Hormonal Control of Sucrose Phosphate Synthase and Sucrose Synthase in Sugar Beet

We studied the effects of synthetic analogs of phytohormones (benzyladenine, IAA, and GA) on the activities of the enzymes catalyzing sucrose synthesis and metabolism, sucrose phosphate synthase (SPS, EC 2.4.1.14) and sucrose synthase (SS, EC 2.4.1.13), and on the content of chlorophyll and protein during the sugar-beet (Beta vulgaris L.) ontogeny. Plant spraying with phytohormonal preparations activated SPS in leaves; direct interaction between phytohormones and the enzyme also increased its activity. The degree of this activation differed during the ontogeny and in dependence on the compound used for treatment. Analogs of phytohormones maintained high protein level in leaves, retarded chlorophyll breakdown, and, thus, prolonged leaf functional activity during development. Phytohormonal preparations practically did not affect the SS activity both after plant treatment and at their direct interaction with the enzyme. It is supposed that the SS activity in sugar-beet roots is controlled by sucrose synthesized in leaves rather than by phytohormones. The effects of hormones on leaf metabolism were mainly manifested in growth activation.     

Enzymes of starch and sucrose metabolism in Zea mays leaves

Mesophyll and bundle sheath cells of maize leaves were separated and enzymes of starch and sucrose metabolism assayed. The starch content and activities of ADPglucose (ADPG) starch synthetase and phosphorylase expressed both on a chlorophyll and a protein basis were much lower in mesophyll cells compared to bundle sheath preparations. Exposure of the leaves to continuous illumination for 2·5 days caused the starch content of mesophyll cells to rise greatly and led to considerable increases in ADPG starch synthetase and phosphorylase activity. In glasshouse grown leaves the bulk of invertase, sucrose phosphate synthetase, sucrose phosphatase, UDPglucose pyrophosphorylase and amylase was situated in the mesophyll layer. Sucrose synthetase, ADPG starch synthetase and phosphorylase were largely confined to the bundle sheath. No enzyme could be completely assigned to one particular cell layer. Upon continuous illumination both ADPG starch synthetase and phosphorylase increased in the mesophyll bythe same relative amount. The mesophyll is likely to be a major site for sucrose synthesis in maize leaves.     

The effect of Albugo Candida (white blister rust) on the photosynthetic and carbohydrate metabolism of leaves of Arabidopsis thaliana

Albugo candida (pers.) O. Kuntze (white blister rust) is a biotrophic fungus which infects cruciferous plants including Arabidopsis thaliana (L) Heynh. We report the effect of this pathogen on the photosynthetic and carbohydrate metabolism of A. thaliana. As infection progressed A. Candida caused a reduction in the rate of photosynthesis when measured at either ambient or saturating concentrations of CO₂. These data suggested that both chlorophyll and Rubisco were lost from regions of infected leaves, and measurements of chlorophyll, Rubisco content and activity supported these observations. The reduction in the rate of photosynthesis was not caused by closure of stomata as transpiration was unaffected by the disease. Infected leaves accumulated both soluble carbohydrates and starch. The activities of sucrose-phosphate synthase, sucrose synthase and ADP glucose pyrophosphorylase did not change in response to infection. However, the activities of both the wall-bound and soluble acid invertases were higher in infected leaves than in controls; a new soluble invertase isoform with a pl of 5-1 appeared in infected leaves. The possible origin of the increase in wall-bound and soluble invertase activities and its effect on the carbohydrate and photosynthetic metabolism of the leaf are discussed.     

