Phenyl indane from acorus calamus

Besides three known compounds, two new compounds, namely Z-3-(2,4,5-trimethoxy phenyl)-2-propenal and a new phenyl indane have been isolated from the rhizomes of Acorus calamus. These compounds have been characterized from their spectral data and by synthesis.     

Energy Metabolism in Rhizomes of Acorus calamus (L.) and in Tubers of Solanum tuberosum (L.) With Regard to their Anoxia Tolerance

Rhizomes of the marsh plant Acorus calamus (L.) and tubers of the flooding-intolerant Solanum tuberosum (L.) var. Bintje, both kept under strict anoxia, differ markedly in their fermentation properties. The fermentation capacities as measured by ADH and LDH activities and their respective product concentrations were estimated. While rhizomes of Acorus calamus, having high ADH and low LDH activities, accumulate mainly ethanol, tubers of Solanum tuberosum tend towards lactic acid fermentation. The total amount of adenine nucleotides is quite stable in Acorus calamus, whereas they show a sharp decline in S. tuberosum during the first 6h of anoxia. The adenylate energy charge of A. calamus recovers after a short initial drop (AEC > 0.8). AEC values of S. tuberosum decrease rapidly and remain at very low values (AEC ～ 0.3). Tuber tissues became soft and lost viability after about 48–72 h of anoxia at 25 °C. This might be due to tissue acidification and impaired energy metabolism, but not to the lack of energy reserves. Energy metabolism of A. calamus is well adapted to anoxia.     

Impact of oxygen stress and energy availability on membrane stability of plant cells

This article reviews the relationship between the energy status of plant cells under O₂ stress (e.g. waterlogging) and the maintenance of membrane intactness, using information largely derived from suspension cultures of anoxia‐intolerant potato cells. Energy‐related parameters measured were fermentation end‐products (ethanol, lactate, alanine), respiratory rate, ATP, adenylate energy charge, nitrate reductase activity and biomass. ATP synthesis rates were calculated from the first four parameters. Reactive oxygen species were estimated from H₂O₂ and superoxide levels, and the enzymatic detoxification potential from the activity levels of catalase and superoxide dismutase. Structure‐related parameters were total fatty acids, free fatty acids (FFAs), lipid hydroperoxides, total phospholipids, N‐acylphosphatidylethanolamine (NAPE) and cell viability. The following issues are addressed in this review: (1) what is the impact of anoxia on membrane lipids and how does this relate to energy status; (2) does O₂ per se play a role in these changes; (3) under which conditions and to what extent does lipid peroxidation occur upon re‐aeration; and (4) can the effects of re‐aeration be distinguished from those of anoxia? The emerging picture is a reappraisal of the relative contributions of anoxia and re‐aeration. Two successive phases (pre‐lytic and lytic) characterize potato cells under anoxia. They are connected by a threshold in ATP production rate, below which membrane lipids are hydrolysed to FFAs, and NAPE increases. Since lipid peroxidation occurs only when cells are reoxygenated during the lytic phase, its biological relevance in an already damaged system is questionable.Key words: Acorus calamus L., energy shortage, free fatty acids, lipid peroxidation, lipolytic acyl hydrolase, lipoxygenase, membrane intactness, N‐acylphosphatidylethanolamine, O₂ stress, reactive oxygen species, Solanum tuberosum L.     

Habitat characteristics of sweet flag (Acorus calamus) and their relationships with sweet flag biomass

The habitat characteristics of Acorus calamus were investigated to obtain data and define criteria that can be applied to restoration, conservation, and management. The investigation was conducted in March, June, and September to span the growth period of A. calamus. While the importance of A. calamus decreased over time, the diversity index (H′) of A. calamus communities increased in June and then decreased in September. Water levels in A. calamus habitats were lowest in March. About two-thirds of the A. calamus communities (68.4%) were found in sandy loam. Although the optimal environmental ranges of the water and soil characteristics for A. calamus changed according to growth stage, the environmental conditions in March were important for seed germination and seedling establishment. The aboveground biomass of A. calamus was 20.4 in March, 349.3 in June, and 267.0 g/m² in September. We also clarified the environmental characteristics of the water and soil at which the aboveground biomass during the growing and senescing seasons peaked. Based on our data, we were able to propose the following guidelines for wetland creation and restoration. (1) To facilitate the seed germination and seedling establishment of A. calamus, managers should seek to achieve conditions that approximate the optimal environment in March. (2) To optimize the growth of A. calamus, the soil pH should be maintained at neutral and the permeability to air should be enhanced. (3) To extend the longevity of A. calamus leaves, the habitat should be managed so as to limit the survival of other species with overlapping niches, and the water depth should be maintained at around 24 cm. Also, the supply of nitrogen components to the soil should not be excessive.     

