Effect of 1-Aminocyclopropane-1-Carboxylic Acid on the Production of Ethylene in Senescing Flowers of Ipomoea tricolor Cav

Application of 1-aminocyclopropane-1-carboxylic acid (ACC) to rib segments excised from flowers of Ipomoea tricolor Cav. resulted in the formation of C₂H₄ in greater quantities than produced under natural conditions. The ability of ACC to enhance C₂H₄ production was independent of the physiological age of the tissue and its capacity to synthesize C₂H₄ without applied ACC. When ACC was fed to rib segments that had been treated with [¹⁴C]methionine, incorporation of radioactivity into C₂H₄ was reduced by 80%. Aminoethoxyvinylglycine and aminooxyacetic acid inhibited C₂H₄ production in rib segments of I. tricolor but had no effect on ACC-enhanced C₂H₄ production. Protoplasts obtained from flower tissue of I. tricolor did not form C₂H₄, even when incubated with methionine or selenomethionine. They produced C₂H₄ upon incubation with ACC, however. ACC-dependent C₂H₄ production in protoplasts was inhibited by n-propyl gallate, AgCl, CoCl₂, KCN, Na₂S, and NaN₃. ACC-dependent C₂H₄ synthesis in rib segments and protoplasts was dependent on O₂, the K_m for O₂ being 1.0 to 1.4% (v/v). These results confirm the following pathway for C₂H₄ biosynthesis in I. tricolor. methionine [selenomethionine] → S-adenosylmethionine [selenoadenosylmethionine] → ACC → C₂H₄.     

Acetylene Metabolism and Stimulation of Denitrification in an Agricultural Soil

The effects of C₂H₂ metabolism on N₂O production were examined in soil slurries. Enrichment of C₂H₂ consumption activity occurred only in aerobic incubations. Rapid disappearance of subsequent C₂H₂ additions, stimulation of CO₂ production, and most-probable-number enumerations of C₂H₂ utilizers indicated enrichment of the population responsible. During C₂H₂ consumption in slurries incubated statically under air, maximal rates of N₂O evolution were 19 times higher than those in anaerobic incubations. After 20 days of enrichment with C₂H₂, the production of N₂O by slurries supplemented with C₂H₂ and nitrate was 10 times higher than that in the unenriched controls. A Nocardia- or Arthrobacter-like bacterium was isolated that grew on C₂H₂ but did not denitrify. The behavior of soil inoculated with this bacterium became similar to that of C₂H₂-enriched soil incubated aerobically. Ethanol, acetate, and acetaldehyde were identified in enrichment experiments, and denitrification in soil slurries was stimulated by addition of the supernatant from a pure culture grown on mineral medium with C₂H₂. These results indicate that denitrification can be stimulated by the actions of an aerobic, nondenitrifying C₂H₂-metabolizing population. Utilization of intermediate metabolites by denitrifiers and enhanced O₂ consumption are two possible mechanisms for this stimulation.     

Ethylene Production and Leaflet Abscission in Mèlia azédarach L

Ethylene production or content was compared to leaflet abscission in detached, compound leaves of Mèlia azédarach L. In late autumn, when abscission was progressing from basal leaves upward, the oldest leaves both produced ethylene at the highest rates and abscised their leaflets first. When C₂H₄ levels were measured in intercellular air removed immediately after leaves were harvested, C₂H₄ levels were also highest in basal leaves and declined progressively in more apical leaves. Levels as high as 1.8 microliters C₂H₄ liter⁻¹ air were observed. Earlier in the season groups of leaves demonstrated a pattern of sequential initiation of abscission from base to apex, but the peak rates of C₂H₄ production followed an opposite trend, being highest in the youngest leaves. Peak rates of C₂H₄ production occurred after the initiation of leaflet abscission and presumably are related to either the auxin content or a climacteric-like, autocatalytic phase of C₂H₄ production not directly involved in the initiation of abscission. In these experiments, the early abscission of the older leaflets reflects their greater sensitivity to C₂H₄, presumably due to lower auxin content. C₂H₄ production rates in all experiments, with rare exceptions, exceeded 3 microliters per kilogram fresh weight per hour at least 24 hours before leaflet abscission reached 10%. This achieving of a threshold internal C₂H₄ level is viewed as an initiating event in leaflet abscission. Hypobaric conditions, to facilitate the escape of endogenous C₂H₄, delayed abscission compared to controls, and termination of hypobaric exposure allowed a normal progression of abscission as well as normal C₂H₄ synthesis rates. All of the data indicate that C₂H₄ initiates leaflet abscission in intact but detached leaves of Mèlia azédarach L. The seasonal patterns observed suggest that C₂H₄, in concert with those hormones which govern sensitivity to C₂H₄, regulate autumn leaf fall in this species.     

