Multiple forms of rat liver mitochondrial DNA polymerase.

Mitochondrial DNA polymerase (DNA polymerase mt) exists in two active forms. DNA polymerase present in crude extract (M-I) and ammonium sulfate precipitate (M-II) stages of purification sediments at 12.1S. The enzyme at the M-II stage of purification has a molecular weight of approximately 250,000 as determined by Sephadex G-200 chromatography in buffers of low ionic strength. In buffers containing 0.15 m NaCl, the enzyme sediments at 9.4S and has a molecular weight of approximately 190,000. When the enzyme is further purified on diethylaminoethyl cellulose (M-III stage of purification), the 9.4S activity predominates. Addition of a polymerase-free fraction from the M-III stage of purification changes the sedimentation coefficient of the enzyme from 9.4 to 12.1S.     

Stability of the annual shoot diameter in Persian walnut: a case study of different morphotypes and years

The basal diameter of the annual shoot (1YD) affects vegetative growth and fruiting of the walnut trees. In order to determine interdependency between the 1YD and the older parent wood, 64 walnut genotypes belonging to four different branching and fruiting habits (morphotypes M-I, M-II, M-III and M-IV) were investigated. Year-to-year stability of 1YD was tested with the architectural analysis of a 3-year-old fruiting branch and its constituents (a 3-year-old bearer + corresponding 2-year-old + annual shoots) during 3 successive years. Based on Pearson's correlation coefficients and the multiple regression analysis of 12 quantitative traits, 12 models (four morphotype in 3 successive years) of 1YD were formed. They were compared with the standard model which was calculated on the basis of 1-year measurements of 1Y with no respect to the branching and fruiting type and comprises three quantitative traits, i.e. basal diameter of a 2-year-old parent shoot (2YD), the length of 2Y shoot (2YL), and the length of annual shoot (1YL). In a single year, the 1YD was influenced by two–five parameters. Five out of 12 models agreed with the standard model: in the lateral fruiting genotypes (M-IV), 1YD was always under the influence of the 2Y diameter, and the 1Y length. In addition, the number of nodes of the 2Y parent shoot had an important influence on 1Y diameter. In the terminal bearers (M-I), the impact of 2YD on the 1YD slightly increased with the tree age, and some other parameters, like 1Ynumber and 1Ynodes, which became to be important for 1YD. In the intermediate genotypes with mezotonic ramification (M-II), the number of vegetative buds per 1Y and angles of 1Y had significant effects on 1YD. In the intermediate bearers with acrotonic ramification (M-III), one to four other parameters were included into the model each year beside the 1Y number. Since the traits of a 2-year-old parent shoot have a great influence on the 1YD, the information from the year N can be used for the prediction of the annual shoot development in the year N+1. Such a prediction is more reliable in M-I and M-IV than in M-II and M-III. When we deal with the intermediate fruiting cultivars, 1Y number has to be considered in prediction of 1Y diameter beside 2YD and 1YL.     

