Higher Electron Transport Rate Research Articles

The Effect of Sulfur Re-Addition on H2 Photoproduction by Sulfur-Deprived Green Algae

Sulfur deprivation of algal cultures selectively and partially inactivates photosystem II (PSII)-catalyzed O(2) evolution, induces anaerobiosis and hydrogenase expression, and results in sustained H(2) photoproduction for several days. We show that re-addition of limiting amounts of sulfate (1-10 microM final concentration) to the cultures during the H(2)-production phase temporarily reactivates PSII photochemical and O(2)-evolution activity and re-establishes higher rates of electron transport through the photosynthetic electron transport chain. The reactivation of PSII occurs by de novo D1 protein synthesis, but does not result in the re-establishment of aerobic conditions in the reactor, detectable by dissolved-O(2) sensors. However, concomitant H(2) photoproduction is inhibited, possibly due to excessive intra-cellular levels of photosynthetically-evolved O(2). The partial recovery of electron transport rates correlates with the re-oxidation of the plastoquinone (PQ) pool, as observed by pulse-amplitude modulated (PAM) and fluorescence-induction measurements. These results show that the presence of a more oxidized PQ pool releases some of the down-regulation of electron transport caused by the anaerobic conditions.

Glucose Suppresses Superoxide Generation in Metabolically Responsive Pancreatic β Cells

High rates of glucose metabolism and mitochondrial electron transport have been associated with increased mitochondrial production of reactive oxygen species (ROS). This mechanism was also proposed as a possible cause for dysfunction and death of pancreatic beta cells exposed to high glucose levels. We examined whether high rates of glucose metabolism increase ROS production in purified rat beta cells. Glucose up to 20 mm did not stimulate H(2)O(2) or superoxide production, whereas it dose-dependently increased cellular NAD(P)H and FADH(2) levels with an EC(50) around 8 mm. On the contrary, glucose concentration-dependently suppressed H(2)O(2) and superoxide formation, with a major effect between 0 and 5 mm, parallel to an increase in cellular NAD(P)H levels. This suppressive effect was more marked in beta cells with higher NAD(P)H responsiveness to glucose; it was not observed in glucagon-containing alpha cells, which lacked a glucose-induced increase in NAD(P)H. Suppression was also induced by the mitochondrial substrates leucine and succinate. Experiments with electron transport chain inhibitors indicate a role of respiratory complex I in ROS production at low mitochondrial activity and low NADH levels. Superoxide production at low glucose is potentially cytotoxic, because scavenging by the superoxide dismutase mimetic agent manganese(III)tetrakis(4-benzoic acid)porphyrin was found to reduce the rate of beta cell apoptosis. Analysis of islets cultured at 20 mm glucose confirmed that this condition does not induce ROS production in beta cells as a result of their increased rates of glucose metabolism. Our study indicates the need of beta cells for basal nutrients maintaining mitochondrial NADH production at levels that suppress ROS accumulation from an inadequate respiratory complex I activity and thus inhibit a potential apoptotic pathway.

Altered photosynthetic performance of a natural Arabidopsis accession is associated with atrazine resistance

Natural variation for photosynthetic traits was studied by determining chlorophyll fluorescence parameters in a collection of Arabidopsis accessions. This screen revealed only one single accession (Ely), exhibiting photosynthetic characteristics markedly different from all others, while a few lines showed small but significant variation. Detailed genetic and physiological analyses showed reduced fitness for Ely compared with the standard laboratory strain Ler for various growth parameters. At low temperature (15 degrees C), Ely had a higher electron transport rate than Ler, indicating increased photosystem II efficiency under this condition, while at high temperature (30 degrees C) the opposite was observed. Ely had a high sensitivity to UV-B radiation compared with Ler and was atrazine resistant. This atrazine-resistance and related chlorophyll fluorescence traits were maternally inherited, pointing towards chloroplast-located gene(s). Definite proof that Ely is atrazine-resistant was obtained by sequencing the psbA gene, encoding the D1 protein of photosystem II, revealing a point mutation causing the same amino acid change as found in other atrazine-resistant species. Additional nuclear encoded genetic variation was also present, as was concluded from the small but significant differences in phenotype between Ely and its reciprocal crosses with Ler. It was concluded that the photosynthetic yield is highly conserved and that only severe selection pressure results in marked variations in photosynthetic performance.

