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Stomatal and mesophyll limitations of photosynthesis in black spruce seedlings during multiple cycles of drought.

Container-grown black spruce (Picea mariana (Mill.) B.S.P.) seedlings were planted in trays containing a sand and peat mixture, and placed in a climate-controlled greenhouse. One group of seedlings was kept well-watered, and another group was subjected to three cycles of drought. Gas exchange analysis showed that mesophyll photosynthetic function was largely unimpaired by drought. In contrast, stomatal conductance was sensitive to drought, although it became less sensitive with each drought cycle. Both stomatal and mesophyll conductances increased with time in control and drought-stressed seedlings, but mesophyll conductance increased with time more rapidly than did stomatal conductance. Limitation of photosynthetic rate was dominated by the mesophyll. In control seedlings, relative stomatal limitation increased from 6 to 16% by the end of the experiment. In drought-stressed seedlings, relative stomatal limitation of photosynthesis reached 40% during the first drought, but decreased to near control values immediately after rewatering. Because the third, most severe drought had only a minor effect on stomatal conductance, relative stomatal limitation of photosynthesis was similar to that in control seedlings by the end of the experiment. Inhibition of ontogenetic change during drought stress may be responsible for the apparent acclimation of mesophyll photosynthetic processes. We conclude that it would be more effective to select for high photosynthetic capacity than for reduced stomatal sensitivity when breeding for increased drought resistance in black spruce seedlings.

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Isoprene emission, photosynthesis, and growth in sweetgum (Liquidambar styraciflua) seedlings exposed to short- and long-term drying cycles.

Isoprene emissions were studied in one-year old sweetgum (Liquidambar styraciflua L.) seedlings during nine drying-rewatering cycles extending over five months. Each drying cycle lasted to the point of leaf wilting. Growth was essentially stopped in response to the first drying cycle, though seedling survival and capacity to recover turgor on rewatering remained high throughout the entire nine cycles. Photosynthetic rates of leaves were inhibited by the drying treatments. Under severe drought, isoprene emission rates of leaves were also inhibited, though isoprene emission was generally less sensitive to drought than photosynthesis. The lower drought sensitivity of isoprene emission compared with photosynthesis resulted in a higher percentage of fixed carbon lost as isoprene as seedlings became more stressed. During the recovery phase of the drying-rewatering cycles, isoprene emission rates in several seedlings were higher than in well-watered control seedlings. Following the ninth drying-rewatering cycle, sustained daily watering resulted in recovery of isoprene emission rates to control values within four days. Photosynthetic rates only recovered to 50% of control values after seven days. We conclude that the mechanisms regulating photosynthetic rate and isoprene emission rate are differentially influenced by limited water supplies. The results are consistent with past studies that predict a protective role for isoprene emission during stress, particularly protection from excessive leaf temperatures during drought.

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Influence of environmental and plant factors on canopy photosynthesis and transpiration of apple trees.

We estimated carbon and water flows, canopy conductance and the assimilation/transpiration ratio of fruiting and non-fruiting apple trees grown in the field, from daily gas exchange measurements taken during the summer with a whole-canopy enclosure device. The relationships between photosynthetic and transpirational responses and environmental conditions were also investigated, as well as the role of canopy conductance in controlling carbon dioxide and water vapor exchange. Light-saturated net photosynthetic rates, which were higher for the fruiting canopy than for the non-fruiting canopy, showed a general decrease in the afternoon, particularly for the non-fruiting canopy, compared with rates in the morning. When light was not limiting, the afternoon decrease in net photosynthesis appeared to be regulated more by non-stomatal factors than by changes in canopy conductance. Canopy conductance, which was higher for the fruiting canopy than for the non-fruiting canopy, may actively regulate photosynthetic activity and may also be modulated by feedback control in response to assimilation capacity. We conclude that adjustments in canopy conductance, which were partially dependent on the vegetative-reproductive status of the tree, control the equilibrium between photosynthesis and transpiration. We also demonstrated that whole-canopy chambers can be used to estimate photosynthetic and transpirational responses thereby overcoming the difficulty of scaling these physiological responses from the leaf to the whole-canopy level.

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Effects of elevated CO(2) concentration and nutrition on net photosynthesis, stomatal conductance and needle respiration of field-grown Norway spruce trees.