Allocation of Organ Biomass in Perfect and Imperfect Flowers

Perianth M_P, gynoecium M_G, and androecium M_A dry-weight biomass(in g) of 39 species of perfect flowers was measured. Thesedata were pooled with published data from an additional 51 speciesand used to determine size-dependent variations in (M_G and M_A)in terms of the hypothesis that the quotient of M_G and M_A exceeds1·0 for out-breeding (xenogamous) species and less than1·0 for in-breeding (autogamous) species. Ordinary leastsquare regression of the pooled data (n = 90) showed M_G = 0·118M^0·916_P (r² = 0·884) and M_A = 0·186 M^0·975_P(r² = 0·865), indicating that the biomass of the gynoeciumproportionally decrease as floral size increases. The exponentsof these regressions indicate that the ratio of gynoecial toandroecial biomass decreased with increasing floral size suchthat comparatively small flowers (M_P < 0·0021 g) hadM_G/M_A > 1·0 (predicted for 'out-breeders') while comparativelylarger flowers (M_P > 0·0021 g) had M_G /M_A < 1·0(predicted for 'in-breeders'). Thus, on average, the type ofbreeding system was a size-dependent phenomenon. To test whether the biomass of a floral organ-type is a legitimateindicator of gender reproductive effort, the biomass (in g)of stamen filaments M_m and anther sacs M_AS of 39 species wasdetermined. Least square regression of these data showed M_AS= 0·188 M^0·854_fil (r² = 0·967), indicatingthat species with larger stamen filaments, on the average, boreproportionally smaller anther sacs and thereby cautioning againstthe uncritical use of the allocation of biomass to floral organ-typeas a strict gauge of gender-function investment. To determine whether the loss of one gender-function resultsin proportional reallocation of biomass to the remaining gender-function,the size-dependency of androecial and gynoecial biomass wasdetermined for a total of 33 perfect and imperfect flowers ofCucumis melo. Regression of the data obtained from perfect flowersyielded M_A = 0·402 M^1·47_P (r² = 0·898)and M_G = 4·63 M^1·36_P (r² = 0·842). SinceM_G/M_A M^0·11_P , the biomass allocation to the gynoeciumrelative to the androecium decreased with increasing floralsize. This result was consistent with the broad interpecificcomparison based on 90 species with perfect flowers . Regressionof the data for imperfect flowers yielded M_A = 0·151M^1·02_P (r² = 0·675) and M_G = 4·68 M^1·47_P(r² = 0·996), indicating a near allometric relation forthe androecium and a strong positive anisometry for the gynoecium.Thus, for flowers of comparable size, a loss of female genderobtains a modest to significant again in androecial biomasswhereas the loss of male gender yields only a slight increasein gynoecial biomass. Collectively, the results of these studies indicate that biomassallocation patterns are size-dependent phenomena whose complexitieshave been largely ignored in the literature.Copyright 1993,1999 Academic Press Allometry, floral biomass, reproduction     

Separation and some properties of two acid phosphatases from suspension cultures ofIpomoea sp.

Two acid phosphatases isolated from culturedIpomoea (moring glory) cells were separated by column chromatography on DEAE-cellulose. The two acid phosphatases have different pH optima (pH 4.8–5.0 and 6.0) and do not require the presence of divalent ions. The enzymes possess high activity toward pyrophosphate,p-nitrophenylphosphate, nucleoside di- and triphosphates, and much less activity toward nucleoside monophosphates and sugar esters. The two phosphatases differ from each other in Michaelis constants, in the degree of inhibition by arsenate, fluoride and phosphate and have quantitative differences of substrate specificity. In addition, they also differ in their response to various ions. Issued as NRCC No. 20658     

Formation of Isodityrosine by Peroxidase Isozymes

Fry, S. C. 1987. Formation of isodityrosine by peroxidase isozymes.—J.exp. Bot. 38: 853–862. Tyrosine residues of extensin are oxidatively coupled in vivoto form isodityrosine bridges, whereas treatment of purifiedextensin with H₂O₂+ peroxidase in vitro yields only dityrosine.Two explanations for the correct mode of coupling in vivo weretested. The first, that the pH of the cell wall is lower thanthat (pH 9-0) at which in vitro experiments have been conducted,provided part of the answer since treatment of L-tyrosine withH₂O₂+peroxidase in vitro at pH 37–5 yielded some isodityrosine.The second, that the wall contains other isozymes of peroxidasethan the basic isozyme usually studied in vitro, appeared unlikelybecause several sharply contrasting isozymes yielded similarisodityrosine: dityrosine ratios from L-tyrosine+ H₂O₂ at anygiven pH. The isozymes were also similar in their ability tooxidize tyrosine-dimers further to higher polymers. It is concludedthat the formation of isodityrosine in vivo is dictated by neighbouringwall molecules, possibly ionically-bound pectins, which modifythe local environment of the tyrosine residues of extensin. Key words: Isodityrosine, peroxidase isozymes, extensin     