Photosynthesis, photosynthetic pigments and mycosporine-like amino acids after exposure of the marine red alga Chondrus crispus (Gigartinales, Rhodophyta) to different light qualities

G. Kräbs and C. Wiencke. 2005. Photosynthesis, photosynthetic pigments and mycosporine-like amino acids after exposure of the marine red alga Chondrus crispus (Gigartinales, Rhodophyta) to different light qualities. Phycologia 44: 95–102.Low light–adapted laboratory-grown thalli of Chondrus crispus were transferred into shallow water and exposed to the natural irradiance prevailing in May–June at Helgoland (German Bight). A set of cut-off filters was used to study the wavelength-dependent response of photosynthesis as well as pigment and mycosporine-like amino acid (MAA) formation. Due to the higher natural irradiance and in some cases additional ultraviolet (UV) radiation, a depression of maximal quantum yield coinciding with a decline in maximal electron transport rate could be observed during the first days of the experiment. Faster recovery of maximal quantum yield and maximal electron transport rate might be related to the increase in lutein and α- and β-carotene concentration. In addition, higher rates of electron transport might be supported by increased chlorophyll a concentration. In parallel, MAA concentration increased resulting in an effective UV sunscreen. Our data suggest that lutein and α- and β-carotene are involved in the recovery of photochemical capacity, whereas MAAs provide a UV sunscreen, which may reduce the light available within the algae to long-wavelength UV-A and photosynthetically active radiation.

Physiological profiles of restricted endemic plants and their widespread congenors in the North Queensland wet tropics, Australia

Little is known about causes of endemic rarity in plants. This study pioneered an approach that determined environmental variables in the rainforest habitat and generated physiological profiles for light, water, and nutrient relations for three endemically restricted versus widespread congeneric species' pairs. We found no overall consistent differences in the physiological variables between the group of restricted species and the group of widespread species, and congeneric species pairs were therefore examined individually. Availability of soil nutrients did not differ between restricted–widespread species sites suggesting that species grow under comparable nutrient conditions. Under ambient and manipulated higher light conditions, widespread Gardenia ovularis had a greater photosynthetic activity than restricted Gardenia actinocarpa suggesting that the two species differ in their photosynthetic abilities. Differences between Xanthostemon species included lower photosynthetic activity, higher transpiration rate, and a higher foliar manganese concentration in restricted Xanthostemon formosus compared to widespread Xanthostemon chrysanthus. It is suggested that X. formosus is restricted by its high water use to its current rainforest creek edge habitat, while X. chrysanthus grows in a range of environments, although naturally found in riparian rainforest. Restricted Archidendron kanisii had higher electron transport rates, greater dissipative capacity for removal of excess light, and more efficient investment of nitrogen into photosynthetic components, than its widespread relative Archidendron whitei. These observations and previous research suggest that restricted Archidendron kanisii is in the process of expanding its range. Physiological profiles suggest a different cause of rarity for each species. This has implications for the conservation strategies required for each species.

The photosynthetic light response of Halophila stipulacea growing along a depth gradient in the Gulf of Aqaba, the Red Sea

Photosynthetic responses to irradiance were measured on the seagrass Halophila stipulacea growing along an extensive depth gradient (7–30 m) in the Gulf of Aqaba on three occasions between January and August 2000. Plant samples were collected for morphological and anatomical (chloroplast clumping) characterizations of the leaves and analysis of carbon and nitrogen content. The highest electron transport rates, calculated firstly from the quantum yield and irradiance and then after accounting for the proportion of incident irradiance absorbed, were found in plants at the upper depth limit of 7 m. At this depth, electron transport rates were highest in summer (35 μmol electrons m −2 s −1) compared to spring and winter (20 μmol electrons m −2 s −1). Relative rates of electron transport at 30 m were 60% lower than those at 7 m in winter and spring and 80% lower in summer. The irradiance at the onset of light saturation was also highest in the shallow growing plants, indicative of successful adaptations to high irradiance. Chloroplast clumping in leaves in situ in shallow water reduced the amount of light absorbed to 55% compared to 85% at 30 m. Despite adequate light for photosynthesis, there was evidence for lower biomass at sampling depths of 7 and 17 m compared to 24 and 30 m. The potential for shallow plants to photosynthesise at high irradiances correlated with their ability to clump chloroplasts, suggesting that factors other than high irradiances are likely to limit growth at shallow depths. Low tissue nitrogen content (<1.8%) indicates that nutrient availability also needs to be considered when determining constraints on growth of H. stipulacea in shallow water in the study region.

