Ammonium formation and assimilation in P(SARK)∷IPT tobacco transgenic plants under low N

Wild Type (WT) and transgenic tobacco plants expressing isopentenyltransferase (IPT), a gene encoding the enzyme regulating the rate-limiting step in cytokinins (CKs) synthesis, were grown under limited nitrogen (N) conditions. We analyzed nitrogen forms, nitrogen metabolism related-enzymes, amino acids and photorespiration related-enzymes in WT and P_SARK∷IPT tobacco plants. Our results indicate that the WT plants subjected to N deficiency displayed reduced nitrate (NO₃⁻) assimilation. However, an increase in the production of ammonium (NH₄⁺), by the degradation of proteins and photorespiration led to an increase in the glutamine synthetase/glutamate synthase (GS/GOGAT) cycle in WT plants. In these plants, the amounts of amino acids decreased with N deficiency, although the relative amounts of glutamate and glutamine increased with N deficiency. Although the transgenic plants expressing P_SARK∷IPT and growing under suboptimal N conditions displayed a significant decline in the N forms in the leaf, they maintained the GS/GOGAT cycle at control levels. Our results suggest that, under N deficiency, CKs prevented the generation and assimilation of NH₄⁺ by increasing such processes as photorespiration, protein degradation, the GS/GOGAT cycle, and the formation of glutamine.     

Ammonia emission from rice leaves in relation to photorespiration and genotypic differences in glutamine synthetase activity

Background and Aims

Rice (Oryza sativa) plants lose significant amounts of volatile NH₃ from their leaves, but it has not been shown that this is a consequence of photorespiration. Involvement of photorespiration in NH₃ emission and the role of glutamine synthetase (GS) on NH₃ recycling were investigated using two rice cultivars with different GS activities.

Methods

NH₃ emission (AER), and gross photosynthesis (P_G), transpiration (Tr) and stomatal conductance (g_S) were measured on leaves of ‘Akenohoshi’, a cultivar with high GS activity, and ‘Kasalath’, a cultivar with low GS activity, under different light intensities (200, 500 and 1000 µmol m⁻² s⁻¹), leaf temperatures (27·5, 32·5 and 37·5 °C) and atmospheric O₂ concentrations ([O₂]: 2, 21 and 40 %, corresponding to 20, 210 and 400 mmol mol⁻¹).

Key Results

An increase in [O₂] increased AER in the two cultivars, accompanied by a decrease in P_G due to enhanced photorespiration, but did not greatly influence Tr and g_S. There were significant positive correlations between AER and photorespiration in both cultivars. Increasing light intensity increased AER, P_G, Tr and g_S in both cultivars, whereas increasing leaf temperature increased AER and Tr but slightly decreased P_G and g_S. ‘Kasalath’ (low GS activity) showed higher AER than ‘Akenohoshi’ (high GS activity) at high light intensity, leaf temperature and [O₂].

Conclusions

Our results demonstrate that photorespiration is strongly involved in NH₃ emission by rice leaves and suggest that differences in AER between cultivars result from their different GS activities, which would result in different capacities for reassimilation of photorespiratory NH₃. The results also suggest that NH₃ emission in rice leaves is not directly controlled by transpiration and stomatal conductance.     

Invasion of <Emphasis Type="Italic">Nipponaclerda biwakoensis</Emphasis> (Hemiptera: Aclerdidae) and <Emphasis Type="Italic">Phragmites australis</Emphasis> die-back in southern Louisiana,USA

Common reed, Phragmites australis (Cav.) Trin. Ex Steud., is the dominant emergent vegetation in the lower Mississippi River Delta (MRD), Louisiana, USA and is comprised primarily of introduced lineages of different phylogeographic origins. Dense stands of P. australis are important for protecting marsh soils from wave action and storm surges. In the Fall of 2016, while investigating symptoms of die-back of Phragmites stands in the lower marsh, a non-native scale was found infesting affected stands in high densities. Identified as Nipponaclerda biwakoensis (Kuwana) (Hemiptera: Aclerdidae), the scale was well established across the lower MRD. This report represents the first recorded population of Nipponaclerda biwakoensis in North America. Intriguingly, there are noticeable differences in die-back symptoms and in scale densities among different lineages of Phragmites in the MRD, with stands of the well-known European invasive lineage appearing healthier and having lower scale densities than other Phragmites lineages. Given its apparent relationship with the die-back syndrome, the scale may have serious implications for the health and stability of Phragmites marsh communities across coastal Louisiana. Efforts are currently underway to investigate the role of the scale and other abiotic stressors in the die-backs and potential management solutions.     