To study the effects of elevated CO(2) on gas exchange, nonstructural carbohydrate and nutrient concentrations in current-year foliage of 30-year-old Norway spruce (Picea abies (L.) Karst.) trees, branches were enclosed in ventilated, transparent plastic bags and flushed with ambient air (mean 370 &mgr;mol CO(2) mol(-1); control) or ambient air + 340 &mgr;mol CO(2) mol(-1) (elevated CO(2)) during two growing seasons. One branch bag was installed on each of 24 selected trees from control and fertilized plots. To reduce the effect of variation among trees, results from each treated branch were compared with those from a control branch on the same whorl of the same tree. Elevated CO(2) increased rates of light-saturated photosynthesis on average by 55% when measured at the treatment CO(2) concentration. The increase was larger in shoots with high needle nitrogen concentrations than in shoots with low needle nitrogen concentrations. However, shoots grown in elevated CO(2) showed a decrease in photosynthetic capacity compared with shoots grown in ambient CO(2). When measured at the internal CO(2) concentration of 200 &mgr;mol CO(2) mol(-1), photosynthetic rates of branches in the elevated CO(2) treatments were reduced by 8 to 32%. The elevated CO(2) treatment caused a 9 to 20% reduction in carboxylation efficiency and an 18% increase in respiration rates. In response to elevated CO(2), starch, fructose and glucose concentrations in the needles increased on average 33%, whereas concentrations of potassium, nitrogen, phosphorus, magnesium and boron decreased. Needle nitrogen concentrations explained 50-60% of the variation in photosynthesis and CO(2) acclimation was greater at low nitrogen concentrations than at high nitrogen concentrations. We conclude that the enhanced photosynthetic rates found in shoots exposed to elevated CO(2) increased carbohydrate concentrations, which may have a negative feedback on the photosynthetic apparatus and stimulate cyanide-resistant respiration. We also infer that the decrease in nutrient concentrations of needles exposed to elevated CO(2) was the result of retranslocation of nutrients to other parts of the branch or tree.

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Interrelationships among light, photosynthesis and nitrogen in the crown of mature Pinus contorta ssp. latifolia.

Scaling leaf-level measurements to estimate carbon gain of entire leaf crowns or canopies requires an understanding of the distribution of photosynthetic capacity and corresponding light microenvironments within a crown. We have compared changes in the photosynthetic light response and nitrogen (N) content (per unit leaf area) of Pinus contorta Dougl. ssp. latifolia Engelm. (lodgepole pine) leaves in relation to their age and light microenvironment. The vertical gradient in integrated daily photosynthetic photon flux density (PPFD) from the upper to the lower crown of lodgepole pine was similar in magnitude to the horizontal gradient in daily PPFD along shoots from young to old leaves. The relationship between light-saturated net photosynthesis (A(max)) and daily PPFD was significant for both young and old leaves. However, old leaves had a lower A(max) than young leaves in a similar daily irradiance regime. For leaves of all ages from throughout the crown, A(max) was linearly related to the estimated daily net carbon gain that leaves could achieve in their natural PPFD environment (estimated A(day)) (r(2) = 0.84, P < 0.001, n = 39), indicating that estimated A(day) may be dominated by carbon fixed when leaves are light-saturated and operating at A(max). Comparison of the PPFD required to achieve A(max) and the PPFD available to the leaves showed that all of the measured leaves (n = 39), regardless of their position in the crown or age, were in light environments that could light-saturate photosynthesis for a similar proportion of the day. For all data pooled, foliar N was weakly correlated with daily PPFD. Analyzing each leaf age class separately showed that foliar N was significantly related to daily PPFD, A(max), and estimated A(day) for the youngest leaves but not for middle-aged or old leaves. Therefore, the general theory that foliar N is allocated within a crown according to total daily light availability was supported only for young (1-4 years old) leaves in this study.

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Magnesium nutrition and photosynthesis in Pinus radiata: clonal variation and influence of potassium.

