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Notes: The influence of the plastid genome (plastome) on the temperature dependence of chlorophyll fluorescence parameters was studied in four Oenothera species from climatically different habitats. Equipped with their natural plastome, the species could be arranged with respect to their low-temperature tolerance in the order Oe‘villosa’ (plastome I) =Oe argillicola (V) 〉 Oe. grandiflora (III) 〉 Oe elata ssp. hookeri (J). Exchanging the naturally occurring plastome with those from other species did not result in changes of the photosynthetic performance of the genotypes, except for decreasing pigment contents and photosynthesis rates (on area basis) in Oe. elata ssp. hookeri with plastome III. Furthermore, plastome exchange in Oe. elata ssp. hookeri and in Oe. grandiflora did not affect kinetic properties of purified ribulose-l,5-bis-phosphate carboxylase/oxygenase (EC 4,1.1.39), indicating that the gene for the large subunit of this enzyme has remained conserved during plastome evolution in Oenothera. It is concluded that the evolution of the plastome in North American Oenothera spp. did not influence the temperature adaptation of the photosynthetic apparatus, but that the latter is governed by effects residing in the nuclear genome.

Notes: Plasma membrane ATPase from the mediterranean halophyte Plantago crassifolia Forskal was analysed in the native and solubilized state. The enzyme revealed a broad pH optimum at 6.3 when analysed in plasma membrane preparations from roots. A further purified preparation of leaf plasma membrane ATPase was obtained by a three-step solubilization method. Final solubilization of the enzyme was achieved with 1% lysophosphatidylcholine. This enzyme showed a narrow pH optimum at 6.3, inhibition by vanadate, fluoride and N,N′-dicyclohexylcarbodiimide and a high specificity for ATP. The preparations contained a major polypeptide of 107 kDa. None of the parameters analysed in this enzyme changed upon transfer to saline conditions, although the leaves accumulated Na+ and Cl− and an enhanced formation of the compatible organic osmoticum, sorbitol, was detected. It is concluded that at least in this halophyte, the plasma membrane ATPase undergoes no changes during the physiological adaptation of the plant to a saline environment.

Notes: Young (16-day-old) Sorghum bicolor plants of a late- and slow-senescing Texas A&M line (B 35) and of an early- and fast-senescing descendant of an Ethiopian landrace (E 36-1) were subjected to drought stress by decreasing the soil water content to 30% field capacity over 6 days. Plant water potentials decreased from − 2 bar (controls) to − 10 to − 18 bar, and this drought stress resulted in: (1) differential phenotypic reactions and (2) differential decreases in photosynthesis rates in the two cultivars. While E 36-1 tended to lose viable leaf area from the leaf tips downwards, B 35 showed a gradual overall drying of the leaf. At the same time, photosynthesis rates decreased from 31.5 ± 1.6 to 12.3 ± 5.0 µmol CO2 m−2 s−1 (E 36-1) and from 30.5 ± 1.6 to 3.3 ± 2.6 µmol CO2 m−2 s−1 (B 35), respectively. In vitro enzyme activities of phosphoenolpyruvate carboxylase (PEPCase), malate dehydrogenase (MDH) and malic enzyme (ME) on a leaf area basis exceeded the photosynthesis rates. Pyruvate phosphate dikinase (PPDK) activity was close to the photosynthesis rates in control plants and higher than the photosynthesis rates in drought-stressed plants. Thus, none of the enzymes appeared to limit photosynthesis under drought stress, and likely bottleneck enzyme activities of the C3 pathway in the bundle-sheath cells, i.e. ribulose-1,5-bisphosphate carboxylase (RubisCO) and stromal fructose-1,5-bisphosphatase (sFBPase), also showed sufficient activities to sustain higher photosynthesis rates than those observed in the stressed plants. However, under drought stress, total leaf malate concentrations were higher in B 35 (up to 33.1 µmol g−1 FW) than in E 36-1 (up to 22.4 µmol g−1 FW). In particular, at the presumed cytosolic pH of 7.0–7.3, S. bicolor PEPCase was strongly inhibited by malate. In contrast with the in vitro PEPCase enzyme activities, the A/Ci curves suggested a stronger decrease in the in vivo activity of the enzyme in B 35 under drought stress than in E 36-1. It is therefore suggested that photosynthesis under drought stress may be inhibited differentially through feedback malate inhibition of PEPCase in S. bicolor.