An overview on traditional uses and pharmacological profile of Acorus calamus Linn. (Sweet flag) and other Acorus species

Acorus calamus (Sweet flag) has a long history of use and has numerous traditional and ethnomedicinal applications. Since ancient times, it has been used in various systems of medicines such as Ayurveda, Unani, Siddha, Chinese medicine, etc. for the treatment of various aliments like nervous disorders, appetite loss, bronchitis, chest pain, colic, cramps, diarrhea, digestive disorders, flatulence, gas, indigestion, rheumatism, sedative, cough, fever, bronchitis, inflammation, depression, tumors, hemorrhoids, skin diseases, numbness, general debility and vascular disorders. Various therapeutic potentials of this plant have been attributed to its rhizome. A number of active constituents from leaves, rhizomes and essential oils of A. calamus have been isolated and characterized. Of the constituents, alpha and beta-asarone are the predominant bioactive components. Various pharmacological activities of A. calamus rhizome such as sedative, CNS depressant, anticonvulsant, antispasmodic, cardiovascular, hypolipidemic, immunosuppressive, anti-inflammatory, cryoprotective, antioxidant, antidiarrheal, antimicrobial, anticancer and antidiabetic has been reported. Genotoxicity and mutagenecity of beta and alpha-asarone is reported, which limits their use at high dosage. Though A. calamus has been used since ancient times, many of its uses are yet to be scientifically validated. In the present review an attempt has been made to explore traditional uses and pharmacological properties of A. calamus.     

Day-night changes of energy-rich compounds in crassulacean acid metabolism (CAM) species utilizing hexose and starch

• Background and Aims Plants with crassulacean acid metabolism (CAM) can be divided into two groups according to the major carbohydrates used for malic acid synthesis, either polysaccharide (starch) or monosaccharide (hexose). This is related to the mechanism and affects energy metabolism in the two groups. In Kalanchoë pinnata and K. daigremontiana, which utilize starch, ATP-dependent phosphofructokinase (tonoplast inorganic pyrophosphatase) activity is greater than inorganic pyrophosphate-dependent phosphofructokinase (tonoplast adenosine triphosphatase) activity, but the reverse is the case in pineapple (Ananas comosus) utilizing hexose. To test the hypothesis that the energy metabolism of the two groups differs, day-night changes in the contents of ATP, ADP, AMP, inorganic phosphate (Pi), phosphoenolpyruvate (PEP) and inorganic pyrophosphate (PPi) in K. pinnata and K. daigremontiana leaves and in pineapple chlorenchyma were analysed.• Methods The contents of energy-rich compounds were measured spectrophotometrically in extracts of tissue sampled in the light and dark, using potted plants, kept for 15 d before the experiments in a growth chamber.• Key Results In the three species, ATP content and adenylate energy charge (AEC) increased in the dark and decreased in the light, in contrast to ADP and AMP. Changes in ATP and AEC were greater in Kalanchoë leaves than in pineapple chlorenchyma. PPi content in the three species increased in the dark, but on illumination it decreased rapidly and substantially, remaining little changed through the rest of the light period. Pi content of Kalanchoë leaves did not change between dark and light, whereas Pi in pineapple chlorenchyma increased in the dark and decreased in the light, and the changes were far greater than in Kalanchoë leaves. Light-dark changes in PEP content in the three species were similar.• Conclusions These results corroborate our hypothesis that day–night changes in the contents of energy-rich compounds differ between CAM species and are related to the carbohydrate used for malic acid synthesis.Key words: Ananas comosus, ATP, chlorenchyma, crassulacean acid metabolism, inorganic pyrophosphate, Kalanchoë daigremontiana, Kalanchoë pinnata, phosphoenolpyruvate     

