A CO2 Concentrating System in Leaves of Higher C3-Plants Predicted by a Model Based on RuBP Carboxylase/Oxygenase Kinetics and 14CO2/12CO2 Exchange

Mächler, F., Lehnherr, B., Schnyder, H. and Nösberger,J. 1985. A CO₂ concentrating system in leaves of higher C₃-plantspredicted by a model based on RuBP carboxylase/oxygenase kineticsand ¹⁴CO₂/¹²CO₂ exchange.–J. exp. Bot. 36: 1542–1550. A model is presented which compares the ratio of the two activitiesof the enzyme nbulose bisphosphate carboxylase/oxygenase asdetermined in vitro with the ratio of photosynthesis to photorespirationin leaves as determined from differential ¹⁴CO₂/¹²CO₂ uptakeor from CO₂ compensation concentration. Discrepancies betweenmeasurements made in vitro and in vivo are attributed to theeffect of a CO₂ concentrating system in the leaf cells. Interferencefrom dark respiration is discussed. A CO₂ concentrating systemis postulated which is efficient mainly at low temperature andlow CO₂ concentration. Key words: —Photosynthesis, photorespiration, ribulose bisphosphate carboxylase/oxygenase     

Fructan contents and dry matter deposition in different tissues of the wheat grain during development

The role of fructan metabolism in the assimilate relations of the grain of wheat (Triticum aestivum L.) was investigated by determination of the dry matter and fructan content of grain components at short intervals during grain filling. During the initial phase of rapid expansion, most of the assimilates entering the grain were partitioned to the outer pericarp. A large fraction of these assimilates were used for the synthesis of fructan. Dry matter deposition and fructan synthesis in the outer pericarp ceased at about 5d after anthesis. At the same time, the endosperm and the inner pericarp and testa started to accumulate dry matter at a fast rate. This was also associated with significant fructan synthesis in the latter tissues. The outer pericarp lost about 45% of its former maximum dry weight between 9 and 19 d after anthesis. This loss was due almost entirely to the near complete disappearance of water-soluble carbohydrates, most of which was fructan. The inner pericarp and testa accumulated dry matter until about mid-grain filling. The fructan contents of the inner pericarp and testa and the endosperm decreased slowly towards the end of grain filling. Most of the fructans in the inner pericarp and testa and the endosperm had a low molecular weight, whereas higher molecular weight fructans predominated in the outer pericarp. The embryo did not contain fructan. The presence of low molecular weight fructans in the endosperm cavity at mid-grain filling was confirmed. It is suggested that fructan synthesis is closely linked to growth-related water deposition in the different tissues of the wheat grain and serves to sequester the surplus of imported sucrose.     

On the 13C/12C isotopic signal of day and night respiration at the mesocosm level

While there is currently intense effort to examine the ¹³C signal of CO₂ evolved in the dark, less is known on the isotope composition of day‐respired CO₂. This lack of knowledge stems from technical difficulties to measure the pure respiratory isotopic signal: day respiration is mixed up with photorespiration, and there is no obvious way to separate photosynthetic fractionation (pure c_i/c_a effect) from respiratory effect (production of CO₂ with a different δ¹³C value from that of net‐fixed CO₂) at the ecosystem level. Here, we took advantage of new simple equations, and applied them to sunflower canopies grown under low and high [CO₂]. We show that whole mesocosm‐respired CO₂ is slightly ¹³C depleted in the light at the mesocosm level (by 0.2–0.8‰), while it is slightly ¹³C enriched in darkness (by 1.5–3.2‰). The turnover of the respiratory carbon pool after labelling appears similar in the light and in the dark, and accordingly, a hierarchical clustering analysis shows a close correlation between the ¹³C abundance in day‐ and night‐evolved CO₂. We conclude that the carbon source for respiration is similar in the dark and in the light, but the metabolic pathways associated with CO₂ production may change, thereby explaining the different ¹²C/¹³C respiratory fractionations in the light and in the dark.     