Notes: A spontaneously occurring chloroplast genome (plastome) mutant of Oenothera, IVβ, was identified as a single point mutation in the Rubisco large subunit gene (G337 → C), leading to an V113L exchange, which topologically occurs at the interface of two adjacent large subunits (LSU). The minor sterical hindrance of dimer formation by this amino acid exchange strongly impairs holoenzyme assembly, leading to an accumulation of a processing precursor of the holoenzyme, the B-complex, consisting of one LSU and 14 units of chaperonine 60 (cpn60). It is associated with very low holoenzyme concentrations in the mutant tissue, but does not affect the kinetic properties of the enzyme once assembled. When grown under moderate or low light, leaf tissue containing the plastome mutant showed decreased Chl contents and Chl a/b ratios, increased relative carotenoid contents and violaxanthin deepoxidation activity, but very low CO2 fixation and O2 evolution rates and was very sensitive to photoinhibition. The light dependence of chlorophyll fluorescence quenching components at low temperature resembled an extremely chilling sensitive Oenothera genotype as compared to the wild-type. The IVβ mutant thus behaves similarly to the Rubisco SSU antisense plants analysed by Stitt and co-workers (summarised by Stitt and Schulze 1994) and gives an example of the possible influence of plastome mutations on the sensitivity of the photosynthetic apparatus to excess light by modifying the capacity of the Calvin cycle.

Notes: The relationship between susceptibility to photoinhibition, zeaxanthin formation and chlorophyll fluorescence quenching at suboptimal temperatures was studied in chilling-sensitive maize and in non-acclimated and cold-acclimated Oxyria digyna, a chilling-tolerant plant of arctic and alpine habitats. In maize, zeaxanthin formation was strongly suppressed by chilling. Zeaxanthin formed during preillumination at 20°C did not protect maize leaves from photoinhibition during a subsequent high-light, low-temperature treatment, as judged from the ratios of variable to maximal fluorescence, Fv/Fm. However, such preillumination significantly increased non-photochemical quenching (qN) at low temperatures, mainly due to an enhancement of the fast-relaxing qN component (i.e., of energy-dependent quenching. qE). In O. digyna, cold-acclimation resulted in an increased zeaxanthin formation in the temperature range of 2.5–20°C. Cold-acclimation substantially decreased the susceptibility towards photoinhibition at 4°C, but qN remained nearly unchanged between 2 and 38°C, as compared to control plants. Effects of cold acclimation on photosynthesis, photochemical quenching and quantum efficiency of photosystem II were small and indicated a slight amelioration only of the function of the photosynthetic apparatus at suboptimal temperatures (2–20°Ct. I) is concluded, that the xanthophyll cycle is strongly influenced by cold acclimation, while effects on the photosynthetic carbon assimilation only play a minor role in O. digyna.

Notes: The properties of NADH-dependent Fe3+-EDTA reductase in plasma membranes (PM) from roots of iron-deficient and -sufficient tomato plants [Lycopersicon esculentum L. (Mill.) cv. Abunda] were examined. Iron deficiency resulted in a 3-fold increase of in vivo root iron-chelate reductase activity with a Km (Fe3+-EDTA) of 230 μM. In purified root PM, average specific activities of ferric chelate reductase of 410 and 254 nmol Fe (mg protein)−1 min−1 were obtained for iron-deficient and -sufficient plants, respectively. In both cases, the PM-bound activity showed a pH optimum at pH 6.8. Activity depended on NADH and not on NADPH and on the presence of detergent. The activity was inhibited 40-50% by superoxide dismutase (EC 1.15.1.1) and ca 30% by oxygen. Kinetic analysis of the membrane-bound enzyme revealed a Km (Fe3+-EDTA) of ca 200 μM for both iron-stressed and -sufficient plants. For NADH, Km values around 230 μM were obtained. The ferric chelate reductase could be solubilised from salt-washed PM with Triton X-100 at a protein:detergent ratio of 1:2.8 (w/w). The Triton-soluble fraction revealed one enzyme-stained band in native polyacrylamide electrophoresis. Although the membranes showed no nitrate reductase (NR; EC 1.6.6.1) activity, anti-spinach NR immunoglobulin G (IgG) recognized a 54 kDa band both in the PM and the Triton-soluble fraction, but not in the enzymatically active material obtained from the native gel. No evidence could be found for the synthesis of a new, biochemically distinct PM-bound ferric chelate reductase under iron deficiency, which might be identified as the so-called Turbo reductase. It is concluded that iron deficiency in tomato induces increased expression of a ferric chelate reductase in root PM, which is already present in iron-sufficient plants and probably also in plants, which do not contain the Turbo reductase, like the grasses. The iron reductase is not identical with the recently reported PM-associated nitrate reductase.

