Katja Hüve

Temperature response of isoprene emission in vivo reflects a combined effect of substrate limitations and isoprene synthase activity: a kinetic analysis.

The responses of isoprene emission rate to temperature are characterized by complex time-dependent behaviors that are currently not entirely understood. To gain insight into the temperature dependencies of isoprene emission, we studied steady-state and transient responses of isoprene emission from hybrid aspen (Populus tremula × Populus tremuloides) leaves using a fast-response gas-exchange system coupled to a proton-transfer reaction mass spectrometer. A method based on postillumination isoprene release after rapid temperature transients was developed to determine the rate constant of isoprene synthase (IspS), the pool size of its substrate dimethylallyldiphosphate (DMADP), and to separate the component processes of the temperature dependence of isoprene emission. Temperature transients indicated that over the temperature range 25°C to 45°C, IspS was thermally stable and operated in the linear range of its substrate DMADP concentration. The in vivo rate constant of IspS obeyed the Arrhenius law, with an activation energy of 42.8 kJ mol−1. In contrast, steady-state isoprene emission had a significantly lower temperature optimum than IspS and higher activation energy. The reversible temperature-dependent decrease in the rate of isoprene emission between 35°C and 44°C was caused by decreases in DMADP concentration, possibly reflecting reduced pools of energetic metabolites generated in photosynthesis, particularly of ATP. Strong control of isoprene temperature responses by the DMADP pool implies that transient temperature responses under fluctuating conditions in the field are driven by initial DMADP pool size as well as temperature-dependent modifications in DMADP pool size during temperature transients. These results have important implications for the development of process-based models of isoprene emission.

Evidence That Light, Carbon Dioxide, and Oxygen Dependencies of Leaf Isoprene Emission Are Driven by Energy Status in Hybrid Aspen

Leaf isoprene emission scales positively with light intensity, is inhibited by high carbon dioxide (CO2) concentrations, and may be enhanced or inhibited by low oxygen (O2) concentrations, but the mechanisms of environmental regulation of isoprene emission are still not fully understood. Emission controls by isoprene synthase, availability of carbon intermediates, or energetic cofactors have been suggested previously. In this study, we asked whether the short-term (tens of minutes) environmental control of isoprene synthesis results from alterations in the immediate isoprene precursor dimethylallyldiphosphate (DMADP) pool size, and to what extent DMADP concentrations are affected by the supply of carbon and energetic metabolites. A novel in vivo method based on postillumination isoprene release was employed to measure the pool size of DMADP simultaneously with the rates of isoprene emission and net assimilation at different light intensities and CO2 and O2 concentrations. Both net assimilation and isoprene emission rates increased hyperbolically with light intensity. The photosynthetic response to CO2 concentration was also hyperbolic, while the CO2 response curve of isoprene emission exhibited a maximum at close to CO2 compensation point. Low O2 positively affected both net assimilation and isoprene emission. In all cases, the variation in isoprene emission was matched with changes in DMADP pool size. The results of these experiments suggest that DMADP pool size controls the response of isoprene emission to light intensity and to CO2 and O2 concentrations and that the pool size is determined by the level of energetic metabolites generated in photosynthesis.

Induction of a longer-term component of isoprene release in darkened aspen leaves: origin and regulation under different environmental conditions

After darkening, isoprene emission continues for 20 to 30 min following biphasic kinetics. The initial dark release of isoprene (postillumination emission), for 200 to 300 s, occurs mainly at the expense of its immediate substrate, dimethylallyldiphosphate (DMADP), but the origin and controls of the secondary burst of isoprene release (dark-induced emission) between approximately 300 and 1,500 s, are not entirely understood. We used a fast-response gas-exchange system to characterize the controls of dark-induced isoprene emission by light, temperature, and CO2 and oxygen concentrations preceding leaf darkening and the effects of short light pulses and changing gas concentrations during dark-induced isoprene release in hybrid aspen (Populus tremula × Populus tremuloides). The effect of the 2-C-methyl-d-erythritol-4-phosphate pathway inhibitor fosmidomycin was also investigated. The integral of postillumination isoprene release was considered to constitute the DMADP pool size, while the integral of dark-induced emission was defined as the “dark” pool. Overall, the steady-state emission rate in light and the maximum dark-induced emission rate responded similarly to variations in preceding environmental drivers and atmospheric composition, increasing with increasing light, having maxima at approximately 40°C and close to the CO2 compensation point, and were suppressed by lack of oxygen. The DMADP and dark pool sizes were also similar through their environmental dependencies, except for high temperatures, where the dark pool significantly exceeded the DMADP pool. Isoprene release could be enhanced by short lightflecks early during dark-induced isoprene release, but not at later stages. Fosmidomycin strongly suppressed both the isoprene emission rates in light and in the dark, but the dark pool was only moderately affected. These results demonstrate a strong correspondence between the steady-state isoprene emission in light and the dark-induced emission and suggest that the dark pool reflects the total pool size of 2-C-methyl-d-erythritol-4-phosphate pathway metabolites upstream of DMADP. These metabolites are converted to isoprene as soon as ATP and NADPH become available, likely by dark activation of chloroplastic glycolysis and chlororespiration.