Magnesium (Mg) nutrition and photosynthesis were studied in clones of Pinus radiata D. Don grown in sand culture for 21 weeks at four Mg concentrations (0.008, 0.04, 0.2 and 0.4 mM) and three potassium (K) concentrations (0.25, 0.5 and 2.5 mM). We found significant clonal variation in Mg nutrition of P. radiata. Plants grown at 0.04 mM [Mg] or less showed pronounced visible symptoms of foliar Mg deficiency. Net photosynthetic rate and leaf conductance were closely related to shoot Mg concentrations below a concentration of 0.6 mg Mg g(DW) (-1). Potassium enhanced the development of visible symptoms of foliar Mg deficiency. At the lowest Mg concentration tested (0.008 mM), the severity of needle chlorosis and necrosis increased with increasing K concentration in the culture solution. With increasing Mg concentration, 2.5 mM [K] in the culture solution markedly increased root Mg concentration, but decreased shoot Mg concentration, suggesting that excessive K inhibited Mg mobilization from roots to shoots. Rates of growth and photosynthesis were both severely inhibited at 0.008 mM [Mg].

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Response of Ulmus americana seedlings to varying nitrogen and water status. 1 Photosynthesis and growth.

Well-watered American elm (Ulmus americana L.) seedlings responded to increased nitrate availability with increased leaf nitrogen (N) concentration and photosynthetic rate, larger and more numerous leaves, greater total growth and greater proportional allocation of carbon to shoot than root. Plasticity of growth and carbon allocation were greater than plasticity of N concentration and photosynthetic capacity. For a given N availability, allocation of N per unit leaf area was positively correlated with dry mass per unit leaf area (specific leaf mass), but these relationships differed with N availability. Rates of net photosynthesis and leaf conductance declined logarithmically with decreasing predawn water status. Increased water stress resulted in a greater relative decline in net photosynthesis and leaf conductance for high-N than low-N plants.

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Influence of crown structural properties on PAR absorption, photosynthesis, and transpiration in Sitka spruce: application of a model (MAESTRO).

The structure of a tree crown can be described by the spatial distribution, inclination, and orientation of all the phytoelements (leaves, twigs, branches, trunk, etc.), and their geometric properties. The following four structural properties have been studied in relation to radiation absorption, photosynthesis, and transpiration using a simulation model named MAESTRO: crown shape, total area of leaves and their spatial distribution within the tree crown, and the leaf inclination angle distribution. It was found that the total area of leaves and their spatial distribution within the tree crown are far more important than the other two properties for radiation absorption, photosynthesis, and transpiration.

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Effects of leaf display on light interception and apparent photosynthesis in two contrasting Populus cultivars during their second growing season.

Effects of the contrasting leaf display of poplar cultivars Eugenei (Populus x euramericana) and Tristis (P. tristis x P. balsamifera) on light interception and photosynthesis were studied in the second year of growth in an irrigated plantation near Rhinelander, Wisconsin, USA (lat. 45 degrees N). Leaves on the current terminal (CT) and on proleptic branches were measured between 0900 and 1500 h on five clear days from June to September 1980. Leaf orientation-based differences between these cultivars were evident as the second growing season progressed and the crowns of the trees in the plantation grew together. Leaves of Eugenei are erectophile or tilted from the horizontal. In this cultivar light penetrated throughout the crown; many leaves on the lowest branches were illuminated as fully as those on the upper CT and had higher photosynthetic rates than equivalent leaves in Tristis. However, by early September many of the lower branches on Eugenei trees had abscised. In the planophile Tristis, adaxial photon flux densities (PPFD) of leaves on the lower portion of the CT and on branches were only a fraction of those measured on the upper CT. This pattern became more extreme as the season progressed. Few of the lower branches of Tristis abscised during the growing season. Photosynthesis rates, especially on a whole-leaf basis, were closely related to incident PPFDs in both cultivars. The ecological significance of these results are discussed, as well as the hypothesized effect of leaf inclination on crop productivity.

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Effects of Water-Deficit Stress on Photosynthesis, Its Components and Component Limitations, and on Water Use Efficiency in Wheat (Triticum aestivum L.).

