Utilization of Photosynthetically Fixed 14CO2 into Alkaloids in Relation to Primary Metabolites in Developing Leaves of Catharanthus roseus

Partitioning of current photosynthates towards primary metabolites and its simultaneous incorporation in leaf alkaloids was investigated in developing leaves of medicinally important Catharanthus roseus. Of the total ¹⁴CO₂ assimilated, the leaves at positions 1–6 fixed 8, 22, 25, 19, 13, and 8 %, respectively, and stem 3 %. Leaf fresh mass, chlorophyll content, and CO₂ exchange rate increased up to the third leaf. The total alkaloid content was highest in young actively growing leaves, which declined with age. Total ¹⁴C fixed and its content in ethanol soluble fraction increased up to the third leaf and then declined. The ¹⁴C content in primary metabolites such as sugars and organic acids was also highest in the 3^rd leaf. The utilization of ¹⁴C assimilates into alkaloids was maximum in youngest leaf which declined with leaf age. Hence the capacity to synthesize alkaloids was highest in young growing leaves and metabolites from photosynthetic pathway were most efficiently utilized and incorporated into alkaloid biosynthetic pathway by young growing leaves.     

Influence of Benzyladenine, Leaf Darkening, and Ringing on Movement of C-labeled Assimilates Into Expanded Leaves of Vitis vinifera L

Leaves of Vitis vinifera L., nearly fully expanded, imported only trace amounts of ¹⁴C following assimilation of ¹⁴CO₂ by a lower leaf on the same shoot, but benzyladenine (BA) application at 4.4 × 10⁻³m caused a marked increase in the movement of ¹⁴C into these leaves. Older leaves near the shoot base were less responsive; BA treatment alone had little effect on import of labeled assimilates from adjacent leaves but when the BA-treated leaves were darkened there was an increased import of labeled materials. When these 2 treatments were combined and applied to leaves on shoots with ringed bases, relatively high levels of radioactivity were detected in the BA-treated leaves but under these conditions darkening, without the application of BA, also resulted in an increased import of ¹⁴C. Accumulation of imported ¹⁴C was found to be restricted to the area of the leaf blade treated with BA. Separation of labeled compounds in ethanol extracts of treated leaves showed a lower percentage of radioactivity present in the sugar fraction from BA-treated leaves and an increased percentage present in the amino acid fraction.     

14C-Studies on Apple Trees. II. Distribution of Photosynthates from Top and Base Leaves from Extension Shoots

The photosynthates from leaves from extension shoots may be used either for new growth of the shoot itself, or for growth in other parts of the tree. An attempt has been made to elucidate this problem by determining the content of ¹⁴C in long shoots to which ¹⁴CO₂ was applied either through fully developed leaves at the base, or through very young leaves at the apex. In the case of ¹⁴C application at the base, 80 per cent or more of the ¹⁴C initially taken up disappears from the shoot, and only a very minor part is translocated upwards in the shoot. When the leaves at the apex are exposed, on the other hand, 80 per cent of the ¹⁴C absorbed is retained in the leaves and shoot components treated as long as there is still considerable terminal growth taking place, although a small percentage is deposited in the lower parts of the shoot. At the same time, a much higher proportion is incorporated into methanol-insoluble components. As terminal growth decreases, a larger proportion of the ¹⁴C activity of the apex leaves also disappears from the shoot. The distribution of activity between the sorbitol, sucrose, glucose and fructose fractions was not significantly different in young and in fully developed leaves. The ¹⁴C labelling in the sugar fraction was highest for sorbitol, then sucrose, but decreases with time compared to glucose and fructose.     

