Ultrastructure,polypeptide composition and photochemical activity of chloroplasts during foliar senescence of a non-yellowing mutant genotype of Festuca pratensis Huds.

A study was made of the structure and function of senescent chloroplasts from a non-yellowing (NY) mutant of Festuca pratensis. Electron microscopy suggested that the stroma matrix was destroyed but that thylakoid membranes persisted in a loose, unstacked condition. By contrast, chloroplasts from the normal (Y) genotype lost both stroma and recognizable thylakoid systems. Fraction 1, the major protein of the stroma, disappeared from Y and NY at similar rates during senescence. The activities of photosystems I and II from NY also declined at a similar rate to Y photosystems. Polypeptides of chloroplast membranes were separated by SDS gel electrophoresis into at least 30 components. There was considerable heterogeneity in rates of breakdown of the different protein species of the membranes. Of the five major polypeptide components, two had kinetics of breakdown similar to those of stroma proteins and were lost from NY and Y at about the same rate, whereas the remaining three (one of which was tentatively identified as the apoprotein of the light-harvesting chlorophyll-protein complex) were more stable in NY than in Y. These results are discussed in relation to the mechanism and function of chloroplast disintegration during leaf senescence.Abbreviations RuDPC ribulose diphosphate carboxylase - NY and Y non-yellowing and normal genotypes of Festuca, respectively - PSI and PSII photosystems I and II, respectively - SDS sodium dodecyl sulphate - MW molecular weight - CF coupling factor     

C4 photosynthesis in Spartina townsendii at low and high temperatures

The metabolism of ¹⁴CO₂ in the cool temperate saltmarsh grass Spartina townsendii was investigated in plants grown in their natural habitats at two temperatures. Both in the spring at 10°C and in the late summer at 25°C radioactivity was initially incorporated into the organic acids malate and aspartate and then transferred to 3-phosphoglycerate in the manner characteristic of the C₄ pathway of photosynthesis. Metabolism was not disrupted at the lower temperature as in some C₄ plants. Radioactivity was transferred more slowly from malate into alanine, glycine and serine at 10°C, but sugars were labelled equally at both temperatures.     

Intact chloroplast electron flow. Effects of ribose 5-phosphate

Additions of ribose 5-phosphate to intact spinach chloroplasts were used to probe the effects of ADP regeneration on pH-gradient formation and electron-transfer reactions. In weakly illuminated chloroplasts, the ATP/ADP ratio dropped by 64% and the transthylakoid pH gradient decreased by a minimum of 0.2 units in response to ribose 5-phosphate. Nitrite reduction increased 2-fold while, under conditions of cyclic electron flow, the half-time for cytochrome f reduction decreased by a factor of two from 4.1 to 1.9 ms. The results suggest that metabolic ATP consumption, during the conversion of ribulose 5-phosphate to ribulose 1,5-bisphosphate, enhances electron transfer between plastohydroquinone and cytochrome f through decreases in the transthylakoid pH gradient caused by phosphorylation of ADP.     

Oxidation and reduction of plastoquinone by photosynthetic and respiratory electron transport in a cyanobacterium Synechococcus sp.

The I-D dip, an early transient of the fluorescence induction, was examined as a means to monitor redox changes of plastoquinone in cells of a cyanobacterium, Synechococcus sp. That the occurrence of the dip depends upon the reduced state of the plastoquinone pool was indicated by observations that 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone and 3-(3,4-dichlorophenyl)-1,1-dimethylurea did not affect the initial rise to I but abolished the subsequent decline from I to D and that illumination of the cells with light 1, prior to fluorescence measurements, eliminated the transient. The I-D dip was prominent in freshly harvested cells containing abundant endogenous substrates, disappeared slowly as the cells were starved by aeration but reappeared on addition of fructose to the starved cells in the dark. The dip that had been induced by a brief illumination of the starved cells with light 2 was rapidly diminished in the dark and KCN inhibited the dark decay of the transient. The results indicate that plastoquinone is reduced with endogenous as well as exogenous substrates and oxidized by a KCN-sensitive oxidase in the dark, thus providing strong support for the view that plastoquinone of photosynthetic electron transport also functions in respiration. In addition, the occurrence of a cyclic pathway of electrons from Photosystem I to plastoquinone, possibly via ferredoxin or NADP, was suggested. Several lines of evidence indicate that, under a strong light 2, Photosystem I-dependent oxidation of plastoquinone predominates over Photosystem II-dependent reduction of the quinone in the cyanobacterium which contains Photosystem I more abundantly than Photosystem II.     

