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  • 1
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: To illustrate the development of the source-to-sink transition in maize leaves during the grain-filling period, an integrated physiological-agronomic approach is presented in this study. The evolution of physiological markers such as total leaf nitrogen (N), chlorophyll, soluble protein, amino acid and ammonium contents was monitored from silking to a period close to maturity in different leaf stages of three maize genotypes grown at high and low levels of N fertilization. In addition, the activities of glutamine synthetase (GS) and glutamate dehydrogenase (GDH), two enzymes known to play a direct or an indirect role during leaf N remobilization, were measured. In the three genotypes examined, we found that a general decrease of most metabolic and enzyme markers occurred during leaf ageing and that this decrease was enhanced when plants were N starved. In contrast, such variations were not observed between different sections of a single leaf even at an advanced stage of leaf senescence. We found that there is a strong correlation between total N, chlorophyll, soluble protein and GS activity, which is not dependent upon the N fertilization level, which indicates the N status of the plant, either in a single leaf or during ageing. In contrast, ammonium, amino acids and GDH activity were not subject to such variations, thus suggesting that they are indicators of the metabolic activity of the whole plant in response to the level of N fertilization. The use of these markers to predict the N status of maize as a function of both plant development and N availability is discussed.
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  • 2
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The terminal step of glutathione (GSH) synthesis is the condensation of γ-glutamyl-cysteine (γ-EC) with glycine. Relatively little information exists concerning the importance of photorespiratory glycine in determining the rate of conversion of γ-EC to GSH. Consequently, the effect of exogenous glycine and of illumination on foliar contents of γ-EC and GSH was studied in excised leaves and leaf discs from untransformed poplar (Populus tremula×P. alba) and poplar overexpressing γ-glutamylcysteine synthetase (γ-ECS; EC 6.3.2.2). Poplars strongly overexpressing γ-ECS (ggs28) had enhanced levels of γ-EC and GSH compared to untransformed poplars. The relationship between γ-EC and GSH contents in ggs28 was light dependent. In illuminated leaves, GSH contents were up to 50-fold higher than γ-EC. On darkening, γ-EC accumulated markedly and GSH declined, so that the GSH:γ-EC ratio was close to 1. These dark-induced changes were prevented by supplying glycine through the petiole or by incubation of leaf discs on glycine. Dark accumulation of γ-EC in leaf discs from untransformed poplar was also prevented by supplying glycine. Supplying cysteine in the dark to discs from untransformed poplar and ggs28 increased γ-EC levels markedly but GSH levels only slightly. Subsequent illumination caused γ-EC to decrease and GSH to increase. Supplying glycine in concert with cysteine had similar effects to illumination. The data suggest that photorespiratory glycine is essential for GSH synthesis, especially under stress conditions, where increased amounts of GSH are required.
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  • 3
    ISSN: 1432-2048
    Keywords: Key words: Glutamate dehydrogenase – Glutamine synthetase – Leaf senescence – Metabolic signals – Nicotiana (N metabolism) – Nitrogen metabolism
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract.  The metabolic, biochemical and molecular events occurring during tobacco (Nicotiana tabacum) leaf ageing are presented, with a particular emphasis on nitrogen metabolism. An integrated model describing the source/sink relationship existing between leaves of different developmental stages along the main plant axis is proposed. The results of our study show that a tobacco plant can be divided into two main sections with regards to sink/source relationships. Sink-to-source transition occurs at a particular leaf stage in which a breakpoint corresponding to an accumulation of carbohydrates and a depletion of both organic and inorganic nitrogen is observed. The sink/source transition is also marked by the appearence of endoproteolytic activities and the induction of both cytosolic glutamine synthetase and NAD(H)-dependent glutamate dehydrogenase transcripts, proteins and activities. The role of the newly induced enzymes and the nature of the potential metabolic and developmental signals involved in the regulation of their expression during leaf senescence are discussed.