Elevated [CO2] magnifies isoprene emissions under heat and improves thermal resistance in hybrid aspen.

Isoprene emissions importantly protect plants from heat stress, but the emissions become inhibited by instantaneous increase of [CO2], and it is currently unclear how isoprene-emitting plants cope with future more frequent and severe heat episodes under high [CO2]. Hybrid aspen (Populus tremula x Populus tremuloides) saplings grown under ambient [CO2] of 380 μmol mol−1 and elevated [CO2] of 780 μmol mol−1 were used to test the hypothesis that acclimation to elevated [CO2] reduces the inhibitory effect of high [CO2] on emissions. Elevated-[CO2]-grown plants had greater isoprene emission capacity and a stronger increase of isoprene emissions with increasing temperature. High temperatures abolished the instantaneous [CO2] sensitivity of isoprene emission, possibly due to removing the substrate limitation resulting from curbed cycling of inorganic phosphate. As a result, isoprene emissions were highest in elevated-[CO2]-grown plants under high measurement [CO2]. Overall, elevated growth [CO2] improved heat resistance of photosynthesis, in particular, when assessed under high ambient [CO2] and the improved heat resistance was associated with greater cellular sugar and isoprene concentrations. Thus, contrary to expectations, these results suggest that isoprene emissions might increase in the future.

Elevated atmospheric CO2 concentration leads to increased whole‐plant isoprene emission in hybrid aspen (Populus tremula × Populus tremuloides)

Effects of elevated atmospheric [CO2] on plant isoprene emissions are controversial. Relying on leaf-scale measurements, most models simulating isoprene emissions in future higher [CO2] atmospheres suggest reduced emission fluxes. However, combined effects of elevated [CO2] on leaf area growth, net assimilation and isoprene emission rates have rarely been studied on the canopy scale, but stimulation of leaf area growth may largely compensate for possible [CO2] inhibition reported at the leaf scale. This study tests the hypothesis that stimulated leaf area growth leads to increased canopy isoprene emission rates. We studied the dynamics of canopy growth, and net assimilation and isoprene emission rates in hybrid aspen (Populus tremula × Populus tremuloides) grown under 380 and 780 μmol mol(-1) [CO2]. A theoretical framework based on the Chapman-Richards function to model canopy growth and numerically compare the growth dynamics among ambient and elevated atmospheric [CO2]-grown plants was developed. Plants grown under elevated [CO2] had higher C : N ratio, and greater total leaf area, and canopy net assimilation and isoprene emission rates. During ontogeny, these key canopy characteristics developed faster and stabilized earlier under elevated [CO2]. However, on a leaf area basis, foliage physiological traits remained in a transient state over the whole experiment. These results demonstrate that canopy-scale dynamics importantly complements the leaf-scale processes, and that isoprene emissions may actually increase under higher [CO2] as a result of enhanced leaf area production.

Acclimation of isoprene emission and photosynthesis to growth temperature in hybrid aspen: resolving structural and physiological controls

Acclimation of foliage to growth temperature involves both structural and physiological modifications, but the relative importance of these two mechanisms of acclimation is poorly known, especially for isoprene emission responses. We grew hybrid aspen (Populus tremula x P. tremuloides) under control (day/night temperature of 25/20 °C) and high temperature conditions (35/27 °C) to gain insight into the structural and physiological acclimation controls. Growth at high temperature resulted in larger and thinner leaves with smaller and more densely packed chloroplasts and with lower leaf dry mass per area (MA). High growth temperature also led to lower photosynthetic and respiration rates, isoprene emission rate and leaf pigment content and isoprene substrate dimethylallyl diphosphate pool size per unit area, but to greater stomatal conductance. However, all physiological characteristics were similar when expressed per unit dry mass, indicating that the area-based differences were primarily driven by MA. Acclimation to high temperature further increased heat stability of photosynthesis and increased activation energies for isoprene emission and isoprene synthase rate constant. This study demonstrates that temperature acclimation of photosynthetic and isoprene emission characteristics per unit leaf area were primarily driven by structural modifications, and we argue that future studies investigating acclimation to growth temperature must consider structural modifications.

