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1.
The molecular mechanism of membrane-associated reactions induced by auxin was investigated in membranes isolated from cultured cells of soybean (Glycine max L.). Auxins increased the activity of phospholipase A2 in microsomes isolated from suspensioncultured soybean cells. The reaction was measured as the accumulation of radioactive lysophosphatidylcholine hydrolyzed from radioactive phosphatidylcholine in membranes which had been prelabelled with [14-C]choline in vivo. Stimulation by auxin was detectable after 1 min and was auxin-specific in that weak auxins had little effect. Auxin concentrations as low as 2·10–8 M and up to 2·10+3 M -naphthaleneacetic acid already stimulated the phospholipase A2 activity. Guanosine and adenosine diphosphate at 100 M, if applied during homogenization of cells, completely abolished the stimulation of phospholipase A2 by auxin and, when applied after homogenization, had no effect. Guanosine and adenosine 5-O-thiotriphosphate, uridine 5-diphosphate, and uridine 5-triphosphate, all at 100 M, had no effect in either treatment, suggesting that only nucleotides entrapped in the vesicles could exert an effect. The effect of auxin on phospholipase A2 had an optimum at pH 5.5 and was abolished completely by an antibody against the membrane-associated auxin-binding protein from maize coleoptiles, applied after homogenization. This antibody recognized a 22-kDa polypeptide in highly purified plasma membranes from cultured soybean cells. This suggests a receptor function for this auxin-binding protein and a role for a cytosolic nucleotide-binding protein in the activation of phospholipase A2 by auxin. It is concluded that phospholipase A2 has a function in plant signal transduction.Abbreviations ABP auxin-binding protein - ATP S adenosine 5-O-thiotriphosphate - 2,4-D 2,4-dichlorophenoxyacetic acid - GTP S guanosine 5-O-(thiotriphosphate) - IgG immunoglobulin G - LPC lysophosphatidylcholine; - -NAA , -naphthaleneacetic acid - PLA2 phospholipase A2 We cordially acknowledge the gift of anti-ABP antibody by D. Klämbt and the help by H. Ordowski (both Botanisches Institut, Universität Bonn) with the immunoblotting experiments. This work was supported by the Deutsche Forschungsgemeinschaft.  相似文献   

2.
Using an immunoblotting technique and goat antihuman C4, we observed five distinct electrophoretic variants of C4 in a panel of 60 random dogs. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of immunoprecipitated C4 showed that dog C4 is composed of three polypeptide subunit chains (, , and ) and that structural variability occurs within the - and -chain regions. Two distinct molecular weight forms of both the C4- ( A and B) and C4-( A and B) chain were detected. The variant forms of C4 and C4 were found in association with particular C4 allotypes.  相似文献   

3.
Summary The spacing between adjacent hairs in vegetative whorls ofAcetabularia acetabulum (formerlyA. mediterranea) was earlier reported as being quantitatively responsive to calcium ion concentration in the culture medium. We here report a quantitative response to the concentration of the calcium-chelator EGTA, in the opposite sense to the effect of calcium. (Increasing [Ca2+] diminishes the spacing; increasing [EGTA] increases it.) The earlier work was interpreted in terms of control of the spacing by a putative reaction-diffusion mechanism in the cell membrane, in which a receptor R was activated by calcium-binding to initiate the process. We extend this interpretation by treating CaEGTA as an uncompetitive inhibitor of the effect of calcium on R. This leads to thermodynamic constants for CaEGTA binding to the CaR complex: H 298 0 =–250 ± 60 kJ/mol; S 298 0 =–820 ± 200 J/mol · K. Consistency of the concentration and temperature dependences reported here with the postulated dynamic mechanism increases the probability that this mechanism is correct.Abbreviations EGTA ethylene glycol bis(-aminoethylether)N,N,N,N-tetraacetic acid - SHEP Shephard's artificial sea-water medium - R postulated membrane-bound calcium receptor - spacing between adjacent hairs in a whorl at first morphological appearance, identified with wavelength in a reaction-diffusion pattern - A, B reactants (morphogen precursors) in a reaction-diffusion mechanism - X, Y reaction intermediates (morphogens) in a reaction-diffusion mechanism  相似文献   