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  • 4
    ISSN: 1432-2048
    Keywords: Nicotiana ; Nitrate nutrition ; Nitrate reductase ; Photosynthesis ; Transgenic plant (tobacco)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Transformed Nicotiana plumbaginifolia plants with constitutive expression of nitrate reductase (NR) activity were grown at different levels of nitrogen nutrition. The gradients in foliar NO 3 − content and maximum extractable NR activity observed with leaf order on the shoot, from base to apex, were much decreased as a result of N-deficiency in both the transformed plants and wild type controls grown under identical conditions. Constitutive expression of NR did not influence the foliar protein and chlorophyll contents under any circumstances. A reciprocal relationship between the observed maximal extractable NR activity of the leaves and their NO 3 − content was observed in plants grown in nitrogen replete conditions at low irradiance (170 μmol photons·m−2 ·s−1). This relationship disappeared at higher irradiance (450 μmol photons·m−2·S−1) because the maximal extractable NR activity in the leaves of the wild type plants in these conditions increased to a level that was similar to, or greater than that found in constitutive NR-expressors. Much more NO 3 − accumulated in the leaves of plants grown at 450 μmol photons·m−2·s−1 than in those grown at 170 μmol photons·m−2·s−1 in N-replete conditions. The foliar NO 3 − level and maximal NR activity decreased with the imposition of N-deficiency in all plant types such that after prolonged exposure to nitrogen depletion very little NO 3 − was found in the leaves and NR activity had decreased to almost zero. The activity of NR decreased under conditions of nitrogen deficiency. This regulation is multifactoral since there is no regulation of NR gene expression by NO 3 − in the constitutive NR-expressors. We conclude that the NR protein is specifically targetted for destruction under nitrogen deficiency. Consequently, constitutive expression of NR activity does not benefit the plant in terms of increased biomass production in conditions of limiting nitrogen.
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  • 5
    ISSN: 1432-2048
    Keywords: Key words: γ-Glutamylcysteine ; γ-Glutamylcysteine synthetase ; Glutathione ; Glycine ; Photorespiration ; Populus (transformed)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract. Glutathione (GSH), γ-glutamylcysteine (γ-EC) and major free amino acids were measured in darkened and illuminated leaves from untransformed poplars (Populus tremula × P. alba) and poplars expressing Escherichia coli genes for γ-glutamylcysteine synthetase (γ-ECS; EC 3.2.3.3) and glutathione reductase (GR; EC 1.6.4.2). In poplars overexpressing γ-ECS, foliar γ-EC contents and GSH contents were markedly enhanced compared to poplars lacking the bacterial gene for the enzyme. However, the quantitative relationship between the foliar pools of γ-EC and GSH in these transformants was markedly dependent on light. In the dark, GSH content was relatively low and γ-EC content high, the latter being higher than the foliar GSH contents of untransformed poplars in all conditions. Hence, this transformation appears to elevate γ-EC from the ranks of a trace metabolite to one of major quantitative importance. On illumination, however, γ-EC content decreased fourfold whereas GSH content doubled. Glutathione was also higher in the light in untransformed poplars and in those overexpressing GR. In these plants, γ-EC was negligible in the light but increased in the dark. Cysteine content was little affected by light in any of the poplar types. No light-dependent changes in the extractable activities of γ-ECS, glutathione synthetase (EC 3.2.3.2) or GR were observed. In contrast, both the activation state and the maximum extractable activity of nitrate reductase (EC 1.6.6.1) were increased by illumination. In all poplar types, glutamate and aspartate were the major amino acids. The most marked light-induced increases in individual amino acids were observed in the glutamine, asparagine, serine and glycine pools. Illumination of leaves from poplars overexpressing γ-ECS at elevated CO2 or low O2 largely abolished the inverse light-dependent changes in γ-EC and GSH. Low O2 did not affect foliar contents of cysteine or glutamate but prevented the light-induced increase in the glycine pool. It is concluded that light-dependent glycine formation through the photorespiratory pathway is required to support maximal rates of GSH synthesis, particularly under conditions where the capacity for γ-EC synthesis is augmented.