It is of theoretical as well as practical interest to identify the components of the photosynthetic machinery that govern variability in photosynthesis rate (A) and water-use efficiency (WUE), and to define the extent by which the component processes limit A and WUE during developing water-deficit stress. For that purpose, leaf exchange of CO(2) and H(2)O was determined in two growth-chamber-grown wheat cultivars (Triticum aestivum L. cv TAM W-101 and cv Sturdy), and the capacity of A was determined and broken down into carboxylation efficiency (c.e.), light- and CO(2)-saturated A, and stomatal conductance (g(s)) components. The limitations on A measured at ambient CO(2) concentration (A(350)) were estimated. No cultivar difference was observed when A(350) was plotted versus leaf water potential (Psi(w)). Light- and CO(2)-saturated A, c.e., and g(s) decreased with decreasing leaf Psi(w), but of the corresponding photosynthesis limitations only those caused by insufficient c.e. and g(s) increased. Thus, reduced stomatal aperture and Calvin cycle activity, but not electron transport/photophosphorylation, appeared to be major reasons for drought stress-induced inhibition of A(350). WUE measured as A(350)/g(s) first increased with stomatal closure down to a g(s) of about 0.25 mol H(2)O m(-2) s(-1) (Psi(w) = -1.6 MPa). However, it was predicted that A(350)/g(s) would decrease with more severe stress due to inhibition of c.e.

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Photosynthesis, Rubisco Activity and Amount, and Their Regulation by Transcription in Senescing Soybean Leaves.

Senescence is a phase of leaf ontogeny marked by declining photosynthetic activity that, in soybean (Glycine max [L.] Merr.), is paralleled by a decline in chloroplast function. Soybean leaves have different patterns of decline in photosynthetic capacity and chloroplast function associated with nodal position and sink activity. The objective of this work was to determine whether leaves from nodes 3 and 6 of soybean, which show these different patterns, are similarly regulated with respect to ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) activity and content and also to ascertain the degree of regulation of Rubisco content by transcription. Leaves from nodes 3 and 6 of field-grown soybean plants were sampled periodically from the time of their unfolding until near death. In situ CO2-exchange rate (CER) increased to a maximal level in both leaves and then declined slowly. For node 3 leaves the decline was progressive, but for node 6 leaves the decline was arrested at about 75% of maximum CER for a period of about 20 d, coincident with the onset of rapid seed growth, before a short period of very rapid decline immediately preceding leaf death. Rubisco activities and Rubisco content were directly correlated with CER in the leaves exhibiting the two different patterns. Rubisco activation ratio was similar for the two leaves and did not change throughout development. The primary regulator of photosynthesis at the physiological level, thus, was the amount of Rubisco protein. Decreases in Rubisco holoenzyme during senescence of both leaves were accompanied by coordinate decreases in the levels of mRNAs for the small and large subunits of Rubisco, suggesting that the decrease in Rubisco enzyme amounts during soybean leaf senescence is due to slower transcription rates and that levels of these mRNAs are coordinately controlled during senescence as they are during chloroplast development. However, plastid DNA template availability and posttranscriptional controls may also influence Rubisco content during senescence of these leaves. We conclude that soybean leaf photosynthesis likely unfolds according to a single developmental program but that modifications can be superimposed upon this program to maximize photosynthetic rates.

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Response of Photochemical Processes of Photosynthesis to Dinitrogen Fixation in Soybean.

Symbiotic N2 fixation activity brings about changes in the photochemical processes of photosynthesis in soybean (Glycine max L. Merr.). For a potential photochemical efficiency ([phi]Po) similar to that obtained with an exclusively mineral nutrition, soybean, at full bloom stage (R2) with a moderate N2 fixation activity, had a better electron transfer quantum yield ([phi]PSII) and a higher photochemical quenching. At the beginning seed stage (R5), corresponding to more intense N2 fixation, the same phenomena were enhanced; in addition, an effect on the photochemical (k2b) and nonphotochemical (Kn-k22) transfer rates and an earlier activation of the electron transfer chain were characterized using a new parameter, the relative induction time of PSII fluorescence (Ap/Fm). The response of the photochemical parameters was related to the N2 fixation level (performance of the host plant-microsymbiont association): the energetic cost of symbiotic N2 fixation appeared to be met by a better photochemical efficiency of photosynthesis coupled with a decrease in thermal dissipation (kn - k22), by faster thylakoid energization, and by faster reopening of photosystem II centers at the time of fluorescence induction, as shown by decreased Ap/Fm.

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Use of Transgenic Plants with Ribulose-1,5-Bisphosphate Carboxylase/Oxygenase Antisense DNA to Evaluate the Rate Limitation of Photosynthesis under Water Stress.