Effect of Mechanical Stress on Sunflower (Helianthus annuus L.) Hypocotyl Growth

Etiolated seedlings of Sunflower (Helianthus annuus L.) weresubjected to mechanical stress by longitudinally compressingthe hypocotyl with approx. 0·05 N force (equivalent to0·025 MPa for a 1·6 mm diameter hypocotyl). Thisrelatively low compressive stress effected an increase in relativegrowth rates (RGR) of the hypocotyl for a period of 1-2 h, followingwhich RGR returned to the pretreatment rate. RGR was also increasedby an equivalent compressive stress treatment (0·025MPa) for 4 h in water or in 10^-10 mol l^-1 IAA. These resultsare discussed in the context of a possible role for mechanicallyinduced stress in the initiation and maintenance of nutationalgrowth movements.Copyright 1994, 1999 Academic Press Helianthus annuus, Sunflower, hypocotyl growth, mechanical stress, seedling growth, nutational growth movements, circumnutation     

The Influence of NO-3 and NH+4 Nutrition on the Gas Exchange Characteristics of the Roots of Wheat (Triticum aestivum) and Maize (Zea mays) Plants

Respiratory oxygen consumption by roots was 1·4- and1·6-fold larger in NH⁺₄-fed than in NO^-₃-fed wheat (Triticumaestivum L.) and maize (Zea mays L.) plants respectively. Higherroot oxygen consumption in NH⁺₄-fed plants than in NO^-₃-fedplants was associated with higher total nitrogen contents inNH⁺4-fed plants. Root oxygen consumption was, however, not correlatedwith growth rates or shoot:root ratios. Carbon dioxide releasewas 1·4- and 1·2-fold larger in NO⁺3-fed thanin NH⁺₄-fed wheat and maize plants respectively. Differencesin oxygen and carbon dioxide gas exchange rates resulted inthe gas exchange quotients of NH^-₄-fed plants (wheat, 0·5;maize, 0·6) being greatly reduced compared with thoseof NO^-₃-fed plants (wheat, 1·0; maize, 1·1). Measuredrates of HCO^-₃ assimilation by PEPc in roots were considerablylarger in 4 mM NH⁺₄-fed than in 4 NO^-₃ plants (wheat, 2·6-fold;maize, 8·3-fold). These differences were, however, insufficientto account for the observed differences in root carbon dioxideflux and it is probable that HCO^-₃ uptake is also importantin determining carbon dioxide fluxes. Thus reduced root extension in NH⁺₄-fed compared with NO^-₃-fedwheat plants could not be ascribed to differences in carbondioxide losses from roots.Copyright 1993, 1999 Academic Press Triticum aestivum, wheat, Zea mays, maize assimilation, ammonium assimilation, root respiration     

Relationship between Leaf Structure and Gas Exchange in Wheat Leaves at Different Insertion Levels

Net photosynthesis rate (P_n), stomatal conductance to CO₂ andresidual conductance to CO₂ were measured in the last six leaves(the sixth or flag leaf and the preceding five leaves) of Triticumaestivum L. cv. Kolibri plants grown in Mediterranean conditions.Recently fully expanded leaves of well-watered plants were alwaysused. Measurements were made at saturating photosynthetic photonflux density, and at ambient CO₂ and O₂ levels. The specificleaf area, total organic nitrogen content, some anatomical characteristics,and other parameters, were measured on the same leaves usedfor gas exchange experiments. A progressive xeromorphic adaptation in the leaf structure wasobserved with increasing leaf insertion levels. Furthermore,mesophyll cell volume per unit leaf area (V_mes/A) decreasedby 52·6% from the first leaf to the flag leaf. Mesophyllcell area per unit leaf area also decreased, but only by 24·5%.However, nitrogen content per unit mesophyll cell volume increasedby 50·6% from the first leaf to the flag leaf. This increasecould be associated to an observed higher number of chloroplastcross-sections per mm² of mesophyll cell cross-sectional areain the flag leaf: values of 23000 in the first leaf and 48000in the flag leaf were obtained. P_n per unit leaf area remainedfairly constant at the different insertion levels: values of33·83±0·93 mg dm^–2 h^–1 and32·32±1·61 mg dm^–2 h^–1 wereobtained for the first leaf and the flag leaf, respectively.Residual conductance, however, decreased by 18·2% fromthe first leaf to the flag leaf. Stomatal conductance increasedby 41·7%. The steadiness in P_n per unit leaf area across the leaf insertionlevels could be mainly accounted for by an opposing effect betweena decrease in V_mes/A and a more closely packed arrangement ofphotosynthetic apparatus. Adaptative significance of structuralchanges with increasing leaf insertion levels and the steadinessin P_n per unit leaf area was studied. Key words: Photosynthesis, structure, wheat     