Transport of foliar applied iron (59Fe) in Vicia faba

Abstract Radioactively labeled iron (59Fe) was used to study iron retranslocation from mature leaves of Broad bean (Vicia faba L. var. Scirocco). Our experiments offered the possibility to detect and quantify the translocation of foliar applied iron by imaging technique in combination with tissue analysis. 59Fe labeled solution was placed as a droplet onto the leafs upper surface of intact plants. Distribution of 59Fe was analyzed after 0.5 h up to 2 days. Iron was translocated acropetally (towards the tip of the treated leaf) as well as basipetally. Movement in the apical direction was predominant, amounting to about 65% of 59Fe translocated from the application site. About 35% of 59Fe were transported basipetally, corresponding to absolute amounts of 2.8–53.6 pmol h−1. After 30 min, it was detectable in the petiole, which included a translocation of 20 mm basipetal from the application site. A mean of 15% of the iron retranslocated from a leaflet was detected in non‐treated leaflets of the same leaf. This iron was supposed to have been exchanged from the phloem into the xylem pathway, probably within the petiole. When the loading rate into the phloem was estimated on basis of the sum of retranslocated 59Fe per time and per area of the leaf treated, a range of 0.031–2.21 pmol h−1 mm−2 (mean: 0.62 pmol h−1 mm−2) was obtained. This was not sufficient to meet an estimated demand for iron in the growing terminal bud, but could cover about 25% of it. In conclusion, average iron retranslocation from leaves of Fe‐sufficient plants was not large enough to meet the iron demand of the growing shoot. This was not due to a limitation in iron availability for transport, as an excess amount of iron was supplied which was not biologically bound, but a limitation due to transport facilities, probably in the phloem, seemed to be more likely in this case.

Determining the timepoint when 14C tracer accurately reflect photosynthate use in the plant-soil system

AbstractBackground and aims Only the carbon (C) isotope pulse labeling approach can provide time-resolved data concerning the input and turnover of plant-derived C in the soil, which are urgently needed to improve the performance of terrestrial C cycle models. However, there is currently very limited information about the point in time after pulse labeling at which the distribution of tracer C accurately represents the usage of photosynthates in different components of the plant-soil system. This should be the case as soon as the tracer has disappeared from the mobile C pool due to respiration, incorporation into the structural C pool of shoot and root tissue and exudation into the soil (rhizodeposition). Methods Following 14CO2

Methodik zur Quantifizierung des Eintrages von Wurzelzellwandresten in den Boden während des Wachstums von Maispflanzen

^{14} {CO}_{2}

Investigations of the mechanisms of long-distance transport and ion distribution in the leaf apoplast of Vicia faba L.

pulse labeling in laboratory and outdoor experiments with spring rye, the 14C dilution rates of soluble fractions and different substances from the structural C pool of the shoot (molecular level), the release of labeled CO2 by belowground respiration (component level), and the 14C kinetics of shoot respiration and 14C remaining in the plant-soil-soil gas continuum (system level) were analyzed during different stages of plant development. Results At all three levels investigated, 14C kinetics indicated that the C tracer levels changed very little between 15 and 21 days after labeling. Results also showed increasing tracer depletion in the mobile C pool. Consequently, only 0.42 % and 0.06 % of all 14C was still available for shoot respiration 15 and 21 days after labeling, respectively. Conclusions The similarities between 14C tracer kinetics at the three investigated levels indicate that tracer disappearance from the mobile pool and distribution throughout the plant-soil system was nearly complete between 15 and 21 days after labeling. Therefore, this appears to be the point at which the pulse labeling approach provides sufficiently precise data concerning the use of C (assimilated during labeling) for root growth, rhizodeposition, root respiration and the microbial turnover of rhizodeposits.