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1.
Levels of activity of the sucrose catabolizing enzymes, acid invertase (EC 3.2.1.26) and sucrose synthase (EC 2.4.1.13), were measured during development of new leaves of Citrus sinensis (L.) Osbeck cv. Shamouti. Soluble acid invertase showed a peak activity of 32 nkat (g fresh weight)−1 at ca 60% of full leaf expansion and rapidly declined toward and after full expansion. There was no concomitant increase in an insoluble form of the enzyme. Sucrose synthase activity, measured in the synthesis direction, declined from 33% of full leaf expansion [10 nkat (g fresh weight)−1] 10, and following, full expansion. Highest sucrose synthase activity, measured in the cleavage direction, was 6 nkat (g fresh weight)−1 and showed little change during development. Acid invertase has a Km of 5 m M for sucrose, while sucrose synthase had a Km of 118 m M for sucrose. Changes in acid invertase activity correlated with changes in the reducing sugar:sucrose ratio. These results suggest that soluble acid invertase activity is the primary enzyme responsible for sucrose catabolism in the expanding Citrus leaf. Changes in leaf expansion rate and invertase activity did not correlate positively with changes in endogenous free IAA level, as determined by enzyme linked immunoassay.  相似文献   

2.
Sugar content and activity of sucrose metabolism enzymes in milled rice grain   总被引:12,自引:1,他引:11  
Most rice (Oryza sativa L.) cultivars grown in the United States were selected for endosperm starch properties and not soluble sugar content. The minor pool of soluble sugar may affect the qualities of rice as a food. Some cultivar variation in soluble sugar content was detected in milled grain, essentially the starchy endosperm, of long grain varieties. Milled grain of cultivars Lemont and Texmati had a soluble sugar content of 0.21 and 0.35% (w/w), respectively, on a fresh weight basis. The dorsal portion of the milled grain contained the greatest amount of soluble sugar, approximately tenfold the amount found in the central core of the grain. Extracts of the milled grain contained sucrose-phosphate synthase (EC 2.4.1.14) and sucrose synthase (EC 2.4.1.13) activities, which were separated by anion exchange chromatography. The presence of sucrose-phosphate synthase in the rice endosperm suggested a mechanism for sucrose accumulation which might be involved in carbon partitioning during grain development.  相似文献   

3.
Aims:  To verify the taxonomic affiliation of bacterium Butyrivibrio fibrisolvens strain A from our collection and to characterize its enzyme(s) responsible for digestion of sucrose.
Methods and Results:  Comparison of the 16S rRNA gene of the bacterium with GenBank showed over 99% sequence identity to the species Pseudobutyrivibrio ruminis . Molecular filtration, native electrophoresis on polyacrylamide gel, zymography and thin layer chromatography were used to identify and characterize the relevant enzyme. An intracellular sucrose phosphorylase with an approximate molecular mass of 52 kDa exhibiting maximum activity at pH 6·0 and temperature 45°C was identified. The enzyme was of inducible character and catalysed the reversible conversion of sucrose to fructose and glucose-1-P. The reaction required inorganic phosphate. The K m for glucose-1-P formation and fructose release were 3·88 × 10−3 and 5·56 × 10−3 mol l−1 sucrose, respectively – while the V max of the reactions were −0·579 and 0·9  μ mol mg protein−1 min−1. The enzyme also released free glucose from glucose phosphate.
Conclusion:  Pseudobutyrivibrio ruminis strain A utilized sucrose by phosphorolytic cleavage.
Significance and Impact of the Study:  Bacterium P. ruminis strain A probably participates in the transfer of energy from dietetary sucrose to the host animal.  相似文献   

