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1.
Aspartate metabolism was investigated in excised cotyledons of radiata pine ( Pinus radiata D. Don). These cotyledons were cultured under shoot-forming (plus N6-benzyladenine, SF), non-shoot-forming (minus N6-benzyladenine, NSF) and unresponsive (plus N6-benzyladenine, OLD) conditions, then incubated with [14C]-aspartate for 3-h pulse treatments followed by 3-h chase treatments with cold aspartate. The majority of label was recovered in the CO2, amino acid, organic acid and pellet fractions. Uptake was greatest in all tissue types early in culture. Most (over 80%) of the [14C]-aspartate taken up by the tissues was converted to CO2 at day 0 in SF and NSF tissues, CO2 accounted for less than 50% of the total radioactivity in other tissues. Greater incorporation into fractions was observed in SF tissues during promeristemoid formation, while in NSF tissues the greatest incorporation was observed during a period of rapid elongation. Generally, less incorporation was observed in OLD cotyledons than in SF and NSF cotyledons. Analysis of the amino acid fraction showed that labelled aspartate was converted to other amino acids, mainly glutamate, glutamine, asparagine and 4-aminobutyric acid.  相似文献   

2.
Abstract: Two samples of the peptide tyrosine-melanocyte-stimulating hormone release-inhibiting factor-1 (Tyr-MIF-1; Tyr-Pro-Leu-Gly-NH2) were tritiated on different amino acids (Tyr or Pro) and incubated together at 37°C with fractions of rat brain. The amount of intact tetrapeptide remaining was determined by HPLC. By 3 min, most of the Tyr-MIF-1 was degraded. Because similar amounts of [3H]Pro and [3H]Tyr appeared after incubation of the Tyr-MIF-1 peptides in brain homogenate, even as early as 30 s, examination of only this crude preparation would misleadingly indicate that Tyr-MIF-1 is not a precursor of melanocyte-stimulating hormone release-inhibiting factor-1 (MIF-1; Pro-Leu-Gly-NH2) in brain tissue. However, incubation of the mitochondrial fractions of brain under the same conditions resulted in more than three times as much [3H]Tyr being formed as [3H]Pro, with accompanying accumulation of MIF-1. Addition of excess MIF-1 to the mitochondrial fraction completely suppressed the formation of MIF-1 and more than doubled the amount of Tyr-MIF-1 remaining intact. When Tyr-MIF-1 tritiated only on the Tyr was added to the mitochondrial fraction, the main peaks of radioactivity appeared only at the positions of Tyr and Tyr-MIF-1, not at the position of Tyr-Pro. The results indicate that Tyr-MIF-1 can serve as a precursor of MIF-1 in brain mitochondria, an effect not evident when crude brain homogenate is used.  相似文献   

3.
Well-nodulated soybean ( Glycine max L. Merr. cv. Akisengoku) plants were allowed to assimilate 13CO2. Plant cytosol and bacteroid fractions were isolated from nodules, and the kinetics of [13C]-labelling of soluble carbohydrates, organic acids and amino acids were investigated.
The concentrations of all metabolites, with the exception of trehalose and 3-hydroxy-butyrate, were 10- to 1000-fold higher in plant cell cytosol than in bacteroids. The major portion of trehalose was found in bacteroids and 3-hydroxybutyrate only in bacteroids. Sucrose was most highly labelled with 13C in nodules, and the levels and time-course of labelling of sucrose were in good agreement with those of respired CO2 from the nodules. The levels and time-courses of labelling of sucrose were closely similar in cytosol and bacteroids. Glucose was less labelled than sucrose and the level of labelling was consistently higher in cytosol than in bacteroids. The levels of [13C]-labelling of organic acids and amino acids in nodules were lower than those of sucrose and of respired CO2. Tricarboxylic acid cycle intermediates, particularly succinate, were considerably less labelled in bacteroids than in the cytosol. All amino acids detected were also much more rapidly labelled in the cytosol. The results are discussed in relation to the utilization and possible compartmentation of carbon substrates in nodule tissues.  相似文献   