Performance and Photosynthetic Ecophysiology of Three Photo‐Types of Dioscorea zingiberensis under Differing Light Intensities

Abstract: The performance and photosynthetic ecophysiology of three photo‐types of Dioscorea zingiberensis were studied. The three types are designated DzTL, DzTM and DzTH, according to their adaptation to low (LL), medium (ML) and high (HL) light intensities, respectively. Under LL (23 ‐ 55 μmol m‐2 s‐1) and simulated natural light (SNL), DzTM grows well with increased longevity, and green leaves which are unspotted; while its leaves became small, light yellow and short‐lived under HL (550 ‐ 850 μmol m‐2 s‐1). In contrast, under LL the leaves of DzTH were very large, spotted, light yellow and short‐lived; while they were small, green and long‐lived under HL. Under HL, DzTH had a much higher chlorophyll content than DzTM. Under LL, DzTM and DzTL had a higher Chl content than DzTH. Among the three types, DzTM had the highest peroxidase activity. DzTL had a higher electron transport rate (ETR), maximal quantum yield (MQY) and effective quantum yield (EQY) than DzTH and DzTL under LL, while DzTH had higher ETR, MQY and EQY than the other two types under ML and HL. Therefore, three different photo‐types can be characterized according to their adaptation to LL, ML and HL: DzTL, DzTM and DzTH, respectively.

Resistance is useful: diurnal patterns of photosynthesis in C3 and crassulacean acid metabolism epiphytic bromeliads.

This paper originates from a presentation at the IIIrd International Congress on Crassulacean Acid Metabolism, Cape Tribulation, Queensland, Australia, August 2001 Diurnal patterns of photosynthesis in response to environmental variables were investigated in an obligate C3 and a facultative C3-crassulacean acid metabolism (CAM) bromeliad species. A midday depression of photosynthesis occurred in both C3 groups, mediated as a decrease in stomatal conductance in response to increased vapour pressure difference. The response was associated with a reduction in Rubisco activation state during the period of maximum photon flux density. In contrast, the switch to CAM resulted in a strong shift in the pattern of Rubisco carbamylation, with full enzyme activation delayed until the midday period. For the first time it is demonstrated that the pattern of Rubisco activation differs between C3 and CAM plants of the same species under identical conditions. Despite large differences in Rubisco content between C3 and CAM plants, neither the amount of Rubisco or enzyme activity is thought to be limiting for photosynthesis, and it is suggested that Rubisco may function as a nitrogen store. Extreme CO2 diffusion limitation resulted in low rates of atmospheric CO2 assimilation that were associated with high rates of photosynthetic electron transport, and it is likely that photorespiration constitutes a significant electron sink over the entire diurnal course. Leaf morphological and physiological adaptations to drought stress are necessary for the epiphytic lifestyle but limit CO2 assimilation and confound the likelihood of high productivity.

Mitochondrial alternative oxidase acts to dampen the generation of active oxygen species during a period of rapid respiration induced to support a high rate of nutrient uptake.

When wild type (wt) tobacco (Nicotiana tabacum L. cv. Petit Havana SR1) suspension cells were grown under phosphate (P) limitation, they contained large amounts of mitochondrial alternative oxidase (AOX). When these cells were resupplied with P, there was a large, immediate and sustained stimulation of respiration to support a period of rapid P uptake. Two lines of evidence suggest that the abundant level of AOX present in wt cells contributed to this stimulated rate of respiration. First, when P-limited transgenic antisense tobacco cells (AS8) lacking AOX were resupplied with P, the stimulation of respiration was much less dramatic even though these cells displayed similar rates of P uptake. Second, while the stimulated rate of respiration in AS8 cells was insensitive (as expected) to the AOX inhibitor n-propyl gallate (nPG), much of the stimulated rate of respiration in wt cells could be inhibited by nPG. Given the non-phosphorylating nature of AOX respiration, wt cells required higher rates of electron transport to O2 than AS8 cells to support similar rates of P uptake. The utilization of AOX by wt cells during P uptake was apparently not occurring because the cytochrome (Cyt) pathway alone could not fully support the rate of P uptake, as the respiration of cells lacking AOX (either untreated AS8 cells or wt cells treated with nPG) supported similar rates of P uptake as wt cells with abundant AOX. Rather, we provide in vivo evidence that the utilization of AOX during the period of high respiration supporting P uptake was to dampen the mitochondrial generation of active oxygen species (AOS).