Amino Acid Transport across the Tonoplast of Vacuoles Isolated from Barley Mesophyll Protoplasts : Uptake of Alanine, Leucine, and Glutamine

Mesophyll protoplasts from leaves of well-fertilized barley (Hordeum vulgare L.) plants contained amino acids at concentrations as high as 120 millimoles per liter. With the exception of glutamic acid, which is predominantly localized in the cytoplasm, a major part of all other amino acids was contained inside the large central vacuole. Alanine, leucine, and glutamine are the dominant vacuolar amino acids in barley. Their transport into isolated vacuoles was studied using ¹⁴C-labeled amino acids. Uptake was slow in the absence of ATP. A three- to sixfold stimulation of uptake was observed after addition of ATP or adenylyl imidodiphosphate an ATP analogue not being hydrolyzed by ATPases. Other nucleotides were ineffective in increasing the rate of uptake. ATP-Stimulated amino acid transport was not dependent on the transtonoplast pH or membrane potential. p-Chloromercuriphenylsulfonic acid and n-ethyl maleimide increased transport independently of ATP. Neutral amino acids such as valine or leucine effectively decreased the rate of alanine transport. Glutamine and glycine were less effective or not effective as competitive inhibitors of alanine transport. The results indicate the existence of a uniport translocator specific for neutral or basic amino acids that is under control of metabolic effectors.     

Respiration and nitrogen and carbohydrate contents in perennial rhizome-forming plants as related to realization of different adaptive strategies

Accumulation of biomass, the respiration rate, and the contents of total nitrogen and nonstructural carbohydrates were studied for 14 perennial long-rhizome-forming species differing in the type of adaptive strategy. Fast-growing species with well expressed competitive-ruderal properties (CR plants) were characterized by a higher productivity, a better nitrogen status, and more intense respiration than slowly growing stress-tolerant species (S plants). The proportion of rhizomes in the weight of the whole plant varied from 30 to 70% and was higher in S species. In CR species, the respiration rate measured in rhizomes at 20°C was equal on the average to 1 mg CO₂/(g dry wt h), which was threefold higher than in S species. In S species, a considerable amount of nitrogen (50%) was present in rhizomes, whereas in CR species, most part of nitrogen (70–80%) was localized in the aboveground organs. The correlation analysis revealed a direct dependence (r = 0.75) between the respiration rate and nitrogen content in leaves; in the rhizomes the correlation between these indices was low (r = 0.39). The content of carbohydrates in the leaves and sink organs, rhizomes, was determined by the type of plant ecological strategy and life duration of their photosynthesizing organs (summergreen, evergreen species). In general, the results obtained demonstrated a close relation between adaptive strategy, ecological confinement, the rhythm of seasonal development, and physiological properties of long-rhizome-forming plants.     

An expanded adenylate kinase gene family in the protozoan parasite Trypanosoma cruzi

Adenylate kinases supply energy routes in cellular energetic homeostasis. In this work, we identified and characterized the adenylate kinase activity in extracts from the flagellated parasite Trypanosoma cruzi, the causative agent of Chagas' disease. Adenylate kinase activity was detected in different subcellular fractions and the cytosolic isoform was biochemically characterized. Cytosolic adenylate kinase specific activity increases continuously during the epimastigote growth and is down-regulated when other soluble phosphotransferase, arginine kinase, is overexpressed. Six different genes of adenylate kinase isoforms were identified and the mRNA expression was confirmed by RT-PCR and Northern Blot. Three open reading frames coding for different enzyme isoforms named TzADK1, TzADK2 and TzADK5 were cloned and functionally expressed in E. coli. This work reports an unusually large number of genes of adenylate kinases and suggests a coordinated regulation of phosphotransferase-mediated ATP regenerating pathways in the unicellular parasite Trypanosoma cruzi.     