Pore water N:P and NH4 +:NO3 ? alter the response of Phragmites australis and Glyceria maxima to extreme nutrient regimes

Phragmites australis and Glyceria maxima are fast-growing littoral grasses often competing for similar wetland habitats. Eutrophication affects their competitiveness, but the outcome is not generally predictable due to the complexity of interrelated factors. We hypotheses that pore water N:P and NH₄ ⁺:NO₃ ^? modify their growth and metabolic responses to the trophic status of the habitat. The hypothesis was tested under standardized conditions of long-term sand cultures. Application of N?+?P up to extreme levels in combination with N:P?<?10 and NH₄ ⁺:NO₃ ^??<?1 triggered positive growth response in both species. In contrast, similar N levels applied in N:P?>?90 and NH₄ ⁺:NO₃ ^??=?4 caused lower productivity, changes in resource allocation, morphology and metabolic relations (e.g. high shoot density, low shoot diameters and heights, reduced root and rhizome growth). Observed signs of stress resembled the factors associated with the reed retreat at the die-back sites. Unbalanced N levels obviously alter plant susceptibility to stresses (altering, e.g. ventilation efficiency, plant anchorage or below-ground storage capacity). The positive effect of sufficient P supply was pronounced in Glyceria. It might therefore favour Glyceria in competition with Phragmites at highly fertile habitats rich in P.     

Physiological regulation of plant-atmosphere ammonia exchange

Plants have a compensation point for NH₃ which ranges from 0.1 to 20 nmol mol^-1, and may be several-fold higher or lower than naturally occurring atmospheric NH₃ concentrations. This implies that NH₃ fluxes over vegetated surfaces are bi-directional and that ammonia exchange with the atmosphere in many cases contributes significantly to the nitrogen economy of vegetation. Physiological regulation of plant–atmosphere NH₃ fluxes is mediated via processes involved in nitrogen uptake, transport and metabolism. A rapid turnover of NH₃ ⁺ in plant leaves leads to the establishment of a finite NH₃ ⁺ concentration in the leaf apoplastic solution. This concentration determines, together with that of H⁺, the size of the NH₃ compensation point. Barley and oilseed rape plants with access to NH₃ ⁺ in the root medium have higher apoplastic NH₃ ⁺ concentrations than plants absorbing NO₃ ^-. Furthermore, the apoplastic NH₃ ⁺ concentration increases with the external NH₃ ⁺ concentration. Inhibition of GS leads to a rapid and substantial increase in apoplastic NH₃ ⁺ and barley mutants with reduced GS activity have higher apoplastic NH₃ ⁺ than wild-type plants. Increasing rates of photorespiration do not affect the steady-state NH₃ ⁺ or H⁺ concentration in tissue or apoplast of oilseed rape, indicating that the NH₃ ⁺ produced is assimilated efficiently. Nevertheless, NH₃ emission increases due to a temperature-mediated displacement of the chemical equilibrium between gaseous and aqueous NH₃ in the apoplast. Sugarbeet plants grown with NO₃ ^- seem to be temporarily C-limited in the light due to a repression of respiration. As a consequence, the activity of chloroplastic GS declines during the day causing a major part of NH₃ ⁺ liberated in photorespiration to be assimilated during darkness when 2-oxoglutarate is supplied in high rates by respiration. This revised version was published online in June 2006 with corrections to the Cover Date.     