Effects of the fungal endophyte, Neotyphodium lolii, on net photosynthesis and growth rates of perennial ryegrass (Lolium perenne) are independent of In Planta endophyte concentration

• Background and Aims Neotyphodium lolii is a fungal endophyteof perennial ryegrass (Lolium perenne), improving grass fitnessthrough production of bioactive alkaloids. Neotyphodium speciescan also affect growth and physiology of their host grasses(family Poaceae, sub-family Pooideae), but little is known aboutthe mechanisms. This study examined the effect of N. lolii onnet photosynthesis (P_n) and growth rates in ryegrass genotypesdiffering in endophyte concentration in all leaf tissues. • Methods Plants from two ryegrass genotypes, Nui D andNui UIV, infected with N. lolii (E+) differing approx. 2-foldin endophyte concentration or uninfected clones thereof (E–)were grown in a controlled environment. For each genotype xendophyte treatment, plant growth rates were assessed as tilleringand leaf extension rates, and the light response of P_n, darkrespiration and transpiration measured in leaves of young (30–45d old) and old (>90 d old) plants with a single-chamber openinfrared gas-exchange system. • Key Results Neotyphodium lolii affected CO₂-limited ratesof P_n, which were approx. 17 % lower in E+ than E– plants(P < 0·05) in the young plants. Apparent photon yieldand dark respiration were unaffected by the endophyte (P >0·05). Neotyphodium lolii also decreased transpiration(P < 0·05), but only in complete darkness. There wereno endophyte effects on P_n in the old plants (P > 0·05).E+ plants grew faster immediately after replanting (P < 0·05),but had approx. 10 % lower growth rates during mid-log growth(P < 0·05) than E– plants, but there was noeffect on final plant biomass (P > 0·05). The endophyteeffects on P_n and growth tended to be more pronounced in NuiUIV, despite having a lower endophyte concentration than NuiD. • Conclusions Neotyphodium lolii affects CO₂ fixation,but not light interception and photochemistry of P_n. The impactof N. lolii on plant growth and photosynthesis is independentof endophyte concentration in the plant, suggesting that theendophyte mycelium is not simply an energy drain to the plant.However, the endophyte effects on P_n and plant growth are stronglydependent on the plant growth phase.     

Evaluation of a Bioassay for Cytokinins Using Soybean Hypocotyl Sections

The dose-response curves of several cytokinins were investigatedin a soybean hypocotyl bioassay. Zeatin riboside, zeatin-O-ß-D-glucoside,dihydrozeatin, and dihydrozeatin riboside produced linear responsesparallel to that for zeatin. The hypocotyl section assay wassuperior to the conventional soybean callus assay because theresponse (log₁₀ transformed data) was linear, exhibited lowvariability, and was more reproducible and more sensitive. Theassay was quicker to perform and required less cytokinin.     

Linear Relations between Carbon Dioxide Concentration and Rate of Development Towards Flowering in Sorghum, Cowpea and Soyabean

Negative linear relations were detected (P < 0·005)between the rate of progress from sowing to panicle initiationand CO₂ concentration (210-720 µmol CO₂ mol^-1 air) fortwo genotypes of sorghum [Sorghum bicolor (L.) Moench]. Relationsbetween CO₂ concentration and the rate of progress from sowingto first flowering were also negative in soyabean [Glycine max(L.) Merrill] (P < 0·025), but positive in cowpea[Vigna unguiculata (L.) Walp.] (P < 0·025), albeitthat in both grain legumes sensitivity was much less than insorghum. Thus CO₂ elevation does not delay flowering in allshort-day species. The considerable effect of CO₂ concentrationon times to panicle initiation resulted in large differencesamong the sorghum plants at this developmental stage; with increasein CO₂ concentration, plants were taller with slightly moreleaves and more pronounced apical extension. At the same timeafter sowing however, sorghum plants were heavier (P < 0·05)at 210 than at 360 µmol CO₂ mol^-1 air. In contrast, relationsbetween the dry masses of the soyabean and cowpea plants andCO₂ concentration were positive and curvilinear (P < 0·05).It is suggested that the impact of global environmental changecould be severe for sorghum production in the semi-arid tropics.Copyright1995, 1999 Academic Press Sorghum bicolor (L.) Moench., sorghum, Glycine max (L.) Merrill, soyabean, Vigna unguiculata (L.) Walp., cowpea, development, flowering, CO₂, dry matter accumulation, environmental change     