ADP-Glucose Transport by the Chloroplast Adenylate Translocator Is Linked to Starch Biosynthesis

In organello starch biosynthesis was studied using intact chloroplasts isolated from spinach leaves (Spinacia oleracea). Immunoblot analysis using a specific antiserum against the mitochondrial adenylate (ADP/ATP) translocator of Neurospora crassa shows the presence of an adenylate translocator protein in the chloroplast envelope membranes, similar to that existing in mitochondria and amyloplasts from cultured cells of sycamore (Acer pseudoplatanus). The double silicone oil layer-filtering centrifugation technique was employed to study the kinetic properties of adenylate transport in the purified chloroplasts; ATP, ADP, AMP, and most importantly ADP-Glc were shown to be recognized by the adenylate translocator. Similar to the situation with sycamore amyloplasts, only ATP and ADP-Glc uptake was inhibited by carboxyatractyloside, an inhibitor of the mitochondrial adenylate translocator. Evidence is presented to show that the ADP-Glc transported into the chloroplast stroma is utilized for starch synthesis catalyzed by starch synthase (ADP-Glc:1,4-α-d-glucan 4-α-d-glucosyltransferase). The high activity of sucrose synthase producing ADP-Glc observed in the extrachloroplastic fractions suggests that starch biosynthesis in chloroplasts may be coupled with the direct import of ADP-Glc from the cytosol.     

Locomotory,ventilatory and metabolic responses of the subterranean Stenasellus virei (Crustacea,Isopoda) to severe hypoxia and subsequent recovery

The locomotory and ventilatory activities and the intermediary and energy metabolism modifications of the hypogean aquatic isopod crustacean Stenasellus virei were investigated in severe hypoxia (Po₂ < 0.03 kPa) and subsequent recovery. The aims of this study were i) to determine why the subterranean species displayed a greater tolerance of hypoxia than numerous other epigean crustaceans, ii) to confirm previous results obtained with four hypogean and epigean crustaceans, iii) to compare the responses to severe hypoxia in hypogean amphipods and isopods, and iv) to better understand the ecological problems of the hypogean organisms survival in subterranean habitats. S. virei responded to experimental long-term, severe hypoxia with classical anaerobic metabolism mainly characterized by a decrease in adenosine triphosphate (ATP) and phosphagen, utilization of glycogen and glutamate, and accumulation of lactate and alanine. Lactate was also largely excreted by this organism, which is unusual for crustaceans in general. Compared to most other epigean crustaceans, the isopod S. virei showed high amounts of stored glycogen and arginine phosphate. These differences in glycogen and phosphagen stores, and the ability to reduce energetic expenditures linked to locomotion and ventilation, extended the survival of S. virei under experimental anaerobiosis. During recovery, the isopod S. virei showed a higher capacity for glyconeogenesis from lactate and a faster and total replenishment of ATP and arginine phosphate levels than epigean crustaceans. Data concerning responses to hypoxia and subsequent recovery in S. virei are similar to those previously obtained with two other hypogean amphipods, except that this isopod did not synthesize succinate in anaerobiosis.     

Transport of arginine and aspartic Acid into isolated barley mesophyll vacuoles

The transport of arginine into isolated barley (Hordeum vulgare L.) mesophyll vacuoles was investigated. In the absence of ATP, arginine uptake was saturable with a K_m of 0.3 to 0.4 millimolar. Positively charged amino acids inhibited arginine uptake, lysine being most potent with a K_i of 1.2 millimolar. In the presence of free ATP, but not of its Mg-complex, uptake of arginine was drastically enhanced and a linear function of its concentration up to 16 millimolar. The nonhydrolyzable adenylyl imidodiphosphate, but no other nucleotide tested, could substitute for ATP. Therefore, it is suggested that this process does not require energy and does not involve the tonoplast ATPase. The ATP-dependent arginine uptake was strongly inhibited by p-chloromercuriphenylsulfonic acid. Furthermore, hydrophobic amino acids were inhibitory (I₅₀ phenylalanine 1 millimolar). Similar characteristics were observed for the uptake of aspartic acid. However, rates of ATP-stimulated aspartic acid transport were 10-fold lower as compared to arginine transport. Uptake of aspartate in the absence of ATP was negligible.     