The role of glutamine synthetase in regulation of nitrogen metabolism within the soil microbial community

Recent advances in our understanding of the enzymology and regulatory systems involved in microbial metabolism of N hold promise to elucidate some of the underlying factors controlling metabolism of N in soil ecosystems. A review of recent work is used to construct a paradigm for N metabolism regulation in soil based on the central role of glutamine synthetase (GS) in such regulation within the soil microbial community. The studies involved use of GS inhibitors to elucidate the role of GS activity in regulation of soil N metabolism. Such studies have shown that the glutamine formed by microbial assimilation of NH₄ ⁺ via GS activity influences the regulatory mechanisms controlling both the production and activity of enzymes involved in N metabolism. For example, these studies showed that the inhibition of GS activity within the soil microbial community relieved the repression of urease production caused by microbial assimilation of inorganic N and blocked the short-term regulation of assimilatory nitrate reductase (ANR) by NH₄ ⁺ assimilation. Other studies have indicated that common environmental factors in soil may influence GS activity in microorganisms and thereby may influence metabolism of N within the soil microbial community. The paradigm for N metabolism regulation in soil that has emerged from such studies should lead to a better understanding of the mechanisms controlling fate of N in soil ecosystems.     

Enhanced tolerance to salt stress in transgenic rice that overexpresses chloroplast glutamine synthetase

The potential role of photorespiration in the protection against salt stress was examined with transgenic rice plants. Oryza sativa L. cv. Kinuhikari was transformed with a chloroplastic glutamine synthetase (GS2) gene from rice. Each transgenic rice plant line showed a different accumulation level of GS2. A transgenic plant line, G39-2, which accumulated about 1.5-fold more GS2 than the control plant, had an increased photorespiration capacity. In another line, G241-12, GS2 was almost lost and photorespiration activity could not be detected. Fluorescence quenching analysis revealed that photorespiration could prevent the over-reduction of electron transport systems. When exposed to 150 mM NaCl for 2 weeks, the control rice plants completely lost photosystem II activity, but G39-2 plants retained more than 90% activity after the 2-week treatment, whereas G241-12 plants lost these activities within one week. In the presence of isonicotinic acid hydrazide, an inhibitor of photorespiration, G39-2 showed the same salt tolerance as the control plants. The intracellular contents of NH₄ ⁺ and Na⁺ in the stressed plants correlated well with the levels of GS2. Thus, the enhancement of photorespiration conferred resistance to salt in rice plants. Preliminary results suggest chilling tolerance in the transformant.     

Organ-specific changes in the activity and subunit composition of glutamine-synthetase isoforms of barley (Hordeum vulgare L.) after growth on different levels of NH+ 4

One cytosolic glutamine synthetase (GS, EC 6.3.1.2) isoform (GS 1a) was active in the germinating seeds of barley (Hordeum vulgare L.). A second cytosolic GS isoform (GS 1b) was separated from the leaves as well as the roots of 10-d-old seedlings. The chloroplastic isoform (GS 2) was present and active only in the leaves. The three GS isoforms were active in N-supplied (NH⁺ ₄ or NO ₃ ^– ) as well as in N-free-grown seedlings. This indicates (i) that a supply of nitrogen to the germinating seeds was not necessary for the induction of the GS isoforms and (ii) that no nitrogen-specific isoforms appeared during growth of seedlings with different nitrogen sources. The activity of GS, however, depended on the seedlings' nitrogen source: the specific activity was much higher in the leaves and much lower in the roots of NH⁺ ₄-grown barley than in the respective organs of NO ₃ ^– -fed or N free-grown plants. With increasing concentrations of NH⁺ ₄ (supplied hydroponically during growth), the specific activity of GS 1b increased in the leaves, but decreased in the roots. The activity of GS 2 (leaf) also increased with increasing NH⁺ ₄ supply, whereas GS 1a activity (leaf and root) was not affected. The changes in the activities of GS 1b and GS 2 were correlated with changes in the subunit compositions of the active holoenzymes: growth at increased levels of external NH⁺ ₄ resulted in an increased abundance of one of the four GS subunits, and of two of the five GS 1b subunits in the leaves. In the roots, however, the abundance of these two GS 1b subunits was decreased under the same growth conditions, indicating an organ-specific difference either in the expression of the genes coding for the respective GS 1b subunits or in the assembly of the GS 1b holoenzymes. Furthermore, growth at different levels of NH⁺ ₄ resulted in changes in the substrate affinities of the isoforms GS 1b (root and leaf) and GS 2 (leaf), presumably due to the changes in the subunit compositions of the active holoenzymes.Abbreviations FPLC fast protein liquid chromatography - GHA -glutamyl hydroxamate - GS glutamine synthetase Dr. Roger Wallsgrove's (Rothamsted Experimental Station, Harpenden, UK) generous gift of GS antiserum is greatly appreciated.     