4.
In the present study it was investigated if a-tocopherol shows protection against in vitro lipid peroxidation of phospholipids located in rod outer segment membranes (ROS). After incubation of ROS in an ascorbate-Fe2+ system, at 37°C during 160 min, the total cpm originated from light emission (chemiluminescence) was found to be lower in those membranes incubated in the presence of -tocopherol. The fatty acid composition of total lipids isolated from rod outer segment membranes was substantially modified when subjected to non-enzymatic lipid peroxidation with a considerable decrease of docosahexaenoic acid (22:6 n-3). The incorporation of -tocopherol (0.35 mol/mg protein) produce a 43.37% inhibition of the lipid peroxidation process evaluated as chemiluminiscence (total cpm originated in 160 min). The phospholipid species containing the highest amount of docosahexaenoic acid: phosphatidyletanolamine and phosphatidylserine were more affected than phosphatidylcholine during the lipid peroxidation process. Not all phospholipids, however, were equally protected after the addition of -tocopherol to the incubation medium. Phosphatidylcholine and phosphatidyletanolamine, were not protected by -tocopherol, the vitamin provides selective antioxidant protection only for phosphatidylserine. These results indicate that -tocopherol may act as antioxidant protecting rod outer segment membranes from deleterious effect by a selective mechanism that diminishes the loss of docosahexaenoic acid from phosphatidylserine.  相似文献   

5.
Five principle monoterpenoid and other constituent volatile chemicals of sunflower heads were combined to resemble two lines of sunflower (Helianthus annuus L.): one U.S.D.A. standard line and one French line which was poorly visited by insects (Etievant et al., 1984). Field trials of attraction to red sunflower seed weevils (Smicronyx fulvus Le Conte, Coleoptera: Curculionidae) showed that one was clearly preferred over the other. The more attractive mixture contained -pinene, -pinene, limonene, camphene and bornyl acetate in a ratio resembling that of Flath et al. (1985) rather than that described by Etievant et al. (1984). One or two volatiles were deleted from the optimal blend but only mixtures of five volatiles showed the highest attraction. Substitution of sabinene, another volatile prominent in sunflower, for one of the five in the optimal blend also decreased attraction of seed weevils. When the monoterpenoid components and green leaf volatiles in the traps resembled the ratios of most of the prominent volatiles of sunflower, attraction was significantly greater than controls.  相似文献   

6.
We studied fatty acid (FA) composition in samples from bodies and intestinal contents of the littoral amphipod Gammarus lacustris Sars, from the Bugach freshwater reservoir. Simultaneously, samples of seston and bottom sediments were also collected from the reservoir during early August. There were no differences in FA composition of gut contents, seston and sediments of pebbly bottom. Seston was the main food source of Gammarus but some FAs Gammarus got from sediments. The FA composition of G. lacustris and seston from the Bugach freshwater reservoir were compared with those of the animals from the saltwater Lake Shira (Siberia). While FA composition of the two Gammarus populations differed significantly, those of seston were practically similar: the composition of long-chain unsaturated fatty acids, 20:53, 22:63 and 20:46, were significantly higher in animals from saline Shira Lake, whereas 16:1 and 16:0 were higher in the freshwater populations of amphipods from the Bugach freshwater reservoir. Taking into account the relevant literature data, we hypothesise that this difference in C16 acid might be a distinguishing characteristic of FA composition of freshwater and saltwater crustaceans.  相似文献   