Glycolytic gene expression in amphibious Acorus calamus L. under natural conditions

Acorus calamus L is an amphibious plant, which is exposed to periods of flooding and consequently hypoxic conditions as a part of its natural life cycle. Previous experiments under laboratory conditions have shown that the plant can survive for two months in the complete absence of oxygen, and that during this period the expression of genes encoding the glycolytic enzymes fructose-1,6-bisphosphate aldolase (ALD), pyruvate decarboxylase (PDC) and alcohol dehydrogenase (ADH) is induced in leaves and rhizomes (Bucher and Kuhlemeier, 1993). Here we studied the expression of ALD and ADH through two years in the natural habitat of A. calamus. Under natural conditions roots and rhizomes were always submerged but newly grown leaves emerged in spring; in autumn the leaves senesced and the whole plant was submerged again. High Ald and Adh mRNA levels in leaf and rhizome were found only in winter when the leaves were entirely submerged. Upon leaf emergence in spring the mRNA levels rapidly declined. Under controlled experimental conditions expression of Ald and Adh was not induced by low temperature. The combination of laboratory and field experiments supports the hypothesis that oxygen deprivation rather than low temperature is a major regulator of glycolytic gene expression in A. calamus. The possible role of other environmental factors is also discussed.Abbreviations ADH alcohol dehydrogenase - Adh gene encoding ADH - ALD cytoplasmic fructose-1,6-bisphosphate aldolase - Ald gene encoding ALD - PDC pyruvate decarboxylase - Pdc gene encoding PDC     

Identification of the allosteric regulatory site in bacterial phosphoribulokinase

Bacterial phosphoribulokinases (PRKs) are octameric members of the adenylate kinase family of enzymes. The enzyme is allosterically activated by NADH and allosterically inhibited by AMP. We have determined the crystal structure of PRK from Rhodobacter sphaeroides bound to the ATP analogue AMP-PCP to a resolution of 2.6 A. The structure reveals that the ATP analogue does not bind to the canonical ATP site found in adenylate kinase family members. Rather, the AMP-PCP binds in two different orientations at the interface of three of the monomers in the octamer. This interface was previously characterized as having an unusually large number of arginine residues. Of the five arginine residues that are near the bound nucleotide, one (Arg 221) is highly conserved in both prokaryotic and eukaryotic (nonallosterically regulated) PRKs, two (Arg 234 and Arg 257) are on a second subunit and conserved in only prokaryotic PRKs, and two (Arg 30 and Arg 31) are on a third subunit with only one of them (Arg 31) conserved in prokaryotic PRKs. Each of these arginine residues was converted by site-directed mutagenesis to alanine. Fluorescence binding data suggest that none of these arginines are involved in active site ATP binding and that Arg 234 and Arg 257 on the second subunit are directly involved in NADH binding, while the other arginines have a minimal effect on NADH binding. While the wild-type enzyme exhibits low maximal activity and hyperbolic kinetics with respect to ATP in the absence of NADH and high maximal activity and sigmoidal kinetics in the presence of NADH, the R31A mutant exhibits identical hyperbolic kinetics with respect to ATP in the presence or absence of NADH. Thus, the transmission of allosteric information from one subunit to another is conducted through a single path that includes NADH and Arg 31.     

Comparison of nitrogen uptake in the roots and rhizomes of <Emphasis Type="Italic">Leymus chinensis</Emphasis>

Leymus chinensis (Trin.) Tzvel is a rhizomatous grass species in the Eastern Eurasian steppe zone that is often limited by low soil nitrogen availability. Although a previous study showed that the rhizomes of L. chinensis have the capacity to take up nitrogen, the importance of such uptake for nitrogen nutrition is unclear. Moreover, little is known regarding the inorganic nitrogen uptake kinetics of roots and rhizomes in response to nitrogen status. Here, we first found that ammonium is preferred over nitrate and glycine for L. chinensis growth. Using the ¹⁵N-labelling method, we found that the rate of ion influx into roots was approximately five-fold higher than into rhizomes under the same nitrogen content, and the ion influxes into roots and rhizomes under 0.05 mM N were greater than in the presence of 3 mM N, especially in the form of NH₄⁺. Using a non-invasive micro-test technique, we characterised the patterns of NH₄⁺ and NO₃^– fluxes in the root mature zone, root tip, rhizome mature zone, and rhizome tip following incubation in the solution with different N compounds and different N concentrations. These results suggest a dynamic balance between the uptake, utilisation, and excretion of nitrogen in L. chinensis.     