Properties of acid phosphatase in the cell wall of tobacco cells cultured in vitro

A cell-wall fraction containing 10.0% dry weight of proteinwas isolated from tobacco (Nicotiana tabacum L.) XD-6 cellscultured in suspension. The fraction possessed high acid phosphataseactivity toward p-nitrophenyl phosphate, phenyl phosphate, inorganicpyrophosphate, adenosine diphosphate, nucleoside triphosphates,and bis- (p-nitrophenyl) phosphate; much less activity towardnaphthyl phosphate, ß-glycerophosphate, glucose-6-phosphate,fructose-6-phosphate, fructose-l,6-diphosphate, thiamine pyrophosphate;and no activity toward glucose-1-phosphate and inositol hexaphosphate.Metallic ions were not required for activation. The enzyme wasextracted from the wall with NaCl solution. The solubilizedenzyme resulted in a single peak on Sephadex G-150 chromatographywith activity toward p-nitrophenyl phosphate, pyrophosphate,and bis(p-nitrophenyl) phosphate. The activity of the solubilizedenzyme toward p-nitrophenyl phosphate was identical with thebound enzyme in its pH optimum, substrate specificity, ion inhibitionand Km value, but it was more sensitive to pH, substrate difference,inhibition, and heat treatment. ¹Present address: Department of Biology, Japan Women's University,Mejiro-dai, Bunkyo-ku, Tokyo, Japan (Received September 13, 1972; )     

Normalizing Development of Cultured Triticum aestivum L. Embryos: I. LOW OXYGEN TENSIONS AND EXOGENOUS ABA

Wheat (Triticum aestivum L.) embryos form in dynamically-regulatedovular environments. Our objectives were to improve developmentof cultured immature wheat embryos by simulating, in vitro,abscisic acid (ABA) levels and O₂ tensions as found in wheatovules during zygotic embryogenesis. We characterized from intactwheat kernels embryo respiration, embryo morphology and embryoand endosperm + ABA levels at 13, 19 and 25 d post-anthesis(DPA). Young (13 DPA) embryos were then excised and culturedin vitro, where they were exposed to 0·2 or 2·Ommol m^–3 ±ABA and 2.·1, 2·5 or 7·4mol m^–3 (6, 7 and 21%, respectively) gaseous O₂. At 6and 12 d in culture, + ABA levels, embryo respiration and embryomorphology were characterized by treatment. Thirteen-day-oldembryos from two different plant populations differed by 17-foldin initial ABA content. However, this difference did not affectprecocious germination in vitro, nor did it affect the amountof exogenous ABA required to reduce precocious germination by40%. In this respect, embryos from both populations were equallysensitive to exogenous ABA. Cavity sap O₂ levels (2·1to 2·5 mol m^–3) were much more effective in preventingprecocious germination of cultured embryos than were cavitysap levels of ABA (0·2 to 2·0 mmol m^–3).The combination of physiological levels of both ABA and O₂ largelynormalized DW accumulation and embryo morphology without alteringendogenous + ABA levels. Residual respiration of cultured embryoswas higher than that of embryos grown in situ, and was not influencedby the exogenous O₂ and ABA treatments Key words: Abscisic acid, embryo development, oxygen tensions, respiration, wheat     

