Marco Frehner

Stimulation of Symbiotic N2 Fixation in Trifolium repens L. under Elevated Atmospheric pCO2 in a Grassland Ecosystem.

Symbiotic N2 fixation is one of the main processes that introduces N into terrestrial ecosystems. As such, it may be crucial for the sequestration of the extra C available in a world of continuously increasing atmospheric CO2 partial pressure (pCO2). The effect of elevated pCO2 (60 Pa) on symbiotic N2 fixation (15N-isotope dilution method) was investigated using Free-Air-CO2-Enrichment technology over a period of 3 years. Trifolium repens was cultivated either alone or together with Lolium perenne (a nonfixing reference crop) in mixed swards. Two different N fertilization levels and defoliation frequencies were applied. The total N yield increased consistently and the percentage of plant N derived from symbiotic N2 fixation increased significantly in T. repens under elevated pCO2. All additionally assimilated N was derived from symbiotic N2 fixation, not from the soil. In the mixtures exposed to elevated pCO2, an increased amount of symbiotically fixed N (+7.8, 8.2, and 6.2 g m-2 a-1 in 1993, 1994, and 1995, respectively) was introduced into the system. Increased N2 fixation is a competitive advantage for T. repens in mixed swards with pasture grasses and may be a crucial factor in maintaining the C:N ratio in the ecosystem as a whole.

Localization of Fructan Metabolism in the Vacuoles Isolated from Protoplasts of Jerusalem Artichoke Tubers (Helianthus tuberosus L.)

Protoplasts were prepared from tubers of Helianthus tuberosus in the developing and in the resting stage of development. Vacuoles were isolated from the protoplasts and purified by sedimentation through a density gradient of glycine betaine. All the fructan (inulin) with a DP ⩾ 3 (i.e. the trisaccharide isokestose and larger polymers of fructose) was found to be located exclusively in the vacuoles whilst sucrose, glucose, and fructose were located only partially there. The vacuoles were also found to be the sole cell compartments containing fructan synthesizing enzyme activities (sucrose-sucrose-fructosyl-transferase and fructan-fructan-fructosyl-transferase) and fructan degrading enzyme activity (fructan exohydrolase), depending on the stage of development of the tubers. The vacuole is therefore proposed to be the cell organelle for fructan storage and metabolism.

Soil mineral nitrogen availability was unaffected by elevated atmospheric pCO2 in a four year old field experiment (Swiss FACE)

The effect of elevated (60 Pa) atmospheric carbon dioxide partial pressure (pCO2) and N fertilisation on the availability of mineral N and on N transformation in the soil of a Lolium perenne L. monoculture was investigated in the Swiss FACE (Free Air Carbon dioxide Enrichment) experiment. The apparent availability of nitrate and ammonium for plants was estimated during a representative, vegetative re-growth period at weekly intervals from the sorption of the minerals to mixed-bed ion-exchange resin bags at a soil depth of 5 cm. N mineralisation was measured using sequential coring and in situ exposure of soil cores in the top 10 cm of the soil before and after the first cut in spring 1997. High amounts of mineral N were bound to the ion exchange resin during the first week of re-growth. This was probably the combined result of the fertiliser application, the weak demand for N by the newly cut sward and presumably high rates of root decay and exudation after cutting the sward. During the first 2 weeks after the application of fertiliser N at the first cut, there was a dramatic reduction in available N; N remained low during the subsequent weeks of re-growth in all treatments. Overall, nitrate was the predominant form of mineral N that bound to the resin for the duration of the experiment. Apparently, there was always more nitrate than ammonium available to the plants in the high N fertilisation treatment for the whole re-growth period. Apparent N availability was affected significantly by elevated pCO2 only in the first week after the cut; under high N fertilisation, elevated pCO2 increased the amount of mineral N that was apparently available to the plants. Elevated pCO2 did not affect apparent net transformation of N, loss of N or uptake of N by plants. The present data are consistent with earlier results and suggest that the amount of N available to plants from soil resources does not generally increase under elevated atmospheric pCO2. Thus, a possible limiting effect of N on primary production could become more stringent under elevated atmospheric pCO2 as the demand of the plant for N increases.