4.
Abstract Turnover times of radioactive glucose were shorter in paddy soil (4–16 min) than in Lake Constance sediment (18–62 min). In the paddy soil, 65–75% of the radioactive glucose was converted to soluble metabolites. In the sediment, only about 25% of the radioactive glucose was converted to soluble metabolites, the rest to particulate material. In anoxic paddy soil, the degradation pattern of position-labelled glucose was largely consistent with glucose degradation via the Embden-Meyerhof-Parnas (EMP) pathway followed by methanogenic acetate cleavage: CO2 mainly originated from C-3,4, whereas CH4 mainly originated from C-1 and C-6 of glucose. Acetate-carbon originated from C-1, C-2 and C-6 rather than from C-3,4 of glucose. In both paddy soil and Lake Constance sediment acetate and CO2 were the most important early metabolites of radioactive glucose. Other early products included propionate, ethanol/butyrate, succinate, and lactate, but accounted each for less than 1–8% of the glucose utilized. The labelling of propionate by [3,4-14C]glucose suggests that it was mainly produced from glucose or lactate rather than from ethanol. Isopropanol and caproate were also detectable in paddy soil, but were not produced from radioactive glucose. Chloroform inhibited methanogenesis, inhibited the further degradation of radioactive acetate and resulted in the accumulation of H2, however, did not inhibit glucose degradation. Since acetate was the main soluble fermentation product of glucose and was produced at a relatively high molar acetate: CO2 ratio (2.5:1), homoacetogenesis appeared to be the most important glucose fermentation pathway.  相似文献   

5.
Winter wheat (Triticum aestivum L. cv. Hereward) plants were grown for 35 d either at 350 μ mol mol–1 CO2 or at 650 μ mol mol–1 CO2. Lipid synthesis was studied in these plants by incubating the 5th leaf on the main stem with [1–14C]acetate. Increased CO2 concentrations did not significantly affect the total incorporation of radiolabel into lipids of whole leaf tissue, but altered the distribution for individual lipid classes. Most noticeable amongst acyl lipids was the reduction in labelling of diacylglycerol and a corresponding increase in the proportion of phosphatidylcholine labelling. In the basal regions, there were similar changes and, in addition, phosphatidylglycerol labelling was particularly increased following growth in an enriched CO2 atmosphere. The stimulation of labelling of the mitochondrial-specific lipid, diphosphatidylglycerol, prompted an examination of the mitochondrial population in wheat plants. Mitochondria were localized in intact wheat sections by immunolabelling for the mitochondrial-specific chaperonin probe. Growth in elevated CO2 doubled the number of mitochondria compared to growth in ambient CO2. Fatty acid labelling was also significantly influenced following growth at elevated CO2 concentrations. Most noticeable were the changes in 16C:18C ratios for the membrane lipids, phosphatidylcholine, phosphatidylglycerol and monogalactosyldiacylglycerol. These data imply a change in the apportioning of newly synthesized fatty acids between the 'eukaryotic' and 'prokaryotic' pathways of metabolism under elevated CO2.  相似文献   