Inhibition of the Conversion of 1-Aminocyclopropane-1-carboxylic Acid to Ethylene by Structural Analogs, Inhibitors of Electron Transfer, Uncouplers of Oxidative Phosphorylation, and Free Radical Scavengers

Cyclopropane carboxylic acid (CCA) at 1 to 5 millimolar, unlike related cyclopropane ring analogs of 1-aminocyclopropane-1-carboxylic acid (ACC) which were virtually ineffective, inhibited C₂H₄ production, and this inhibition was nullified by ACC. Inhibition by CCA is not competitive with ACC since there is a decline, rather than an increase, in native endogenous ACC in the presence of CCA. Similarly, short-chain organic acids from acetic to butyric acid and α-aminoisobutyric acid inhibited C₂H₄ production at 1 to 5 millimolar and lowered endogenous ACC levels. These inhibitions, like that of CCA, were overcome with ACC. Inhibitors of electron transfer and oxidative phosphorylation effectively inhibited ACC conversion to C₂H₄ in pea and apple tissues. The most potent inhibitors were 2,4-dinitrophenol (DNP) and carbonyl cyanide m-chlorophenylhydrazone (CCCP) which virtually eliminated ACC-stimulated C₂H₄ production in both tissues. Still other inhibitors of the conversion of ACC to C₂H₄ were putative free radical scavengers which reduced chemiluminescence in the free radical-activated luminol reaction. These inhibitor studies suggest the involvement of a free radical in the reaction sequence which converts ACC to C₂H₄. Additionally, the potent inhibition of this reaction by uncouplers of oxidative phosphorylation (DNP and CCCP) suggest the involvement of ATP or the necessity for an intact membrane for C₂H₄ production from ACC. In the latter case, CCCP may be acting as a proton ionophore to destroy the membrane integrity necessary for C₂H₄ production.     

Variation in nitrogenase and hydrogenase activity of alaska pea root nodules

Hydrogenase activity of root nodules in the symbiotic association between Pisum sativum L. and Rhizobium leguminosarum was determined by incubating unexcised nodules with tritiated H₂ and measuring tissue HTO. Hydrogenase activity saturated at 0.50 millimolar H₂ and was not inhibited by the presence of 0.10 atmosphere C₂H₂, which prevented H₂ evolution from nitrogenase. Total H₂ production from nitogenase was estimated as net H₂ evolution in air plus H₂ exchange in 0.10 atmosphere C₂H₂. Although such an estimate of nitrogenase function may not be quantitatively exact, due to uncertain relationships between H₂ exchange and H₂ uptake activity of hydrogenase, differences observed in H₂ exchange under various conditions represent an indication of changes in hydrogenase activity. Hydrogenase activity was lower in associations grown under higher photosynthetic photon flux densities and decreased relative to total H₂ production by nitrogenase. Total H₂ production and hydrogenase activity were maximum 28 days after planting. Thereafter, hydrogenase activity and H₂ production declined, but the potential proportion of nitrogenase-produced H₂ recovered by the uptake hydrogenase system increased. Of five R. leguminosarum strains tested two possessed hydrogenase activity. Strains which had the potential to reassimilate H₂ had significantly higher rates of N₂ reduction than those which did not exhibit hydrogenase activity.     

