorylation(10^–M), inhibited slow change but did not affect rapidchange. The rapid change was inhibited by higher concentrationsof Cl-CCP (10–4 M) and by o-phenanthroline in the concomitantpresence of an uncoupling concentration of Cl-CCP. The rapidand slow changes have midpoint potentials of 440 and 420 mv,respectively; as calculated from absorption changes in the presenceof ferri- and ferrocyanide mixtures. Relationships between absorptionspectrum changes and other reactions in the chromatophores wereanalyzed. Slow absorption change is closely related to the highenergy intermediate, or state, of photophosphorylation. Rapidabsorption change, with a midpoint potential of 440 mv, is relatedto the electron flow mediated by P870; although it does notdirectly reflect the state of P870, itself. ¹ This article is the second of a series published under thesame title in this journal, volume 11 (1970) p. 519–530. ² Present address: Department of Biology, Faculty of Science,Toho University, Narashino City, Chiba 275, Japan. (Received October 30, 1970; )     

Highly Repetitive Tandem Arrays of a Short 32 bp Unit in the Pharbitis nil Genome: the RsaI Family

A highly repetitive DNA sequence of Pharbitis nil, designatedthe RsaI family, was cloned, sequenced and analyzed with respectto its genomic organization. The RsaI family is arranged intandem arrays and composed of a 32 bp repeat unit, which isthe shortest unit thus far reported for plant repetitive sequences.The RsaI family represents 3% of the total genomic DNA and thecopy number of the 32 bp unit is estimated to be about 1 ? 10⁶per haploid genome. We suggest the existence of a higher orderrepeat unit, which may be composed of hundreds of the 32 bpunits and be repeated many times in the genome. (Received May 12, 1988; Accepted July 28, 1988)     

Purification and Characterization of Soluble and Membrane-Associated DNA Polymerases from a Virus PBCV-1 Infected Green Alga Chlorella NC64A

DNA polymerases were purified several hundred-fold from the10 000 x g soluble (polymerase I) and particulate (polymeraseIII) fractions prepared from virus PBCV-1 infected ChlorellaNC64A extracts. Both DNA polymerases exhibited optimal activitywith activated calf thymus DNA at pH 8.5. DNA polymerase I required3.0 mol m^–3 MgSO₄ and 150 to 250 mol m^–3 KCl foroptimum activity whereas, DNA polymerase III required 2.0 molm^–3 MgSO₄ and 150 mol m^–3 KCl. Both enzymes wereinhibited by pyrophosphate, actinomycin D, ethidium bromide,dideoxythymidine triphosphate, and N-ethylmaleimide but wererelatively insensitive to aphidicolin. DNA polymerase I differedfrom DNA polymerase III in its response to cations (particularlyNH₄Cl), elution from a DEAE cellulose column, and molecularweight. Key words: Algal virus, DNA polymerase, Chlorella     

Some Experimental and Theoretical Observations Concerning Mass Flow in the Vascular Bundles of Heracleum

The theory and practice of applying the thermodynamics of irreversibleprocesses to mass-flow theories is presented. Onsager coefficientswere measured on cut and uncut phloem and cut xylem strandsof Heracleum muntegazzimum. In 0.3 N sucrose + 1 mN KC1 theyare as follows. In phloem, L_EE = 5 ? 10–⁴ mho cm^–1,L_pE = 9 ? 10^–6 cm³ s^–1 cm^–2 volt^–1 cm,and L_PP = 0.16 cm³ s^–1 cm^–2 (J cm^–3)^–1cm. In uncut phloem strands L_EE is about 1 ? 10^–3 mhocm^–1. In xylem in 2 x 10^–3 N KCI, L_pp = 50 to 225,L_PE = 2 ? 10^–4, and L^EE = 4 ? 10^–3. The measurementsare tentative since the blockage of the sieve plates is an interferingfactor, but if they are valid they lead to the conclusion thatneither a pressure-flow nor an electro-kinetic mechanism envisaginga ‘long distance’ current pathway can be the majormotive ‘force’ for transport in mature phloem. Measurementsof biopotentials along conducting but laterally detached phloembundles of Heracleum suggest, nevertheless, that there may bea small electro-osmotic component of at least 0.1 mV cm^–1endogenous in the phloem.     