7.
Palma-Silva  C.  Albertoni  E.F.  Esteves  F.A. 《Plant Ecology》2000,148(2):157-164
This investigation of the aquatic macrophyte Eleocharis mutata (L.) Roem. Et Schult. was carried out in Imboassica lagoon, a coastal lagoon in Macaé (22°50S; 44°42W), in the northern part of Rio de Janeiro, Brazil. The sandbar separating this lagoon from the ocean has been opened several times for flood control and to allow the entrance of marine species of commercial interest. The barrier bar has been breached without appropriate planning, and the consequences of breaching for the lagoon ecosystem are poorly understood. These openings drastically affect the structure and functioning of the lagoon, but there are no data on possible effects on macrophyte communities. In this project, we obtained data on the increase of the distribution, biomass accumulation and production rates of E. mutata in the establishment of a new stand, in an effort to relate effects of sandbar breach events to the expansion of this species. During 22 months of sampling, 4 breachings of the sandbar occurred, and E. mutata increased its area of coverage by about 8104 m2, or 2.5% of the total area of the lagoon. The total aerial biomass reached a maximum of 1515 g DW m–2, and the underground biomass reached 583 g DW m–2. During the establishment and development of the stand, both dead and live aerial biomass and underground biomass tended to accumulate. Aerial net primary production (ANPP) was quite variable, reaching a maximum of 18.9 g DW m–2 d–1. We conclude that the sudden variations in water level caused by breaches in the sandbar were beneficial to the expansion of this species in Imboassica lagoon.  相似文献   

8.
pHrMA4a-Z is a recombinant plasmid in which about 1.4 kb of the 5 flanking region of a gene for muscle actin HrMA4a from the ascidian Halocynthia roretzi is fused with the coding sequence of a bacterial gene for -galactosidase (lac-Z). In this study, we examined the expression of the fusion gene construct when it was introduced into eggs of another ascidian, namely Ciona savignyi. When a moderate amount of linearized pHrMA4a-Z was introduced into fertilized Ciona eggs, the expression of the reporter gene was evident in muscle cells of the larvae, suggesting that both species share a common machinery for the expression of muscle actin genes. The 5 upstream region of HrMA4a contains several consensus sequences, including a TATA box at -30, a CArG box at -116 and four E-boxes within a region of 200 bp. A deletion construct, in which only the 216-bp 5 flanking region of HrMA4a was fused with lac-Z, was expressed primarily in larval muscle cells. However, another deletion construct consisting of only the 61-bp upstream region of HrMA4a fused with lac-Z was not expressed at all. When pHrMA4a-Z or pHrMA4a-Z (–216) was injected into each of the muscle-precursor blastomeres of the 8-cell embryo, expression of the reporter gene was observed in larval muscle cells in a lineage-specific fashion. However, expression of the reporter gene was not observed when the plasmid was injected into non-muscle lineage. Therefore, the expression of the reporter gene may depend on some difference in cytoplasmic constituents between blastomeres of muscle and non-muscle lineage in the 8-cell embyo.  相似文献   

9.
The mechanism of ammonia assimilation in nitrogen fixing bacteria   总被引:1,自引:0,他引:1  
Summary Enzymatic and genetic evidence are presented for a new pathway of ammonia assimilation in nitrogen fixing bacteria: ammonium glutamine glutamate. This route to the important glutamate-glutamine family of amino acids differs from the conventional pathway, ammonium glutamate glutamine, in several respects. Glutamate synthetase [(glutamine amide-2-oxoglutarate aminotransferase) (oxidoreductase)], which is clearly distinct from glutamate dehydrogenase, catalyzes the reduced pyridine nucleotide dependent amination of -ketoglutarate with glutamine as amino donor yielding two molecules of glutamate as product. The enzyme is completely inhibited by the glutamine analogue DON, whereas glutamate dehydrogenase is not affected by this inhibitor; the glutamate synthetase reaction is irreversible. Glutamate synthetase is widely distributed in bacteria; the pyridine nucleotide coenzyme specificity of the enzyme varies in many of these species.The activities of key enzymes are modulated by environmental nitrogenous sources; for example, extracts of N2-grown cells of Klebsiella pneumoniae form glutamate almost exclusively by this new route and contain only trace amounts of glutamate dehydrogenase activity whereas NH3-grown cells possess both pathways. Also, the biosynthetically active form of glutamine synthetase with a low K m for ammonium predominates in the N2-grown cell.Several mutant strains of K. pneumoniae have been isolated which fail to fix nitrogen or to grow in an ammonium limited environment. Extracts of these strains prepared from cells grown on higher levels of ammonium have low levels of glutamate synthetase activity and contain the biosynthetically inactive species of glutamine synthetase along with high levels of glutamate dehydrogenase. These mutants missing the new assimilatory pathway have serious defects in their metabolism of many inorganic and organic nitrogen sources; utilization of at least 20 different compounds is effected. We conclude that the new ammonia assimilatory route plays an important role in nitrogenous metabolism and is essential for nitrogen fixation.Abbreviation DON 6-diazo-5-oxo-l-norleucine  相似文献   