Effect of High Temperature on Plant Growth and Carbohydrate Metabolism in Potato

This study was undertaken to determine the role of sucrose-metabolizing enzymes in altered carbohydrate partitioning caused by heat stress. Potato (Solanum tuberosum L.) genotypes characterized as susceptible and tolerant to heat stress were grown at 19/17[deg]C, and a subset was transferred to 31/29[deg]C. Data were obtained for plant growth and photosynthesis. Enzyme activity was determined for sucrose-6-phosphate synthase (SPS) in mature leaves and for sucrose synthase, ADP-glucose pyrophosphorylase, and UDP-glucose pyrophosphorylase in developing tubers of plants. High temperatures reduced growth of tubers more than of shoots. Photosynthetic rates were unaffected or increased slightly at the higher temperature. Heat stress increased accumulation of foliar sucrose and decreased starch accumulation in mature leaves but did not affect glucose. SPS activity increased significantly in mature leaves of plants subjected to high temperature. Changes in SPS activity were probably not due to altered enzyme kinetics. The activity of sucrose synthase and ADP-glucose pyrophosphorylase was reduced in tubers, albeit less quickly than leaf SPS activity. There was no interaction of temperature and genotype with regard to the enzymes examined; therefore, observed differences do not account for differences between genotypes in heat susceptibility.     

Up-regulation of sucrose metabolizing enzymes in Oncidium goldiana grown under elevated carbon dioxide

Experiments were conducted in controlled growth chambers to evaluate how increase in CO₂ concentration affected sucrose metabolizing enzymes, especially sucrose phosphate synthase (SPS; EC 2.4.1.14) and sucrose synthase (SS; EC 2.4.1.13), as well as carbon metabolism and partitioning in a tropical epiphytic orchid species (Oncidium goldiana). Response of ribulose‐1,5‐bisphosphate carboxylase/oxygenase (Rubisco; EC 4.1.1.39) to elevated CO₂ was determined along with dry mass production, photosynthesis rate, chlorophyll content, total nitrogen and total soluble protein content. After 60 days of growth, there was a 80% and 150% increase in dry mass production in plants grown at 750 and 1 100 μl l^?1 CO₂, respectively, compared with those grown at ambient CO₂ (about 370 μl l^?1). A similar increase in photosynthesis rate was detected throughout the growth period when measured under growth CO₂ conditions. Concomitantly, there was a decline in leaf Rubisco activity in plants in elevated CO₂ after 10 days of growth. Over the growth period, leaf SPS and SS activities were up‐regulated by an average of 20% and 40% for plants grown at 750 and 1100 μl l^?1 CO₂, respectively. Leaf sucrose content and starch content were significantly higher throughout the growth period in plants grown at elevated CO₂ than those at ambient CO₂. The partitioning of photosynthetically fixed carbon between sucrose and starch appeared to be unaffected by the 750 μl l^?1 CO₂ treatment, but it was favored into starch under the 1 100 μl l^?1 CO₂ condition. The activities of SPS and SS in leaf extracts were closely associated with photosynthetic rates and with partitioning of carbon between starch and sucrose in leaves. The data are consistent with the hypothesis that the up‐regulation of leaf SPS and SS might be an acclimation response to optimize the utilization and export of organic carbon with the increased rate of inorganic‐carbon fixation in elevated CO₂ conditions.     

Phosphorus Nutrition Influence on Starch and Sucrose Accumulation, and Activities of ADP-Glucose Pyrophosphorylase and Sucrose-Phosphate Synthase during the Grain Filling Period in Soybean

Several lines of evidence indicate that the partitioning of photosynthate between starch and sucrose is influenced by the relative concentrations of inorganic phosphate (Pi) in the cytosol and chloroplast. Two greenhouse experiments were conducted to determine the influence of long-term differences in soil P levels, ranging from deficient to supraoptimum, on leaf starch and sucrose concentrations, and activities of adenosine diphosphate glucose (ADPG) pyrophosphorylase and sucrose-phosphate synthase (SPS) during the grain filling period in soybean (Glycine max [L.] Merr.). It was hypothesized that, compared with optimum P nutrition, leaf starch and sucrose concentrations would be increased and decreased, respectively, for P deficiency and visa versa for supraoptimum P nutrition. Relative to the optimum soil P level, leaf Pi concentration was not altered by P deficiency but was increased two- to fourfold for the supraoptimum soil P treatment. The concentrations of leaf starch and sucrose were not markedly affected by any of the P fertility treatments and were not closely related to the activities of ADPG pyrophosphorylase and SPS. P deficiency resulted in increased activity of both enzymes in one of the experiments. The results indicated that long-term soil P treatments, that caused either large decreases in plant growth (P deficiency) or large increases in leaf Pi concentration (supraoptimum P), did not markedly alter starch and sucrose metabolism. Furthermore, it can be inferred that the method of plant culture and/or imposition of the P treatments is a critical factor in interpreting results of P nutrition studies.     