6.
The number, size, solubility in chloroform/methanol and some aspects of the formation of the components labeled by radioactive amino acids in isolated mitochondria of rat liver and Zajdela hepatoma were studied. Isolated mitochondria were labeled with radioactive amino acids under various conditions, and the distribution of radioactivity in sodium dodecylsulfate-polyacrylamide gels after electrophoresis of mitochondrial membrane fraction was analysed. 1. Isolated mitochondria of rat liver and Zajdela hepatoma incroporated radioactive amino acids almost exclusively into the membrane fraction. Electrophoretic analysis of this fraction revealed the presence of 15 distinct peaks of radioactivity with corresponding apparent molecular weights of 10 000 to 58 000. The electrophoretic mobility of the labeled components was identical and the general pattern of the radioactivity distribution in the gel for the rat liver and the tumour mitochondria was very similar. 2. Components of the membrane fraction of rat liver mitochondria labeled in vitro displayed an unequal solubility in acidic (2 mM HC1) chloroform/methanol (2/1) mixture; as detected by sodium dodecylsulfate-polyacrylamide gel electrophoresis a single labeled component with apparent molecular weight of 10 000 was soluble in neutral chloroform/methanol. 3. Inverse relation was observed between amino acid incorporation activity of isolated mitochondria and the portion of the label incorporated into the component with apparent molecular weight 10 000. The identity of this component with that soluble in neutral chloroform/methanol mixture has been indicated. 4. The rate of incorporation of [3H]leucine by isolated Zajdela hepatoma mitochondria into the components with lower (10 000-25 000) apparent molecular weights decreased with time, whereas that into components with higher (above 25 000) apparent molecular weight remained approximately constant within the time interval tested (30 min). 5. From the total radioactivity incorporated into the membrane fraction during 5-min pulse labeling of isolated Zajdela hepatoma mitochondria by [3H]leucine up to 25% was recovered in the region of the gel corresponding to a component with apparent molecular weight 10 000. After 25 min chase the radioactivity in this region decreased about 3.5 times while the specific radioactivity of the total membrane fraction did not change significantly. The pattern of radioactivity distribution observed after the pulse was preserved by chloramphenicol. 6. Unlabeled sonicated mitochondria or postribosomal supernatant from rat liver regenerating in the presence of chloramphenicol were incubated with neutral chloroform/methanol extract of in vitro with [14C]leucine labeled rat liver mitochondria. After this incubation several labeled components with apparent molecular weights above 10 000 were recovered in the electrophoreograms of the originally unlabeled fractions.  相似文献   

7.
Differences in mitochondrial membrane composition and ultrastructure were studied after storage of cauliflower ( Brassica oleracea , L., Botrytis group) for 5 days at 25°C in air or under controlled atmospheres: 3% O2, 21% O2+ 15% CO2 or 3% O2+ 15% CO2. In air, postharvest senescence involved a 20% decrease in mitochondrial phospholipid content. A large reduction in the relative abundance of phosphati-dylcholine (PC) and in the degree of unsaturation of PC and phosphatidyl ethanolamine (PE) was observed. However, the degree of unsaturation increased in cardiolipin (CL). Storage under 3% O2 did not prevent phospholipid breakdown. Low O2 prevented the relative decrease in PC observed during storage in air and the loss of linoleic acid from PC, but not from PE. This relative protection offered by the low O2 atmosphere was lost under 3% O2+ 15% CO2. The high CO2 atmospheres caused twice as much loss in phospholipids as that observed during storage in air. Extensive loss of mitochondrial protein, a marked decrease in phospholipid to protein ratio, and electron micrograph observations suggest structural alterations in the presence of high CO2.  相似文献   

8.
Abstract: The effect of an inhibitor of N -glycosylation of glycoproteins, tunicamycin, on synthesis of PNS myelin proteins was investigated in vitro by using chopped sciatic nerves or spinal roots of 21-day-old Wistar rats. Tunicamycin when incubated with these nerves in the presence of 3H-labeled fucose, mannose, or glucosamine inhibited the uptake of radioactivity into myelin proteins including some high-molecular-weight proteins, P0, 23K protein, and 19K protein by amounts ranging from 42 to 79%. Uptake of 14Camino acid mixture was inhibited much less by tunicamycin, but a new radioactive protein peak appeared when the protein mixtures had been separated by electrophoresis on polyacrylamide gels in the presence of sodium dodecyl sulfate. This protein ran directly in front of the P0 peak, did not correspond to any bands stained by Fast green, and was not labeled by fucose. This peak appeared in increasing larger proportions with progressive time of incubation of nerves with 3H amino acids in the presence of tunicamycin. The new protein, which cross-reacts with P0 antiserum, was tentatively identified as a nonglycosylated P0 protein that appears to be almost as well incorporated as P0 into the subcellular fraction containing myelin. At this time it is not possible to determine whether the unglycosylated P0 is actually assembled into a site and configuration like that of P0.  相似文献   