4.
Multiple activity peaks of neutral invertase (EC 3.2.1.26) were found in chicory roots ( Cichorium intybus L. var. foliosum cv. Flash). The main activity peak was purified by a combination of anion-exchange chromatography, hydrophobic interaction chromatography, chromatofocusing and gel filtration. This protocol produced a 77-fold purification and a specific activity of 1.6 μmol (mg protein)−1 min−1. The mass of the enzyme was 260 kDa as estimated by gel filtration and 65 kDa on SDS-PAGE. Optimal activity was found between pH 7 and 7.5. The purified enzyme exhibited hyperbolic saturation kinetics with a Km between 10 and 20 mM for sucrose. No other products than glucose and fructose could be detected. Raffinose was hydrolyzed at a rate of 2.4% relative to sucrose whereas the enzyme did not hydrolyze maltose, cellobiose, trehalose, 1-kestose, 1.1-nystose or inulin. Neutral invertase activity was completely inhibited by HgCl2 and AgNO3 and partially inhibited by CoCl2, and ZnSO4 (1 mM). Pyridoxal phosphate (Ki≅ 500 μ M ), Tris (Ki≅ 1.2 m M ), glucose and fructose (Ki≅ 16 m M ) were strong inhibitors of the enzyme. Fructose and Tris behaved as competitive inhibitors. A possible role for the enzyme's activity in vivo is discussed.  相似文献   

5.
Anaerobic production of succinate, a common feature in animals able to sustain anoxia, has seldom been reported in plants. By the use of 1H-nuclear magnetic resonance spectroscopy we show here that succinate is produced by rice seedlings (Oryza sativa L. cv. Arborio) subjected to anoxic conditions. Starting from levels below I μmol (g fresh weight)−1 in air, after 48 h of anoxia the levels of alanine, succinate and lactate had increased to 23.8, 5.2 and 1.0 μmol (g fresh weight) −1, respectively, in shoot tissues. Succinate was accumulated in shoots, notably in the coleoptiles, but not in roots of the rice seedlings, suggesting its involvement in rice coleoptile elongation under anoxia. Other possible functions of succinate production in rice seedling, an organism highly tolerant to anoxia, are discussed.  相似文献   

6.
Previously 'frozen' Tulipa gesneriana L. bulbs cv. Apeldoorn, were planted and grown at higher temperatures to study the role of invertase (EC 3.2.1.26) in the cold-induced elongation of the flower stalk internodes. After planting, flower stalks were left intact, or, the leaves and flower bud were both removed to inhibit internode elongation. In intact flower stalks, elongation of the internodes was accompanied by an accumulation of glucose and an initial decrease in the sucrose content g,−1 dry weight. Insoluble invertase activity g,−1 dry weight hardly changed, but soluble invertase activity showed a peak pattern, that was related, at least for the greater part, to the changes in the sugar contents. Peak activities of soluble invertase were found during (lower- and uppermost internodes) or around the onset of the rapid phase of internode elongation (middle internodes). Internode elongation and glucose accumulation immediately ceased when the leaves and flower bud were removed. Insoluble invertase activity g,−1 dry weight remained at its initial level (lowermost internode) or increased more towards the upper internodes. Soluble invertase activity did not further increase (uppermost internode) or decreased abruptly to a low level. It is concluded that soluble invertase may be one of the factors contributing to glucose accumulation and internode elongation in the tulip flower stalk.  相似文献   

7.
Salt (NaCl) tolerance of 3 eucalypt species ( Eucalyptus alba Reinw. ex Bl., E. camaldulensis Dehnh and E. microtheca f.v. Muell.) was studied: three-month-old seedlings grown in a greenhouse were watered by a saline solution (up to 700 m M ) for 1 month. Mineral, water and sugar contents were highly affected by the saline stress. Sodium, K and Ca were accumulated in the leaves. No significant differences were found between E. camaldulensis and E. microtheca , the tolerant species, in water and mineral contents. Sugar contents were greater in E. microtheca . In E. microtheca Na was highly accumulated in roots [up to 910 μmol (g fresh weight)−1], less in stems [up to 580 μmol (g fresh weight)-1] and leaves [up to 410 μmol (g fresh weight)−r]. Chloride was also accumulated, its content was greater than the total content of Na and K, especially in the salt-tolerant provenance. Potassium and Ca contents were variously affected by the saline stress whereas soluble protein, amino acid and sugar contents were increased. Just after the saline stress, plants showed a large increase in the Na content of the leaf while the decrease in the K content of the stem and leaf continued. Plants were killed by the stress, probably because of too high accumulation of Na in leaves or roots according to the provenance. Osmoregulation and especially the participation of Na are discussed.  相似文献   