Chlorophyll fluorescence as a selection tool for cold tolerance of photosynthesis in maize (Zea mays L.)

The possibility of using quenching analysis of chlorophyll a fluorescence as a selection tool for improving the cold tolerance of maize was investigated in six genotypes differing greatly in the ability to develop a competent photosynthetic apparatus at low temperature. Upon gradual cooling, measurements of the quantum yield of electron transport (Φ PSII ) indicated that leaves of tolerant genotypes, that developed at suboptimal temperature (15°C), maintained higher rates of electron transport than leaves of sensitive genotypes. This difference was largely due to the ability of the tolerant plants to keep higher efficiency of excitation energy capture by open photosystem II reaction centres (F' v /F' m ). The absence of genotypic differences in leaves that developed at optimal temperature indicates that the trait is not expressed constitutively, but relies on adaptation mechanisms. Furthermore, the genotypic difference was not expressed under increasing illumination at 15°C and 25°C suggesting that the trait is also low-temperature-specific and is not expressed solely in response to increasing excess light energy. Applying the method to flint and dent breeding population led to a substantial increase (up to 31%) in the photosynthetic capacity of hybrids between selected F 3 inbreeding families grown at suboptimal temperature, demonstrating that the method is an efficient selection tool for improving the cold tolerance of maize through breeding.

Ecophysiology of the C3-CAM intermediate Clusia minor L. in Trinidad: seasonal and short-term photosynthetic characteristics of sun and shade leaves

The seasonal changes in crassulacean acid metabolism (CAM) activity in response to daily integrated photon flux density (PFD) and precipitation were compared in sun and shade leaves of the C3-CAM intermediate tree Clusia minor L. Measurements of CAM activity (ΔH+) showed that maximum leaf acidity consistently occurred 4 h after dawn, suggesting that new sampling procedures need to be adopted in order to quantify CAM in Clusia species. Whilst exposed leaves responded to intermittent dry conditions, shaded leaves showed a clear induction of CAM activity as conditions became drier. The magnitude of CAM activity correlated well with daily integrated PFD, such that the extent of decarboxylation of organic acids was consistently associated with increased acidification during the subsequent dark period. Over two sampling days, both sun and shade leaves exhibited the four phases of CAM, although PEPc remained active throughout phase II with the result that 50% of the maximum leaf acidity in shade leaves was accumulated during this time. During phase III, internal CO2 supply was augmented by substantial citrate decarboxylation, in addition to malic acid. Chlorophyll fluorescence characteristics were dominated by high rates of PSII electron transport, together with an extremely high potential for thermal dissipation, such that excess light was maintained within safe limits at times of maximum PFD. Photochemical stability was maintained by matching supply and demand for internal CO2: in the morning, C3 and C4 carboxylation processes were regulated by extended PEPc activity, so that decarboxylation was delayed until temperature and light stress were highest at midday.

Ecophysiology of the C3-CAM intermediate Clusia minor L. in Trinidad: seasonal and short-term photosynthetic characteristics of sun and shade leaves

