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21.
Protoplasts isolated from beetroot tissue took up glucose preferentially whereas sucrose was transported more slowly. The 14C-label from [14C]glucose and [14C]sucrose taken up by the cells could be detected rapidly in phosphate esters and, after feeding of [14C]glucose was found also in sucrose. The temperature-dependent uptake process (activation energy EA about 50 kJ · mol–1) seems to be carrier mediated as indicated by its substrate saturation and, for glucose, by competition experiments which revealed positions C1, C5 and C6 of the D-glucose molecule as important for effective uptake. The apparent Km(20° C) for glucose (3-O-methylglucose) was about 1 mM whereas for sucrose a significantly lower apparent affinity was determined (Km about 10 mM). When higher concentrations of glucose (5 mM) or sucrose (20 mM) were administered, the uptake process followed first-order kinetics. Carrier-mediated transport was inhibited by N,N-dicyclohexylcarbodiimide, Na-orthovanadate, p–chloromercuribenzenesulfonic acid, and by uncouplers and ionophores. The uptake system exhibited a distinct pH optimum at pH 5.0. The results indicate that generation of a proton gradient is a prerequisite for sugar uptake across the plasma membrane. Protoplasts from the bundle regions in the hypocotyl take up glucose at higher rates than those derived from bundle-free regions. The results favour the idea that apoplastic transport of assimilates en route of unloading might be restricted to distinct areas within the storage organ (i.e. the bundle region) whereas distribution in the storage parenchyma is symplastic.Abbreviations CCCP
Carbonylcyanide m–chlorophenylhydrazone
- DCCD
N,N-dicyclohexylcarbodiimide
- DOG
deoxyglucose
- Mes
2-(N-morpholino)ethanesulfonic acid
- 3-OMG
3-O-methylglucose
- PCMBS
p–chloromercuribenzenesulfonic acid
- SDS
Sodium dodecyl sulfate
- Tris
2-amino-2-(hydroxymethyl)-1,3-propanediol 相似文献
22.
The nitrate reductase (NR, EC 1.6.6.1) activity in root nodules formed by hydrogenase positive (Hup+ ) and hydrogenase negative (Hup− ) Rhizobium leguminosarum strains was examined in symbioses with the pea cultivar Alaska ( Pisum sativum L.), Rates of activity were determined by the in vivo assay in nodules from plants that were only N2 -dependent or grown in the presence of 2 m M KNO3 . The rates varied widely among strains, regardless of the Hup phenotype of the R. leguminosarum strain used for inoculation, but the overall results indicated that nodules formed by Hup− strains accumulated more nitrite in the incubation medium than did those with Hup− phenotypes. Total plant dry weight and reduced nitrogen content of pea plants grown in the presence of 2 m M KNO3 and inoculated with single Hup+ and Hup− R. leguminosarum strains were statistically different among some strains. These observations suggest that the possible advantages derived from the presence of the Hup system on whole plant growth may be counteracted by the higher rates of NR activity in the Hup− strains in the R. leguminosarum -pea symbiosis. 相似文献
23.
The fate of15N labeled nitrogen applied to mature citrus trees 总被引:1,自引:0,他引:1
Summary The efficiency and balance of nitrogen from one year's application was studied in a long-term fertigation experiment. Enriched
nitrogen fertilizer, K15NO3, was applied to a 22-year-old Shamouti orange tree with a history of high N applications (N3) and to an N-starved tree (N1). The distribution of N in the different parts of the trees and in the soil was determined after the experimental trees were
excavated.
Similar total recovery of the labeled fertilizer N was found in the trees and soil in both treatments (N1−61.7% N3−56%). However, the distribution between tree and soil was different. The amount of recovered residual fertilizer in the soil
was much larger in the N3 treatment than in N1. The highest percentage of fertilizer N was found in the new organs,i.e. fruits, twigs and leaves. The roots and branches took up only 6–14% from the labeled fertilizer.