Photoreactions of cytochrome b6 in systems using resolved chloroplast electron-transfer complexes

Photoreactions of cytochrome b₆ have been studied using resolved chloroplast electron-transfer complexes. In the presence of Photosystem (PS) II and the cytochrome b₆-f complex, photoreduction of the cytochrome can be observed. No soluble components are required for this reaction. Cytochrome b₆ photoreduction was found to be inhibited by quinone analogs, which inhibit at the Rieske iron-sulfur center of the cytochrome complex, by the addition of ascorbate and by depletion of the Rieske center and bound plastoquinone from the cytochrome complex. Photoreduction of cytochrome b₆ can also be demonstrated in the presence of the cytochrome complex and PS I. This photoreduction requires plastocyanin and a low-potential electron donor, such as durohydroquinone. Cytochrome b₆ photoreduction in the presence of PS I is inhibited by quinone analogs which interact with the Rieske iron-sulfur center. These results are discussed in terms of a Q-cycle mechanism in which plastosemiquinone serves as the reductant for cytochrome b₆ via an oxidant-induced reductive pathway.     

The influence of carotenoids on the conformation of chlorophyll-protein complexes isolated from the cyanobacterium Plectonema boryanum. Absorption and circular dichroism study

Thylakoid membranes of the cyanobacterium Plectonema boryanum solubilized with Triton X-100 can be resolved into three fractions of pigment-protein complexes (Hladík, J. and Sofrová, D. (1981) Photosynthetica 15, 490–503). Fraction I contained relatively the highest amount of carotenoids as well as monomeric forms of chlorophyll a, Fractions II and III contained chlorophyll-protein complexes with a characteristic exciton-split circular dichroism in the red region. It has been shown that fraction III is an oligomeric form of the chlorophyll-protein complex of fraction II. Circular dichroism spectra indicate that, different from fraction II, fraction III contains specifically oriented and space-fixed molecules of carotenoids. Thermal dissociation of fracion III to fraction II is accompanied by disappearance of the positive circular dichroism effect of carotenoids in the 500–550 nm region, thus causing deorganization of the carotenoids, proceeding in parallel to the geometrical rearrangement of chlorophyll molecules. Extraction of the carotenoids of fraction III with heptane is acompanied by dissociation of fraction III. We assume that the observed effects are due to binding of the two pigments to the protein component of the complex and that carotenoids can mediate a part of the interactions which stabilize the structure of pigment-protein complexes. Thus, besides the light-harvesting and protective functions, carotenoids can also play a structural role.     

Light scattering,chlorophyll fluorescence and state of the adenylate system in illuminated spinach leaves

Scattering of green light and chlorophyll fluorescence by spinach leaves kept in a stream of air or nitrogen were compared with leaf adenylate levels during illumination with blue, red or far-red light. Energy charge and ATP-ADP ratios exhibited considerable variability in different leaves both in the dark and in the light. Variability is explained by different possible states of the reaction oxidizing triose phosphate or reducing 3-phosphoglycerate. Except when oxygen levels were low, there was an inverse relationship between light scattering and chlorophyll fluorescence during illumination with blue or red light. When CO₂ was added to a stream of CO₂-free air, chlorophyll fluorescence increased, sometimes after a transient decrease, and both light scattering and leaf $\text{ATP}\text{ADP}$ ratios decreased. Similar observations were made when air was replaced by nitrogen under blue or high-intensity red light. Under these conditions, over-reduction caused inhibition of electron transport and phosphorylation in chloroplasts. However, when air was replaced by nitrogen during illumination with low-intensity red light or far-red light, light scattering increased instead of decreasing. Under these light conditions, $\text{ATP}\text{ADP}$ ratios were maintained in the light. They decreased drastically only after darkening. Although $\text{ATP}\text{ADP}$ ratios responded faster than light scattering or the slow secondary decline of chlorophyll fluorescence due to illumination, it appeared that in the steady state, light scattering and chlorophyll fluorescence are useful indicators of the phosphorylation state of the leaf adenylate system at least under aerobic conditions, when chloroplast and extrachloroplast adenylate systems can effectively communicate.     