The seasonal changes in crassulacean acid metabolism (CAM) activity in response to daily integrated photon flux density (PFD) and precipitation were compared in sun and shade leaves of the C 3 -CAM intermediate tree Clusia minor L. Measurements of CAM activity (ΔH + ) showed that maximum leaf acidity consistently occurred 4 h after dawn, suggesting that new sampling procedures need to be adopted in order to quantify CAM in Clusia species. Whilst exposed leaves responded to intermittent dry conditions, shaded leaves showed a clear induction of CAM activity as conditions became drier. The magnitude of CAM activity correlated well with daily integrated PFD, such that the extent of decarboxylation of organic acids was consistently associated with increased acidification during the subsequent dark period. Over two sampling days, both sun and shade leaves exhibited the four phases of CAM, although PEPc remained active throughout phase II with the result that 50% of the maximum leaf acidity in shade leaves was accumulated during this time. During phase III, internal CO 2 supply was augmented by substantial citrate decarboxylation, in addition to malic acid. Chlorophyll fluorescence characteristics were dominated by high rates of PSII electron transport, together with an extremely high potential for thermal dissipation, such that excess light was maintained within safe limits at times of maximum PFD. Photochemical stability was maintained by matching supply and demand for internal CO 2 : in the morning, C 3 and C 4 carboxylation processes were regulated by extended PEPc activity, so that decarboxylation was delayed until temperature and light stress were highest at midday.

Photosynthesis and photoinhibition in a tropical alpine giant rosette plant, Lobelia rhynchopetalum.

Carbodioxide uptake, oxygen evolution and chlorophyll fluorescence of leaves of Lobelia Lobelia rhynchopetalum Hemsl., a giant rosette plant of the tropical alpine regions of Ethiopia, were studied under field conditions at 4000 m above sea level. Our objective was to investigate the photosynthetic adaptation to the combination of wide fluctuation in diurnal temperature, high photon flux densities (PFD) and low CO2 partial pressure encountered in these regions. At an ambient CO2 partial pressure of c. 17 Pa, maximal rates of CO2 uptake were low, ranging between 4 and 6 μmol m-2 s-1 . Such rates, however, required high PFDs and were observed only at levels of 1500 μmol photons m-2 s-2 . Carbon dioxide uptake was significantly inhibited when PFD was ≤ 2000 μmol photons m-2 s-1 . On the other hand, at saturating CO2 levels, maximal photosynthetic oxygen evolution was higher (30 μmol C2 m-2 s-1 ). saturating at the same PFD as CO2 uptake. Quantum efficiency of CO2 uptake (0.006 mol CO2 mol photons-1 , at high altitude and a low CO, partial pressure of 17 Pa) and even of oxygen evolution under CO2 -saturating conditions in the leaf O2 electrode (0.05 mol O2 mo) photons-1 ) indicated reduced photosynthetic efficiency. Electron transport rate (ETR) was strongly correlated with the leaf temperature. Non-photochemical quenching (NPQ) responded inversely to leaf temperature and stomatal conductance. The results indicated that in the morning, when the sun irradiates the partly frozen leaves with closed stomata, NPQ is the principal mechanism by which Lobelia leaves protect their photosynthetic apparatus. However, during the day, the predominant upright inclination of the leaves significantly contributes to protecting the leaves from excess light absorption. A comparison of the chlorophyll fluorescence of young and old leaves revealed that the former had high ETR and quantum efficiency of photosynthetic electron transport but a lower capacity for NPQ. Extremely high NPQ values but low ETR and low quantum efficiency were recorded for the old leaves. Thus, in the course of maturation the leaves apparently lose photosynthetic efficiency but increase their capability for protective non-photochemical quenching.

Photosynthetic Characteristics of Sun Versus Shade Plants of Encelia farinosa as Affected by Photosynthetic Photon Flux Density, Intercellular CO2 Concentration, Leaf Water Potential, and Leaf Temperature

Limitations to photosynthesis were examined for Encelia farinosa Toney et A.Gray, a common C3 sub-shrub in arid regions of south-westem United States, for plants grown in full sunlight and those shaded to 40% of full sunlight. The initial slopes of CO2 assimilation (A) versus intercellular CO2 concentration curves were similar for sun and shade plants at low photosynthetic photon flux density (PPFD) but higher for sun plants as the PPFD increased, indicating a greater limitation by carboxylation capacity in shade plants. Sun plants had higher electron transport rates but a lower ratio of electron transport capacity to carboxylation capacity (Vmax); the ratio was inversely proportional to mesophyll conductance for both sun and shade plants. Dark respiration decreased with decreasing leaf water potential (Ψ1) in sun plants but remained unchanged in shade plants; day respiration was little affected by PPFD for both sun and shade plants. Stomatal conductance (gs) was similar for sun and shade plants under high soil-moisture conditions but higher in sun plants as Ψ1 decreased; for all data considered together, changes in the leaf-air vapour pressure difference accounted for 71% of the variation in gs. The lower A for shade plants of E. farinosa apparently resulted from a lower Vmax, as well as a lower gs when plants were under water stress.