Diffusion of Oxyleghaemoglobin

The diffusion of oxyleghaemoglobin, prepared from soybean rootnodules, was measured at 24°C in agar and agarose gels ofvarious strengths, or in 1% agarose containing 0-18% (w/v) bovineserum albumin, to simulate the protein content of the cytoplasmof root nodule cells. Values of D_p, the diffusion coefficient,were unaffected (D_p = 11·8 x 10^-11 m² s^-1; s.e.m. 0·3x 10^-11) until the protein concentration exceeded 6%, abovewhich D_p declined sharply. With 18% bovine serum albumin, theconcentration of total soluble protein calculated to be presentin the cytoplasm of infected cells, where most of the leghaemoglobinis located in vivo, D_p was 5·9 x 10^-11 m² s^-1. Theseresults are discussed in relation to leghaemoglobin-facilitateddelivery of O₂ to the respiring N₂-fixing bacteroids in rootnodule cells.Copyright 1993, 1999 Academic Press Bacteroids, diffusion, Glycine max, N₂ fixation, oxyleghaemoglobin, soybean, root nodules     

Photosynthesis of Stands of Tomato, Cucumber and Sweet Pepper Measured in Greenhouses under Various CO2-concentrations

The rate of net photosynthesis (P) of whole plant stands oftomato (Lycopersicon esculentum Mill.), cucumber (Cucumis sativusL.) and sweet pepper (Capsicum annuum L.) was measured in sixlong-term experiments in large greenhouses under normal operatingconditions and CO₂-concentrations between 200 and 1200 µmolmol^-1. The objective was to quantify the responses to lightand carbon dioxide and to obtain data sets for testing simulationmodels. The method of measuring canopy photosynthesis involvedan accurate estimation of the greenhouse CO₂ balance, usingnitrous oxide (N₂O) as tracer gas to determine, on-line, theexchange rate between greenhouse and outside air. The estimatedrelative error in the observed P was about ± 10%, exceptthat higher relative errors could occur under particular conditions. A regression equation relating P to the photosynthetically activeradiation, the CO₂ concentration and the leaf area index explained83-91% of the variance. The main canopy photosynthesis characteristicscalculated with the fitted regression equations were: canopyP_max 5-9 g m^-2 h^-1 CO₂ uptake; ratio P_max/LAI 1·5-3 gm^-2 h^-1; light compensation point 32-86 µmol s^-1 m^-2;light use efficiency (quantum yield) at low light 0·06-0·10µmol µmol^-1 and CO₂ compensation point 18-54 µmolmol^-1. The results were related to the prevailing conditions.Copyright1994, 1999 Academic Press Canopy photosynthesis, Capsicum annuum L., carbon dioxide, CO₂, CO₂ balance, CO₂ use efficiency, cucumber, _{Cucumis sativus} L., glasshouse, greenhouse, light use efficiency, Lycopersicon esculentum Mill., sweet pepper, tomato, tracer gas     

Enhancement of Gibberellic Acid-stimulated Hypocotyl Elongation of Lettuce (Lactuca sativa L. cv. Grand Rapids) by Phlorizin and Light

The interaction of light, gibberellic acid (GA₃), and phlorizinin the growth of lettuce (Lactuca sativa L. cv. ‘GrandRapids’) hypocotyls was investigated. At all concentrationsof GA₃, phlorizin enhanced GA₃-induced growth at luminous intensitiesabove 50 ft-c (continuous light). Without GA₃, phlorizin hadno effect on hypocotyl growth in the light but it inhibitedgrowth in the dark. Both seedlings and hypocotyl sections respondedto phlorizin in the presence of GA₃. There was no iteractionbetween phlorizin and KCl. Water-growth was severly inhibitedby light. GA₃,-induced growth was slightly inhibited by light,and then only at luminous intensities above 50 ft-c. Thus, relativeto H₂O-growth, GA₃-induced growth increased with increasingluminous intensity up to 450 ft-c, where it reached saturation.It seems that a synergism may exist between light and GA₃ aswell as between phlorizin and GA₃. Lactuca sativa L, lettuce, hypocotyl elongation, gibberellic acid, phlorizin, light     