Decomposition of leaf and root tissue of three perennial grass species grown at two levels of atmospheric CO2 and N supply

Leaf and root tissue of Lolium perenne L., Agrostis capillaris L. and Festuca ovina L. grown under ambient (350 μl l^-1 CO₂) and elevated (700 μl l^-1) CO₂ in a continuously ¹⁴C-labelled atmosphere and at two soil N levels, were incubated at 14°C for 222 days. Decomposition of leaf and root tissue grown in the low N treatment was not affected by elevated [CO₂], whereas decomposition in the high N treatment was significantly reduced by 7% after 222 days. Despite the increased C/N ratio (g g^-1) of tissue cultivated at elevated [CO₂] when compared with the corresponding ambient tissue, there was no significant correlation between initial C/N ratio and ¹⁴C respired. This finding suggests that the CO₂-induced changes in decomposition rates do not occur via CO₂-induced changes in C/N ratios of plant materials. We combined the decomposition data with data on ¹⁴C uptake and allocation for the same plants, and give evidence that elevated [CO₂] has the potential to increase soil C stores in grassland via increasing C uptake and shifting C allocation towards the roots, with an inherent slower decomposition rate than the leaves. An overall increase of 15% in ¹⁴C remaining after 222 days was estimated for the combined tissues, i.e., the whole plants; the leaves made a much smaller contribution to the C remaining (+6%) than the roots (+26%). This shows the importance of clarifying the contribution of roots and leaves with respect to the question whether grassland soils act as a sink or source for atmospheric CO₂. This revised version was published online in June 2006 with corrections to the Cover Date.     

Reversal of the Direction of Photosynthate Allocation in Urtica dioica L. Plants by Increasing Cytokinin Import into the Shoot

The natural cytokinin import from the root into the shoot of Urtica dioica plants was enhanced by supplying zeatin riboside (ZR) solutions of various concentrations to a portion less than 10 % of the root system after removal of their tips. After 6 h ZR pretreatment of the plants, ¹⁴CO₂ was supplied for 3 h to a mature (source) leaf or to an expanding leaf and the ¹⁴C-distribution in the whole plant was determined after a subsequent dark period of 14 h. ZR substantially increased ¹⁴C fixation by the expanding leaves and also enhanced export of carbon and transport to the shoot apex. The effect of the hormone treatment was, however, more pronounced when the ¹⁴CO₂ was supplied to a mature leaf. In the control plants these leaves exported carbon only to the roots: When the amount of the natural daily ZR input from the roots to the shoot was enhanced by 20%, the bulk of the ¹⁴C exported from a mature leaf moved to the shoot apex and only a minor portion of ¹⁴C was still detected in the root fraction. A several-fold increase of the natural daily ZR input into the shoot resulted in a flow of ¹⁴C only to the growing parts of the shoot. The results suggest control of the sink strength of the shoot apex by ZR in Urtica diocia.     

Effect of nitrates supplied with the transpiration flow on assimilate translocation

Solutions of nitrates (0.5% KNO₃, 0.2% NH₄NO₃) or urea (0.15%) were fed under the pressure of 10⁴ Pa to 50–60-cm-long detached shoots of common flax (Linum usitatissimum L.). One hour after the start of supplying the solutions, an assimilation clip chamber was fastened to the middle part of the shoot (¹⁴C source area), and ¹⁴CO₂ was blown through in the light for 2.5 min. The analysis of distribution of ¹⁴C among the labeled products of photosynthesis produced by source leaves showed that nitrates reduced the incorporation of the label into sucrose. At the same time, the ratio of labeled sucrose to labeled hexoses decreased, and the incorporation of the label into serine greatly increased. Urea did not produce such effects. The pattern of distribution of ¹⁴C within the plant 3 h after the assimilation of ¹⁴CO₂ points to the suppression of assimilate efflux from the leaves of plants fed with nitrates. In plants supplied with water or urea, 17–20% of labeled carbon was found below the ¹⁴C source area of the shoot, in nitrate type of treatment, only 3–5% was found there. In plants supplied with nitrates, the cortex tissue below the source leaf contained more ¹⁴C in proteins and less in low-molecular substances. In the wood tissue, such a correlation was not observed. When the shoot was supplied with water or urea, the content of ¹⁴C in sucrose in the source leaves in 3 h declined from 55–60% to 38–42%. When the shoot was fed with nitrates, the share of label in sucrose increased from 50 to 62–73%. Autoradiography of the source leaves showed that, in plants supplied with water or urea, the label was predominantly accumulated in large vascular bundles, and in nitrate type of treatment, it was accumulated outside large bundles. Electron microscopy showed that, in nitrate plants, the companion cells of phloem endings were very much vacuolated.     