Elevated sucrose-phosphate synthase activity in transgenic tobacco sustains photosynthesis in older leaves and alters development

Constitutive over-expression of a maize sucrose-phosphate synthase (SPS) gene in tobacco (Nicotiana tabacum) had major effects on leaf carbohydrate budgets with consequences for whole plant development. Transgenic tobacco plants flowered earlier and had greater flower numbers than wild-type plants. These changes were not linked to modified source leaf carbon assimilation or carbon export, although sucrose to starch ratios were significantly higher in leaves expressing the transgene. The youngest and oldest leaves of plants over-expressing SPS had up to 10-fold wild-type maximal extractable SPS activity, but source leaf SPS activities were only 2-3 times greater in these lines than in the wild type. In the oldest leaves, where the expression of the transgene led to the most marked enhancement in SPS activity, photosynthesis was also increased. It was concluded that these increases in the capacity for sucrose synthesis and carbon assimilation, particularly in older leaves, accelerate the whole plant development and increase the abundance of flowers without substantial changes in the overall shoot biomass.     

Regulation of leaf photosynthetic rate correlating with leaf carbohydrate status and activation state of Rubisco under a variety of photosynthetic source/sink balances

There is evidence suggesting that in plants changes in the photosynthetic source/sink balance are an important factor that regulates leaf photosynthetic rate through affects on the leaf carbohydrate status. However, to resolve the regulatory mechanism of leaf photosynthetic rate associated with photosynthetic source/sink balance, information, particularly on mutual relationships of experimental data that are linked with a variety of photosynthetic source/sink balances, seems to be still limited. Thus, a variety of manipulations altering the plant source/sink ratio were carried out with soybean plants, and the mutual relationships of various characteristics such as leaf photosynthetic rate, carbohydrate content and the source/sink ratio were analyzed in manipulated and non-manipulated control plants. The manipulations were removal of one-half or all pods, removal of one-third or two-third leaves, and shading of one-third or one-half leaves with soybean plants grown for 8 weeks under 10 h light (24 degrees C) and 14 h darkness (17 degrees C). It was shown that there were significant negative correlations between source/sink ratio (dry weight ratio of attached leaves to other all organs) and leaf photosynthetic rate; source/sink ratio and activation ratio (percentage of initial activity to total activity) of Rubisco in leaf extract; leaf carbohydrate (sucrose or starch) content and photosynthetic rate; carbohydrate (sucrose or starch) content and activation ratio of Rubisco; amount of protein-bound ribulose-1,5-bisphosphate (RuBP) in leaf extract and leaf photosynthetic rate; and the amount of protein-bound RuBP and activation ratio of Rubisco. In addition, there were significant positive correlations between source/sink ratio and leaf carbohydrate (sucrose or starch) content; source/sink ratio and the amount of protein-bound RuBP; carbohydrate (sucrose or starch) content and amount of protein-bound RuBP and the activation ratio of Rubisco and leaf photosynthetic rate. The plant water content, leaf chlorophyll and Rubisco contents were not affected significantly by the manipulations. There is a previous report in Arabidopsis thaliana that the amount of protein-bound RuBP in leaf extract correlates negatively with the activation ratio of Rubisco in the leaf extract. Therefore, the results obtained from the manipulation experiments indicate that there is a regulatory mechanism for the leaf photosynthetic rate that correlates negatively with leaf carbohydrate (sucrose and starch) status and positively with the activation state of Rubisco under a variety of photosynthetic source/sink balances.     