Effect of adenine nucleotides on the reaction catalyzed by pyruvate,orthophosphate dikinase in maize

The effect of adenine nucleotides in pyruvate, orthophosphate dikinase (EC 2.7.9.1, ATP, pyruvate, orthophosphate phosphotransferase)_was studied with the enzyme furified from maize, and with the enzyme obtained from mesophyll chloroplast extracts during assay in the direction of pyruvate conversion to phosphoenolpyruvate. (1) In studies with the purified enzyme, the relationship of initial velocity to ATP concentrations follows Michaelis-Menten kinetics, and the K_m value for ATP was 22.8 μM (± 5.1 μM, n = 5). (2) AMP was a competitive inhibitor with respect to ATP, and its K_i value was 35.8 μM (± μM, n = 4). There was no inhibition of catalysis by ADP up to a concentration of 460 μM. (3) The theoretical response of the enzyme to change in the adenylate energy charge was calculated from the kinetic constants for ATP and AMP. The experimentally obtained values were similar to the theoretical response when varying energy charge was generated by addition of appropriate amounts of ATP, ADP and AMP in assays with the purified enzyme. The response of the enzyme to energy charge at different pH values (pH 7.0, 7.5, and 8.0) was similar, although the activity of the enzyme at pH 7.0 was about 40% of that at pH 8.0. (4) When mesophyll chloroplast extracts of maize, which contain high levels of adenylate kinase, were used as the source of the enzyme and the adenylate energy charge was generated by addition of different concentrations of ATP and AMP, the influence on catalysis was similar to that with the purified enzyme. (5) The data show that the effect of varying energy chage on the activity of the dikinase is not typical of a U-type enzyme, in contrast to phosphoglycerate kinase (EC 2.7.2.3, ATP: 3-phospho-D-glycerate 1-phosphotransferase), which is more strongly regulated. (6) Evidence is presented for competition between the dikinase and phosphoglycerate kinase for ATP in mesophyll chloroplast extracts of maize. (7) When the effect of adenylate energy charge on the state of activation and the direct effect on catalysis of the dikanase are combined, the total capacity for catalysis is very dependent on the energy charge.     

The relationship between leaf nitrogen,nitrogen metabolites and herbivory in two species of Nyctaginaceae from the Brazilian Cerrado

Guapira graciliflora and Neea theifera are taxonomically related species of the tribe Pisoneae. Both species are found in the same environment, the Brazilian Cerrado, and therefore, are subjected to similar selective pressures. These species occur in oligotrophic environments, yet contain high concentrations of nitrogen in their leaves. The present study was carried out to investigate the ecological role of nitrogen in herbivory on these species. The differences in the N content, compositions of secondary N-metabolites, mechanical resistance, and water content between their leaves indicate that these species have different adaptations as defense mechanisms. In both species, their high nitrogen content seems to promote herbivory. The presence of secondary nitrogen metabolites does not prevent the species from suffering intense damage by herbivores on their early leaves. The herbivory rates observed were lower for mature leaves of both species than for young leaves. In G. graciliflora, nutritional content and leaf hardness are the most important variables correlated with reduction of herbivory rates, whereas in N. theifera, N compounds are also correlated with herbivory rates. Despite the differences in the strategies of these two species, they exhibit a similar efficiency of protection against natural enemies because their total herbivory rates are similar. The difference in their N defense allocation may imply benefits for survival under Cerrado conditions. We briefly discuss the oligotrophic habitat conditions of the studied plants and possible advantages of their strategies of N accumulation and metabolic uses.     