Glutamic Acid metabolism and the photorespiratory nitrogen cycle in wheat leaves: metabolic consequences of elevated ammonia concentrations and of blocking ammonia assimilation

The effects of methionine sulfoximine and ammonium chloride on [¹⁴C] glutamate metabolism in excised leaves of Triticum aestivum were investigated. Glutamine was the principal product derived from [U¹⁴C]glutamate in the light and in the absence of inhibitor or NH₄Cl. Other amino acids, organic acids, sugars, sugar phosphates, and CO₂ became slightly radioactive. Ammonium chloride (10 mm) increased formation of [¹⁴C] glutamine, aspartate, citrate, and malate but decreased incorporation into 2-oxoglutarate, alanine, and ¹⁴CO₂. Methionine sulfoximine (1 mm) suppressed glutamine synthesis, caused NH₃ to accumulate, increased metabolism of the added radioactive glutamate, decreased tissue levels of glutamate, and decreased incorporation of radioactivity into other amino acids. Methionine sulfoximine also caused most of the ¹⁴C from [U-¹⁴C]glutamate to be incorporated into malate and succinate, whereas most of the ¹⁴C from [1-¹⁴C]glutamate was metabolized to CO₂ and sugar phosphates. Thus, formation of radioactive organic acids in the presence of methionine sulfoximine does not take place indirectly through “dark” fixation of CO₂ released by degradation of glutamate when ammonia assimilation is blocked. When illuminated leaves supplied with [U-¹⁴C] glutamate without inhibitor or NH₄Cl were transferred to darkness, there was increased metabolism of the glutamate to glutamine, aspartate, succinate, malate, and ¹⁴CO₂. Darkening had little effect on the labeling pattern in leaves treated with methionine sulfoximine.     

Combined effects of elevated CO2 and soil drought on carbon and nitrogen allocation of the desert shrub Caragana intermedia

Impacts of either elevated CO₂ or drought stress on plant growth have been studied extensively, but interactive effects of these on plant carbon and nitrogen allocation is inadequately understood yet. In this study the response of the dominant desert shrub, Caragana intermedia Kuanget H.c.Fu, to the interaction of elevated CO₂ (700 ± 20 μmol mol⁻¹) and soil drought were determined in two large environmental growth chambers (18 m²). Elevated CO₂ increased the allocation of biomass and carbon into roots and the ratio of carbon to nitrogen (C:N) as well as the leaf soluble sugar content, but decreased the allocation of biomass and carbon into leaves, leaf nitrogen and leaf soluble protein concentrations. Elevated CO₂ significantly decreased the partitioning of nitrogen into leaves, but increased that into roots, especially under soil drought. Elevated CO₂ significantly decreased the carbon isotope discrimination (Δ) in leaves, but increased them in roots, and the ratio of Δ values between root and leaf, indicating an increased allocation into below-ground parts. It is concluded that stimulation of plant growth by CO₂ enrichment may be negated under soil drought, and under the future environment, elevated CO₂ may partially offset the negative effects of enhanced drought by regulating the partitioning of carbon and nitrogen.     

Populus tremuloides photosynthesis and crown architecture in response to elevated CO2 and soil N availability