Rhythmic growth and carbon allocation in Quercus robur. Sucrose metabolizing enzymes in leaves

The high sucrose phosphate synthase (SPS) capacity and the low soluble acid invertase activity of mature leaves of the first flush of leaves remained stable during second flush development. Conversely, fluctuations of sucrose synthase (SS) activity were in parallel with the sucrose requirement of the second flush. Sucrose synthase activity (synthesis direction) in first flush leaves could increase in 'response' to sink demand constituted by the second flush growth. Only the ptotosynthates provided by flush mature leaves were translocated for a current flush, while the starch content of these leaves remained stable. After their emergence, second flush leaves showed an increase in SPS and SS (Synthetic direction) activities. The high sucrose synthesis in second flush leaves was used for leaf expansion. When young leaves were 30% fully expanded (stage II20), SPS activity showed little change whereas SS activity declined rapidly toward and after full leaf expansion. The starch accumulation in the young leaves occured simultaneously with their expansion. Developing leaves showed a high level of acid invertase activity until maximum leaf expansion (stage II1). In first and second flush leaves, changes in acid invertase activity correlated positively with changes in reducing sugar concentrations. Alkaline invertase and sucrose synthase (cleavage direction) activities showed similar changes with low values when compared with those of acid invertase activity, especially in second flush leaves. The present results suggest that soluble acid invertase was the primary enzyme responsible for sucrose catabolism in the expanding common oak leaf.     

Sucrose metabolism in Sorghum vulgare at ripening

Enzymes associated with sucrose metabolism in root, stem, leaf and grain of Sorghum vulgare Pers. (cv. JS 263) were studied at the ripening stage. Sucrose phosphate synthetase was dominating in the leaf and sucrose synthetase in the grain. Invertases were more active in leaf, root and stem tissues than in grains. The maximum activities of ADPG pyrophosphorylase and UDPG pyrophosphorylase were found in grains and leaves, respectively. Sucrose synthetase from grains catalyses both synthesis and cleavage of sucrose but the two activities differed in their responses to the effect of temperature, pH and type of buffer. The Km values of the enzyme for UDPG, ADPG, GDPG, TDPG and CDPG were 8.5, 5.3, 16.8 2.2 and 10.7 mM, and for UDP and ADP they were 17.2 and 55.0 mM respectively.     

Carbohydrate metabolism in Musa paradisiaca

Considerable variations exist in the content of glucose, fructose, sucrose, starch and protein and in the activities of enzymes involved in carbohydrate metabolism between different parts of the banana plant (Musa paradisiaca). Sucrose synthetase is present in the highest concentration in rootstock and fruit pulp, and sucrose phosphate synthetase in the pseudostem. The highest ratio of the activity of sucrose phosphate synthetase to sucrose synthetase is found in leaves. Acid invertase is present in leaves, leaf-sheath and fruit pulp and is not demonstrable in rootstock and pseudostem. Neutral invertase activity is high in pseudostem and leaf-sheath. Starch phosphorylase is largely concentrated in fruit pulp and rootstock. The maximum activity of ATP:d-phosphoglucose (ADPG) pyrophosphorylase is found in rootstock. β-Amylase is not demonstrable in rootstock and is largely concentrated in leaf-sheath. Hexokinase is most active in rootstock and the lowest in leaves. Acid phosphatase and alkaline phosphatase activity is highest in fruit pulp and pseudostem. Glucosephosphate isomerase is most active in the rootstock and lowest in the leaves.     