Hair of grazing cattle provides an integrated measure of the effects of site conditions and interannual weather variability on δ13C of temperate humid grassland

The carbon isotope composition (δ¹³C) of C3 ecosystems is sensitive to water availability, and provides important information for the assessment of terrestrial carbon (C) sink/source activity. Here, we report the effects of plant available soil water (PAW) on community ¹³C signatures of temperate humid grassland. The 5‐year study was conducted on pastures exhibiting a large range of PAW capacity that were located on two site types: peat and mineral soils. The data set included the centennial drought year 2003, and data from wet years (2000 and 2002). Seasonal variation of PAW was modeled using PAW capacity of each pasture, precipitation inputs and evapotranspiration estimates. Community ¹³C signatures were derived from the δ¹³C of vegetation and segments of tail switch hair of cattle grown while grazing pastures. Hair ¹³C signatures provided an assimilation‐weighted ¹³C signal that integrated both spatial (paddock‐scale) and temporal (grazing season) variation of ¹³C signatures on a pasture. The δ¹³C of hair and vegetation increased with decreasing modeled PAW in the same way on mineral and peat soils. But, at a given PAW, the δ¹³C of hair was 2.6‰ less negative than that of vegetation, reflecting the diet‐hair isotopic shift. Furthermore, the δ¹³C of hair and vegetation on peat soil pastures was 0.5‰ more negative than on pastures situated on mineral soil. This may have resulted from a ～10 ppm CO₂ enrichment of canopy air derived from ongoing peat mineralization. Community‐scale season‐mean ¹³C discrimination (Δ) exhibited a saturation‐type response towards season‐mean modeled PAW (r²=0.78), and ranged between 19.8‰ on soils with low PAW capacity during the drought year of 2003, and 21.4‰ on soils with high PAW capacity in a wet year. This indicated relatively small variation in season‐mean assimilation‐weighted p_i/p_a (0.68–0.75) between contrasting sites and years. However, this range is similar to that reported in other studies, which encompass the range from subtropical arid to humid temperate grassland. Furthermore, the tight relationship between season‐mean Δ and modeled mean PAW suggests that PAW may be used as proxy for Δ.     

C-isotope composition of CO2 respired by shoots and roots: fractionation during dark respiration?

The CO₂ respired by leaves is ¹³C-enriched relative to leaf biomass and putative respiratory substrates (Ghashghaie et al., Phytochemistry Reviews 2, 145–161, 2003), but how this relates to the ¹³C content of root, or whole plant respiratory CO₂ is unknown. The C isotope composition of respiratory CO₂ (δ_R) from shoots and roots of sunflower (Helianthus annuus L.), alfalfa (Medicago sativa L.), and perennial ryegrass (Lolium perenne L.) growing in a range of conditions was analysed. In all instances plants were grown in controlled environments with CO₂ of constant concentration and δ¹³C. Respiration of roots and shoots of individual plants was measured with an open CO₂ exchange system interfaced with a mass spectrometer. Respiratory CO₂ from shoots was always ¹³C-enriched relative to that of roots. Conversely, shoot biomass was always ¹³C-depleted relative to root biomass. The δ-difference between shoot and root respiratory CO₂ was variable, and negatively correlated with the δ-difference between shoot and root biomass (r² = 0.52, P = 0.023), suggesting isotope effects during biosynthesis. ¹³C discrimination in respiration (R) of shoots, roots and whole plants (e_Shoot, e_Root, e_Plant) was assessed as e = (δ_Substrate − δ_R)/(1 + δ_R/1000), where root and shoot substrate is defined as imported C, and plant substrate is total photosynthate. Estimates were obtained from C isotope balances of shoots, roots and whole plants of sunflower and alfalfa using growth and respiration data collected at intervals of 1 to 2 weeks. e_plant and e_Shoot differed significantly from zero. e_plant ranged between −0.4 and −0.9‰, whereas e_Shoot was much greater (−0.6 to −1.9‰). e_Root was not significantly different from zero. The present results help to resolve the apparent conflict between leaf- and ecosystem-level ¹³C discrimination in respiration.     