Effect of gamma radiation on the ripening of bartlett pears

Gamma radiation at doses of 300 Krad or more inhibits the ripening of Bartlett pears (Pyrus communis L.). Immediately after irradiation there is a transitory burst of C₂H₄, which subsequently declines in fruits subjected to inhibitory doses. Ethylene production associated with ripening begins at the same time in unirradiated fruits and those subjected to noninhibitory doses, but the latter produces much more C₂H₄ at the climacteric peak. Fruits subjected to inhibitory doses produce low levels of C₂H₄ unless subjected to exogenously applied C₂H₄, whereupon they produce enough of the gas to induce ripening in unirradiated fruits.

Pears subjected to 300 and 400 Krad of gamma rays did not ripen even when held in a flowing atmosphere containing 1000 ppm of C₂H₄ for 8 days at 20°. It is concluded that the action of gamma rays on Bartlett pears involves both an inhibition of C₂H₄ production and a decreased sensitivity of the fruit to the ripening action of the gas. Ripening of Bartlett pears is inhibited by gamma radiation only when applied to preclimacteric fruit.

     

Use of a laser-driven photoacoustic detection system for measurement of ethylene production in cymbidium flowers

A laser-based photoacoustic method was used for determination of ethylene (C₂H₄) production of emasculated orchid (Cymbidium) flowers in a flow-through system. The laser photoacoustic equipment consisted of a line-tuneable CO₂ laser in conjunction with a single-pass resonant acoustic cell. The minimum detection limit of the system for C₂H₄ in air was 0.03 nanoliter per liter. C₂H₄ production of intact Cymbidium (cv Mary Pinchess `Del Rey') flowers was very low (0.015 nanoliter per gram per hour) and showed an increase within 3 hours following emasculation (removal of pollinia plus anthercap). Production peaked (0.14 nanoliter per gram per hour) 8 hours after emasculation and decreased thereafter. Production again increased 45 hours after emasculation. Coloration of the labellum appeared shortly after the first peak; wilting of the petals and sepals appeared during the second rise in ethylene production. The use of the laser photoacoustic technique in plant physiological studies is discussed.     

Biohydrogen production in a three-phase fluidized bed bioreactor using sewage sludge immobilized by ethylene–vinyl acetate copolymer

Ethylene–vinyl acetate (EVA) copolymer was used to immobilize H₂-producing sewage sludge for H₂ production in a three-phase fluidized bed reactor (FBR). The FBR with an immobilized cell packing ratio of 10% (v/v) and a liquid recycle rate of 5 l/min (23% bed expansion) was optimal for dark H₂ fermentation. The performance of the FBR reactor fed with sucrose-based synthetic medium was examined under various sucrose concentration (C_so) and hydraulic retention time (HRT). The best volumetric H₂ production rate of 1.80 ± 0.02 H₂ l/h/l occurred at C_so = 40 g COD/l and 2 h HRT, while the optimal H₂ yield (4.26 ± 0.04 mol H₂/mol sucrose) was obtained at C_so = 20 g COD/l and 6 h HRT. The H₂ content in the biogas was stably maintained at 40% or above. The primary soluble metabolites were butyric acid and acetic acid, as both products together accounted for 74–83% of total soluble microbial products formed during dark H₂ fermentation.     

Increase in Dry Weight and Total Nitrogen Content in Zea mays and Setaria italica Associated with Nitrogen-fixing Azospirillum spp

The association between nitrogen-fixing bacteria from the genus Azospirillum and the grasses Zea mays and Setaria italica was investigated in sterilized Leonard-jar assemblies. Nitrogen-fixing bacteria isolated from Cynodon dactylon roots in Israel and Azospirillum brasilense (Sp-7, Sp-80, and Cd) were examined. C₂H₂ reduction activity was detected in systems containing 0.0 to 0.08 but not in those containing 0.16 gram per liter NH₄NO₃. The organisms tested significantly increased plant dry weight (50-100%), total N content of leaves (50-100%) and C₂H₄ production (300-1000 nanomoles C₂H₄ per plant per hour). Highest C₂H₂ reduction activities were obtained above 30 C and with high light intensities. Significant increases in S. italica dry weight (DW) and nitrogen (N) content were observed in sand (DW = 80%, N = 150%), sandy loam soil (DW = 80%, N = 75%) and loess (DW = 37%, N = 25%). The results obtained in this work clearly demonstrate the potential benefit of inoculating grasses with Azospirillum.     

