ALBERT

Notes: Abstract The effect of nitrogen supply during growth on the contribution of ribulose-1,5-bisphosphate carboxylase-oxygenase (Rubisco; EC 4.1.1.39) to the control of photosynthesis was examined in tobacco (Nicotiana tabacum L.). Transgenic plants transformed with antisense rbcS to produce a series of plants with a progressive decrease in the amount of Rubisco were used to allow the calculation of the flux-control coefficient of Rubisco for photosynthesis (CR). Several points emerged from the data: (i) The strength of Rubisco control of photosynthesis, as measured by CR, was altered by changes in the short-term environmental conditions. Generally, CR was increased in conditions of increased irradiance or decreased CO2. (ii) The amount of Rubisco in wild-type plants was reduced as the nitrogen supply during growth was reduced and this was associated with an increase in CR. This implied that there was a specific reduction in the amount of Rubisco compared with other components of the photosynthetic machinery. (iii) Plants grown with low nitrogen and which had genetically reduced levels of Rubisco had a higher chlorophyll content and a lower chlorophyll a/b ratio than wild-type plants. This indicated that the nitrogen made available by genetically reducing the amount of Rubisco had been re-allocated to other cellular components including light-harvesting and electron-transport proteins. It is argued that there is a “luxury” additional investment of nitrogen into Rubisco in tobacco plants grown in high nitrogen, and that Rubisco can also be considered a nitrogen-store, all be it one where the opportunity cost of the nitrogen storage is higher than in a non-functional storage protein (i.e. it allows for a slightly higher water-use efficiency and for photosynthesis to respond to temporarily high irradiance).

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.

Notes: Abstract Tobacco (Nicotiana tabacum L.) plants transformed with ‘antisense’ rbcS to decrease the expression of ribulose-1,5-bisphosphate carboxylase-oxygenase (Rubisco) have been used to investigate the contribution of Rubisco to the control of photosynthesis in plants growing at different irradiances. Tobacco plants were grown in controlled-climate chambers under ambient CO2 at 20°C at 100, 300 and 750 μmol·m−2·s−1 irradiance, and at 28°C at 100, 300 and 1000 μmol·m−2·s−1 irradiance. (i) Measurement of photosynthesis under ambient conditions showed that the flux control coefficient of Rubisco (C infRubisco supA ) was very low (0.01–0.03) at low growth irradiance, and still fairly low (0.24–0.27) at higher irradiance. (ii) Short-term changes in the irradiance used to measure photosynthesis showed that C infRubisco supA increases as incident irradiance rises, (iii) When low-light (100 μmol·m−2·s−1)-grown plants are exposed to high (750–1000 μmol·m−2·s−1) irradiance, Rubisco is almost totally limiting for photosynthesis in wild types. However, when high-light-grown leaves (750–1000 μmol·m−2·s−1) are suddenly exposed to high and saturating irradiance (1500–2000 μmol·m−2·s−1), C infRubisco supA remained relatively low (0.23–0.33), showing that in saturating light Rubisco only exerts partial control over the light-saturated rate of photosynthesis in “sun” leaves; apparently additional factors are co-limiting photosynthetic performance, (iv) Growth of plants at high irradiance led to a small decrease in the percentage of total protein found in the insoluble (thylakoid fraction), and a decrease of chlorophyll, relative to protein or structural leaf dry weight. As a consequence of this change, high-irradiance-grown leaves illuminated at growth irradiance avoided an inbalance between the “light” reactions and Rubisco; this was shown by the low value of C infRubisco supA (see above) and by measurements showing that non-photochemical quenching was low, photochemical quenching high, and NADP-malate dehydrogenase activation was low at the growth irradiance. In contrast, when a leaf adapted to low irradiance was illuminated at a higher irradiance, Rubisco exerted more control, non-photochemical quenching was higher, photochemical quenching was lower, and NADP-malate dehydrogenase activation was higher than in a leaf which had grown at that irradiance. We conclude that changes in leaf composition allow the leaf to avoid a one-sided limitation by Rubisco and, hence, overexcitation and overreduction of the thylakoids in high-irradiance growth conditions, (v) ‘Antisense’ plants with less Rubisco contained a higher content of insoluble (thylakoid) protein and chlorophyll, compared to total protein or structural leaf dry weight. They also showed a higher rate of photosynthesis than the wild type, when measured at an irradiance below that at which the plant had grown. We propose that N-allocation in low light is not optimal in tobacco and that genetic manipulation to decrease Rubisco may, in some circumstances, increase photosynthetic performance in low light.