Regional-scale patterns of δ<Superscript>13</Superscript>C and δ<Superscript>15</Superscript>N associated with multiple ecosystem functions along an aridity gradient in grassland ecosystems

Background and aims

Nitrogen (N) deficiency and drought are two key limiting factors for rice production worldwide, but the relationship of drought stress with N homeostasis in rice is rarely advanced. The aim of this study was to dissect the physiological effects of drought stress on rice growth that coupled unbalanced N metabolism.

Results

Water-deficient stress (WD) limited stomatal aperture function and activity of Rubisco carboxylase to photosynthesis. The rate of total electron transport (J_t) and the electron to carboxylation (J_c) were considerably decreased, whereas the proportion of e^? flow to photorespiration was stimulated by WD, especially at 1600 μmol m^?2 s^?1 PPFD. Concurrently, the expressions of glycolate oxidase genes (GOX1, GOX5) and glycine decarboxylase complex (GDCH, GDCP and GDCT) were significantly induced in leaves of WD treatment, which led to the accumulation of reactive oxygen species in leaves. With the photosynthetic change, nitrate uptake and reduction were suppressed. Moreover, the enhanced photorespiration generated excess NH₃ accumulation in leaves and stimulated the expressions of GS1;1, GS1;2 and GS2, which were tightly coupled with the expressions of PEPC1 and PEPC2 under WD stress.

Conclusions

Our results suggest that the inhibited nitrate reduction associated with diminished electron transport rate, and the photorespiration-associated accumulation of hydrogen peroxide and NH₃ were critical in the drought-induced rice growth inhibition.

     

A comparison of Phragmites australisin freshwater and brackish marsh environments in North America

This paper compares the available North Americanliterature and data concerning several ecologicalfactors affecting Phragmites australisin inlandfreshwater, tidal fresh, and tidal brackish marshsystems. We compare aboveground productivity, plantspecies diversity, and sediment biogeochemistry; andwe summarize Phragmiteseffects on faunalpopulations in these habitats. These data suggest thatPhragmitesaboveground biomass is higher thanthat of other plant species occurring in the samemarsh system. Available data do not indicate anysignificant difference in the aboveground Phragmitesbiomass between marsh types, nor doesthere appear to be an effect of salinity on height.However, Phragmitesstem density wassignificantly lower in inland non-tidal freshwatermarshes than in tidal marshes, whether fresh orbrackish. Studies of the effects of Phragmiteson plant species richness suggest that Phragmitesdominated sites have lower diversity.Furthermore, Phragmiteseradication infreshwater sites increased plant diversity in allcases. Phragmitesdominated communities appearto have different patterns of nitrogen cyclingcompared to adjacent plant communities. Abovegroundstanding stocks of nitrogen (N) were found to behigher in Phragmitessites compared to thosewithout Phragmites. Porewater ammonium(NH₄ ⁺) did not differ among plant covertypes in the freshwater tidal wetlands, but inbrackish marshes NH₄ ⁺was much higher inSpartinaspp. than in neighboring Phragmitesstands. Faunal uses of Phragmitesdominated sites in North America were found to vary bytaxa and in some cases equaled or exceeded use ofother robust emergent plant communities. In light ofthese findings, we make recommendations for futureresearch.     

Root and Nodule Enzymes of Ammonia Assimilation in Two Plant-Conditioned Symbiotically Ineffective Genotypes of Alfalfa (Medicago sativa L.)

Biochemical and physiological parameters associated with nitrogen metabolism were measured in nodules and roots of glasshouse-grown clones of two symbiotically ineffective alfalfa (Medicago sativa L.) genotypes supplied with either NO₃⁻ or NH₄⁺. Significant differences were observed between genotypes for nodule soluble protein concentrations and glutamine synthetase (GS) and glutamate synthase (GOGAT) specific activities, both in untreated controls and in response to applied N. Nodule soluble protein of both genotypes declined in response to applied N, while nodule GS, GOGAT, and glutamate dehydrogenase (GDH) specific activities either decreased or remained relatively constant. In contrast, no genotype differences were observed in roots for soluble protein concentrations and GS, GOGAT, and GDH specific activities, either in untreated controls or in response to applied N. Root soluble protein levels and GS and GOGAT specific activities of N-treated plants increased 2- to 4-fold within 4 days and then decreased between days 13 and 24. Root GDH specific activity of NH₄⁺-treated plants increased steadily throughout the experiment and was 50 times greater than root GS or GOGAT specific activities by day 24.     