Nitrogen plays a major role in leaves when source-sink relations change: C and N metabolism in Lolium perenne growing under free air CO2 enrichment

Swards of Lolium perenne L. were grown in the field in a long-term free air CO2 enrichment (FACE) facility. The CO2 treatment was combined with two levels of N fertilization and regular defoliation, which resulted in plants with a wide range of source-sink relations. C and N metabolism were investigated to assess the role of carbohydrate and nitrogenous compounds in leaves in indicating source-sink relations. Sucrose exhibited the largest changes in contents during the day-night cycle; therefore, it was identified as the main short-term storage compound for night-time export. Fructan accumulation indicated the degree of surplus C supply in the source compared to C use in sinks. Nitrate content depended mainly on N fertilization, and was reduced under elevated pCO2. Nitrate appeared to indicate a current surplus of available N relative to the need for growth. Amino acid content responded strongly to N fertilization but decreased only slightly under elevated pCO2. Protein content, however, decreased significantly under elevated pCO2. The patterns of diurnal changes of C or N compounds did not differ between CO2 treatments. Down-regulation of photosynthesis appeared to occur when plants were extremely N-limited as under elevated pCO2, low N and at a late regrowth stage.

Partitioning of P and the activity of root acid phosphatase in white clover (Trifolium repens L.) are modified by increased atmospheric CO2 and P fertilisation

The growth response of white clover (Trifolium repens L.) to the expected increase in atmospheric partial pressure of CO2 (pCO2) may depend on P availability. A decrease in the rate of transpiration due to increased pCO2 may reduce the amount of P transported to the shoot, thereby causing a change in the partitioning of P between the root and shoot. To test these hypotheses, four concentrations of P in the nutrient solution, combined with two pCO2 treatments, were applied to nodulated white clover plants. Compared to ambient pCO2 (35 Pa), twice ambient pCO2 (70 Pa) reduced the rate of transpiration but did not impair the total P uptake per plant. However, at twice ambient pCO2 and a moderate to high supply of P, concentrations of structural P and soluble P (Pi) were lower in the leaves and higher in the roots. The activity of root acid phosphatase was lower at twice ambient pCO2 than at ambient pCO2; it depended on the Pi concentration in the roots. At the highest P concentration, twice ambient pCO2 stimulated photosynthesis and the growth rate of the plant without affecting the concentration of nonstructural carbohydrates in the leaves. However, at the lower P concentrations, plants at twice ambient pCO2 lost their stimulation of photosynthesis in the afternoon, they accumulated nonstructural carbohydrates in the leaves and their growth rate was not stimulated; indicating C-sink limitation of growth. P nutrition will be crucial to the growth of white clover under the expected future conditions of increased pCO2.

Does the response of perennial ryegrass to elevated CO2 concentration depend on the form of the supplied nitrogen

To test whether different nitrogen form (nitrate or ammonium) in substrate can alter the response to elevated partial pressure of CO2 (pCO2) plants of perennial ryegrass (Lolium perenne cv. Bastion) were grown from seeds in growth chambers under pCO2 of either 35 Pa (ambient, CA) or 70 Pa (elevated, CE) in a hydroponic system (with nutrient and pH control) for 24 d. Nitrogen was supplied as ammonium, nitrate or an equimolar mixture of both N forms. Under CE plants grew faster than their counterparts under CA during the first 14 d but after 23 d of cultivation stimulation disappeared. Despite the strong positive effect of mixed forms of N on plant growth, the beneficial effect of CE was similar to that in the other two N treatments. However, the almost alike final growth response to CE had different underlying mechanisms in different N treatments. Plants supplied with nitrate as a sole source of nitrogen had lower leaf mass ratio but much higher specific leaf area compared to plants supplied with ammonium. The decrease in the content of leaf organic N (per unit of structural dry mass) under CE was found only in leaves of plants supplied with ammonium on day 14. Nevertheless, the available form of N evidently contributes to changes of leaf N content under CE. The high levels of N and non-structural saccharides in plants supplied with ammonium at CE suggest that the CO2 response of these plants was controlled by factors other than amount of available carbon and nitrogen.