We tested the hypothesis that elevated CO₂ would stimulate proportionally higher photosynthesis in the lower crown of Populus trees due to less N retranslocation, compared to tree crowns in ambient CO₂. Such a response could increase belowground C allocation, particularly in trees with an indeterminate growth pattern such as Populus tremuloides. Rooted cuttings of P. tremuloides were grown in ambient and twice ambient (elevated) CO₂ and in low and high soil N availability (89 ± 7 and 333 ± 16 ng N g⁻¹ day⁻¹ net mineralization, respectively) for 95 days using open-top chambers and open-bottom root boxes. Elevated CO₂ resulted in significantly higher maximum leaf photosynthesis (A _max) at both soil N levels. A _max was higher at high N than at low N soil in elevated, but not ambient CO₂. Photosynthetic N use efficiency was higher at elevated than ambient CO₂ in both soil types. Elevated CO₂ resulted in proportionally higher whole leaf A in the lower three-quarters to one-half of the crown for both soil types. At elevated CO₂ and high N availability, lower crown leaves had significantly lower ratios of carboxylation capacity to electron transport capacity (V _cmax/J _max) than at ambient CO₂ and/or low N availability. From the top to the bottom of the tree crowns, V _cmax/J _max increased in ambient CO₂, but it decreased in elevated CO₂ indicating a greater relative investment of N into light harvesting for the lower crown. Only the mid-crown leaves at both N levels exhibited photosynthetic down regulation to elevated CO₂. Stem biomass segments (consisting of three nodes and internodes) were compared to the total A _leaf for each segment. This analysis indicated that increased A _leaf at elevated CO₂ did not result in a proportional increase in local stem segment mass, suggesting that C allocation to sinks other than the local stem segment increased disproportionally. Since C allocated to roots in young Populus trees is primarily assimilated by leaves in the lower crown, the results of this study suggest a mechanism by which C allocation to roots in young trees may increase in elevated CO₂. Received: 12 August 1996 / Accepted: 12 November 1996     

Metabolic Pathway of alpha-Ketoglutarate in Citrus Leaves as Affected by Phosphorus Nutrition

The uptake and metabolism of α-[5-¹⁴C]ketoglutarate by phosphorus-deficient and full nutrient (control) lemon (Citrus limon) leaves were studied over various time intervals. After 45 minutes in P-deficient leaves, the bulk of incorporated ¹⁴C appeared in organic acids and much less in amino acids, while in the control leaves, the ¹⁴C contents of organic and amino acids were equal. In P-deficient leaves, after longer incubation times the ¹⁴C content of organic acids and amino acids increased, while that of CO₂ and residue fractions remained low. In full nutrient leaves the ¹⁴C content of amino acids and organic acids decreased after longer incubation time and increased in the insoluble residue and CO₂. In full nutrient leaves the organic and amino acid metabolism were closely related and accompanied by protein synthesis and CO₂ release, while in P-deficient leaves an accelerating accumulation of arginine and citric acid was linked together with inhibition of protein synthesis and CO₂ liberation.     

The effect of lead on photosynthesis and respiration in detached leaves and in mesophyll protoplasts of Pisum sativum

Photosynthesis and transpiration rate of detached leaves of pea (Pisum sativum L. cv. Iłowiecki) exposed to solution of Pb(NO₃)₂ at 1 or 5 mmol·dm⁻³ concentrations were inhibited. The higher concentration of this toxicant decreased photosynthesis and transpiration rates 2 and 3 times respectively, and increased respiration by about 20 %, as measured after 24 hours of treatment. Similarly to Pb(NO₃)₂, glyceraldehyde solution, an inhibitor of phosphoribulokinase, at 50 mmol·dm⁻³ concentration decreased the rates of photosynthesis and transpiration during introduction into pea leaves. The rate of dark respiration, however, remained unchanged during 2 hours of experiment. The potential photochemical efficiency of PS II (Fv/Fm) and the activity of Rubisco (EC 4.1.1.39) at 5 mmol·dm⁻³ of Pb(NO₃)₂ were lowered by 10 % and 20 % respectively, after 24 hours. Neither changes in the activity of PEPC (EC 4.1.1.31) or protein and pigment contents were noted in Pb-treated leaves. The photosynthetic activity of protoplasts isolated from leaves treated for 24 or 48 hours with Pb(NO₃)₂ at 5 mmol·dm⁻³ concentration was decreased 10 % or 25 %, whereas, the rate of dark respiration was stimulated by about 40 % and 75 %, respectively. The content of abscisic acid, a hormone responsible for stomatal closure, in detached pea leaves treated for 24 h with 5 mmol·dm⁻³ of Pb(NO₃)₂ solution was increased by about 3 times; a longer (48h) treatment led to further increase (by about 7 times) in the amount of this hormone. The results of our experiments provide evidences that CO₂ fixation in detached pea leaves, at least up to 24 hours of Pb(NO₃)₂ treatment, was restricted mainly by stomatal closure.     