8.
In lyophilized needles of Norway spruce ( Picea abies [L.] Karsten) and starting from bud break, we determined enzyme activities (sucrose phosphate synthase [SPS; EC 2.4,1.14]. sucrose synthase [SS; EC 2.4,1.13]. acid invertase [AI; EC 3.2,1.26]) and intermediates (starch, sucrose, glucose, fructose; fructose 6-phosphate, fructose 2.6-bisphosphate [F26BP]) of carbohydrate metabolism together with needle weight, shoot length, chlorophyll and protein. For up to 110 days after bud break, samples were taken twice a week from about 25-year-old trees under field conditions. At least three periods can be distinguished during needle maturation. During the first period (up to 45 days after bud break) Al showed the highest extractable activity. This coincided with very high levels of F26BP (up to 11 pmol [mg dry weight]−1) and a transient increase of starch in parallel to a decrease of sucrose. The interval between 45 and 70 days after bud break was characterized by high SS activity (ratio of fructose/glucose >1), much decreased levels of F26BP (down to below 1 pmol [mg dry weight]−1), and a pronounced increase in the dry weight/fresh weight ratio. In parallel, starch declined and soluble carbohydrates increased. Finally, needle maturation was characterized by decreasing SS and continuously increasing SPS activities, so that the ratio of SPS/SS increased more than 6-fold. AI. however, did not decline with maturation. Changes in pool sizes of metabolites and enzyme activities (AI. SPS) are consistent with current concepts on sink/source transition. SS is obviously important with regard to the synthesis of structural polysaccharides.  相似文献   

9.
Apoplastic mobility of sucrose in storage parenchyma of sugar beet   总被引:2,自引:0,他引:2  
The apoplastic movement of sucrose through storage parenchyma discs (2.4 mm thick) from roots of sugar beet ( Beta vulgaris var. altissima ) was investigated in order to evaluate the suitability of the apoplast for transcellular sugar transport. The sucrose permeability of the discs (P = 5.7 × 10−8 cm s−1 at 25°C) was more than two orders of magnitude lower than that of an equally thick layer of unstirred water. This is due to the small volume fraction of free space (3.1%) and the decreased diffusion coefficient D of sucrose in the cell walls. The effective diffusion coefficient of the apoplast (6 to 9 × 10−7 cm2 s−1 at 25°C) was determined independently of the cross sectional area of free space by treating the time course of fluxes according to Fick's second law. The high diffusion resistance of the apoplast has to be considered in models of native parenchyma transport.  相似文献   

10.
Yearling brown trout, Salmo trutta , were exposed to low mineral content water (nominal concentrations of 20μmol 1−1 magnesium, 7.7 μmol 1−1 potassium, 44 μmol 1−1 sodium) over a pH range of 4.0–5.2 with ambient calcium concentrations of 2.5–60 μmol 1−1. All fish died at pH 4.0 and 4.2 irrespective of ambient calcium concentration and also at pH 4.4 with only 2–3 μmol 1 −1 calcium (that is calcium-free water except for that leached from the diet or excreted by the fish). Good growth rates were obtained over the remaining treatments which extended down to pH 4.4 with as little as 7 μmol 1−1 calcium. When starved, weight loss was inversely correlated with pH. Effects on plasma chloride, percentage dry weight and calcium, potassium sodium, and phosphorus contents of skin, muscle and bone tissue were also investigated. These demonstrated pH effects on mineral metabolism in starved fish, but no effects were detected in fed fish.  相似文献   

11.
The nectar of Strelitzia reginae Ait. was analysed using enzymatic methods and found to contain glucose, fructose and sucrose. Sugar composition changed considerably over the nectar producing period: there was an increase in the amount of glucose (41%) and fructose (32%) between the early and middle stage of secretion and thereafter a decrease of 13 and 24%, respectively, towards the end of secretion. Although the amount of sucrose secreted was initially as much as the glucose and fructose combined, it subsequently decreased, first by 14% and then by 70% at the end of the secretory period so that, whereas in the initial stages of secretion sucrose was quantitatively the dominant sugar, glucose and fructose made up the major part of the nectar as secretion reached its conclusion.
The amounts of potassium and sodium remained at the same low level (around 150 and 30 μg g-1 [w/v) respectively) throughout secretion, while calcium (initially 18 μg g-1) and magnesium (initially 8.0 μg g-1) increased by 47 and 56% respectively, between the early and late stages of secretion. No free amino acids, inorganic phosphate or iron could be detected. Enzymatic analysis revealed only a trace amount of starch. Transmission electron micrographs from both immature and mature plants, however, showed starch grains among other cytoplasmic remnants in the nectary lumen. Mitochondria, vesicles, lipid droplets and ribosomes could also be identified among the luminal cytoplasmic remnants.  相似文献   