Seasonal nitrogen speciation in temperate seagrass Posidonia oceanica (L.) Delile

To better understand some basic aspects of the nitrogen economy in Posidonia oceanica and, specifically, the seasonality of the processes of storage, translocation and assimilation, we examined nitrogen speciation into soluble compounds, both inorganic (nitrates, nitrites and ammonium) and organic (free amino acids, FAA, and total soluble protein, TSP), and the nitrogen assimilation potential (through the glutamine synthetase activity measurement) in the leaves, rhizomes and roots of P. oceanica over a 1-year cycle. Only a limited amount of inorganic nitrogen was found, accounting for less than 3.3% of the total nitrogen content, and it was mostly in the form of ammonium. Nitrate and nitrite concentrations were very low, always below 7.2 μmol g⁻¹ dw in annual average. Among the organic soluble fractions, FAAs were the most abundant, accounting for up to 50% of N pools. Rhizomes were the organs in which FAA concentrations reached their maximum value. The leaves showed higher nitrogen assimilation potential than the roots and this assimilation potential was highest during and after the period of maximum leaf growth, probably corresponding to the assimilation of both new and recycled nitrogen. Our results suggest that 5% of the total nitrogen assimilation occurs in roots and 79% in leaves on an annual average. In addition, rhizomes contributed to the total shoot nitrogen assimilation by 32-54% between autumn and spring. Rhizomes appear as key organs in the nitrogen economy of the plant, not only as a major site for nitrogen assimilation but also as an organ for nitrogen storage. This storage, mostly in the form of FAA, occurs during periods of high availability and low demand (winter). This stored nitrogen can supply up to 33% of plant demands during the moment of maximum leaf growth (i.e. late spring).     

Efficient nitrogen economy: Key to the success ofPolygonum bistorta in an abandoned mountain meadow

The dominance ofPolygonum bistorta in abandoned mountain meadows was studied. Successional and seasonal changes in biomass, nitrogen concentration and content were studied at the community level. During the successionP. bistorta increases its biomass both in absolute and mainly in relative values compared to the surrounding species. Nitrogen stored in its rhizomes seems to be quickly translocated into above-ground parts at the beginning of the growing season. High amounts of nitrogen allow rapid growth of both vegetative and generative plant parts. The success ofP. bistorta is probably the result of efficient use of stored nitrogen, suppression of surrounding vegetation by above-ground cover of leaves and by decrease in the level of soil nitrogen, and the effect of a large amount of litter. Management consisting in the combination of mowing and fertilization is proposed.     

The Effect of Adenine Nucleotides on Purified Phosphoenolpyruvate Carboxylase from the CAM Plant Crassula argentea

The effects of adenine nucleotides on phosphoenolypyruvate carboxylase were investigated using purified enzyme from the CAM plant, Crassula argentea. At 1 millimolar total concentration and with limiting phosphoenolpyruvate, AMP had a stimulatory effect, lowering the K_m for phosphoenolpyruvate, ADP caused less stimulation, and ATP decreased the activity by increasing the K_m for phosphoenolpyruvate. Activation by AMP was not additive to the stimulation by glucose 6-phosphate. Furthermore, AMP increased the K_a for glucose 6-phosphate. Inhibition by ATP was competitive with phosphoenolpyruvate. In support of the kinetic data, fluorescence binding studies indicated that ATP had a stronger effect than AMP on phosphoenolpyruvate binding, while AMP was more efficient in reducing glucose 6-phosphate binding. As free Mg²⁺ was held constant and saturating, these effects cannot be ascribed to Mg²⁺ chelation. Accordingly, the enzyme response to the adenylate energy charge was basically of the “R” type (involving enzymes of ATP regenerating sequences) according to D. E. Atkinson's (1968 Biochemistry 7: 4030-4034) concept of energy charge regulation. The effect of energy charge was abolished by 1 millimolar glucose 6-phosphate. Levels of glucose 6-phosphate and of other putative regulatory compounds of phosphoenolpyruvate carboxylase were determined in total leaf extracts during a day-night cycle. The level of glucose 6-phosphate rose at night and dropped sharply during the day. Such a decrease in glucose 6-phosphate concentration could permit an increased control of phosphoenolpyruvate carboxylase by energy charge during the day.