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  • Nicotiana (transformed with antisense DNA)  (2)
  • δ15N  (2)
  • 1995-1999
  • 1990-1994  (4)
  • 1
    ISSN: 1432-2048
    Keywords: Nicotiana (transformed with antisense DNA) ; Photosynthesis ; Ribulose-1,5-bisphosphate carboxylase-oxygenase ; Transgenic plant (antisense)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Experiments were carried out to determine how decreased expression of ribulose-1,5-bisphosphate carboxylase-oxygenase (Rubisco) affects photosynthetic metabolism in ambient growth conditions. In a series of tobacco (Nicotiana tabacum L.) plants containing progressively smaller amounts of Rubisco the rate of photosynthesis was measured under conditions similar to those in which the plants had been grown (310 μmol photons · m−2 · s−1, 350 μbar CO2, 22° C). (i) There was only a marginal inhibition (6%) of photosynthesis when Rubisco was decreased to about 60% of the amount in the wildtype. The reduced amount of Rubisco was compensated for by an increase in Rubisco activation (rising from 60 to 100%), with minor contributions from an increase of its substrates (ribulose-1,5-bisphosphate and the internal CO2 concentration) and a decrease of its product (glycerate-3-phosphate). (ii) The decreased amount of Rubisco was accompanied by an increased ATP/ADP ratio that may be causally linked to the increased activation of Rubisco. An increase of highenergy-state chlorophyll fluorescence shows that thylakoid membrane energisation and high-energy-state-dependent energy dissipation at photosystem two had also increased. (iii) A further decrease of Rubisco (in the range of 50–20% of the wildtype level) resulted in a strong and proportional inhibition of CO2 assimilation. This was accompanied by a decrease of fructose-1,6-bisphosphatase activity, coupling-factor 1 (CF1)-ATP-synthase protein, NADP-malate dehydrogenase protein, and chlorophyll. The chlorophyll a/b ratio did not change, and enolase and sucrose-phosphate synthase activity did not decrease. It is argued that other photosynthetic enzymes are also decreased once Rubisco decreases to the point at which it becomes strongly limiting for photosynthesis. (iv) It is proposed that the amount of Rubisco in the wildtype represents a balance between the demands of light, water and nitrogen utilisation. The wildtype overinvests about 15% more protein in Rubisco than is needed to avoid a strict Rubisco limitation of photosynthesis. However, this “excess” Rubisco allows the wildtype to operate with lower thylakoid energisation, and decreased high-energy-state-dependent energy dissipation, hence increasing light-use efficiency by about 6%. It also allows the wildtype to operate with a lower internal CO2 concentration in the leaf and a lower stomatal conductance at a given rate of photosynthesis, so that instantaneous water-use efficiency is marginally (8%) increased.
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  • 2
    ISSN: 1432-2048
    Keywords: Flux control (photosynthesis) ; Nicotiana (transformed with antisense DNA) ; Ribulose-1,5-bisphosphate carboxylase-oxygenase (control of photosynthesis) ; Transgenic plant (antisense)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Transgenic tobacco (Nicotiana tabacum L.) plants transformed with ‘antisense’ rbcS to produce a series of plants with a progressive decrease in the amount of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) have been used to investigate the contribution of Rubsico to the control of photosynthesis at different irradiance, CO2 concentrations and vapour-pressure deficits. Assimilation rates, transpiration, the internal CO2 concentration and chlorophyll fluorescence were measured in each plant. (i) The flux-control coefficient of Rubisco was estimated from the slope of the plot of Rubisco content versus assimilation rate. The flux-control coefficient had a value of 0.8 or more in high irradiance, (1050 μmol·m−2·s−1), low-vapour pressure deficit (4 mbar) and ambient CO2 (350 μbar). Control was marginal in enhanced CO2 (450 μbar) or low light (310 μmol·m−2·s−1) and was also decreased at high vapour-pressure deficit (17 mbar). No control was exerted in 5% CO2. (ii) The flux-control coefficients of Rubisco were compared with the fractional demand placed on the calculated available Rubisco capacity. Only a marginal control on photosynthetic flux is exerted by Rubisco until over 50% of the available capacity is being used. Control increases as utilisation rises to 80%, and approaches unity (i.e. strict limitation) when more than 80% of the available capacity is being used. (iii) In low light, plants with reduced Rubisco have very high energy-dependent quenching of chlorophyll fluorescence (qE) and a decreased apparent quantum yield. It is argued that Rubisco still exerts marginal control in these conditions because decreased Rubisco leads to increased thylakoid energisation and high-energy dependent dissipation of light energy, and lower light-harvesting efficiency. (iv) The flux-control coefficient of stomata for photosynthesis was calculated from the flux-control coefficient of Rubisco and the internal CO2 concentration, by applying the connectivity theorem. Control by the stomata varies between zero and about 0.25. It is increased by increased irradiance, decreased CO2 or decreased vapour-pressure deficit. (v) Photosynthetic oscillations in saturating irradiance and CO2 are suppressed in decreased-activity transformants before the steady-state rate of photosynthesis is affected. This provides direct evidence that these oscillations reveal the presence of “excess” Rubisco. (vi) Comparison of the flux-control coefficients of Rubisco with mechanistic models of photosynthesis provides direct support for the reliability of these models in conditions where Rubisco has a flux-control coefficient approach unity (i.e. “limits” photosynthesis), but also indicates that these models are less useful in conditions where control is shared between Rubisco and other components of the photosynthetic apparatus.