12.
The effects of water hardness (9 and 220 mgl−1 as CaCO3) upon zinc exchange in brown trout exposed to 0.77 μmol Zn 1−1 have been investigated using artificial soft water (<49.9 μmol Ca l-1, <40.1 μmol Mg 1−1) and mains hard water (1671.7 μmol Ca 1−1, 493.6 μmol Mg 1−1) of known composition. Both hard and soft water-adapted fish exhibited a bimodal pattern of net zinc influx. Net zinc influxes during both fast and slow uptake phases were significantly greater ( P <0.001) in soft (82.9 and 6.2 μmol Zn 100 g−1 h−1) than in hard water (46.3 and 2.4 μmol Zn 100 g h−1). Zinc efflux (- 0.2 μmol Zn 100 g−1 h−1) was enhanced only in hard water during the slow net influx phase.
Brown trout exposed to zinc in hard water and placed in metal-free media exhibited a greater net efflux (- 25.6 μmol Zn 100 g−1 h−1) of the metal than did fish in soft water (-4.2 μmol Zn 100 g−1 h−1) treated in the same manner. Tissue 65Zn activities reflected both the differences in uptake and excretion rates of the metal between hard and soft water fish. During zinc exposure (0.77 μmol Zn 1−1) high water hardness reduced tissue burdens of the metal by reducing net branchial influx, and enhancing efflux of the metal in hard water fish.  相似文献   

13.
Sucrose accumulation in developing peach fruit   总被引:35,自引:0,他引:35  
Uptake of 14C-sugars and activities of sucrose metabolizing enzymes were determined in order to study the mechanism(s) of sucrose accumulation in developing peach fruit. Mesocarp of young peach fruit contained glucose and fructose but little sucrose. Starting 88 days after anthesis (DAA) the sucrose concentration increased greatly. The mechanism of sucrose accumulation was studied by measuring 14C-sucrose and 14C-glucose uptake rates at three different stages of fruit development, and by assaying weekly the activity of enzymes involved in the hydrolysis and/or synthesis of the soluble sugars. Uptake of 0.5–100 m M 14C-sucrose and 14C-glucose by mesocarp tissue slices showed a complex pattern at the first stage of fruit development (62 DAA). During the subsequent growth stages the pattern of sugar uptake changed and was approximately monophasic at the third stage of fruit development.
At 10 m M , glucose was taken up more rapidly than sucrose at the first and second stage of fruit development. Uptake was partially inhibited by the uncoupler carbonylcyanide m -chlorophenylhydrazone (CCCP) at 25 μ M. These results, together with the presence of a putative extracellular invertase, suggest an apoplastic route for sucrose uptake which is dependent, at least in part, on energy supply.
Activities of sucrose hydrolyzing enzymes (insoluble acid invertase, soluble acid invertase, neutral invertase, sucrose synthase) were high in young fruits and declined sharply with fruit development concomitantly with accumulation of sucrose. The storage of the sugar was not accompanied by a rise in synthetic activities (sucrose synthase, sucrose phosphate synthase), suggesting that sucrose could, at least in part enter the carbohydrate pool directly.  相似文献   