9.
Application of anaerobic conditions with CO2 or N2 atmospheres to remove astringency from harvested persimmon fruit ( Diospryros kaki L. cv. Triumph), caused production of more acetaldehyde under CO2 than under N2, 14CO2 applied in a 100% CO2 atmosphere, for 48 h to astringent persimmon fruits was incorporated mainly into malate and very little into other metabolites, such as carbohydrate or amino acids. Application of malate or pyruvate to pulp discs of astringent persimmons caused an immediate rise in acetaldehyde production. The higher levels of acetaldehyde produced by whole fruits held in a CO2 atmosphere, than by fruits held in a N2 atmosphere, can be explained through fixation of atmospheric CO2 into malate, leading to acetaldehyde production.  相似文献   

10.
Ananas comosus L. (Merr.) (pineapple) was grown at three day/night temperatures and 350 (ambient) and 700 (elevated) μ mol mol–1 CO2 to examine the interactive effects of these factors on leaf gas exchange and stable carbon isotope discrimination ( Δ ,‰). All data were collected on the youngest mature leaf for 24 h every 6 weeks. CO2 uptake (mmol m–2 d–1) at ambient and elevated CO2, respectively, were 306 and 352 at 30/20 °C, 175 and 346 at 30/25 °C and 187 and 343 at 35/25 °C. CO2 enrichment enhanced CO2 uptake substantially in the day in all environments. Uptake at night at elevated CO2, relative to that at ambient CO2, was unchanged at 30/20 °C, but was 80% higher at 30/25 °C and 44% higher at 35/25 °C suggesting that phosphoenolpyruvate carboxylase was not CO2-saturated at ambient CO2 levels and a 25 °C night temperature. Photosynthetic water use efficiency (WUE) was higher at elevated than at ambient CO2. Leaf Δ -values were higher at elevated than at ambient CO2 due to relatively higher assimilation in the light. Leaf Δ was significantly and linearly related to the fraction of total CO2 assimilated at night. The data suggest that a simultaneous increase in CO2 level and temperature associated with global warming would enhance carbon assimilation, increase WUE, and reduce the temperature dependence of CO2 uptake by A. comosus .  相似文献   

11.
12.
Abstract: Chains of lumbar sympathetic ganglia from 15-day-old chicken embryos were incubated for 4 h at 36°C in a bicarbonate-buffered salt solution equilibrated with 5% CO2-95% O2. Glucose (1–10 m M ), lactate (1–10 m M ), [U-14C]glucose, [1-14C]glucose, [6-14C]glucose, and [U-14C]lactate were added as needed. 14CO2 output was measured continuously by counting the radioactivity in gas that had passed through the incubation chamber. Lactate reduced the output of CO2 from [U-14C]glucose, and glucose reduced that from [U-14C]lactate. When using uniformly labeled substrates in the presence of 5.5 m M glucose, the output of CO2 from lactate exceeded that from glucose when the lactate concentration was >2 m M . The combined outputs at each concentration tested were greater than those from either substrate alone. The 14CO2 output from [1-14C]glucose always exceeded that from [6-14C]glucose, indicating activity of the hexose monophosphate shunt. Lactate reduced both of these outputs, with the maximum difference between them during incubation remaining constant as the lactate concentration was increased, suggesting that lactate may not affect the shunt. Modeling revealed many details of lactate metabolism as a function of its concentration. Addition of a blood-brain barrier to the model suggested that lactate can be a significant metabolite for brain during hyperlactemia, especially at the high levels reached physiologically during exercise.  相似文献   

13.
Five species of sap-feeding homoptera were studied on Fagus sylvatica and Acer pseudoplatanus and exposed to elevated concentrations of carbon dioxide (600 μL L–1). The concentration of total soluble amino acids in foliage of F. sylvatica was unaffected by growing saplings in elevated atmospheric CO2 concentrations. Although experiments on individual aphids indicated poorer performance of Phyllaphis fagi (fewer, smaller nymphs produced), resultant populations did not differ from those in ambient (350 μL L–1) conditions. The area of beech foliage stippled by the leafhopper Fagocyba cruenta was similar at ambient and elevated CO2 concentrations. The concentration of total amino acids and that of serine of A. pseudoplatanus foliage were significantly lower at elevated CO2 concentrations. However, the relative growth rates of two aphid species Drepanosiphum platanoidis and Periphyllus testudinaceus and one leafhopper Ossiannilssonola callosa were not significantly different in elevated CO2. No evidence was found that, under the conditions of these experiments, populations of aphids and leafhoppers will change as concentrations of CO2 increase.  相似文献   