The effects of elevated atmospheric [CO<Subscript>2</Subscript>] on Norway spruce needle parameters

Studies of selected morphological needle parameters were carried out on young (17–19 year old) Norway spruce trees cultivated inside glass domes at ambient (A, 370 μmol (CO₂) mol⁻¹) and elevated (E, 700 μmol (CO₂) mol⁻¹) atmospheric CO₂ concentrations [CO₂] beginning in 1997. Annual analyses performed from 2002 to 2004 revealed higher values for needle length (especially for current needles, up to 18%) and projected needle area (up to 13%) accompanied by lower values for specific needle area (up to 15% lower, as quantified by needle mass to projected area ratio) in the E treatment compared to the A treatment. Statistically significant differences for most of the investigated morphological parameters were found in young needles in the well irradiated sun-adapted crown parts, particularly under water-limiting soil conditions in 2003. This was likely a result of different water relations in E compared to A trees as investigated under temperate water stress (Kuper et al. in Biol Plantarum 50:603–609, 2006). Furthermore, E trees had much higher absorbing root area, which modified and enhanced root:shoot as well as root:conductive stem area proportions. These hydraulic properties and early seasonal stimulation of photosynthesis forced advanced needle development in E trees, particularly under limited soil water conditions. The number of needles per unit shoot length was found to be unaffected by elevated [CO₂].     

Postphotosynthetic Utilization of Labeled Assimilates in Fiber Flax

The distribution of ¹⁴C in various tissues of fiber flax was assayed 1, 17, and 21 days after 30-min assimilation of ¹⁴CO₂ by the whole rapidly growing plant. Polymeric photosynthetic products were largely hydrolyzed in the ¹⁴C-donor part of the shoot, and the hydrolysates were transported upward. The content of ¹⁴C in pigments and lipids of the donor leaves (that absorbed ¹⁴CO₂) was significantly higher than that in the ¹⁴C-acceptor leaves. Additional nitrogen nutrition decreased the labeled sucrose-to-hexose ratio and inhibited transport of the assimilates from both ¹⁴C-donor and acceptor leaves. ¹⁴C transported to the shoot tip was largely used for the synthesis of poorly soluble proteins (extractable with alkali and Triton X-100) in the acceptor tissues. In the donor part of the shoot, particularly in the bast, cellulose was mainly synthesized from the new assimilates.     

Xeromorphy increases in shoots of Pseudotsuga menziesii (Mirb.) Franco seedlings with exposure to elevated temperature but not elevated CO2

Seedling structure influences tree structure and function, ultimately determining the potential productivity of trees and their competitiveness for resources. We investigated changes in shoot structure for seedlings of Pseudotsuga menziesii (Douglas-fir) grown under climate change scenarios of ambient or elevated CO₂ (+180 mol mol^–1) plus ambient or elevated temperature (+3.5°C), for 4 years in outdoor, sunlit chambers. Mass allocation and allometry were measured for buds, leaves, branches, and stems, and anatomy was evaluated for leaves and stems. Seedlings became more xeromorphic with elevated temperature: allocation of total mass to branches over stems and leaves increased, sapwood area to height ratio increased, number of growing points relative to seedling size increased, and stem and branch length and mass decreased for sections initiated during the three full CO₂ and temperature seasons. Neither stem nor leaf anatomy was affected by elevated temperature. Elevated CO₂ increased specific mass of leaves, but had few other effects on mass allocation, allometry, or anatomy for any shoot organ. There were no CO₂ × temperature interactions for any important parameter. Thus, under realistic simulated field environmental conditions representative for in at least some P. menziesii forests (i.e., OR, USA, forests with limited soil nitrogen and summer soil moisture), elevated temperature, but not elevated CO₂, may affect seedling shoot structure and, hence, function.     