Variation in the DNA Content of Glycine Species

Nuclei were isolated from cotyledons of a range of accessionsfrom 14 species of Glycine. These were stained with ethidiumbromide and the relative fluorescence for each genotype wasmeasured by flow cytometry. The DNA content was estimated bycomparison of relative fluorescence with that from nuclei fromseedling leaves of Allium cepa, whose DNA content has been calculatedpreviously by chemical assay. The 4C amounts for diploid Glycineranged from 3.80 to 6.59 pg. Two groups of diploid species appearedfrom the analysis. The first consisted of species with amountsranging from 3.80 to 5.16 pg and included G. canescens (AA),G. argyrea (A₁ A₁), G. clandestina (A²A²), G. microphylla(BB),G. latifolia (B₁B₁), G. tabacina 2n=40 (B²B²), G. tomentella2n=38 (EE) and 2n=40 (DD), G. max and G. soja (GG), G. arenariaand G. latrobeana. A second group had higher DNA contents rangingfrom 5.27 to 6.59 pg, and consisted of G. curvata, G. cyrtoloba(CC), and G. falcata (FF). The polyploid species, G. tabacina2n=80 (AABB, BBB¹B¹), G. tomentella 2n=78 and 2n=80 (AAEE andDDEE, respectively) contained amounts approximating to the sumsof the respective parental diploid species thought to have givenrise to these allotetraploids. Intraspecific variation was detectedin the DNA content of G. canescens. Within the overall distributionof DNA amounts found in A genome species, each genome containeda range of DNA contents specific to that species. This phenomenonwas also detected amongst B genome species.     

Ethylene and 1 -Aminocyclopropane-1-Carboxylic Acid Production in flacca, a Wilty Mutant of Tomato, Subjected to Water Deficiency and Pre-treatment with Abscisic Acid

Neill, S. J., McGaw, B. A. and Horgan, R. 1986. Ethylene and1-aminocyclopropane-l-carboxylic acid production in flacca,a wilty mutant of tomato, subjected to water deficiency andpretreatment with abscisic acid —J. exp. Bot. 37: 535–541. Plants of Lycoperstcon esculentum Mill. cv. Ailsa Craig wildtype and flacca (flc) were sprayed daily with H²O or 2?10^–2mol m^–3 abscisic acid (ABA). ABA treatment effected apartial phenotypic reversion of flc shoots; leaf areas wereincreased and transpiration rates decreased. Leaf expansionof wild type shoots was inhibited by ABA. Indoleacetic acid (IAA), ABA and l-aminocyclopropane-l-carboxylicacid (ACC) concentrations were determined by combined gas chromatography-massspectrometry using deuterium-labelled internal standards ABAtreatment for 30 d resulted in greatly elevated internal ABAlevels, increasing from 1?0 to 4?3 and from 0?45 to 4?9 nmolg^–1 fr. wt. in wild type and flc leaves respectively.Endogenous IAA and ACC concentrations were much lower than thoseof ABA. IAA content ranged from 0?05 to 0?1 nmol g^–1 andACC content from 0?07 to 0?24 nmol g^–1 Ethylene emanationrates were similar for wild type and flc shoots. Wilting of detached leaves induced a substantial increase inethylene and ACC accumulation in all plants, regardless of treatmentor type. Ethylene and ACC levels were no greater in flc leavescompared to the wild type. ABA pretreatment did not preventthe wilting-induced increase in ACC and ethylene synthesis. Key words: ABA, ACC, ethylene, wilting, wilty mutants     

Potassium Transport Across the Membranes of Chara: I. THE RELATIONSHIP BETWEEN RADIOACTIVE TRACER INFLUX AND ELECTRICAL CONDUCTANCE