Properties of peroxidases from tobacco cell suspension culture

An enzyme preparation from suspension cultured tobacco cells oxidized IAA only in the presence of added cofactors, Mn²⁺ and 2,4-dichlorophenol, and showed two pH optima for the oxidation at pH 4·5 and 5·5. Effects of various phenolic compounds and metal ions on IAA oxidase activity were examined. The properties of seven peroxidase fractions separated by column chromatography on DEAE-cellulose and CM-Sephadex, were compared. The peroxidases were different in relative activity toward o-dianisidine and guaiacol. All the peroxidases catalysed IAA oxidation in the presence of added cofactors. The pH optima for guaiacol peroxidation were very similar among the seven isozymes, but the optima for IAA oxidation were different. The anionic and neutral fractions showed pH optima near pH 5·5, but the cationic isozymes showed optima near pH 4·5. With guaiacol as hydrogen donor, an anionic peroxidase (A-1) and a cationic peroxidase (C-4) were very different in H₂O₂ concentration requirements for their activity. Peroxidase A-1 was active at a wide range of H₂O₂ concentrations, while peroxidase C-4 showed a more restricted H₂O₂ requirement. Gel filtration and polyacrylamide gel studies indicated that the three cationic peroxidases have the same molecular weight.     

The isozymic nature and kinetic properties of glutamate dehydrogenase from safflower seedlings

Levels of glutamate dehydrogenase (GDH) [L-glutamate: NAD oxidoreductase(deaminating), EC 1.4.1.2 [EC] ] from safflower roots and cotyledonsincreased (?2.7) and decreased ( ?5.7), respectively, as a functionof seedling age. No significant changes in enzyme levels weredetected during hypocotyl development. GDH preparations of thedifferent organs were resolved by polyacrylamide gel electrophoresisinto 2 to 4 isozymes. The isozymic pattern was influenced byseedling age and organ tested. The slowest moving isozyme (No.1) appears to be responsible for the changes in GDH levels observedin cotyledons and roots. We isolated isozyme 1 and GDH fractionchiefly containingisozyme 2, by DEAE-cellulose chromatography. GDH was purified approximately 53-fold from the particulatefraction of cotyledons. The pH optima for NADH and NAD activitieswere 8.2 and 8.9, respectively. Michaelis constants were foundto be: -ketoglutarate, 8mM; glutamate, 4 mM; ammonium, 35.4mM; NAD, 0.26 mM; NADH, 0.065 mM. Km values of isozymes 1 and2 were similar. The binding order of substrates in die reductiveamination reaction was NADH, -ketoglutarate and NH₄⁺. (Received July 17, 1972; )     

Effect of Sodium on Phosphate Uptake in Unicellular and Filamentous Cyanobacteria

The effect of Na⁺ on phosphate uptake was studied in four strainsof cyanobacteria: Synechococcus PCC 7942, Gloeothece PCC 6501,Phormidium sp. and Chlorogloeopsis PCC 6912. Phosphate uptakewas stimulated by Na⁺ in all cases. Li⁺ and K⁺ acted as partialanalogues for Na⁺. Half-saturation [K_1/2(Na⁺)] of phosphateuptake was reached with Na⁺ concentrations ranging from 317µM in Chlorogloeopsis to 659 µM in Phormidium. Theconcentration of phosphate required to reach half-saturationof phosphate uptake [K_1/2(P_i)]was not changed by the presenceof Na⁺. (Received April 11, 1994; Accepted July 5, 1994)     

Flavine nucleotide nitrate reductase from broad bean leaves

Hydrosulfite-reduced FMN served as an electron donor for nitratereductase purified from broad bean leaves. FMN was successfullyreplaced with BV. The flavine nucleotide nitrate reductase hadits pH optima at about 7.8 with phosphate buffer and at about7.4 with Tris-HCl buffer. The Km's for nitrate and FMN were3.7 ? 10^–4 M and 3.7 ? 10^–5 M, respectively. NADH₂: nitrate reductase activity was completely inhibited by0.1 mM p-CMB, whereas FMNH₂: nitrate reductase activity wasnot. Inhibited activity was restored by the addition of cysteine.A sulfhydryl enzyme is involved in the NADH₂: nitrate reductasesystem but not in the FMNH₂ : nitrate reductase system. NADH₂and FMNH₂ probably feed electrons into the electron transportchain at different sites. The nitrate reductase preparationhad an NADH₂-specific diaphorase activity which was almost completelyinhibited by 0.1 mM p-CMB. The NADH₂-specific diaphorase mayform the sulfhydryl enzyme which mediates electron transferbetween NADH₂ and nitrate. (Received May 6, 1969; )