Only 20.9% of the leaf N and 23.4% of the fruit N in the N3 tree originated in the labeled fertilizer, indicating translocation of N from older parts of the tree to new growth.
Evidence was found of storage of N in the wooded branches, while the roots contained a surprisingly small part of labeled
fertilizer.
Contribution 1599E. 相似文献
24.
Growth of 2-month-old nonnodulatedHippophaë rhamnoides seedlings supplied with combined N was compared with that of nodulated seedlings grown on zero N. Plant growth was significantly better with combined N than with N2 fixation and, although not statistically significant for individual harvests, tended to be highest in the presence of NH 4 + , a mixture of NH 4 + and NO 3 ? producing the highest yields. Growth was severely reduced when solely dependent on N2 fixation and, unlike the combined-N plants, shoot to root ratios had only slightly increased after an initial decrease. An apparently insufficient nodule mass (nodule weight ratio <5 per cent) during the greater part of the experimental period is suggested as the main cause of the growth reduction in N2-fixing plants. Thein vivo nitrate reductase activity (NRA) of NO 3 ? dependent plants was almost entirely located in the roots. However, when grown with a combination of NO 3 ? and NH 4 + , root NRA was decreased by approximately 85 per cent.H. rhamnoides demonstrated in the mixed supply a strong preference for uptake of N as NH 4 + , NO 3 ? contributing only for approximately 20 per cent to the total N assimilation. Specific rates of N acquisition and ion uptake were generally highest in NO 3 ? +NH 4 + plants. The generation of organic anions per unit total plant dry weight was approximately 40 per cent less in the NH 4 + plants than in the NO 3 ? plants. Measured extrusions of H+ or OH? (HCO 3 ? ) were generally in good agreement with calculated values on the basis of plant composition, and the acidity generated with N2 fixation amounted to 0.45–0.55 meq H+. (mmol Norg)?1. Without acidity control and in the presence of NH 4 + , specific rates of ion uptake and carboxylate generation were strongly depressed and growth was reduced by 30–35 per cent. Growth of nonnodulatedH. rhamnoides plants ceased at the lower pH limit of 3.1–3.2 and deterioration set in; in the case of N2-fixing plants the nutrient solution pH stabilized at a value of 3.8–3.9 without any apparent adverse effects upon plant performance. The chemical composition of experimental and field-growing plants is being compared and some comments are made on the nitrogen supply characteristics of their natural sites. 相似文献
25.
Activity of key nitrogen assimilating enzymes was studied in developing grains of high-lysine opaque sorghum P-721 and normal
sorghum CSV-5. The higher percentage of protein in opaque sorghum was mainly due to lower starch content since protein per
grain was less than in CSV-5. During grain development, albufn and globulin decreased while prolafne and glutelin increased.
Prolafne content in CSV-5 was higher than in opaque sorghum. Average nitrate reductase activity in flag and long leaf were
similar in both the varieties. The nitrate reductase activity decreased during grain development. Glutamate dehydrogenase
activity was higher during early development and lower at later stages in opaque sorghum than in CSV-5. Glutamate oxaloacetate
transaminase activity was higher and glutamine synthetase lower in opaque sorghum than in CSV-5 grains during development.
Glutamate synthase activity was higher in opaque sorghum up to day 20 and lower thereafter than in CSV-5. It is suggested
that reduced activities of glutamine synthetase as well as glutamate synthase in opaque sorghum as compared to CSV-5 during
later stages of development may restrict protein accumulation in the former. 相似文献
26.