The biochemical lesion that causes impaired chloroplast metabolism (and, hence, photosynthetic capacity) in plants exposed to water deficits is still a subject of controversy. In this study we used tobacco (Nicotiana tabacum L.) transformed with "antisense" ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) DNA sequences to evaluate whether Rubisco or some other enzymic step in the photosynthetic carbon reduction cycle pathway rate limits photosynthesis at low leaf water potential ([psi]w). These transformants, along with the wild-type material, provided a novel model system allowing for an evaluation of photosynthetic response to water stress in near-isogenic plants with widely varying levels of functional Rubisco. It was determined that impaired chloroplast metabolism (rather than decreased leaf conductance to CO2) was the major cause of photosynthetic inhibition as leaf [psi]w declined. Significantly, the extent of photosynthetic inhibition at low [psi]w was identical in wild-type and transformed plants. Decreasing Rubisco activity by 68% did not sensitize photosynthetic capacity to water stress. It was hypothesized that, if water stress effects on Rubisco caused photosynthetic inhibition under stress, an increase in the steady-state level of the substrate for this enzyme, ribulose 1,5-bisphosphate (RuBP), would be associated with stress-induced photosynthetic inhibition. Steady-state levels of RuBP were reduced as leaf [psi]w declined, even in transformed plants with low levels of Rubisco. Based on the similarity in photosynthetic response to water stress in wild-type and transformed plants, the reduction in RuBP as stress developed, and studies that demonstrated that ATP supply did not rate limit photosynthesis under stress, we concluded that stress effects on an enzymic step involved in RuBP regeneration caused impaired chloroplast metabolism and photosynthetic inhibition in plants exposed to water deficits.

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On the Function of Mitochondrial Metabolism during Photosynthesis in Spinach (Spinacia oleracea L.) Leaves (Partitioning between Respiration and Export of Redox Equivalents and Precursors for Nitrate Assimilation Products).

The functioning of isolated spinach (Spinacia oleracea L.) leaf mitochondria has been studied in the presence of metabolite concentrations similar to those that occur in the cytosol in vivo. From measurements of the concentration dependence of the oxidation of the main substrates, glycine and malate, we have concluded that the state 3 oxidation rate of these substrates in vivo is less than half of the maximal rates due to substrate limitation. Analogously, we conclude that under steady-state conditions of photosynthesis, the oxidation of cytosolic NADH by the mitochondria does not contribute to mitochondrial respiration. Measurements of mitochondrial respiration with glycine and malate as substrates and in the presence of a defined malate:oxaloacetate ratio indicated that about 25% of the NADH formed in vivo during the oxidation of these metabolites inside the mitochondria is oxidized by a malate-oxaloacetate shuttle to serve extramitochondrial processes, e.g. reduction of nitrate in the cytosol or of hydroxypyruvate in the peroxisomes. The analysis of the products of the oxidation of malate indicates that in the steady state of photosynthesis the activity of the tricarboxylic acid cycle is very low. Therefore, we have concluded that the mitochondrial oxidation of malate in illuminated leaves produces mainly citrate, which is converted via cytosolic aconitase and NADP-isocitrate dehydrogenase to yield 2-oxoglutarate as the precursor for the formation of glutamate and glutamine, which are the main products of photosynthetic nitrate assimilation.

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The Unsaturation of Membrane Lipids Stabilizes Photosynthesis against Heat Stress.

The effect of the unsaturation of glycerolipids of thylakoid membranes on the heat tolerance of the photosynthetic evolution of oxygen was studied in vivo by mutation and transformation of fatty-acid desaturases in the cyanobacterium Synechocystis PCC6803. The experimental results indicate that elimination of dienoic lipid molecules decreases, to a small but distinct extent, the heat tolerance of photosynthetic oxygen evolution, but that elimination of trienoic lipid molecules has no effect on the heat tolerance. This conclusion contrasts with the previous hypothesis that the heat tolerance of photosynthesis is enhanced upon an increase in the level of saturation of membrane lipids. It is also shown that light does not affect the nature of the effect of lipid unsaturation on the heat tolerance of photosynthesis.

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Nitrogen Source Regulation of Growth and Photosynthesis in Beta vulgaris L.