Changes in Mulberry Leaf Metabolism in Response to Water Stress

A series of experiments were conducted to characterize the water stress-induced changes in the activities of RuBP carboxylase (RuBPCO) and sucrose phosphate synthase (SPS), photosystem 2 activity, and contents of chlorophylls, carotenoids, starch, sucrose, amino acids, free proline, proteins and nucleic acids in mulberry (Morus alba L. cv. K-2) leaves. Water stress progressively reduced the activities of RuBPCO and SPS in the leaf extracts, the chlorophyll content, and PS2 activity in isolated chloroplasts. Plants exposed to drought showed lower content of starch and sucrose but higher total sugar content than control plants. While the soluble protein content decreased under water stress, the amino acid content increased. Proline accumulation (2.5-fold) was noticed in stressed leaves. A reduction in the contents of DNA and RNA was observed. Reduced nitrogen content was associated with the reduction in nitrate reductase activity. SDS-PAGE protein profile showed few additional proteins (78 and 92 kDa) in the water stressed plants compared to control plants.     

Responses of carboxylating enzymes,sucrose metabolizing enzymes and plant hormones in a tropical epiphytic CAM orchid to CO2 enrichment

After exposure to a doubled CO₂ concentration of 750 µL L^?1 for 2 months, average relative growth rate (RGR) of Mokara Yellow increased 25%. The two carboxylating enzymes, ribulose‐1,5‐bisphosphate carboxylase/oxygenase (Rubisco) and phosphoenolpyruvate carboxylase (PEPCase), responded differently to CO₂ enrichment. There was a significant daytime down‐regulation in Rubisco activity in the leaves of CO₂‐enriched plants. However, PEPCase activity in CO₂‐enriched plants was much higher in the dark period, although it was slightly lower during the daytime than that at ambient CO₂. Leaf sucrose–phosphate synthase (SPS) and sucrose synthase (SS) activities in CO₂‐enriched plants increased markedly, along with a night‐time increase in total titratable acidity and malate accumulation. There was a remarkable increase in the levels of indole‐3‐acetic acid (IAA), gibberellins A₁ and A₃ (GA₁₊₃), isopentenyladenosine (iPA) and zeatin riboside (ZR) in the expanding leaves of plants grown at elevated CO₂. It is suggested that (1) the down‐regulation of Rubisco and up‐regulation of SPS and SS are two important acclimation processes that are beneficial because it enhanced both photosynthetic capacity at high CO₂ and reduced resource investment in excessive Rubisco capacity; (2) the increased levels of plant hormones in CO₂‐enriched M. Yellow might play an important role in controlling its growth and development.     

Activities of chlorophyllase, phosphoenolpyruvate carboxylase and ribulose-1,5-bisphosphate carboxylase in the primary leaves of soybean during senescence and drought

The effects of senescence and drought on the levels and activities of chlorophyllase (EC 3.1.1.14), phosphoenolpyruvate carboxylase (PEPC, EC 4.1.1.31) and ribulose-1,5-bisphosphate carboxylase (Rubisco, EC 4.1.1.39) in the intact primary leaves of soybean ( Glycine max L. cv. Jackson) were monitored. Plants were grown either (1) for 2 to 8 weeks and the primary leaves harvested every week or (2) for 2 weeks and the plants subjected to drought stress and compared to control plants that were watered daily. In the senescence experiment, chlorophyllase activity changed in parallel with water content, leaf chlorophyll and total protein per unit dry weight of leaf tissue, with all factors increasing in concert during expansion of the primary leaves in the first 4 to 5 weeks of seedling development. Thereafter, all factors, including chlorophyllase activity, declined reaching markedly reduced values at weeks 7 and 8 when the primary leaves were yellow and ready to abscise. PEPC and Rubisco activities peaked in the third week, i.e. well before full leaf expansion, and then declined. In contrast to its response during senescence, chlorophyllase activity per unit leaf dry weight did not change during drought stress, but the specific activity of the enzyme rose and showed an inverse relationship to total leaf chlorophyll and protein content. Rubisco activity was highly sensitive to drought, with decrements observed in the activity and in levels of the large subunit within 2 days of withholding water and before significant changes in leaf water content were detected.     