Last‐century changes of alpine grassland water‐use efficiency: a reconstruction through carbon isotope analysis of a time‐series of Capra ibex horns

The ecophysiological response of an alpine grassland to recent climate change and increasing atmospheric CO₂ concentration was investigated with a new strategy to go back in time: using a time‐series of Capra ibex horns as archives of the alpine grasslands' carbon isotope discrimination (¹³Δ). From the collection of the Natural History Museum of Bern, horns of 24 males from the population of the Augstmatthorn–Brienzer Rothorn mountains, Switzerland, were sampled covering the period from 1938 to 2006. Samples were taken from the beginning of each year‐ring of the horns, representing the beginning of the horn growth period, the spring. The horns' carbon ¹³C content (Δ¹³C) declined together with that of atmospheric CO₂ over the 69‐year period, but ¹³Δ increased slightly (+0.4‰), though significantly (P<0.05), over the observation period. Estimated intercellular CO₂ concentration increased (+56 μmol mol^?1) less than the atmospheric CO₂ concentration (+81 μmol mol^?1), so that intrinsic water‐use efficiency increased by 17.8% during the 69‐year period. However, the atmospheric evaporative demand at the site increased by approximately 0.1 kPa between 1955 and 2006, thus counteracting the improvement of intrinsic water‐use efficiency. As a result, instantaneous water‐use efficiency did not change. The observed changes in intrinsic water‐use efficiency were in the same range as those of trees (as reported by others), indicating that leaf‐level control of water‐use efficiency of grassland and forests followed the same principles. This is the first reconstruction of the water‐use efficiency response of a natural grassland ecosystem to last century CO₂ and climatic changes. The results indicate that the alpine grassland community has responded to climate change by improving the physiological control of carbon gain to water loss, following the increases in atmospheric CO₂ and evaporative demand. But, effective leaf‐level water‐use efficiency has remained unchanged.     

Intrinsic water‐use efficiency of temperate seminatural grassland has increased since 1857: an analysis of carbon isotope discrimination of herbage from the Park Grass Experiment

A 150‐year‐long record of intrinsic water‐use efficiency (W_i) was derived from community‐level carbon isotope discrimination (¹³Δ) in the herbage of the unfertilized, unlimed control treatment (plot 3) of the Park Grass Experiment at Rothamsted (England) between 1857 and 2007. ¹³Δ during spring growth (first cut harvested in June) averaged 21.0‰ (±0.5‰ SD) and has not shown a long‐term trend (P=0.5) since 1857. ¹³Δ of summer/autumn growth (second cut harvested between September and November) increased from 21.3‰ to 22.0‰ (P < 0.001) between 1875 and 2007. W_i during spring growth has therefore increased by 33% since the beginning of the experiment, and W_i of summer/autumn growth has increased by 18%. The variation in ¹³Δ was mainly related to weather conditions. Plant available soil water explained 51% and 40% of the variation in spring growth ¹³Δ and summer/autumn growth ¹³Δ, respectively. In the 1857–2007 period yields have not increased, suggesting that community‐level photosynthesis has not increased either. Therefore, the increased W_i probably resulted from a decreased stomatal conductance. Vapour pressure deficit (VPD) during spring growth (March–June) has not changed since 1915, meaning that instantaneous water‐use efficiency (W_t) in spring time has increased and transpiration has probably decreased, provided that leaf temperature followed air temperature. Conversely, VPD in the months between the first and second cut has increased by 0.07 kPa since 1915, offsetting the effect of increased W_i on W_t during summer and early autumn. Our results suggest that vegetation has adjusted physiologically to elevated CO₂ by decreasing stomatal conductance in this nutrient‐limited grassland.     

Trophic interactions among invertebrates in termitaria in the African savanna: a stable isotope approach

Abstract 1. Termites (Isoptera) in tropical savannas are known as ecosystem engineers, affecting the spatial and temporal distribution of water, carbon, cations, and nutrients through their mound structures. Their mounds, however, also offer habitation to diverse taxa and feeding guilds of other invertebrates; a keystone role that has not been properly quantified. 2. The aim of this study was to explore the ecosystem role of termitaria in determining invertebrate diversity and their potential trophic interactions. We used stable isotopes to distinguish termite‐feeding invertebrates from invertebrates merely living in termite mounds under field conditions. 3. The results suggest that inquiline spiders (Arachnida) do not feed on termites directly, but on other invertebrates within the termitaria that are termitophagous, elevating the spiders three trophic levels higher than the termites. 4. This study is the first to demonstrate food web interactions among inquiline invertebrates with a stable isotope approach. It provides evidence that termites play a keystone role in the system by providing habitat for various, trophically interacting invertebrates. These results illustrate a rather unexplored ecosystem property of savanna termites.     