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  • 6
    ISSN: 1432-2048
    Keywords: Key words: Carbon dioxide enrichment ; Carbon/nitrogen ratio ; Glutamine ; Nicotiana (C and N metabolism) ; Nitrate ; Nitrate reductase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract. Transformed plants of Nicotiana plumbaginifolia Viv. constitutively expressing nitrate reductase (35S-NR) or β-glucuronidase (35S-GUS) and untransformed controls were grown for two weeks in a CO2-enriched atmosphere. Whereas CO2 enrichment (1000 μl · l−1) resulted in an increase in the carbon (C) to nitrogen (N) ratio of both the tobacco lines grown in pots with vermiculite, the C/N ratio was only slightly modified when plants were grown in hydroponic culture in high CO2 compared to those grown in air. Constitutive nitrate reductase (NR) expression per se did not change the C/N ratio of the shoots or roots. Biomass accumulation was similar in both types of plant when hydroponic or pot-grown material, grown in air or high CO2, were compared. Shoot dry matter accumulation was primarily related to the presence of stored carbohydrate (starch and sucrose) in the leaves. In the pot-grown tobacco, growth at elevated CO2 levels caused a concomitant decrease in the N content of the leaves involving losses in NO− 3 and amino acid levels. In contrast, the N content and composition were similar in all plants grown in hydroponic culture. The 35S-NR plants grown in air had higher foliar maximum extractable NR activities and increased glutamine levels (on a chlorophyll or protein basis) than the untransformed controls. These increases were maintained following CO2 enrichment when the plants were grown in hydroponic culture, suggesting that an increased flux through nitrogen assimilation was possible in the 35S-NR plants. Under CO2 enrichment the NR activation state in the leaves was similar in all plants. When the 35S-NR plants were grown in pots, however, foliar NR activity and glutamine content fell in the 35S-NR transformants to levels similar to those of the untransformed controls. The differences in NR activity between untransformed and 35S-NR leaves were much less pronounced in the hydroponic than in the pot-grown material but the difference in total extractable NR activity was more marked following CO2 enrichment. Foliar NR message levels were decreased by CO2 enrichment in all growth conditions but this was much more pronounced in pot-grown material than in that grown hydroponically. Since β-glucuronidase (GUS) activity and message levels in 35S-GUS plants grown under the same conditions of CO2 enrichment (to test the effects of CO2 enrichment on the activity of the 35S promoter) were found to be constant, we conclude that NR message turnover was specifically accelerated in the 35S-NR plants as well as in the untransformed controls as a result of CO2 enrichment. The molecular and metabolic signals involved in increased NR message and protein turnover are not known but possible effectors include NO3 −, glutamine and asparagine. We conclude that plants grown in hydroponic culture have greater access to N than those grown in pots. Regardless of the culture method, CO2 enrichment has a direct effect on NR mRNA stability.
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  • 7
    ISSN: 1432-2048
    Keywords: Nicotiana (nitrate reductase) ; Nitrate ; Nitrate reductase (activation state) ; Protein phosphorylation ; Transgenic plants ; Zea (nitrate reductase)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Maize (Zea mays L.) grown on low (0.8 mM) NO 3 - , as well as untransformed and transformed Nicotiana plumbaginifolia constitutively expressing nitrate reductase (NR), was used to study the effects of NO 3 - on the NR activation state. The NR activation state was determined from the relationship of total activity extracted in the presence of ethylenediaminetetracetic acid to that extracted in the presence of Mg2+. Light activation was observed in both maize and tobacco leaves. In the tobacco lines, NO 3 - did not influence the NR activation state. In excised maize leaves, no correlation was found between the foliar NO 3 - content and the NR activation state. Similarly, the NR activation state did not respond to NO 3 - . Since the NR activation state determined from the degree of Mg2+-induced inhibition of NR activity is considered to reflect the phosphorylation state of the NR protein, the protein phosphatase inhibitor microcystin LR was used to test the importance of protein phosphorylation in the NO 3 - -induced changes in NR activity. In-vivo inhibition of endogenous protein phosphatase activity by microcystin-LR decreased the level of NR activation in the light. This occurred to the same extent in the presence or absence of exogenous NO 3 - . We conclude that NO 3 - does not effect the NR activation state, as modulated by protein phosphorylation in either tobacco (a C3 species) or maize (a C4 species). The short-term regulation of NR therefore differs from the NO 3 - -mediated responses observed for phosphoenolpyruvate carboxylase and sucrose phosphate synthase.
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  • 8
    Publication Date: 1982-01-01
    Print ISSN: 0249-5627
    Electronic ISSN: 1297-9643
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Published by EDP Sciences
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  • 9
    Publication Date: 1994-07-01
    Print ISSN: 0031-9422
    Electronic ISSN: 1873-3700
    Topics: Biology , Chemistry and Pharmacology
    Published by Elsevier
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  • 10
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