Alteration of the iron-sulfur cluster composition of Escherichia coli dimethyl sulfoxide reductase by site-directed mutagenesis.

We have used site-directed mutagenesis to alter the [Fe-S] cluster composition of Escherichia coli dimethyl sulfoxide (DMSO) reductase (DmsABC). The electron-transfer subunit (DmsB) of this enzyme contains 16 Cys residues arranged in 4 groups (I-IV) which provide ligands to 4 [4Fe-4S] clusters [Cammack, R., & Weiner, J. H. (1990) Biochemistry 29, 8410-8416]. Strong homologies exist between these Cys groups and the four Cys groups of the electron-transfer subunit (NarH) of E. coli nitrate reductase (NarGHJI), which contains a [3Fe-4S] cluster in addition to multiple [4Fe-4S] clusters. The Cys group primarily involved in providing ligands to the [3Fe-4S] cluster of NarH has a Trp residue at a position equivalent to Cys102 of DmsB. We have mutated Cys102 to Trp, Ser, Tyr, and Phe and have investigated the altered enzymes in terms of their enzymatic activities and EPR properties. The mutant enzymes do not support electron transfer from menaquinol to DMSO, although they retain high rates of electron transport from reduced benzyl viologen to DMSO. The mutations cause major changes in the EPR properties of the enzyme in the fully reduced and oxidized states. In the oxidized state, new species are observed in all the mutants; these have spectral features comprising a peak at g = 2.03 (gz) and a peak-trough at g = 2.00 (gxy). The temperature dependencies, microwave power dependencies, and spin quantitations of these species are consistent with the Trp102, Ser102, Phe102, and Tyr102 mutations causing conversion of one of the [4Fe-4S] clusters present in the wild-type enzyme into [3Fe-4S] clusters in the mutant enzymes.

Streptomycin accumulation by Bacillus subtilis requires both a membrane potential and cytochrome aa3

Cytochrome aa3 concentrations in the cytoplasmic membrane of Bacillus subtilis were altered by growth conditions, and the effects on the membrane potential (delta psi) in whole cells were measured. When cytochrome aa3 was absent, the magnitude of delta psi was not diminished by comparison with the delta psi measured in cells containing normal cytochrome aa3 concentrations. In addition, the energy-dependent uptake of proline and glutamate was comparable at both cytochrome aa3 concentrations. However, in the cytochrome aa3-deficient cell preparation, accumulation of the aminoglycoside antibiotic streptomycin was much lower than that of the cytochrome aa3-sufficient cells. When cells were cultured under conditions that stimulated higher than normal concentrations of cytochrome aa3, delta psi was also increased, and enhanced streptomycin accumulation was observed. Phenazine methosulfate-ascorbate was used both in delta psi measurements and in uptake studies to provide high rates of electron transport and maximal delta psi values. These results, taken together with those previously published (A. S. McEnroe and H. W. Taber, Antimicrob. Agents Chemother. 26:507-512, 1984) suggest that the uptake of streptomycin by B. subtilis requires adequate levels both of delta psi and cytochrome aa3.

Influence of detergent polar and apolar structure upon the temperature dependence of beef heart cytochrome c oxidase activity.