The influence of altered gravity on carbohydrate metabolism in excised wheat leaves

We developed a system to study the influence of altered gravity on carbohydrate metabolism in excised wheat leaves by means of clinorotation. The use of excised leaves in our clinostat studies offered a number of advantages over the use of whole plants, most important of which were minimization of exogenous mechanical stress and a greater amount of carbohydrate accumulation during the time of treatment. We found that horizontal clinorotation of excised wheat leaves resulted in significant reductions in the accumulation of fructose, sucrose, starch and fructan relative to control, vertically clinorotated leaves. Photosynthesis, dark respiration and the extractable activities of ADP glucose pyrophosphorylase (EC 2.7.7.27), sucrose phosphate synthase (EC 2.4.4.14), sucrose sucrose fructosyltransferase (EC 2.4.1.99), and fructan hydrolase (EC 3.2.1.80) were unchanged due to altered gravity treatment.     

A role for brassinosteroids in the regulation of photosynthesis in Cucumis sativus

The effects of 24-epibrassinolide (EBR) spray application on gas-exchange, chlorophyll fluorescence characteristics, Rubisco activity, and carbohydrate metabolism were investigated in cucumber (Cucumis sativus L. cv. Jinchun No. 3) plants grown in a greenhouse. EBR significantly increased the light-saturated net CO(2) assimilation rate (A(sat)) from 3 h to 7d after spraying, with 0.1 mg l(-1) EBR proving most effective. Increased A(sat) in EBR-treated leaves was accompanied by increases in the maximum carboxylation rate of Rubisco (V(c,max)) and in the maximum rate of RuBP regeneration (J(max)). EBR-treated leaves also had a higher quantum yield of PSII electron transport (phi(PSII)) than the controls, which was mainly due to a significant increase in the photochemical quenching (q(P)), with no change in the efficiency of energy capture by open PSII reaction centres (F'(v)/F'(m)). EBR did not influence photorespiration. In addition, significant increases in the initial activity of Rubisco and in the sucrose, soluble sugars, and starch contents were observed followed by substantial increases in sucrose phosphate synthase (SPS), sucrose synthase (SS), and acid invertase (AI) activities after EBR treatment. It was concluded that EBR increases the capacity of CO(2) assimilation in the Calvin cycle, which was mainly attributed to an increase in the initial activity of Rubisco.     

Evidence for the sucrose-binding protein role in carbohydrate metabolism and transport at early developmental stage

Despite the large amount of data regarding sucrose-binding proteins (SBP), their functions remain largely unknown and controversial. In this investigation we performed a detailed temporal and spatial characterization of the phenotypes related to photosynthesis, sucrose exudation and carbohydrate metabolism in SBP antisense plants to gain insights into the physiological role of SBP. Significant reductions in net photosynthesis and in stomatal conductance were observed in the SBP antisense lines but were restricted to the vegetative phase, and persisted during a daily time course at this phase. Photosynthesis was saturated at a substantially lower irradiance in source leaves of the antisense lines, suggesting that light utilization is decreased in these plants. A slight reduction in soluble sugars was observed throughout the development of source leaves, partially overlapping a decrease in sucrose synthase activity (EC 2.4.1.13); whereas a transient increase in starch and adinosine diphosphate (ADP)-glucose pyrophosphorylase activity (EC 2.7.7.27) as well as decreased leaf sucrose exudation were detected in the beginning of the vegetative phase. These changes in source leaves were accompanied by reductions in sucrose and starch in sink leaves, hexoses and sucrose in roots and hexoses in shoot apex, which were observed before the occurrence of a significant reduction in height and in leaf number in the transgenic lines. These alterations in growth parameters did not persist throughout the development, but were associated with a delay in flowering time and leaf senescence in the SBP antisense lines. A likely involvement of SBP in sink strength is discussed.