14.
Abstract— Glucose metabolism in the superior cervical ganglion for calves has been studied by incubating slices with [1-14C]-, [6-14C]- and [U-14C]-labelled glucose at 37°C and pH 7.4. Glucose utilization and the metabolic partitioning of glucose carbon in products during different incubation periods ranging from 5 to 60 min were determined by isotopic methods.
Separation and identification of labelled compounds have been achieved by anion and cation exchange chromatography as well as by TLC and enzymatic analyses.
From the data obtained a carbon balance could be constructed showing lactate to be the major product of glucose metabolism followed by CO2 and amino acids. Measuring the release of 14CO2 from differently 4C-labelled glucose, the existence of an active pentose phosphate pathway in the ganglion could be demonstrated although this pathway seems to contribute only to a small extent to glucose metabolism. The marked decrease of the C-U: C-6 and the C-U:C-1 ratios in 14CO2 observed in the course of incubation is discussed in terms of a time-dependent change in the rate of synthesis of amino acids which are directly connected with intermediates of the citric acid cycle.  相似文献   

15.
Root and mycelial exudation contributes significantly to soil carbon (C) fluxes, and is likely to be altered by an elevated atmospheric carbon dioxide (CO2) concentration and nitrogen (N) deposition. We quantified soluble, low-molecular-weight (LMW) organic compounds exuded by ectomycorrhizal plants grown under ambient (360 p.p.m.) or elevated (710 p.p.m.) CO2 concentrations and with different N sources. Scots pine seedlings, colonized by one of five different ectomycorrhizal or nonmycorrhizal fungi, received 70 μM N, either as NH4Cl or as alanine, in a liquid growth medium. Exudation of LMW organic acids (LMWOAs), dissolved monosaccharides and total dissolved organic carbon were determined. Both N and CO2 had a significant impact on exudation, especially of LMWOAs. Exudation of LMWOAs was negatively affected by inorganic N and decreased by 30–85% compared with the organic N treatment, irrespective of the CO2 treatment. Elevated CO2 had a clear impact on the production of individual LMWOAs, although with very contrasting effects depending on which N source was supplied.  相似文献   

16.
Effects of carbon dioxide in anther cultures   总被引:1,自引:0,他引:1  
In anther cultures of Anemone canadensis L., Anemone dichotoma L., Anemone hupehensis Lemoine, Clematis viticella L. and Papaver setigerum DC. a positive relationship between incubation in 2% CO2 and the production of microspore-derived embryos was observed. In anther cultures of Nicotiana tabacum L., Anemone hupehensis and Clematis viticella a combination of cold treatment (7°C) and incubation in 2% CO2 increased embryo production. In Anemone canadensis cold treatment increased the number of proembryos, whereas incubation in 2% CO2 had no effect. In Anemone hupehensis 5% CO2 increased embryo production by more than 2%. In Anemone dichotoma and Papaver setigerum 2% CO2 was the more efficient level. CO2 had no significant effect on pH in the culture medium in anther cultures of Anemone canadensis.  相似文献   

17.
Over a period of several days, rhythmic changes in extracellular NH+4 concentration take place in cultures of the cyanobacterium Microcystis firma (Bré et Lenorm.) Schmidle, strain Gromov/St. Petersb. 398, under conditions of restricted CO2 supply and light/dark alternation. The changes are enhanced by nitrate supply. Among the various processes generating intracellular NH+4 (NH44 uptake, NO3 reduction, protein and amino acid degradation, photorespiration), NO3 reduction appears as the one most important. This can be concluded from experiments with and without nitrate and/or ammonium in the medium. In the presence of saturating CO2, continuous light, or continuous darkness, rhythmic NH+44 oscillations are not induced. Studies of the incorporation of NH+4 nitrogen by in vivo 15N-NMR show that if CO2 is supplied, 15N is accumulated in several components with the following time course: in the first hour in Gln (δ), in the second hour in the α-amino groups of most nonbranched amino acids, in the third hour in γ-aminobutyric acid (GABA), Orn (δ) and Lys (ε), and in the sixth hour in Ala. Carbon limitation, however, results in accumulation of label in the amide nitrogen of glutamine only.  相似文献   