C. S. Ramarao S. R. Acharya K. S. Krishnan U. W. Kenkare 《Journal of biosciences》1987,11(1-4):119-135
Preparations having properties resembling those of synaptosomes have been isolated from whole fly homogenates ofDrosophila melanogaster using ficoll gradient floatation technique. These have been characterized by marker enzymes and electron microscopy and binding of muscarinic antagenist3H Quinuclidinyl benzilate. An uptake system for neurotransmitter, ã-Aminobutyric acid has been demonstrated in these preparations. A high affinity uptake system for L-glutamate has also been studied in these subcellular fractions. This uptake of glutamate is transport into an osmotically sensitive compartment and not due to binding of glutamate to membrane components. The transport of glutamate has an obligatory requirements for either sodium or potassium ions. Kinetic experiments show that two transport systems, withK m values 0.33 X 10-6M and 2.0 X 10-6M, respectively, function in the accumulation of glutamate. ATP stimulates lower affinity transport of glutamate. Inhibition of glutamate uptake by L-aspartate but not by phenylalanine and tyrosine indicates that a common carrier mediates the transport of both glutamate and aspartate. β-N-oxalyl-L-β β-diamino propionic acid and kainic acid, both inhibitors of glutamate transport in mammalian brain preparations, strongly inhibited transport of glutamate inDrosophila preparations Comparison with uptake of ã-aminobutyric acid and glutamate in isolated larval brain is presented to show that the synaptosome-like preparations we have isolated are rich in central nervous system derived structures, and presynaptic endings from neuromuscular junctions. 相似文献
27.
Effect of rainstorms on heterotrophic bacterial activity in a hypertrophic African lake 总被引:1,自引:1,他引:0
Richard D. Robarts 《Hydrobiologia》1987,148(3):281-286
Hartbeespoort Dam is a hypertrophic man-made lake which is located in the Transvaal Province of South Africa. This region has recently experienced its most severe drought of the century. However, on three occasions in the summer rainy seasons of 1984 and 1985, major rainfalls (> 50 mm) occurred which caused large inflows to the lake. Inflowing river water entered as a density current causing marked silting of the water. Within the epilimnion (0–10 m) prior to these rainfalls there was usually no variation of bacterial numbers with depth, but heterotrophic bacterial activity (glucose uptake) decreased with depth concomitant with primary production. With the increased river inflow bacterial numbers did not increase but bacterial activity at the bottom of the epilimnion (10 m) increased to as high as 2.7 µg C l–1 h–1 in January 1985, reversing the depth profile of bacterial activity within the epilimnion. This resulted in decreased glucose concentrations (Kt + Sn) and turnover times. Heterotrophic activity per cell increased by between 2.5 and 5 times. These data demonstrate that storm events are important phenomena causing short-term changes in the metabolic activity of planktonic heterotrophic bacteria in lakes. 相似文献
28.
Temperature Dependence of Drug Interaction with the Platelet 5-Hydroxytryptamine Transporter: A Clue to the Imipramine Selectivity Paradox 总被引:2,自引:2,他引:0
Although [3H]imipramine is a selective radioligand for the 5-hydroxytryptamine (5-HT) transporter in human platelets, its affinity for binding to the 5-HT transporter complex at 0 degrees C (0.6 nM) is significantly higher than its potency for inhibition of [3H]5-HT uptake at the physiological temperature of 37 degrees C (Ki = 29 nM). As this apparent discrepancy could be related to the assay temperature, we studied the thermodynamics of drug interaction with the 5-HT transporter at assay temperatures between 0 degrees C and 37 degrees C, using as radioligands [3H]imipramine (0 degrees C and 20 degrees C) and [3H]paroxetine (20 degrees C and 37 degrees C), a newly available probe for the 5-HT transporter. At 20 degrees C, Ki values of 14 tricyclic and nontricyclic drugs for inhibition of [3H]imipramine and [3H]paroxetine binding to human platelet membranes were highly significantly correlated (r = 0.98, p less than 0.001), validating the use of these two radioligands to study the 5-HT transporter over a temperature range larger than was previously possible with [3H]imipramine alone. The affinity of imipramine for the 5-HT transporter is progressively enhanced with decreasing incubation temperature, thus favoring the selectivity of [3H]imipramine for the 5-HT transporter at 0 degrees C. At 37 degrees C, the Ki of imipramine for inhibition of [3H]paroxetine binding is 32 nM, and equals its Ki value for inhibition of 5-HT uptake into human platelets. With the exception of chlorimipramine, other tricyclic 5-HT uptake inhibitors showed a temperature sensitivity in their interaction with the 5-HT transporter similar to that of imipramine.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
29.