Etude du vieillissement d’Anagallis arvensis L.: Action régulatrice de la lumière,du saccharose et de l’acide indolyl-acétique sur la capacité rhizogènique des feuilles et des rameaux axillaires

The rooting capacity of leaves isolated from a vegetative clone ofAnagallis arvensis L. exposed to 9 hours of light (75 W m⁻²) at 22 °C and 15 hours of dark at 12 °C a day is significant only in F₁ young leaves and not in adult ones. The rooting capacity of the young leaves and of the vegetative shoots is greater in longer photoperiods. The leaves make roots even under weak (14 W m⁻²) irradiance. The rooting capacity of the leaves is diminished and even suppressed by exogenous sucrose (14,60 ×10⁻³M). This inhibition may be counteracted by IAA (10⁻⁶M). When leaves and shoots are taken from clones under long (16 h) photoperiods, or in constant irradiance, they progressively lose their rooting capacity during the treatment. Rooting capaoity is regained if the clones are returned again to “short day” (9 h) condition.      

Nutrition and the ozone sensitivity of birch (Betula pendula)

 Cuttings of a single birch clone (Betula pendula) were grown in field fumigation chambers throughout the growing season in either filtered air (control) or 90/40 nl O₃ l^–1 (day/night). Both regimes were split into plants under high and low nutrient supply (macro- and micronutrients). The stomatal density of leaves was increased by ozone but was lowered at high nutrition, while the inner air space was hardly affected by the treatments. Ozone induced macroscopic leaf injury regardless of nutrition, but leaf shedding was delayed in the low-fertilized plants, despite O₃ uptake being similar to that in high-fertilized plants. The leaf turn-over was enhanced in the O₃-exposed high-fertilized plants, but length growth and leaf formation of stems were not affected by ozone in either nutrient regime. Leaves of high-fertilized plants showed O₃-caused decline in photosynthetic capacity, water-use efficiency, apparent carbon uptake efficiency and quantum yield earlier as compared with low-fertilized plants, whereas chlorophyll fluorescence (F_V/F_M) and leaf nitrogen concentration were rather stable. CO₂ uptake rate and rubisco activity of young leaves compensated for the O₃ injury in the ageing leaves of the low-fertilized plants. In 8-week-old leaves, however, the O₃-induced decline in CO₂ uptake did not differ between the nutrient regimes and was associated with increased dark respiration rather than changed photorespiration. The balance between CO₂ supply and demand was lost, as was stomatal limitation on CO₂ uptake. High nutrition did not help leaves to maintain a high photosynthetic capacity and life span under O₃ stress. Received: 6 July 1996 / Accepted: 4 June 1997     

Pathways of carbohydrate oxidation in leaves of Pisum sativum and Triticum aestivum

The aim of this work was to establish the pathways of carbohydrate oxidation used in the dark by leaves of Pisum sativum and Triticum aestivum. Segments of young and mature leaves of pea released the carbons of glucose-[¹⁴C] as ¹⁴CO₂ in the order 3,4 > 1 > 2 > 6 whereas in segments of young and mature leaves of wheat the order was 3,4 > 1 > 6 > 2. The detailed labelling of the constituents of mature leaves of wheat by glucose-[1-¹⁴C], -[2-¹⁴C], -[3,4-¹⁴C], and -[6-¹⁴C] was determined and showed that the high yield of CO₂ from C-6 relative to that from C-2 was due to release of C-6 during pentan synthesis. Estimates were made of the maximum catalytic activities of phosphofructokinase and glucose-6-phosphate dehydrogenase in pea and wheat leaves of three ages. The results of all the above investigations strongly indicate that both pea and wheat leaves in the dark oxidize carbohydrate via glycolysis and the pentose phosphate pathway with the latter accounting for no more than a third of the total. No evidence was obtained of any major change in the relative activities of the two pathways during the development of either type of leaf.     

Comparison of leaf water use efficiency of oak and sycamore in the canopy over two growing seasons