Does Ethylene Play a Role in the Release of Lateral Buds (Tillers) from Apical Dominance in Oats?

The growth of lateral buds (tillers), which are undergoing release from apical dominance, was measured in upright and gravistimulated intact Avena sativa L. cv. `Victory' (oat) shoots as well as in isolated Avena stem segments treated with kinetin and sucrose. During release, the tiller bud initially shows a slow rate of elongation accompanied by swelling. It is followed by a more rapid rate of elongation. Ethylene (C₂H₄) production in shoot segments containing a tiller bud was found to occur at the onset of tiller swelling during gravistimulation as well as during inflorescence emergence. Exogenous application of indoleacetic acid or C₂H₄ inhibits kinetin-induced tiller bud swelling and elongation. However, stem segments pulsed for 24 hours in C₂H₄ or the C₂H₄ biosynthesis precursor, 1-amino-cyclopropane-1-carboxylic acid (ACC) and then transferred to kinetin and sucrose, showed a significant increase in swelling elongation as compared with segments maintained under the same conditions but without C₂H₄ or ACC in the pulse. Segments pulsed for 24 hours with kinetin and sucrose plus the ACC biosynthesis inhibitor, aminoethoxyvinylglycine, or the C₂H₄ action inhibitor, CO₂, then transferred to kinetin and sucrose medium, showed inhibition of tiller swelling during the pulse and of subsequent elongation. These results indicate that C₂H₄ plays a role in promoting tiller swelling during the onset of tiller release from apical dominance and may act as a modulator hormone in promoting tiller elongation in the presence of cytokinin.     

Comparing Time Course Profiles of Immediate Acetylene Reduction by Grasses and Legumes

The time course profiles of C₂H₂ reduction by intact Scirpus olneyi (bulrush), Oryza sativa (rice) and Spartina alterniflora (cordgrass) with roots in atmospheres of N₂ and 30-day-old Glycine max (soybean) in air were all immediately linear. This is the first report of immediately linear rates of C₂H₂ reduction by grass roots removed from soil. The immediately linear profile of C₂H₂ reduction by soil-free grass roots was achieved by preventing contact between the roots and air. Roots of soybeans and S. olneyi receiving pretreatments of O₂ above normal environmental levels for 15 min before assay exhibited a short delay in C₂H₂ reduction. These initially nonlinear rates of C₂H₂ reduction are attributable to transient O₂ inhibition of nitrogenase. Initial nonlinear rates of C₂H₂ reduction were also observed with immature soybean plants and with intact plant assays of O. sativa and S. olneyi in which C₂H₂ was injected into cylinders surrounding the plant tops. These results indicate that, apart from O₂ inhibition of nitrogenase, the diffusion of C₂H₂ and C₂H₄ between the nitrogen-fixing sites and the sampling ports may cause initial nonlinear rates of C₂H₂ reduction. We conclude that in situ plant-associated nitrogenase activity should result in immediate reduction of C₂H₂ and that linear rates are observed when the proper assay conditions are used. Our data suggest that nitrogen fixation is closely associated with the roots of S. olneyi, O. sativa, and S. alterniflora growing in salt marsh sediment.     

Novel ruthenium(II) diamine complexes

The complex cations [Ru(C₇H₁₆N₂)(C₁₀H₁₄)Cl]⁺, [Ru(C₇H₁₆N₂)(C₆H₆)Cl]⁺, [Ru(C₉H₁₈N₂)(C₆H₆)Cl]⁺, [Ru(C₉H₁₈N₂)(C₁₀H₁₄)Cl]⁺ and [Ru(C₁₄H₁₆N₂)(C₁₀H₁₄)Cl]⁺ have been synthesised from the reaction between the ruthenium-arene complexes [with C₆H₆ (benzene) or C₁₀H₁₄ (p-cymene)] and the respective chiral diamines [C₇H₁₆N₂=(S)-(−)-2-aminomethyl-1-ethylpyrrolidine, C₉H₁₈N₂=(S)-(+)-2-(pyrrolidinylmethyl)-pyrrolidine, or C₁₄H₁₆N₂=(1R,2R)-(+)-1,2-diphenylethylenediamine], isolated and characterised as chloride salts using single-crystal X-ray diffraction. All complexes were fully characterised by elemental analysis, mass spectrometry, ¹³C and ¹H NMR, and also found to exhibit catalytic activity in the transfer hydrogenation of acetophenone to 1-phenylethanol at 50 °C (enantiomeric excesses range from ca. 25% to 60%, and conversions from ca. 30% to 50%).     