Photorespiration Process and Nitrogen Metabolism in Lettuce Plants (Lactuca sativa L.): Induced Changes in Response to Iodine Biofortification

Iodine is vital to human health, and iodine biofortification programs help improve human intake through plant consumption. There is no research on whether iodine biofortification influences basic plant physiological processes. Because nitrogen (N) uptake, utilization, and accumulation are determining factors in crop yield, the aim of this work was to establish the effect of the application of different doses (20, 40, and 80 μM) and forms of iodine (iodate [IO₃ ⁻] vs. Iodide [I⁻]) on N metabolism and photorespiration. For this study we analyzed shoot biomass and the activities of nitrate reductase (NR), nitrite reductase (NiR), glutamine synthetase (GS), glutamate synthase (GOGAT), aspartate aminotransferase (AAT), glutamate dehydrogenase (GDH), glycolate oxidase (GO), glutamate:glyoxylate aminotransferase (GGAT), serine:glyoxylate aminotransferase (SGAT), hydroxypyruvate reductase (HR) and catalase (CAT), nitrate (NO₃ ⁻), ammonium (NH₄ ⁺), organic and total N, amino acids, proteins, serine (ser), malate, and α-ketoglutaric acid in edible lettuce leaves. Application of I⁻ at doses of at least 40 μM reduced the foliar concentration of NO₃ ⁻ with no decrease in biomass production, which may improve the nutritional quality of lettuce plants. In contrast, the application of 80 μM of I⁻ is phytotoxic for lettuce plants, reducing the biomass, foliar concentration of organic N and NO₃ ⁻, and NR and GDH activities. HR activity is significantly inhibited with all doses of I⁻; the least inhibition was at 80 μM. This may involve a decrease in the incorporation of carbonated skeletons from photorespiration into the Calvin cycle, which may be partially associated with the biomass decrease. Finally, the application of IO₃ ⁻ increases biomass production, stimulates NO₃ ⁻ reduction and NH₄ ⁺ incorporation (GS/GOGAT), and optimizes the photorespiratory process. Hence, this appears to be the most appropriate form of iodine from an agronomic standpoint.     

Effect of altitude on the primary products of photosynthesis and the associated enzymes in barley and wheat

There is little information available on the primary products of photosynthesis and the change in the activity of the associated enzymes with altitude. We studied the same in varieties of barley and wheat grown at 1300 (low altitude, LA) and 4200 m (high altitude, HA) elevations above mean sea level in the western Himalayas. Plants at both the locations had similar photosynthetic rates, leaf water potential and the chlorophyll fluorescence kinetics. The short-term radio-labelling experiments in leaves showed appearance of ¹⁴CO₂ in phosphoglyceric acid and sugar phosphates in plants at both the LA and HA, suggesting a major role of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) in CO₂ fixation in the plants at two altitudes, whereas the appearance of labelled carbon in aspartate (Asp) and glutamate (Glu) at HA suggested a role of phosphoenolpyruvate carboxylase (PEPCase) in photosynthesis metabolism. Plants at HA had significantly higher activities of PEPCase, carboxylase and oxygenase activity of Rubisco, aspartate aminotransferase (AspAT), and glutamine synthetase (GS). However, the activities of malate dehydrogenase, NAD-malic enzyme and citrate synthase were similar at the two locations. Such an altered metabolism at HA suggested that PEPCase probably captured CO₂ directly from the atmosphere and/or that generated metabolically e.g. from photorespiration at HA. Higher oxygenase activity at HA suggests high photorespiratory activity. OAA thus produced could be additionally channelised for Asp synthesis using Glu as a source of ammonia. Higher GS activity ensures higher assimilation rate of NH₃ and the synthesis of Glu through GS-GOGAT (glutamine:2-oxoglutarate aminotransferase) pathway, also as supported by the appearance of radiolabel in Glu at HA. Enhanced PEPCase activity coupled with higher activities of AspAT and GS suggests a role in conserving C and N in the HA environment.     