10.
A new method for testing the strength of cells against fluid shear stress by using a long capillary column was proposed. The trajectories of cells in the column were simulated by introducing the Brownian motion model. The Brownian motion was performed by the generation of random numbers. The mean exposure time to shear stress and the mean shear stress acting on the surface of cells were discussed by the result of computer simulation. The mean shear stress acting on the surface of cells flowing in the capillary column was estimated as 4/3-fold of the shear stress at the column wall provided that the ratio of the cell radius to the column radius does not exceed 0.08. The effectiveness of this new method for testing the strength of cells against fluid shear stress was shown.List of Symbols a m radius of cell - c constant - E distribution function - L m length of capillary column - M number of division - N number of division - p probability - Q m3/s flow rate - R m radius of capillary column - r m radial position - t s time - T s exposure time - T m s mean exposure time - T 0 s mean residence time - m/s axial velocity - u m m/s cross-sectional flow velocity - z m axial position - s–1 shear rate - w s–1 shear rate at wall - Pa s viscosity - spherical coordinate - spherical coordinate - Pa shear stress - m Pa mean shear stress - w Pa shear stress at wall  相似文献   

11.
Identification of global biodiversity hotspots is a leading conservationpriority ( Hotspots.Conservation International, Washington, DC; Precious Heritage: The Statusof Biodiversity in the United States. Oxford University Press, Oxford, UK), but weknow little about the nature and structure of such hotspots. Botanical richnessin one recently identified hotspot, the California Floristic Province, has longbeen attributed by evolutionists to edaphic and climatic heterogeneity. Here we demonstrate that the flora restricted (endemic) toserpentine soil, the state's most botanically distinctive substrate, showsunusually high ' diversity' – i.e., spatial variation inspecies composition – among regions, although ''diversity within any given region is low. Conservation of an endemic-rich florarequires a network of sites to capture this among-region or spatial component ofdiversity.  相似文献   

12.
Summary Biomass harvested from batch and single-stage continuous cultures of the fungus Geotrichum candidum, growing on the spent wash from an Irish malt whiskey distillery, was analysed for macromolecular components. The true protein content was on average more than 47% (w/w), while crude protein levels of as much as 61% were found. The protein was not deficient in any amino acid when compared to the FAO standard. The effects of varying growth temperature and dilution rate on the composition of the organism are discussed, and also the problem of its accumulation of copper from the medium.  相似文献   

13.
Glycosides were synthesized from soluble starch and hydroxybenzylalcohols (o, m and p) by the transglycosylation activity of amyloglucosidase from Rhizopus sp. in a water system. The synthesized glycosides were identified as hydroxybenzyl--glucosides, of which one molecule of hydroxybenzylalcohol was bound to 1-OH of glucose. The transglycosylation reaction was performed in a reaction system containing 50 mg starch, 50 mg hydroxybenzylalcohol, and 10 units of enzyme in pH 5.0 at 45°C. The glycoside was rapidly hydrolyzed to glucose and hydroxybenzylalcohol by the -glucosidase.  相似文献   

14.
The locus for Tyrosinase-Positive Oculocutaneous Albinism (ty-pos OCA) has not yet been localised. The search for the ty-pos OCA locus has included a search for linkage to candidate pigment loci and a candidate chromosomal region, as well as a random search using highly polymorphic markers in 42 families, including 271 individuals of whom 79 are affected. The lod scores for the tyrosinase (TYR) locus (11q14–q21), homologous to the albino locus, c, in the mouse and the CAS2/TRP1 locus (9p22-pter), homologous to the brown locus, b, in the mouse were -5.89 and -7.22, respectively, at a recombination fraction of =0.01, thus excluding them from being the ty-pos OCA locus. In the candidate chromosomal region, 11p, four loci (probes) were tested, SAA (pSAA82), CALC (pHC36), HBB (Gamma-globin haplotype) and an AC repeat polymorphism at the Wilm's Tumour locus (WT1). A portion of 11p was excluded with the following lod scores: pSAA82 lod=-2.05 at =0.10; pHC36 lod=-3.87 at =0.05; gamma-globin haplotype lod=-2.80 at =0.10; and WT1 lod=-2.34 at =0.10. Thirty-three polymorphic markers randomly distributed on 13 different chromosomes were all excluded from close linkage to ty-pos OCA.  相似文献   