Notes: Brüggemann, W. and Moog, P. R. 1989. NADH-dependent Fe3+EDTA and oxygen reduction by plasma membrane vesicles from barley roots.Biochemical properties of pyridine-dinucleotide-dependent Fe3+-EDTA reductase were analysed in purified plasma membranes (PM) from barley (Hordeum vulgare L. cv. Marinka) roots. The enzymatic activity preferred NADH over NADPH as electron donor and it was 3-fold increased in the presence of detergent. The reductase showed a pH optimum of 6.8 and saturable kinetics for NADH with Km (NADH) of 125 μM and Vmax of 143 nmol Fe (mg protein)-1 min-1 in the presence of 500 μM Fe3+EDTA. For the dependence of the reaction rate on the iron compound, Km(Fe3+EDTA) of 120 μM and Vmax of 184 nmol (mg protein)-1 min-1 were obtained. The activity was insensitive to superoxide dismutase (SOD; EC 1.15.1.1), catalase (EC 1.11.1.6) and antimycin A, but stimulated by an oxygen-free reaction medium. It could be solubilized by 0.25% (w/v) Triton X-100. The solubilized enzyme revealed one band in native polyacrylamide gel electrophoresis (PAGE) and in isoelectric focussing (IEF) at pl 7.4 by enzyme staining. Major polypeptides with molecular weights of 94, 106, 120 and 205 kDa corresponded to the enzyme-stained band from native PAGE.Analysis of oxygen consumption by the membranes revealed the existence of NADH:CK oxidoreductase activity, which was stimulated by salicylhydroxamic acid (SHAM), chinhydron, Fe3+EDTA and Fe3+EDTA but not by K3 [Fe(CN)6] or K4[Fe (CN)6). The stimulating effect of the iron chelates on oxygen consumption was due to Fe2+ and could be suppressed by bathophenanthroline disulfonate (BPDS), SOD and p-chloromercurophenylsulfonic acid (PCMS). The results are discussed with respect to the nature of the stimulation.

Notes: Abstract. Results from a control integration and time-dependent greenhouse warming experiments performed with a coupled ocean-atmosphere model are analysed in terms of their signal-to-noise properties. The aim is to illustrate techniques for efficient description of the space-time evolution of signals and noise and to identify potentially useful components of a multivariate greenhouse-gas ”fingerprint". The three 100-year experiments analysed here simulate the response of the climate system to a step-function doubling of CO2 and to the time-dependent greenhouse-gas increases specified in Scenarios A (”Business as Usual") and D (”Draconian Measures") of the Intergovernmental Panel on Climate Change (IPCC). If signal and noise patterns are highly similar, the separation of the signal from the natural variability noise is difficult. We use the pattern correlation between the dominant Empirical Orthogonal Functions (EOFs) of the control run and the Scenario A experiment as a measure of the similarity of signal and noise patterns. The EOF 1 patterns of signal and noise are least similar for near-surface temperature and the vertical structure of zonal winds, and are most similar for sea level pressure (SLP). The dominant signal and noise modes of precipitable water and stratospheric/tropospheric temperature contrasts show considerable pattern similarity. Despite the differences in forcing history, a highly similar EOF 1 surface temperature response pattern is found in all three greenhouse warming experiments. A large part of this similarity is due to a common land-sea contrast component of the signal. To determine the degree to which the signal is contaminated by the natural variability (and/or drift) of the control run, we project the Scenario A data onto EOFs 1 and 2 of the control. Signal contamination by the EOF 1 and 2 modes of the noise is lowest for near-surface temperature, a situation favorable for detection. The signals for precipitable water, SLP, and the vertical structure of zonal temperature and zonal winds are significantly contaminated by the dominant noise modes. We use cumulative explained spatial variance, principal component time series, and projections onto EOFs in order to investigate the time evolution of the dominant signal and noise modes. In the case of near-surface temperature, a single pattern emerges as the dominant signal component in the second half of the Scenario A experiment. The projections onto EOFs 1 and 2 of the control run indicate that Scenario D has a large common variability and/or drift component with the control run. This common component is also apparent between years 30 and 50 of the Scenario A experiment, but is small in the 2×CO2 integration. The trajectories of the dominant Scenario A and control run modes evolve differently, regardless of the basis vectors chosen for projection, thus making it feasible to separate signal and noise within the first two decades of the experiments. For Scenario D it may not be possible to discriminate between the dominant signal and noise modes until the final 2–3 decades of the 100-year integration.