The seasonal trends in water use efficiency of sun and shade leaves of mature oak (Quercus robur) and sycamore (Acer pseudoplatanus) trees were assessed in the upper canopy of an English woodland. Intrinsic water use efficiency (net CO₂ assimilation rate/leaf conductance, A/g) was measured by gas exchange and inferred from C isotope discrimination (δ¹³C) methods. Shade leaves had consistently lower δ¹³C than sun leaves (by 1–2‰), the difference being larger in sycamore. Buds had distinct sun and shade isotopic signatures before bud break and received an influx of ¹³C-rich C before becoming net autotrophs. After leaf full expansion, δ¹³C declined by 1–2‰ gradually through the season, emphasising the importance of imported carbon in the interpretation of leaf δ¹³C values in perennial species. There was no significant difference between the two species in the value of intrinsic water use efficiency for either sun or shade leaves. For sun leaves, season-long A/g calculated from δ¹³C (72–78 μmol CO₂ [mol H₂O]⁻¹) was 10–16% higher than that obtained from gas exchange and in situ estimates of leaf boundary layer conductance. For shade leaves, the gas exchange–derived values were low, only 10–18% of the δ¹³C-derived values. This is ascribed to difficulties in obtaining a comprehensive sample of gas exchange measurements in the rapidly changing light environment.     

Insights of carbon assimilation and allocation in young cork oak (Quercus suber L.) plants using Carbon-14

¹⁴C methods were applied to young, woody, branched and well-watered cork oak (Quercus suber L.) plants to determine carbon assimilation and its distribution among plant organs. Carbon assimilation rates by attached leaves clamped in a foliar ¹⁴CO₂ assimilation chamber containing 3.7 × 10⁴ Bq of a portable ventilated diffusion porometer were measured at different ¹⁴CO₂ pulse-labeling periods (15, 30, 45, 60 and 120 s) in summer. Allocation of recently fixed C by attached leaves within plants was evaluated 7 days after a 60-min of 5.6 MBq of ¹⁴CO₂ pulse-labeling in late winter. ¹⁴CO₂ pulse-labeling was separately induced on leaves of a lower branch, two opposite branches at the same lower level, a middle branch and a top branch. ¹⁴C activity incorporated into the plants was measured by liquid scintillation and autoradiography. Our results show the optimum ¹⁴CO₂ pulse-labeling period is between 15 and 30 s, which corresponds to 9.81 ± 0.15 and 9.16 ± 0.12 µmol m⁻² s⁻¹ C assimilation rates in summer, respectively. The investment of current assimilates ranged from 18 to 29% in leaves, 1 to 7% in lateral branches, 0 to 3% in the stem and over 65% in roots, in late winter. Roots displayed the greatest sink strength for the total ¹⁴C recovered by whole-plants. These results were expected because the trial was done in winter, when cork oak does not produce their leaves. Our results highlight the contribution of current assimilates for growth and maintenance of roots, in young woody plants under Mediterranean climate.     

Effects of the interaction between ozone and carbon dioxide on gas exchange,photosystem II and antioxidants in rice leaves

To understand the interactive effects of O₃ and CO₂ on rice leaves; gas exchange, chlorophyll (Chl) fluorescence, ascorbic acid and glutathione were examined under acute (5 h), combined exposures of O₃ (0, 0.1, or 0.3 cm³ m⁻³, expressed as O₀, O_0.1, or O_0.3, respectively), and CO₂ (400 or 800 cm³ m⁻³, expressed as C⁴⁰⁰ or C⁸⁰⁰, respectively) in natural-light gas-exposure chambers. The net photosynthetic rate (P _N), maximum (F_v/F_m) and operating (F_q′/F_m′) quantum efficiencies of photosystem II (PSII) in young (8^th) leaves decreased during O₃ exposure. However, these were ameliorated by C⁸⁰⁰ and fully recovered within 3 d in clean air (O⁰ + C⁴⁰⁰) except for the O^0.3 + C⁴⁰⁰ plants. The maximum PSII efficiency at 1,500 μmol m⁻² s⁻¹ PPFD (F_v′/F_m′) for the O^0.3 + C₄₀₀ plants decreased for all measurement times, likely because leaves with severely inhibited P _N also had a severely damaged PSII. The P _N of the flag (16^th) leaves at heading decreased under O₃ exposure, but the decline was smaller and the recovery was faster than that of the 8^th leaves. The F_q′/F_m′ of the flag leaves in the O^0.3 + C⁴⁰⁰ and O^0.3 + C⁸⁰⁰ plants decreased just after gas exposure, but the F_v/F_m was not affected. These effects indicate that elevated CO₂ interactively ameliorated the inhibition of photosynthesis induced by O₃ exposure. However, changes in antioxidant levels did not explain the above interaction.