Smith, J. R., Smith, F. A. and Walker, N. A. 1987. Potassiumtransport across the membrane of Chara. I. The relationshipbetween radioactive tracer influx and electrical conductance.—J.exp. Bot. 38:731–751. The ⁴²K influx () and the electrical conductance (G_m) were measured simultaneously for the ‘membrane’of internodal cells of Chara australis as a function of theexternal [KCl] (K?. In bathing solutions of pH = 5?0, progressively increased from 20?5to 430?60 nmol m^–2 s^–1 and G_m increased from 0?36?0?02to 3?8?0?8 S m^–2 when K? was increased from 0?1 to 10mol m^–3. The resting membrane potential difference (p.d.)was approximately -135 mV for low K? and approached the expectedNernst equilibrium p.d. for K⁺ ions when K? > 1?0 mol m^–3.Measurements of ³⁶Cl influx suggested that the ⁴²K influx waspredominantly electrogenic. The equivalent Goldman permeabilityto K⁺ ions (P_k) was approximately 20–30 nm s^–1 anddid not vary significantly with increasing K?. The equivalentconductance attributable to the electrogenic transport of K⁺ ions was calculated from assuming passive, independent diffusionof K⁺ ions and the ratio was found to be typically close to one. It was also found that themagnitudes of and G_m measuredsimultaneously for each individual cell were also well correlatedfor K? 1?0 mol m^–3, and that the slope of the line ofbest fit was close to one. For each K? it was found that theconductance not attributable to K⁺ translocation and presumablyassociated primarily with the transport of protons or theirequivalents was typically 0?2–0?5 Sm^–2. For K? >1?0 mol m^–3 the results indicated that the transport ofK⁺ ions was essentially independent, i.e. there was no evidencefor flux interactions. The results also indicated that the equivalentconductance derived from the measured ⁴²K influx could usefullyindicate the fraction of the electrical conductance attributableto the translocation of K⁺ ions. Key words: Potassium, conductance, influx     

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; )     

Root Hydraulic Conductivity of Two Cactus Species in Relation to Root Age, Temperature, and Soil Water Status

The effects of root age, temperature, and soil water statuson root hydraulic conductivity (L_P) were investigated for twocactus species, Ferocactus acanthodes and Opuntia ficus-indica.The volumetric flux density of water was measured for excisedroot segments, either using negative hydrostatic pressures appliedto the proximal end or using reverse flow of water from theroot to the soil. For both species, L_P at 20 ?C increased withroot age, average values reaching a maximum of 3.9 ? 10^–7m s^–1 MPa^–1 for F. acanthodes and 5.2 ? 10^–7m s^–1 MPa^–1 for O.ficus-indica at 11 to 17 weeksof age; L_P subsequently declined with increasing root age forboth species. L_P was maximal at a temperature of about 10 ?Cfor the youngest roots (1–3 weeks), this optimum shiftingto 40 ?C for 8-week-old roots of both species. For older roots(up to 1.5-years-old), L_P increased with temperature from 0?C to 50 ?C, with a Q₁₀ of 1.3 between 20 ?C and 30 ?C. At asoil water potential (_soil) of –0.016 MPa, root L_P wasindependent of the direction of water flow for both species.Depending on root age, L_P declined 45- to 500-fold for F. acanthodesand 90- to 800-fold for O.ficus-indica as _soil was reduced from–0.016 to –1.06 MPa, consistent with a rectifier-likebehaviour with respect to water movement between soil and roots.Incorporation of such responses into water uptake models shouldlead to a better understanding of root function. Key words: Ferocactus acanthodes, Opuntia ficus-indica, water potential, tension, reverse flow     

Effect of Temperature and External pH on the Cytoplasmic pH of Chara corallina

Cytoplasmic pH (pH_c) in Chara corallina was measured (from [¹⁴C]stribution)as a function of external pH (pH₀)and temperature. With pH₀near 7, pH_c at 25?C is 7.80; pH_cincreases by 0.005 pH units?C^–1 temperature decrease, i.e. pH_c at 5 ?C is 7.90. WithpH? near 5.5, the increase in pH_c with decreasing temperatureis 0.015 units ?C^–1 between 25 and 15?C, but 0.005 units?C^–1 between 15 and 5?C. This implies a more precise regulationof pH_c with variations in pH_o at 5 or 15 ?C compared with 25?C. The observed dp H_c/dT is generally smaller than the –0.017units ?C^–1 needed to maintain a constant H⁺/OH^–1,or a constant fractional ionization of histidine in protein,with variation in temperature. It is closer to that needed tomaintain the fractional ionization of phosphorylated compoundsor of CO₂–HCO₃^– The value of dpH_c/dT has importantimplications for several regulatory aspects of cell metabolism.These include (all as a function of temperature) the rates ofenzyme reactions, the _H+ at the plasmalemma(and hence the energy available for cotransport processes),and the mechanism for pH_c regulation by the control of bidirectionalH⁺ fluxes at the plasmalemma.     