14.
Seed dimorphism in Salicornia europaea: Nutrient reserves   总被引:1,自引:0,他引:1  
Median and lateral seeds of Salicornia europaea L. were separately analysed for their sizes and nutrient reserves. The mean air-dry weight of a single median and lateral seed was 0.31 and 0.25 mg, respectively. The composition as well as the concentration of the nutrient reserves were similar in both seed types. The bulk of the cations was derived from K+, followed by Mg2+, Na+ and Ca2+. The chloride content was somewhat higher than the sodium content, and phosphate was equalled by acid soluble Ca2+ and Mg2+. Starchy compounds and sucrose were present in equal amounts, each of them accounted for about 50% of the carbohydrates. Glucose and fructose were less than 1%. Protein-nitrogen (ethanol-insoluble N) was about 34 g (kg dry seeds)−1. About 7 g (kg dry seeds)−1 was ethanol-soluble nitrogen, of which 10% was derived from amino acids. The total lipid content was more than 290 g (kg dry seeds)−1, 65% were calculated to be glycerides. More than 90% of the fatty acids consisted of linoleic and oleic acids, the majority (72%) of which was linoleic acid.  相似文献   

15.
Abstract. Seedlings of Phaseolus vulgaris were exposed to solutions containing Cd2+ in the range 0 to 1 molm−3. Ethylene formation started following 3 h of exposure to 10−2, 10−1 and 1 mol m−3 Cd2+, peaked at 18 h and returned to a relatively low rate after 24 h. Cadmium-induced ethylene formation depended on the formation of 1-aminocyclopropane-1-carboxylic acid (ACC). Aminoethoxyvinylglycine (AVG, 0.1 mol m−3) inhibited ACC accumulation and ethylene production during exposure to 0.2 mol m−3 Cd2+.
Activity of soluble and ionically-bound peroxidase increased after 18 h of exposure to Cd2+ concentrations above 10−3 mol m−3 due to an increase in activity of cathodic isoperoxidases. Stimulation of soluble and ionically-bound peroxidase by 0.2 mol m−3 Cd2+ was reduced in the presence of 0.1 mol m−3 AVG.
Accumulation of soluble and insoluble ('ligninlike') phenolics was found in plants exposed to Cd2+ (10−2 mol m−3 or above) in the presence or absence of AVG. Deposition of insoluble (autofluorescing) material occurred in cell walls around vessels and was associated with reduced expansion and water content of leaves.  相似文献   

16.
Abstract. Transverse slices through developing grains of Triticum aestivum cv. SUN 9E 16 d after anthesis were incubated in simple defined media with various radioactive labels. In some enzymic assays slices were pretreated with 2.5% Triton X-100 or with 5% butanol to remove cellular membranes and endogenous substrates.
Endogenous potassium leaked from endosperm slices into 30mol m−3 sucrose while sucrose was converted partly into starch. Exogenous alkali-ions, except Li+, stimulated conversion of sucrose to insoluble matter, specifically to starch with K+. Starch synthetase activity of Triton-pretreated slices was stimulated by K+ at both high and low substrate ADPG concentration, but was not affected by phosphate (25 mol m−3).
Phosphate in the medium had no effect on incorporation of sucrose or glucose into alcohol-insoluble material or starch in fresh slices (internal inorganic phosphate (P,) concentration was about 11 mol m−3). Three- to four-fold contrasts in internal Pi level, achieved by prolonged preincubations in different media, did not show an inhibition of starch synthesis by Pi. However, phosphate (25mol m−3) inhibited starch synthesis, that was mediated by ADPG pyrophosphorylase in butanol-pretreated endosperm slices by 15–18%.
It is concluded that starch synthesis in wheat endosperm is not regulated directly by apoplastic Pi; level.  相似文献   

17.
Sugar beets ( Beta vulgaris L. cv. Monohill) grown in a complete nutrient solution, were treated with Cd2+ (5 or 50 μ M ) and/or EDTA (10 or 100 μ M ) in different combinations. The Cd contents of five-week-old roots and shoots were determined by atomic absorption spectrophotometry, and the sucrose, glucose and fructose contents were measured enzymatically. The Cd2+ uptake in both roots and shoots shows a linear relationship to the concentration of free Cd2+ in the nutrient solution. This uptake is diminished in the presence of EDTA, suggesting that the Cd-EDTA complex is unable to penetrate the membranes. The contents of glucose, fructose and sucrose in both roots and shoots decrease with increasing uptake of free Cd2+. This may be a secondary effect caused by the inhibition of photosynthesis in the presence of Cd2+. EDTA reduces the inhibition of Cd2+ on sugar formation and accumulation. In the presence of EDTA alone the sugar content increases somewhat. EDTA slightly influences the dry weights of whole plants. The ratio roots:whole plants increases. Cd2+ (≤ 50 μ M ) increases the dry matter portion of roots by ca 30%, but not that of shoots.  相似文献   