Azolla-Anabaena azollae Relationship: V. N(2) Fixation, Acetylene Reduction, and H(2) Production

In order to characterize the reactions catalyzed by nitrogenase in the Azolla-Anabaena association, ¹⁵N₂ fixation, C₂H₂ reduction, and ATP-dependent H₂ production were measured in both the Azolla-Anabaena complex and in the alga isolated from the complex.     

Ethylene biosynthesis: Methionine as an in-vivo precursor of ethylene in auxin-treated mungbean hypocotyl segments

Summary Ethylene production was induced in excised hypocotyl segments of etiolated mungbean seedlings in response to exogenous auxin. 1,2,3,4-¹⁴C-Methionine was efficiently incorporated into C₂H₄, although cold methionine added at substrate level did not enhance C₂H₄ production. Incorporation of labeled glucose into C₂H₄ was reduced when hypocotyl segments were incubated with cold methionine and homoserine, but the rate of labeling of CO₂ was not affected. Feeding of labeled glucose to segments resulted in production of labeled methionine both in the presence and the absence of auxin, and auxin did not affect rate of methionine synthesis from glucose. The decrease in the amount of endogenous methionine during auxin treatment approximated the amount of C₂H₄ produced. The timecourse pattern of incorporation of radioactivity from labeled methionine into C₂H₄ during removal or re-addition of auxin was very similar to that of C₂H₄ production. The role of endogenous methionine as a C₂H₄ precursor is discussed.     

Role of uptake hydrogenase in providing reductant for nitrogenase in Rhizobium leguminosarum bacteroids

The role of uptake hydrogenase in providing reducing power to nitrogenase was investigated in Rhizobium leguminosarum bacteroids from nodules of Pisum sativum L. (cv. Homesteader). H₂ increased the rate of C₂H₂ reduction in the absence of added substrates. Malate also increased nitrogenase (C₂H₂) activity while decreasing the effect of H₂. At exogenous malate concentrations above 0.05 mM no effect of H₂ was seen. Malate appeared to be more important as a source of reductant than of ATP. When iodoacetate was used to minimize the contribution of endogenous substrates to nitrogenase activity in an isolate in which H₂ uptake was not coupled to ATP formation, H₂ increased the rate of C₂H₂ reduction by 77%. In the presence of iodoacetate, an ATP-generating system did not enhance C₂H₂ reduction, but when H₂ was also included, the rate of C₂H₂ reduction was increased by 280% over that with the ATP-generating system alone. The data suggest that, under conditions of substrate starvation, the uptake hydrogenase in R. leguminosarum could provide reductant as well as ATP in an isolate in which the H₂ uptake is coupled to ATP formation, to the nitrogenase complex.     

Effect of carbon dioxide and ethylene on tuberization of isolated potato stolons cultured in vitro

Carbon dioxide stimulates tuberization of isolated potato (Solanum tuberosum L.) stolons cultured in vitro. The stimulatory effect is inhibited by C₂H₄ which is by itself also inhibitory of tuberization. Furthermore, C₂H₄ inhibits kinetin-induced tuber initiation. Both the formation and elongation of roots are inhibited by C₂H₄. The antagonistic actions of CO₂ and C₂H₄ on tuberization are discussed.     