Yutaka Nagata Masato Ando Mitsuyoshi Iwata Atsushi Hara Tamotsu Taketomi 《Journal of neurochemistry》1987,49(1):201-207
The effects of some gangliosides on active uptake of nonmetabolizable alpha-aminoisobutyric acid (AIB) and Na+, K+-ATPase and Ca2+, Mg2+-ATPase activities in superior cervical ganglia (SCG) and nodose ganglia (NG) excised from adult rats were examined during aerobic incubation at 37 degrees C for 2 h. In NG, amino acid uptake was greatly accelerated with the addition of galactosyl-N-acetylgalactosaminyl-[N-acetylneuraminyl]-galactosylgluc osyl ceramide (GM1) (85%) and also with N-acetylgalactosaminyl-[N-acetylneuraminyl]-galactosylglucosyl ceramide (GM2) or [N-acetylneuraminyl]-galactosyl-N-acetylgalactosaminyl-[N-acetyl- neuraminyl]-galactosylglucosyl ceramide (GD1a) (43% each) compared with a nonaddition control at a 5 nM concentration. Under identical conditions, Na+, K+-ATPase activity was strongly stimulated with GM1 (180%) and GD1a (93%), whereas Ca2+, Mg2+-ATPase activity showed no change. In SCG, on the other hand, AIB uptake was apparently inhibited (-27%) by addition of GM1, with a slight decrease in Na+, K+-ATPase but no change in Ca2+, Mg2+-ATPase activity in the tissue. Both asialo-GM1, in which N-acetylneuraminic acid is deficient, and Forssman glycolipid, which is not present in nervous tissue, failed to produce any significant increase in both SCG and NG not only in amino acid uptake, but also in Na+, K+-ATPase activity. A kinetic study of active AIB uptake showed that GM1 ganglioside produced an increase in Km with no change in Vmax in SCG, whereas it caused a decrease in Km with a slight increase in Vmax in NG. Treatment of NG and SCG with neuraminidase from Vibrio cholerae, an enzyme that split off sialic acid from polysialoganglioside, leaving GM1 intact, caused little inhibition of the amino acid uptake.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
30.
[3 H]Dipyridamole Binding to Guinea Pig Brain Membranes: Possible Heterogeneity of Central Adenosine Uptake Sites 总被引:1,自引:0,他引:1
The binding of [3H]dipyridamole ([3H]DPR) to guinea pig brain membranes is described and compared to that of [3H]nitrobenzylthioinosine ([3H]NBI). The binding of [3H]DPR is saturable, reversible, and specific with pharmacologic evidence indicating that this ligand is binding to the adenosine uptake site. Compared to [3H]NBI the binding of [3H]DPR is of higher capacity (Bmax = 208 +/- 16 fmol/mg protein for [3H]NBI and 530 +/- 40 fmol/mg protein for [3H]DPR) and lower affinity (KD = 0.35 +/- 0.02 nM for [3H]NBI and 7.6 +/- 0.7 nM for [3H]DPR). The adenosine uptake inhibitors are the most potent inhibitors of binding (Ki of 10(-8)-10(-7) M) whereas adenosine receptor ligands such as cyclohexyladenosine, 2-chloroadenosine, and various methylxanthines are several orders of magnitude less potent (Ki 10(-5)-10(-2). The inhibition of [3H]DPR binding by NBI is biphasic, with only 40% of binding being susceptible to inhibition of NBI concentrations less than 10(-5) M. The tissue distribution of [3H]DPR binding parallels that of [3H]NBI although in most cases significantly more sites are observed with [3H]DPR. Calcium channel blocking agents such as nifedipine, nimodipine, and verapamil are also inhibitors of [3H]DPR binding with potencies in the micromolar range. The data are consistent with [3H]DPR being a useful additional ligand for the adenosine uptake site and provide evidence that multiple uptake binding sites exist of which only about 40% are NBI-sensitive. 相似文献