18.
Even in the presence of glucose the growth of Marchantia polymorpha L. (cell line HYH-2F) requires light, and growth is more sensitive to 10−6 M 3-(3, 4-dichlorophenyl)-1, 1-dimethylurea than to 10−4 Antimycin A. The inability of the cells to grow in the dark is due to the low level of respiration. The respiration rate under light increased to four times the dark value. The values of the compensation ratio (the photosyntehtic rate/the respiration rate) for the oxygen exchange were below 1.0 daring the growth period, although oxygen evolution was found. At the early exponential phase, oxygen evolution was 0.373 μmol (mg cell dry weight)−1 h−1 [61.7 μmol (mg chlorophyll)−1 h−1]. M. polymorpha cells are unable to grow anaerobically in the light without a supply of carbon dioxide. When 1% carbon dioxide in nitrogen is supplied, photochemically produced oxygen and energy are sufficient for sustained growth although at significantly reduced yields in both cell dry weight and chlorophyll. Photosyntehtic CO2 assimilation rate was 0.13 μmol (mg cell dry weight)−1 h−1[11.3 μmol (mg chlorophyll)−1 h−1]. At least one-third of the carbon atoms in cellular constituents seem to be derived from atmospheric carbon dioxide, which indicates that M. polymorpha cells grow photomixotrophicaily.  相似文献   

19.
When Phaseolus vulgaris L. cv. Kentucky Wonder plants were supplied with various levels of nitrate for 34 days, nodule weight (plant)−1, acetylene reduction activity (g nodule)−1, and sugar concentration in nodules were depressed >60% (7.5 m M nitrate vs nil nitrate). Starch concentration in nodules was more than double the sugar concentration and declined only slightly in response to nitrate level. At the highest level of nitrate, sugar concentration in nodules was 50% greater than that in roots and nodule starch was about 6-fold greater than root starch on a fresh weight basis. When plants were grown with 1 m M nitrate and then supplied with 12 m M nitrate for 7 days, the rapid decline in acetylene reduction activity coincided with a decline in sucrose concentration. However, glucose and fructose concentrations declined only after the largest decrease in acetylene reduction had occurred, and the quantitative decrease in glucose and fructose in nodules was small relative to sucrose. Other results showed that the magnitude of the effect of nitrate on some nodule carbohydrate compounds depends on Rhizobium phaseoli strain and on whether plants were grown with or without nitrate prior to experimental treatments. Some of the results are consistent with the carbohydrate-deprivation hypothesis for inhibition of legume nodules by nitrate. However, there are several complications involved in the interpretation of results of this type, and other possible explanations for the results are suggested.  相似文献   

20.
Abstract The anoxygenic phototrophic purple sulfur bacterium Thiocapsa roseopersicina was grown in illuminated continuous cultures with thiosulfate as growth limiting substrate. Aeration resulted in completely colorless cells growing chemotrophically, whereafter the conditions were changed to a 23 h oxic/1 h anoxic regime. After 11 volume changes at a dilution rate of 0.031 h−1 (35% of μmax) a time dependent equilibrium was established. During the 23 h oxic periods bacteriochlorophyll a synthesis (BChl a ) was not observed, whereas during the 1 h anoxic periods synthesis was maximal (i.e. 1.1 μg (mg protein)−1 h−1). As a result the BChl a concentration gradually increased from zero to an average value over 24 h of 1.9 μg (mg protein)−1. Concomitantly, the protein concentration increased from 13.9 mg 1−1 during continuous oxic conditions to 28.8 mg 1−1. For comparison, the protein concentration during fully phototrophic growth at an identical thiosulfate concentration in the inflowing medium was 53.7 mg 1−1. The specific respiration rate was 8 μmol O2 (mg protein)−1 h−1 during full chemotrophic growth and gradually decreased to 3.5 μmol O2 (mg protein)−1 h−1 after 11 volume changes at the regime employed. These data show that T. rosepersicina is able to simultaneously utilize light and aerobic respiration of thiosulfate as sources of energy. The ecological relevance of the data is discussed.  相似文献   

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