15.
Eric A. Kwei 《Hydrobiologia》1977,56(2):157-174
Two types of lagoon which occur along the West Coast of Africa described as open and closed, were studied using oceanographic methods. The parameters were temperature, salinity, dissolved oxygen and productivity. It was observed that rainfall, evaporation and the tides have a combined effect on the variations in the temperature and the salinity. The opening of the lagoons into the sea affects the level of the productivity and the rate at which salinity increases with time.Minimum temperatures at the heads of the lagoons were found to be lower than those near the ocean, while the maximum temperatures were higher at the heads. During the dry period, due to continuous evaporation of water, the salinity in the closed lagoon usually rises steeply, while those in the open lagoon rise gently.The levels of dissolved oxygen concentration in the open lagoon are higher than those in the closed lagoon. This is the result of the fact that phytoplankton count in the open lagoon is higher than that in the closed lagoon. There is also a greater diversity of phytoplankton and more live phytoplankton cells per cubic centimetre of water in the open lagoon than in the closed lagoon.As a result of all this, the main organic productivity in the open lagoon is higher than those in the closed lagoon.  相似文献   

16.
Summary T-type calcium channels (I T channels) were studied in cell-attached patch electrode recordings from the ventricular cell membrane of 14-day embryonic chick heart. All experiments were performed in the absence of Ca2+ with Na+ (120mm) as the charge carrier.I T channels were distinguished from L-type calcium channels (I L) by their more negative activation and inactivation potential ranges; their smaller unitary slope conductance (26 pS), and their insensitivity to isoproterenol or D600. Inactivation kinetics were voltage dependent. The time constant of inactivation was 37 msec when the membrane potential was depolarized 40 mV from rest (R+40 mV), and 20 msec atR+60 mV. The frequency histogram of channel open times 0 was fit by a single-exponential curve while that of closed times c was biexponeintial. o was the same atR+40 mV andR+60 mV whereas c was shortened atR+60 mV. The open-state probability (P o) increased with depolarization: 0.35 atR+40 mV, 0.8 atR+60 mV and 0.88 atR+80 mV. This increase inP o at depolarized potentials could be accounted for by the decrease in c.  相似文献   

17.
A preparation of rod outer segments has been used to study the polypeptides characteristic of an early-onset retinal dystrophy in cats (Rdy) by two-dimensional gel electrophoresis. Comparison of 2-D gels of rod outer segment preparations from retinas of normal and Rdy animals shows several differences. In particular, a polypeptide of Mr 51 kDa and pI 7.5 is present at increased levels in preparations from Rdy cats at 6 weeks, 9 weeks and 12.5 weeks of age but not at 3.5 weeks.Abbreviations cGMP Guanosine 3 - 5 Cyclic Monophosphate - SDS-PAGE Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis - TRIS 2-amino-2(hydroxymethyl)-1,3 propandiol - PMSF Phenylmethylsulphonylfluoride - IEF Isoelectrofocussing  相似文献   

18.
Laboratory studies on predation, reproduction and development were conducted on three phytoseiid species,Typhlodromus pomi (Parrott),Phytoseius macropilis (Banks) andAmblyseius finlandicus (Oudemans). These species were dominant phytoseiid mites on the abandoned apple trees with which ecological experiments were carried out in southern Ontario, Canada. Throught the studies, a rust mite species (Diptacus gigantorhynchus sp. complex), a single consistent prey in the orchard, was used as food. Results showed that the three predators developed well with the rust mites as food, and thatA. finlandicus developed most rapidly and had the highest predation rate. However, this species showed a poor tolerance to low prey densities. These observations suggest that this species is a predacious species and the other two species were more or less general feeders. All species showed the same pre-mating behaviour (Amblyseius-Typhlodromus type), but they exhibited a variety of mating and oviposition behaviours.  相似文献   