Notes: Abstract Short-term (hours) changes in plant water status were induced in hazel (Corylus avellana L.) by changing the evaporative demand on a major portion of the shoot while maintaining a branch in a constant environment. Stomatal conductance of leaves on the branch was influenced little by these short-term changes in water status even with changes in leaf water potential as great as 8 bars. Long-term (days) changes in plant water status were imposed by soil drying cycles. Stomatal conductance progessively decreased with increases in long-term water stress. Stomata still responded to humidity with long-term water stress but the range of the conductance response decreased. Threshold responses of stomata to leaf water potential were not observed with either short-term or long-term changes in plant water status even when leaves wilted. It is suggested that concurrent measurements of plant water status may not be sufficient for explaining stomatal and other plant responses to drought.

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.

Notes: Summary Net photosynthesis of Picea abies was measured in a spruce forest in northern Germany with temperature- and humidity-controlled cuvettes in 4 different crown layers on shoots of different ages. These measurments were performed such that temperature and humidity either followed ambient conditions or were kept constant. Annual courses of light-, temperature-, and humidity-related net photosynthesis were determined. Spruce had a remarkably constant rate of CO2 uptake from April to September for 1-year and older needles. Light saturation was achieved at 25 klx. Current year needles had the highest rates of CO2 uptake in early summer, but these rates decreased by autumn. Photosynthetic capacity decreased with needle age and, on a dry weight basis, it was higher in the shade than in the sun crown. The temperature optimum was between 13 and 23° C. Photosynthesis in spruce decreased when air humidity was low. The effect of the natural weather conditions on photosynthetic capacity was determined. The habitat is characterized by a high frequency of low light intensities (75% of total daytime below 20 klx) and cool temperatures (80% of daytime between 9 and 21° C). Low air humidity was only present when light intensities were high. The major limiting factor for production was low light intensities, which reduced photosynthetic capacity in the sun crown to 42% below maximum possible rates. Adverse temperatures reduced CO2 uptake by 28% and large water vapor pressure deficits reduced rates by only 2% compared with maximum possible rates. The limited adaptation to light is discussed.

Notes: Abstract Natural abundances of nitrogen isotopes, δ15N, indicate that, in the same habitat, Alaskan Picea glauca and P. mariana use a different soil nitrogen compartment from the evergreen shrub Vaccinium vitis-idaea or the deciduous grass Calamagrostis canadensis. The very low δ15N values (-7.7 ‰) suggest that (1) Picea mainly uses inorganic nitrogen (probably mainly ammonium) or organic N in fresh litter, (2) Vaccinium (-4.3 ‰) with its ericoid mycorrhizae uses more stable organic matter, and (3) Calamagrostis (+0.9 ‰) exploits deeper soil horizons with higher δ15N values of soil N. We conclude that species limited by the same nutrient may coexist by drawing on different pools of soil N in a nutrient-deficient environment. The differences among life-forms decrease with increasing N availability. The different levels of δ15N are associated with different nitrogen concentrations in leaves, Picea having a lower N concentration (0.62 mmol g−1) than Vaccinium (0.98 mmol g−1) or Calamagrostis (1.33 mmol g−1). An extended vector analysis by Timmer and Armstrong (1987) suggests that N is the most limiting element for Picea in this habitat, causing needle yellowing at N concentrations below 0.5 mmol g−1 or N contents below 2 mmol needle−1. Increasing N supply had an exponential effect on twig and needle growth. Phosphorus, potassium and magnesium are at marginal supply, but no interaction between ammonium supply and needle Mg concentration could be detected. Calcium is in adequate supply on both calcareous and acidic soils. The results are compared with European conditions of excessive N supply from anthropogenic N depositions.