Effect of NH4+/NO3− availability on nitrate reductase activity and nitrogen accumulation in wetland helophytes Phragmites australis and Glyceria maxima

The effect of NH₄⁺/NO₃⁻ availability on nitrate reductase (NR) activity in Phragmites australis and Glyceria maxima was studied in sand and water cultures with the goal to characterise the relationship between NR activity and NO₃⁻ availability in the rhizosphere and to describe the extent to which NH₄⁺ suppresses the utilization of NO₃⁻ in wetland plants.The NR activity data showed that both wetland helophytes are able to utilize NO₃⁻. This finding was further supported by sufficient growth observed under the strict NO₃⁻ nutrition. The distribution of NR activity within plant tissues differed between species. Phragmites was proved to be preferential leaf NO₃⁻ reducer with high NR activity in leaves (NR_max > 6.5 μmol NO₂⁻ g dry wt⁻¹ h⁻¹) under all N treatments, and therefore Phragmites seems to be good indicator of NO₃⁻ availability in flooded sediment. In the case of Glyceria the contribution of roots to plant NO₃⁻ reduction was higher, especially in sand culture. Glyceria also tended to accumulate NO₃⁻ in non-reduced form, showing generally lower leaf NR activity levels. Thus, the NR activity does not necessarily correspond with plant ability to take up NO₃⁻ and grow under NO₃⁻-N source. Moreover, the species differed significantly in the content of compounds interfering with NR activity estimation. Glyceria, but not Phragmites, contained cyanogenic glycosides releasing cyanide, the potent NR inhibitor. It clearly shows that the use of NR activity as a marker of NO₃⁻ utilization in individual plant species is impossible without the precise method optimisation.     

外源CaCl2对NaCl胁迫下酸枣幼苗氮代谢的影响

为了研究CaCl₂对NaCl胁迫下酸枣幼苗根、茎、叶的氮代谢影响,探索钙缓解幼苗NaCl胁迫的作用途径。该研究以酸枣幼苗为试验材料,检测不同浓度CaCl₂(0、5、10、20 mmol/L)对NaCl(150 mmol/L)胁迫下幼苗叶片H₂O₂、O^-·₂含量,根、茎、叶中硝酸还原酶(NR)、谷氨酰胺合成酶(GS)、谷氨酸合酶(GOGAT)活性及游离氨基酸、可溶性蛋白、硝态氮含量的影响,并采用主成分分析法筛选出评价CaCl₂缓解NaCl胁迫效应的生理指标。结果表明：与NaCl胁迫相比,盐胁迫幼苗叶片的H2O₂、O^-·₂积累量在5、10 mmol/L CaCl₂处理下显著减少;GOGAT活性在5、10 mmol/L CaCl₂处理下的植株根和茎内以及各浓度 CaCl₂处理的叶内均显著升高, GS、NR活性在10、20 mmol/L CaCl₂处理的根内和10 mmol/L CaCl₂处理的茎内以及5、10、20 mmol/L CaCl₂处理的叶内均显著升高;可溶性蛋白含量在5、10、20 mmol/L CaCl₂处理的根、茎、叶内均显著升高,游离氨基酸含量在10、20 mmol/L CaCl₂处理的根和茎内以及10 mmol/L CaCl₂处理的叶内均显著升高,硝态氮含量在10 mmol/L CaCl₂处理的根和茎内以及5、10、20 mmol/L CaCl₂处理的叶内均显著升高。研究发现,150 mmol/L NaCl胁迫对酸枣幼苗造成明显过氧化伤害,抑制了体内氮代谢;外源CaCl₂可通过促进幼苗根和茎内GS/GOGAT循环对NH₄⁺的同化作用,提高叶片NR活性,加快硝态氮的转化速率,从而增强幼苗对NaCl胁迫的适应性,并以10 mmol/L CaCl₂处理缓解效果最佳;游离氨基酸、GOGAT、NR可以作为CaCl₂缓解幼苗NaCl胁迫伤害的评价指标。     

Prolonged root hypoxia effects on enzymes involved in nitrogen assimilation pathway in tomato plants