Morphological and physiological characterization of IAA-less-sensitive and IAA-sensitive phases in rooting of Azukia cuttings

In disbudded epicotyl cuttings taken from light grown 5-dayold Azukia angularis Phaseolus angularis) seedlings, all adventitiousrootlets appeared on the second day of incubation. No root primordiawere observed within the first 24 hr and no increase in thenumber of roots occurred after 48 hr. Puromycin (5.5?10^–5M), p-fluorophenylalanine (1?10^–3M),2-thiouracil (2.3?10^–4M) and 2,6-diaminopurine (2?10^–5M)inhibited rooting when applied to cuttings on the second day,but showed no inhibition when applied on the first day. Unlike these inhibitors, pyrithiamine (7.2?10^–5M) inhibitedrooting when it was applied to cuttings on the first day. A rooting promoting effect was observed with actinomycin D (2.4?10^–6M),2,4-dinitrophenol (3?10^–5M) and p-fluorophenylalanine(1?10^–4M) applied to the cuttings on the first day, whereasindoleacetic acid (1.7?10^–4M) showed its promoting effectmost effectively on the second day. ¹Contribution No. 17 from the Botanical Gardens, Faculty ofScience, University of Tokyo, Tokyo, Japan. (Received June 4, 1969; )     

Relationships between Acetylene Reduction Activity, Hydrogen Evolution and Nitrogen Fixation in Nodules of Acacia spp.: Experimental Background to Assaying Fixation by Acetylene Reduction under Field Conditions

Hansen, A. P., Pate, J. S. and Atkins, C. A. 1987. Relationshipsbetween acetylene reduction activity, hydrogen evolution andnitrogen fixation in nodules of Acacia spp.: Experimental backgroundto assaying fixation by acetylene reduction under field conditions.—J.exp. Bot. 38: 1–12 Glasshouse grown, symbiotically-dependent seedlings of Acaciaalata R.Br., .A. extensa Lindl., and A. pulchella R.Br. wereexamined for acetylene reduction in closed assay systems usingundisturbed potted plants, excavated whole plants, nodulatedroots or detached nodules. Nitrogenase activity declined sharplyover the first hour after exposure of detached nodules to acetylene(10% v/v in air), less steeply or not at all over a 3 h periodin assays involving attached nodules. Using detached nodules,rates of acetylene reduction, nitrogen (¹⁵N₂) fixation, andhydrogen evolution in air (¹⁵N₂) and acetylene-containing atmosphereswere measured in comparable 30 min assays. Total electron flowthrough nitrogenase in air was determined from rates of nitrogen(¹⁵N₂) fixation ( ? 3) plus hydrogen evolution, that in thepresence of acetylene from rates of acetylene reduction andhydrogen evolution in air: acetylene. Values for the ratio ofelectron flow in air: acetylene to that in air ranged from 0?43to 0?83 in A. pulcheila, from 0?44 to 0?66 in A. alala and from0?37 to 0?70 in A. extensa, indicating substantial inhibitionof electron flow through nitrogenase of detached nodules byacetylene. Relative efficiencies of nitrogenase functioningbased on hydrogen evolution and acetylene reduction were from0?15 to 0?79, those based on nitrogen (¹⁵N₂) fixation and hydrogenevolution from 0?53 to 0?87. Molar ratios of acetylene reducedto nitrogen (¹⁵N₂) fixed were 2?82 ? 0?24, 201 ? 0?15, and 1?91? 0?11 (?s.e.; n = 7) for A. pulcheila,A. extensa and A. alata respectively A standard 5–10 min acetylene reduction assay, conductedon freshly detached unwashed nodules in daytime (12.00–14.00h), was calibrated for field use by comparing total N accumulationof seedlings with estimated cumulative acetylene reduction overa 7-week period of glasshouse culture. Molar ratios for acetylenereduced: nitrogen fixed using this arbitrary method were 3?58for A. alata, 4?82 for A. extensa and 1?60 for A. pulchella.The significance of the data is discussed. Key words: Acacia spp, nitrogenase functioning     

Some properties of the amine oxidase in Vicia faba seedlings

Growth of the Vicia faba seedling is accompanied by a rapid15-day increase in amine oxidase activity of the apical parts.Cotyledons and roots were found to be devoid of activity. Thepartially purified enzyme from leaves readily oxidized putrescine,cadaverine, agmatine and spermidine, while dopamine (3-hydroxytyramine)and L- and D-lysine were oxidized more slowly. The Km valueswere 1.9?10^–3 M for cadaverine, 3.7?10^–5 M for putrescine,7.8?10^–4 M for spermidine, and 5.9?10^–3 M for dopamine.Carbonyl reagents and copper-binding agents were effective inhibitorsof Vicia faba amine oxidase. The diethyldithiocarbamate-treatedenzyme could be reactivated specifically by cupric copper. (Received May 25, 1977; )