Notes: Summary The photosynthesizing branches of Hammada scoparia, one of the typical dwarf shrubs of the Negev desert, undergo a seasonal change from succulent to xeromorphic anatomy. This trend is accompanied by a marked decrease of water content and of total water Ψ plant and osmotic Ψ π plant potential. Irrigated plants do not show such transitions. The daily courses of Ψ plant and Ψ π plant showed minima around noon and a tendency for maxima before sunrise. Turgor pressure Ψ p plant reached minima around noon and became negative (until ca.-10 bars). Generally, Ψ plant decreases with increasing water vapour concentration difference between plant and air (WD) in the first half of the day, and in the second half the reversal of this trend occurs. Mostly smaller increments of Ψ plant were correlated with larger increases in WD which lead to the conclusion that stomates closed enough to maintain transpiration at a constant value. Non-irrigated and irrigated plants showed different hysteresis loops of relation between Ψ plant and WD. Regulatory reduction of transpiration appears largely independently of Ψ plant which is in spring and with irrigated plants on a high level, with non-irrigated plants in summer on a low level. In summer the continous but decreasing drop of Ψ plant with increasing WD was interpreted as caused by a change in soil or root resistance. Independent of the seasonal state and of the Ψ plant level, H. scoparia regulates its water status within limited ranges of Ψ p plant changes: the irrigated plants on a higher level, the non-irrigated on a lower level of Ψ p plant . The water contents of the tissues of H. scoparia are linearily related to Ψ plant as well as Ψ p plant . Steeper slopes with non-irrigated plants in summer than with spring palnts and with irrigated plants during the whole season signify that in the latter a certain increment in turgor pressure corresponds to a large gain in water content while in the non-irrigated summer plants it varies only little for an identical change in Ψ p plant . This behaviour of non-irrigated wild plants apparently is due to the change of the elastic properties of the tissues in the assimilating branches.

Notes: Summary Measurements of CO2 and water vapor exchange were performed on Prunus armeniaca L. with humidity- and temperature-controlled chambers under the climatic conditions of a desert habitat. In apricot, the stomatal response to changes in temperature and water-vapor concentration difference between leaf and air (WD) significantly determined the rates of gas exchange during the day (parts I and II). The effect of climate-controlled stomatal response on the transpiration/net photosynthesis (T/P)-ratio was analyzed and simulated using experiments conducted at constant temperature and/or humidity conditions for input parameters. The measured values of the T/P-ratio at naturally varying conditions of humidity and temperature were compared with calculated results of a model in which it was assumed, (1) that stomata and photosynthetic activity are not affected by air humidity and temperature, (2) that the stomata only respond with a constant photosynthetic activity to changes in WD, and (3) that the stomata respond to both, leaf temperature and air humidity with a constant photosynthetic activity. These simulations facilitated an analysis of the naturally observed changes in the T/P-ratio. The calculated T/P-ratios were very small if the simulation assumed that stomata only respond to WD at a constant photosynthetic activity. These low predicted values of the T/P-ratio were not obtained under natural conditions, since an increase in WD during the day was correlated with a temperature rise which tended to open stomata and change the photosynthetic activity. Humidity induced stomatal closure did appear to substantially reduce T/P-ratios. The measured T/P-ratio changed considerably during the year. The lowest T/P-ratios were obtained in the middle of the dry season at a time when stomata responded strongly to air humidity and when optimum of photosynthesis was reached at high temperatures. The daily average T/P-ratio calculated from the daily sum of P and T showed little change during the seasons. A high T/P-ratio was also observed at reduced rates of gas exchange. The T/P-ratios of apricot were compared with different species in different environments.

Notes: Summary The gas exchange of the apricot (Prunus armeniaca L.) growing in the runoff farm at Avdat (Negev, Israel) was measured during its growing period using temperature- and humidity-controlled chambers. Water potentials of the xylem were measured with a pressure bomb, and the mesophyll internal CO2 concentration was calculated from simultaneous measurements of net photosynthesis and transpiration. The daily changes in water potential Ψ had only little effect on the daily course of stomatal resistance. The early morning peak of CO2 uptake was reached when Ψ had already dropped to very low values. On dry days, Ψ and the relative water content of the leaf were improved at noon during the time of stomatal closure. On humid days, Ψ dropped to very low values (43.5 bar) at a high transpiration rate without causing stomatal closure, as much as on the dry days when stomata where more closed at less water stress. The observed changing sensitivity of the stomata to changes in air humidity during the season is related to the water status in the plant. This change is possibly caused by a long-term effect of stress in this habitat. The daily changes in stomatal diffusion resistance did not consistently correlate with changes of the CO2 concentration in the intercellular air spaces. In the morning a decreasing internal CO2 concentration was even inversely correlated to the stomatal response. In the afternoon the effect of an increasing internal CO2 concentration and the effect of external climate on stomatal response could be additive. However, at the time, when CO2 uptake reached a second peak in the afternoon the same value of diffusion resistance is reached at very different levels of internal CO2 concentration as compared to the morning. For the regulation of the diffusion resistance in apricot under the natural conditions, the effects of plant internal control mechanisms are overruled and/or modified by the external climatic factors of air humidity and temperature. The significance of the climate-controlled stomatal response for the existence and cultivation of this plant species in an arid habitat is discussed.