The temperature dependence of lipid-depleted beef heart cytochrome c oxidase activity was studied in a series of chemically homogeneous detergents. The detergents that were tested included C10 to C18 maltosides, C8 to C12 glucosides, C8 to C16 Zwittergents, and C12 poly(oxyethylene) ethers. The observed rates of electron transport were dependent upon the structure of the polar head group and the length of the hydrocarbon tail. Of the detergents tested, the alkyl maltosides were the best in terms of both high rates of electron transport and superior enzyme stability. With the maltosides, changing the length of the alkyl tail affected the activity of cytochrome c oxidase in a manner quite similar to that reported with synthetic phosphatidylcholines and phosphatidylethanolamines [Vik, S. B., & Capaldi, R. A. (1977) Biochemistry 16, 5755-5759], suggesting that the alkyl maltosides can mimic some of the features of the membrane environment. In each of the detergents, the activation enthalpy (determined from the slope of an Arrhenius plot) was nearly identical, suggesting that the same electron-transfer step within cytochrome c oxidase is rate limiting. This result has been interpreted as evidence for the existence of two or more conformers of cytochrome c oxidase, one of which is significantly more active than the other(s). The enzyme turnover number, which changes by 2 orders of magnitude depending upon the structure of the bound detergent, may reflect the ability of each detergent to alter the equilibrium between the active and nearly inactive conformers.

Characterization of a spinach photosystem II core preparation isolated by a simplified method

A photosystem II core complex from spinach exhibiting high rates of electron transport was obtained rapidly and in high yield by treatment of a Tris-extracted, O 2-evolving photosystem II preparation with the detergent dodecyl-β- d-maltoside. The core complex was essentially free of light-harvesting chlorophyll-protein and photosystem I polypeptides, and was highly enriched in the polypeptides associated with the photosystem II reaction center (45 and 49 kDa), cytochrome b 559, and three polypeptides in the region 32–34 kDa. The photosystem II core complex contained two chlorophyll-proteins which had a slightly higher apparent molecular mass than CPa-1 and CPa-2. Additionally, a high-molecular-mass chlorophyll-protein complex termed CPa ∗ was observed, which exhibited a low fluorescence yield when illuminated with ultraviolet light. This observation suggests that CPa ∗ contains a functionally efficient quencher of chlorophyll fluorescence, possibly P680.

Photosynthesis in Tall Fescue

Previous work in our laboratory (Krueger, Miles 1981 Plant Physiol 68: 1110-1114) indicated that a decaploid genotype (I-16-2) of tall fescue (Festuca arundinacea Schreb.) which exhibits unusually high net photosynthesis rates also had high potential rates of photosynthetic electron transport through photosystem I (PSI) compared to the typical hexaploid genotype (V6-802). Analysis of electron transport activity revealed that the oxidizing side of PSI as the major site of difference. Examination of the whole thylakoids and subchloroplast particle protein components of the common hexaploid and the decaploid genotypes had major polypeptide differences at 30, 21, and 12.5 kilodaltons. These differences could not be assigned to a specific physiological function in PSI. The decaploid had increased P(700) and plastocyanin content on a chlorophyll basis. Antibodies raised against fescue plastocyanin were used to quantitate plastocyanin in crude (Triton X-100) solubilized extracts of plant material. Results showed that the decaploid had 16% and 40% more plastocyanin on a weight and area basis, respectively. The antibodies did not inhibit electron transport (diaminodiurene to methyl viologen) in isolated thylakoids strengthening the hypothesis of plastocyanin as an internal mobile electron shuttle. The trend of inhibition of plastocyanin by KCN was similar in the two genotypes but the decaploid had 15 to 20% higher rates of electron flow under nearly all inhibiting conditions.

Photosynthesis in Fescue

Chloroplasts isolated from tall fescue, Festuca arundinacea Schreb., showed high rates of electron transport, comparable to rates observed for spinach chloroplasts.Chloroplasts were well coupled and rates of electron transport from water to methyl viologen (photosystem II and I) were increased two to five times when ADP and inorganic phosphate or methylamine (uncoupler) were added to the reaction mixture. Ratios of P:2e for photosystem II plus I were found to be near 1.2. Electron transport rates from water to p-phenylenediamine or 2,6-dichlorobenzoquinone (photosystem II) were over 300 micromoles O(2) per hour per milligram chlorophyll, while P:2e ratios were found to be over 0.5. The highest rates of electron transport were found in electron flow from diaminodurene to methyl viologen (photosystem I) and P:2e ratios remained near 0.5.Light intensity saturation curves for photosystem II and I, as well as the photosystems independently, resembled curves for spinach, with saturation of electron transport in photosystem I and photosystem II separately occurring at 35% of the available light intensity (6000 microeinsteins per square meter per second). Photosystem II and I in sequence were saturated at about half this light intensity.