19.
Acetone degradation by cell suspensions of Desulfobacterium cetonicum was CO2-dependent, indicating initiation by a carboxylation reaction. Degradation of butyrate was not CO2-dependent, and acetate accumulated at a ratio of 1 mol acetate per mol butyrate degraded. In cultures grown on acetone, no CoA transfer apparently occurred, and no acetate accumulated in the medium. No CoA-ligase activities were detected in cell-free crude extracts. This suggested that the carboxylation of acetone to acetoacetate, and its activation to acetoacetyl-CoA may occur without the formation of free acetoacetate. Acetoacetyl-CoA was thiolytically cleaved to two acetyl-CoA, which were oxidized to CO2 via the acetyl-CoA/carbon monoxide dehydrogenase pathway. The measured intracellular acyl-CoA ester concentrations allowed the calculation of the free energy changes involved in the conversion of acetone to acetyl-CoA. At in vivo concentrations of reactants and products, the initial steps (carboxylation and activation) must be energy-driven, either by direct coupling to ATP, or coupling to transmembrane gradients. The G of acetone conversion to two acetyl-CoA at the expense of the energetic equivalent of one ATP was calculated to lie very close to 0kJ (mol acetone)-1. Assimilatory metabolism was by an incomplete citric acid cycle, lacking an activity oxidatively decarboxylating 2-oxoglutarate. The low specific activities of this cycle suggested its probable function in anabolic metabolism. Succinate and glyoxylate were formed from isocitrate by isocitrate lyase. Glyoxylate thus formed was condensed with acetyl-CoA to form malate, functioning as an anaplerotic sequence. A glyoxylate cycle thus operates in this strictly anaerobic bacterium. Phosphoenolpyruvate (PEP) carboxykinase formed PEP from oxaloacetate. No pyruvate kinase activity was detected. PEP presumably served as a precursor for polyglucose formation and other biosyntheses.Abbreviations MV 2+ Oxidized methyl viologen - PEP Phosphoenolpyruvate - PHB Poly--hydroxybutyrate  相似文献   

20.
Transitions in the growth limiting factor from light (I) to nitrogen (N) and vice versa caused changes in geosmin production, protein and carbohydrate content, and the synthesis of pigments such as chlorophyll a (Chl a), phycobiliproteins (PBPs), and -carotene of the cyanobacterium Oscillatoria brevis. Following IN transition the first 150h, the decrease in protein content was compensated for by an increase of carbohydrates, and thereby, a constant biomass level was maintained in this period. Thereafter, biimass dropped to 15% of its initial level. A decrease in geosmin and pigment content was observed during transition from IN-limited growth. However, geosmin increased relative to phytol (Chl a) and -carotene which may indicate that a lowered demand for phytol and -carotene during N-limited growth allows isoprenoid precursors to be directed to geosmin rather than to pigment synthesis. Synthesis of Chl a and -carotene at the expense of geosmin was suggested for the observed start of increase in geosmin production only at the time that Chl a and -carotene had reached their I-limited steady state. Transition from nitrogen to light limited growth caused an acceleration of metabolism shown by a rapid decrease in carbohydrate content accompanied by an increase in protein content. The growth rate of the organisms temporarily exceeded the dilution rate of the culture and the biomass level increased 6-fold. Due to the only modest changes in geosmin production (2-fold) compared to changes in biomass level (6-fold) during I-or N-limited growth, environmental factors seem to have limited effect on geosmin production.Abbreviations Chl a chlorophyll a - dry wt dry weight; - I-limited light-limited - N-limited nitrogen-limited - PBP phycobiliprotein This research was performed at the Department of Microbiology, University of Amsterdam, with finacial support provided by the Royal Norwegian Ministry of Foreign Affairs and the Royal Norwegian Council for Scientific and Industrial Research  相似文献   

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