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  • 3
    ISSN: 1573-5036
    Keywords: atmospheric deposition ; δ15N ; δ34S ; forest decline ; nitrogen ; Picea abies ; stable isotopes ; sulfur
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Concentrations and natural isotope abundance of total sulfur and nitrogen as well as sulfate and nitrate concentrations were measured in needles of different age classes and in soil samples of different horizons from a healthy and a declining Norway spruce (Picea abies (L.) Karst.) forest in the Fichtelgebirge (NE Bavaria, Germany), in order to study the fate of atmospheric depositions of sulfur and nitrogen compounds. The mean δ15N of the needles ranged between −3.7 and −2.1 ‰ and for δ34S a range between −0.4 and +0.9 ‰ was observed. δ34S and sulfur concentrations in the needles of both stands increased continuously with needle age and thus, were closely correlated. The δ15N values of the needles showed an initial decrease followed by an increase with needle age. The healthy stand showed more negative δ15N values in old needles than the declining stand. Nitrogen concentrations decreased with needle age. For soil samples at both sites the mean δ15N and δ34S values increased from −3 ‰ (δ15N) or +0.9 ‰ (δ34S) in the uppermost organic layer to about +4 ‰ (δ15N) or +4.5 ‰ (δ34S) in the mineral soil. This depth-dependent increase in abundance of 15N and 34S was accompanied by a decrease in total nitrogen and sulfur concentrations in the soil. δ15N values and nitrogen concentrations were closely correlated (slope −0.0061 ‰ δ15N per μmol eq N gdw −1), and δ34S values were linearly correlated with sulfur concentrations (slope −0.0576 ‰ δ34S per μmol eq S gdw −1). It follows that in the same soil samples sulfur concentrations were linearly correlated with the nitrogen concentrations (slope 0.0527), and δ34S values were linearly correlated with δ15N values (slope 0.459). A correlation of the sulfur and nitrogen isotope abundances on a Δ basis (which considers the different relative frequencies of 15N and 34S), however, revealed an isotope fractionation that was higher by a factor of 5 for sulfur than for nitrogen (slope 5.292). These correlations indicate a long term synchronous mineralization of organic nitrogen and sulfur compounds in the soil accompanied by element-specific isotope fractionations. Based on different sulfur isotope abundance of the soil (δ34S=0.9 ‰ for total sulfur of the organic layer was assumed to be equivalent to about −1.0 ‰ for soil sulfate) and of the atmospheric SO2 deposition (δ34S=2.0 ‰ at the healthy site and 2.3 ‰ at the declining site) the contribution of atmospheric SO2 to total sulfur of the needles was estimated. This contribution increased from about 20 % in current-year needles to more than 50 % in 3-year-old needles. The proportion of sulfur from atmospheric deposition was equivalent to the age dependent sulfate accumulation in the needles. In contrast to the accumulation of atmospheric sulfur compounds nitrogen compounds from atmospheric deposition were metabolized and were used for growth. The implications of both responses to atmospheric deposition are discussed.
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  • 4
    ISSN: 1432-1939
    Keywords: δ13C ; δ15N ; Nitrogen assimilation ; Forest decline ; Picea abies ; Stable isotopes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Natural carbon and nitrogen isotope ratios were measured in different compartments (needles and twigs of different ages and crown positions, litter, understorey vegetation, roots and soils of different horizons) on 5 plots of a healthy and on 8 plots of a declining Norway spruce (Picea abies (L.) Karst.) forest in the Fichtelgebirge (NE Bavaria, Germany), which has recently been described in detail (Oren et al. 1988a; Schulze et al. 1989). The δ13C values of needles did not differ between sites or change consistently with needle age, but did decrease from the sun-to the shade-crown. This result confirms earlier conclusions from gas exchange measurements that gaseous air pollutants did no long-lasting damage in an area where such damage was expected. Twigs (δ13C between-25.3 and-27.8‰) were significantly less depleted in 13C than needles (δ13C between-27.3 and-29.1‰), and δ13C in twigs increased consistently with age. The δ15N values of needles ranged between-2.5 and-4.1‰ and varied according to stand and age. In young needles δ15N decreased with needle age, but remained constant or increased in needles that were 2 or 3 years old. Needles from the healthy site were more depleted in 15N than those from the declining site. The difference between sites was greater in old needles than in young ones. This differentiation presumably reflects an earlier onset of nitrogen reallocation in needles of the declining stand. δ15N values in twigs were more negative than in needles (-3.5 to-5.2‰) and showed age- and stand-dependent trends that were similar to the needles. δ15N values of roots and soil samples increased at both stands with soil depth from-3.5 in the organic layer to +4‰ in the mineral soil. The δ15N values of roots from the mineral soil were different from those of twigs and needles. Roots from the shallower organic layer had values similar to twigs and needles. Thus, the bulk of the assimilated nitrogen was presumably taken up by the roots from the organic layer. The problem of separation of ammonium or nitrate use by roots from different soil horizons is discussed.
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