Role of external carbonic anhydrase in light-dependent alkalization byFucus serratus L. andLaminaria saccharina (L.) Lamour. (Phaeophyta)

It has been proposed that many marine macroalgae are able to utilize HCO ₃ ^– for photosynthesis and growth, and that energy-dependent ion pumping is involved in this process. We have therefore studied the light-dependent alkalization of the surrounding medium by two species of marine macroscopic brown algae,Fucus serratus L. andLaminaria saccharina (L.) Lamour. with the aim of investigating the role of extracellular carbonic anhydrase (EC 4.2.1.1.) in the assimilation of inorganic carbon from the seawater medium. In particular, the influence of membrane-impermeable or slowly permeable carbonic-anhydrase inhibitors on the rate of alkalization of the seawater has been investigated. Inhibition of the alkalization rate occurred in both species at an alkaline pH (pH 8.0) but no inhibition was observed at an acidic pH (pH 6.0). The alkalization was found to be light-dependent and inhibited by 3-(3,4-dichlorophenyl)-1, 1-dimethylurea and, thus, correlated with photosynthesis. Alkalization by macroalgae has previously been shown to be proportional to inorganiccarbon uptake. We suggest that alkalization of the medium at alkaline pH in both of the species examined is mainly the consequence of an extracellular reaction. The reaction is catalyzed by extracellular carbonic anhydrase which converts HCO ₃ ^– to OH^– and CO₂; CO₂ is then taken up through the plasmalemma. However, we do not exclude the involvement of other mechanisms of inorganic-carbon uptake.Abbreviations AZ acetazolamide - CA carbonic anhydrase - CAext extracellular carbonic anhydrase - C_i inorganic carbon - DBS dextran-bound sulfonamide - DCMU 3-(3,4-dichloro-phenyl)-1,1-dimethylurea - PPFD photosynthetic photon flux density This study was carried out with financial support by SAREC (Swedish Agency for Research Cooperation with Developing Countries), Carl Trygger's Fund for Scientific Research (Sweden), SJFR (Swedish Council for Forestry and Agricultural Research) and CICYT (Spain). Z. Ramazanov is an invited professor of Ministerio de Educación y Ciencia, Spain.     

Canopy structure,nitrogen distribution and whole canopy photosynthetic carbon gain in growing and flowering stands of tall herbs

Using a combination of mathematical modeling and field studies we showed that in dense stands of growing herbaceous plants the vertical pattern of leaf nitrogen distribution resembles the pattern of mean light attenuation in the stand and hence tends to maximize total daily photosynthetic carbon gain of the whole stand. Flowering represents a strong sink of nitrogen away from the photosynthetic apparatus and in herbs like Solidago altissima it induces leaf shedding. We studied both the effect of nitrogen reallocation and leaf shedding on the whole canopy photosynthesis and changes in leaf nitrogen distributions in stands moving from the growing to the flowering stage. Despite a decrease in leaf area index and total nitrogen available for photosynthesis in the flowering stand, the leaf nitrogen distribution here also leads to an almost maximum canopy photosynthesis. In both the growing and the flowering stands the leaf area index was higher than calculated optimum values. It is pointed out that this should not necessarily be interpreted as non-adaptive.     

ESR and ENDOR of primary reactants in photosynthesis

The primary reactants in photosynthesis are defined as the chemical entities on which charges are generated and stabilized after capture of a photon by the photochemical trap: PIX ^hv P ^* IX P ⁺ I ^– X P ⁺ IX ^–, where P stands for the primary electron donor, P ^* for its excited singlet state, I for the first (ESR-detectable) electron acceptor and X for the secondary acceptor complex. The ESR and ENDOR experiments which have played a rÔle in the identification and characterization of P, I, and X in the bacterial and plant photosystems are comprehensively reviewed. The structural and kinetic information obtained with magnetic resonance techniques are integrated with results obtained with optical spectroscopy to give a unified picture of the pathway of primary photochemistry in photosynthesis. Nomenclature of Primary Reactants In the interest of uniformity this review introduces a nomenclature of the primary reactants that deviates in some respects from the commonly used labels. The nametags used here and listed below are abbreviations of the molecules that are identified as primary reactants, with the exception of the donors, for which I have retained the commonly accepted designation. Photosystems: PS 1, photosystem 1 of plants; PS 2, photosystem 2 of plants; pBPS, the photosystem of purple bacteria; gBPS, ditto of green bacteria. P: Primary donors: P700 (PS 1), P680 (PS 2), P860 (generic label for BChl a containing purple bacteria), P960 (generic label for BChl b containing purple bacteria), P840 (generic name for green bacteria). I: First acceptors: Chl a (PS 1), Ph a (PS 2), BPh a,b (pBPS). X: Secondary acceptors: F _x (PS 1), pQ ₁ (PS 2), uQ ₁ or mQ ₁ (pBPS), B (gBPS). Tertiary acceptors: F _A,B (PS 1), pQ ₂ (PS 2), uQ ₂ (pBPS), F ₁ (gBPS).This paper is based on a lecture given at the Joint Meeting of the Belgium, German (FRG), and Netherlands Societies for Biophysics, Aachen 1980