AN EXPERIMENTAL SEPARATION OF OXYGEN LIBERATION FROM CARBON DIOXIDE FIXATION IN PHOTOSYNTHESIS BY CHLORELLA

Using intact cells of Chlorella pyrenoidosa it is possible to obtain oxygen by the reduction of certain reducible materials other than carbon dioxide. Of these, benzaldehyde was studied in some detail. This reduction does not involve the production of carbon dioxide from the benzaldehyde. Stoichiometrical relationships as expressed by the following equation: 2C₆H₅CHO + 2H₂O → 2C₆H₅CH₂OH + O₂ are somewhat difficult to obtain because the benzaldehyde can disappear from the reaction mixtures by dark reactions. The technique is now available which permits detailed studies of the oxygen-liberating mechanisms in photosynthesis.     

A convenient synthesis of meso-substituted porphyrins

Acid-catalyzed condensations of 1,19-diunsubstituted 1,19-dideoxybiladiene-ac dihydrobromides with aldehydes, R · CHO, afford the corresponding meso-substituted porphyrins (R = C₆H₅, p-Me·C₆H₄, p-MeO·C₅H₄, p-O₂N·C₆H₄, p-HOC·C₆H₄, p-(MeO)₂CH·C₆H₄, Me, n-Pr, CO₂Et), mostly in good yield.     

Biohydrogen production in alkalithermophilic conditions: Thermobrachium celere as a case study

In the present work the hydrogenesis in the anaerobic alkalithermophilic bacterium Thermobrachium celere was studied. The impact of several factors on hydrogen production during glucose fermentation was investigated in batch conditions. The optimal hydrogen production occurred at pH_67 °C 8.2 with phosphate buffer concentration of 50 mM. Hydrogen yield reached the highest value of 3.36 mol H₂/mol glucose when the partial pressure in the gas headspace was reduced. Supplementation of nitrogen sources and iron affected hydrogen production. Under optimized conditions, the maximum H₂ accumulation and H₂ production rate were estimated to be respectively 124.3 mmol H₂/l culture and 20.7 mmol H₂/l/h. Considering the efficient and rapid hydrogen evolution, and the ability to grow in extreme environments, T. celere might be a good candidate for biohydrogen production in open (non-sterile) bioprocess system.     

m-Terphenylphosphines: Synthesis, structures and coordination properties

A series of m-terphenylphosphines TerphPCl₂, TerphPH₂ and TerphPMe₂ (Terph = 2,6-Mes₂C₆H₃-, 2,6-(4-t-BuC₆H₄)₂C₆H₃-, 2,6-(3,5-Me₂C₆H₃)₂C₆H₃-, 2,6-(2,6-Et₂C₆H₃)₂C₆H₃-, and 2,6-(2,6-i-Pr₂C₆H₃)₂C₆H₃-; Mes = 2,4,6-Me₃C₆H₂-) was prepared and fully characterized. The structural investigation by X-ray crystallography and density functional theory revealed significant distortions in the environment of the ipso carbon and phosphorus centers. These can be traced back to steric interactions and repulsions of the chlorine and methyl substituents on phosphorus with one of the flanking arenes of the m-terphenyl substituents. The primary phosphine 2,6-Mes₂C₆H₃PH₂, 6, and the dimethylphosphine 2,6-(3,5-Me₂C₆H₃)₂C₆H₃PMe₂, 9, readily form complexes with the Cl₂Ru(p-cymene) complex fragment, whereas the larger phosphine 2,6-Mes₂C₆H₃PMe₂, 8, does not. Heating of the complexes TerphPR₂Ru(Cl₂)(p-cymene) 11 and 12 and the mixture of 8 and {(p-cymene)RuCl₂}₂ lead to expulsion of the p-cymene ligand and intramolecular η⁶ coordination of one of the flanking arene rings to the ruthenium center to afford the complexes Cl₂RuP(H₂)C₆H₃-2-η⁶-Mes-6-Mes, 13, Cl₂RuP(Me₂)C₆H₃-2-η⁶-Mes-6-Mes, 14, and Cl₂RuP(H₂)C₆H₃-2-η⁶-(3,5-Me₂C₆H₃)-6-(3,5-Me₂C₆H₃), 15. All complexes were characterized by NMR spectroscopy and complexes 14 and 15 also by X-ray crystallography.