Sugar beets (Beta vulgaris L. cv F58-554H1) were grown hydroponically in a 16-h light, 8-h dark period at a photosynthetic photon flux density of 0.5 mmol m-2 s-1 for 4 weeks in half-Hoagland culture solution containing only nitrate-nitrogen. Half of the plants were then transferred to half-Hoagland solution with ammonium-nitrogen (7.35mM), while the other half continued on 7.5 mM nitrate. Growth analysis was carried out by sampling the plants at 3-d intervals over a period of 21 d. Compared to plants supplied with nitrate, ammonium initially slowed the growth of shoots more than roots. Ammonium reduced both the area expansion of individual leaves and the relative water content of these leaves, but increased the amount of dry matter/area. The increase in specific leaf weight in ammonium-grown leaves was associated with a doubling of chloroplast volume, as much as a 62% rise in chlorophyll content, and a 4.3-fold higher accumulation of soluble protein. Ammonium nutrition substantially decreased the rate of expansion of photosynthetic (leaf) surface but did not decrease the rate of photosynthesis per area; in fact, net photosynthetic CO2 exchange rates were slightly higher than in nitrate plants, due to the build-up in stromal enzymes of the Calvin cycle, several of which increased in total extractable activity on a leaf area basis, e.g. ribulose-1,5- biphosphate carboxylase oxygenase, sedoheptulose-1,7-biphosphatase. Nitrogen source had no effect on stomatal conductance. Rates of photosynthesis per chlorophyll were decreased slightly in ammonium-grown leaves, possibly due to an increased CO2-diffusion resistance associated with the enlarged chloroplasts.

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Carbon Partitioning in Eelgrass (Regulation by Photosynthesis and the Response to Daily Light-Dark Cycles).

Diel variations in rates of C export, sucrose-phosphate synthase (SPS) and sucrose synthase (SS) activity, and C reserves were investigated in Zostera marina L. (eelgrass) to elucidate the environmental regulation of sucrose formation and partitioning in this ecologically important species. Rates of C flux and SPS activity increased with leaf age, consistent with the ontogenic transition from sink to source status. Rates of C export and photosynthesis were low but quantitatively consistent with those of many terrestrial plant species. The Vmax activity of SPS approached that of maize, but substrate-limited rates were 20 to 25% of Vmax, indicating a large pool of inactive SPS. SPS was unresponsive to the day/night transition or to a 3-fold increase in photosynthesis generated by high [CO2] and showed little sensitivity to inorganic phosphate. Consequently, regulation of eelgrass SPS appeared similar to starch- rather than to sugar-accumulating species even though eelgrass accumulates sucrose. Leaf [sucrose] was constant and high throughout the diel cycle, which may contribute to the down-regulation of SPS. Root sucrose synthase activity was high but showed no response to nocturnal anoxia. Root [sucrose] also showed no diel cycle. The temporal stability of [sucrose] confers an ability for eelgrass to buffer the effects of prolonged light limitation that may be key to its survival and ecological success in environments subject to periods of extreme light limitation and chaotic daily variation in light availability.

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Abolition of Posttranscriptional Regulation of Nitrate Reductase Partially Prevents the Decrease in Leaf NO3- Reduction when Photosynthesis Is Inhibited by CO2 Deprivation, but Not in Darkness.

The activity of nitrate reductase (NR) in leaves is regulated by light and photosynthesis at transcriptional and posttranscriptional levels. To understand the physiological role of these controls, we have investigated the effects of light and CO2 on in vivo NO3- reduction in transgenic plants of Nicotiana plumbaginifolia lacking either transcriptional regulation alone or transcriptional and posttranscriptional regulation of NR. The abolition of both levels of NR regulation did not modify the light/dark changes in exogenous 15NO3- reduction in either intact plants or detached leaves. The same result was obtained for 15N incorporation into free amino acids in leaves after 15NO3- was supplied to the roots, and for reduction of endogenous NO3- after transfer of the plants to an N-deprived solution. In the light, however, deregulation of NR at the posttranscriptional level partially prevented the inhibition of leaf 15NO3- reduction resulting from the removal of CO2 from the atmosphere We concluded from these observations that in our conditions deregulation of NR in the transformants investigated had little impact on the adverse effect of darkness on leaf NO3- reduction, and that posttranscriptional regulation of NR is one of the mechanisms responsible for the short-term coupling between photosynthesis and leaf NO3- reduction in the light.

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