18.
Effects of the current (38 Pa) and an elevated (74 Pa) CO2 partial pressure on root and shoot areas, biomass accumulation and daily net CO2 exchange were determined for Opuntia ficus-indica (L.) Miller, a highly productive Crassulacean acid metabolism species cultivated worldwide. Plants were grown in environmentally controlled rooms for 18 weeks in pots of three soil volumes (2 600, 6 500 and 26 000 cm3), the smallest of which was intended to restrict root growth. For plants in the medium-sized soil volume, basal cladodes tended to be thicker and areas of main and lateral roots tended to be greater as the CO2 level was doubled. Daughter cladodes tended to be initiated sooner at the current compared with the elevated CO2 level but total areas were similar by 10 weeks. At 10 weeks, daily net CO2 uptake for the three soil volumes averaged 24% higher for plants growing under elevated compared with current CO2 levels, but at 18 weeks only 3% enhancement in uptake occurred. Dry weight gain was enhanced 24% by elevated CO2 during the first 10 weeks but only 8% over 18 weeks. Increasing the soil volume 10-fold led to a greater stimulation of daily net CO2 uptake and biomass production than did doubling the CO2 level. At 18 weeks, root biomass doubled and shoot biomass nearly doubled as the soil volume was increased 10-fold; the effects of soil volume tended to be greater for elevated CO2. The amount of cladode nitrogen per unit dry weight decreased as the CO2 level was raised and increased as soil volume increased, the latter suggesting that the effects of soil volume could be due to nitrogen limitations.  相似文献   

19.
The oxygen requirement for stomatal opening in maize plants ( Zea mays L. hybrid INRA 508) was studied at different CO2 concentrations and light intensities. In the absence of CO2, stomatal opening always required O2, but this requirement decreased with increasing light intensity. In darkness, the lowest O2 partial pressure needed to obtain a weak stomatal movement was about 50 Pa. This value was lowered to ca 10 Pa in light (320 μmol m−2 s−1).
On the other hand. in the absence of O2, CO2enabled stomatal opening to occur in the light, presumably due to the evolved photosynthetic O2. Thus, CO2, which generally reduced stomatal aperture, could induce stomatal movement in anoxia and light. The effect of CO2 on stomatal opening was closely dependent on O2 concentration and light intensity. Stomatal aperture appeared CO2-independent at an O2 partial pressure which was dependent on light intensity and was about 25 Pa at 320 umol m−2 s−1.
The presence of a plasmalemma oxidase, in addition to mitochondrial oxidase, might explain the differences in the O2 requirement at various light intensities. The possible involvement of such a system in relation to the effect of CO2 is discussed.  相似文献   

20.
Abstract— [2-14C]Propionate injected into rats was metabolized into [14C]glucose and 14C-labelled aspartate, glutamate, glutamine and alanine. The results are consistent with the conversion of propionate into succinate and the oxidation of succinate into oxaloacetate, the precursor of labelled amino acids and the substrate for gluconeogenesis.
The ratio of the specific radioactivity of glutamine to glutamate was greater than 1 during the 30 min period in the brain, indicating that propionate taken up by the brain was metabolized mainly in the 'small glutamate compartment' in the brain. The results, therefore, support the previous conclusion (G aitonde , 1975) that the labelling of amino acids by [14C]propionate formed from [U-14C>]-threonine in thiamin-deficient rats was metabolized in the 'large glutamate compartment' of the brain.
The specific radioactivity ratio of glutamine to glutamate in the liver was less than 1 during the 10 min period but greater than 1 at 30min. These findings which gave evidence against metabolic compartments of glutamate in the liver, were interpreted as indicative of the entry of blood-borne [14C]glutamine synthesized in other tissues, e.g. brain. The labelling of amino acids when compared to that after injection of [U-14C]glucose showed that [2-14C]propionate was quantitatively a better source of amino acids in the liver. The concentration of some amino acids in the brain and liver was less in the adult than in the young rats, except for alanine and glutathione, where the liver content was more than double that in the adult.  相似文献   

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