Kinetics and relative significance of remobilized and current C and N incorporation in leaf and root growth zones of Lolium perenne after defoliation: assessment by 13C and 15N steady-state labelling

The contribution of pre-defoliation reserves and current assimilates to leaf and root growth was examined in Lolium perenne L. during regrowth after defoliation. Differential steady-state labelling with ¹³C (CO₂ with δ¹³C = -0.0281 and -0.0088) and ¹⁵N (NO₃^? with 1.0 and 0.368 atom percentage, i.e. δ¹⁵N = 1.742 and 0.0052, respectively) was applied for 2 weeks after defoliation. Rapidly growing tissues were isolated, i.e. the basal elongation and maturation zones of the most rapidly expanding leaves and young root tips, with a biomass turnover rate > 1 d^?1. C and N weights of the elongation zone showed a transient decline. The dry matter and C concentration in fresh biomass of leaf growth zones transiently decreased by up to 25% 2 d after defoliation, while the N concentration remained constant. This ‘dilution’ of growth zone C indicates a decreased net influx of carbohydrates relative to growth-related influx of water and N in expanding cells, immediately after defoliation. Recovery of the total C and N weights of the leaf elongation zone coincided with net incorporation of currently absorbed C and N, as shown by the kinetics of δ¹³C and atom percentage ¹⁵N in the growth zones after defoliation. C isotope discrimination (Δ¹³C) in leaf growth zones was about 23‰, 1–2‰ higher than the Δ in root tips. Δ¹⁵N in the leaf and root growth zones was 10±3‰. The leaf elongation zones (at 0–0.03 m from the tiller base) and the distant root tips (about 0.2 m from the base) exhibited similar kinetics of current C and N incorporation. The amount of pre-defoliation C and N in the growth zones, expressed as a fraction of total C and N, decreased from 1.0 to 0.5 at 3 (C) and 5 (N) d after defoliation, and to 0.1 at 5 (C) and 14 (N) d after defoliation. Thus, the dependence of growth zones on current assimilate supply was significant, and stronger for C than for N. The important roles of current assimilates (as compared to pre-defoliation reserves) and ‘dilution’ of dry matter in regrowth after defoliation are discussed in relation to the method of labelling and the functional and morphological heterogeneity of shoot tissues.     

Influence of Inorganic Phosphate on Photosynthesis of Wheat Chloroplasts: I. PHOTOSYNTHESIS AND ASSIMILATE EXPORT AT 5 ?C AND 25 ?C

Chloroplasts were isolated from 10 d old wheat seedlings andilluminated at 5 ?C or 25 ?C in various concentrations of . Photosynthetic oxygen evolution, ATP content, andexport of triose phosphates and 3-phosphoglycerate were measured.Incorporation of ¹⁴C from NaH¹⁴CO₃ into pentose monophosphates,fructose monophosphate, and glucose monophosphates was determined. The ATP content in illuminated chloroplasts decreased when the concentration in the medium was low. The ATP content increased when the concentration was increased. A higher . concentration in the medium was needed to increase the ATP at 5 ?C than at 25?C. This would suggest that deficiency occurs more readily at low than at high temperatures. More ¹⁴C wasincorporated into photosynthetic metabolites within the chloroplastsat 5 ?C than at 25 ?C, indicating decreased assimilate exportwhen the temperature was low. Dihydroxyacetone phosphate waspreferentially exported when the concentration enabled a high rate of photosynthesis at 25 ?C. However, underconditions of deficiency, either due to low concentration in the medium or due to low temperature, 3-phosphoglycerate was preferred for export. The results suggest that the relatively high photosyntheticrates at low temperature are due to increased concentrationsof photosynthetic metabolites. The assimilate export at lowtemperature seems to be decreased due to decreased concentrationsof dihydroxyacetone phosphate in the stroma. Preferential exportof 3-phosphoglycerate at low temperature or at low concentration in the medium may be a consequence of high stromalconcentrations of this metabolite. On the other hand, it couldalso be due to decreased stromal pH. Key words: Chloroplast, Photosynthesis, Phosphate, Temperature, Translocation, ATP, Wheat