Notes: Abstract The uptake of 59Fe from FeCl3, ferric (Fe3+) citrate (FeCitr) and Fe3+-EDTA (FeEDTA) was studied in leaf mesophyll of Vigna unguiculata (L.) Walp. Uptake rates decreased in the order FeCl3〉FeCitr≫FeEDTA, and uptake depended on an obligatory reduction step of Fe3+ to Fe2+, after which the ion could be taken up independently of the chelator, citrate. Uptake was strongly increased by photosynthetically active light (λ〉630 nm), and kinetic analysis revealed saturation kinetics with a K m (FeCitr) of 80–110 μM. In the presence of an external Fe2+ scavenger, bathophenanthroline disulfonate, the mesophyll also reduced external FeCitr with a K m of approx. 50–60 μM. The reduction rates for FeCitr were five-to eightfold higher than necessary for uptake. Purified plasma membranes from leaves revealed an NADH-dependent FeCitr- and FeEDTA-reductase activity, which had a pH optimum of 6.5–6.8 and a K m of approx. 20 μM for NADH. Under anaerobic conditions, a K m of 130–170 μM for ferric chelates was obtained, while in the presence of oxygen a K m (FeCitr) of approx. 100 μM was found. It is concluded that the leaf plasma membrane provides a ferric-chelate-reductase activity, which plays a crucial role in iron uptake of leaf cells. Under in-vivo conditions, however, reactive oxygen species or strong (blue) light may also contribute to the obligatory reduction of Fe3+ prior to uptake.

Notes: Abstract The properties of two Calvin-cycle key enzymes, i.e. stromal fructose-1,6-bisphosphatase (sFBPase) and ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) were studied in the cultivated tomato (Lycopersicon esculentum Mill.) and in four lines of a wild tomato (L. peruvianum Mill.) from different altitudes. During chilling for 14 d at 10°C and low light, the activation energy (EA) of the reaction catalyzed by sFBPase decreased by 5–10 kJ·mol−1 in L. esculentum and the three L. peruvianum lines from high altitudes. In L. peruvianum, no loss or only small losses of enzyme activity were observed during the chilling. Together with the change in EA, this indicates that the latter species is able to acclimate its Calvin-cycle enzymes to low temperatures. In L. esculentum, the chilling stress resulted in the irreversible loss of 57% of the initial sFBPase activity. Under moderately photoinhibiting chilling conditions for 3 d, the L. peruvianum line from an intermediate altitude showed the largest decreases in both the ratio of variable to maximum chlorophyll fluorescence (Fv/Fm) and the in-vivo activation state of sFBPase, while the other L. peruvianum lines showed no inhibition of sFBPase activation. Ribulose-1,5-bisphosphate carboxylase/oxygenase was isolated by differential ammonium-sulfate precipitation and gel filtration and characterized by two-dimensional electrophoresis. The enzyme from L. esculentum had three isoforms of the small subunit of Rubisco, each with different isoelectric points. Of these, the L. peruvianum enzyme contained only the two more-acidic isoforms. Arrhenius plots of the specific activity of purified Rubisco showed breakpoints at approx. 17°C. Upon chilling, the specific activity of the enzyme from L. esculentum decreased by 51%, while EA below the breakpoint temperature increased from 129 to 189 kJ·mol−1. In contrast, Rubisco from the L. peruvianum lines from high altitudes was unaffected by chilling. We tested several possibile explanations for Rubisco inactivation, using two-dimensional electrophoresis, analytical ultracentrifugation, gel filtration and inhibitor tests. No indications were found for differential expression of the subunit isoforms, proteolysis, aggregation, subunit disassembly, or inhibitor accumulation in the enzyme from chilled L. esculentum. We suggest that the activity loss in the L. esculentum enzyme upon chilling is the result of a modification of sulfhydryl groups or other sidechains of the protein.