In order to investigate the effects of root hypoxia (1–2% oxygen) on the nitrogen (N) metabolism of tomato plants (Solanum lycopersicum L. cv. Micro-Tom), a range of N compounds and N-assimilating enzymes were performed on roots and leaves of plants submitted to root hypoxia at the second leaf stage for three weeks. Obtained results showed that root hypoxia led to a significant decrease in dry weight (DW) production and nitrate content in roots and leaves. Conversely, shoot to root DW ratio and nitrite content were significantly increased. Contrary to that in leaves, glutamine synthetase activity was significantly enhanced in roots. The activities of nitrate and nitrite reductase were enhanced in roots as well as leaves. The higher increase in the NH₄⁺ content and in the protease activities in roots and leaves of hypoxically treated plants coincide with a greater decrease in soluble protein contents. Taken together, these results suggest that root hypoxia leaded to higher protein degradation. The hypoxia-induced increase in the aminating glutamate dehydrogenase activity may be considered as an alternative N assimilation pathway involved in detoxifying the NH₄⁺, accumulated under hypoxic conditions. With respect to hypoxic stress, the distinct sensitivity of the enzymes involved in N assimilation is discussed.Key words: tomato, hypoxia, nitrogen, glutamine synthetase, protease, glutamate dehydrogenase     

Carbon and Nitrogen Metabolism in Leaves and Roots of Dwarf Bamboo (Fargesia denudata Yi) Subjected to Drought for Two Consecutive Years During Sprouting Period

Dwarf bamboo is an ecologically and economically important forest resource that is widespread in mountainous regions of eastern Asia and southern America. Fargesia denudata, one of the most important dwarf bamboos, is a staple food of the giant panda, but our knowledge about how F. denudata copes with drought stress is very limited. The objective of this study was to determine the responses of carbon (C) and nitrogen (N) metabolism to drought in leaves and roots of F. denudata plants. Plants were subjected to three water treatments, well-watered [WW, 85 % relative soil water content (RSWC)], moderate drought (MD, 50 % RSWC), and severe drought (SD, 30 % RSWC), for two consecutive years during the sprouting period. Plant growth parameters, levels of carbohydrates and N compounds, and activities of key enzymes involved in C and N metabolism were analyzed. In young leaves, C metabolism was in balance after drought stress, but nitrate (NO₃ ^?) reduction and ammonium (NH₄ ⁺) assimilation were accelerated. In old leaves, drought stress decreased carbohydrate contents by spurring the activities of the main enzymes that participate in C metabolism, whereas N metabolism was enhanced only under SD. Roots showed unchanged C metabolism parameters under MD, together with stable NO₃ ^? reduction and the key enzymes related to NH₄ ⁺ assimilation, whereas they were stimulated by SD. Hydrolysates of carbohydrates in old leaves could be transferred into roots, but only to meet MD. Meanwhile, roots could allocate more N nutrition to young leaves and less to old leaves. These changes regulated the overall metabolic balance of F. denudata. Consequently, the results indicate that different organs with various response strategies will be well adapted to different drought intensities for ensuring regular growth of F. denudata plants at the whole-plant level.     

High irradiance increases NH(4)(+) tolerance in Pisum sativum: Higher carbon and energy availability improve ion balance but not N assimilation

The widespread use of NO₃⁻ fertilization has had a major ecological impact. NH₄⁺ nutrition may help to reduce this impact, although high NH₄⁺ concentrations are toxic for most plants. The underlying tolerance mechanisms are not yet fully understood, although they are thought to include the limitation of C, the disruption of ion homeostasis, and a wasteful NH₄⁺ influx/efflux cycle that carries an extra energetic cost for root cells.In this study, high irradiance (HI) was found to induce a notable tolerance to NH₄⁺ in the range 2.5-10 mM in pea plants by inducing higher C availability, as shown by carbohydrate content. This capacity was accompanied by a general lower relative N content, indicating that tolerance is not achieved through higher net N assimilation on C-skeletons, and it was also not attributable to increased GS content or activity in roots or leaves. Moreover, HI plants showed higher ATP content and respiration rates. This extra energy availability is related to the internal NH₄⁺ content regulation (probably NH₄⁺ influx/efflux) and to an improvement of the cell ionic balance.The limited C availability at lower irradiance (LI) and high NH₄⁺ resulted in a series of metabolic imbalances, as reflected in a much higher organic acid content, thereby suggesting that the origin of the toxicity in plants cultured at high NH₄⁺ and LI is related to their inability to avoid large-scale accumulation of the NH₄⁺ ion.