Pigment-protein complexes of illuminated etiolated leaves

Photoconversion of protochlorophyllide in etiolated leaves of Avena sativa L., var. Pennal or Peniarth and Phaseolus vulgare L., var. `The Prince' results in the sequential appearance of spectrally distinct chlorophyllide complexes (Chlide 678, 684, and 672). This paper reports on the generation of similar forms in vitro, under controlled conditions, using well characterized etioplast membranes enriched in the enzyme protochlorophyllide reductase. Excess NADP<sup>+</sup> and NADPH stabilize complexes related to Chlide 678 and Chlide 684, respectively, whereas addition of exogenous Pchlide induces formation of a species related to Chlide 672. Evidence is provided to support the suggestion that Chlide 678 and Chlide 684 represent ternary complexes of the enzyme protochlorophyllide reductase, with Chlide and either NADP<sup>+</sup> (Chlide 678) or NADPH (Chlide 684). Chlide 672 is seen as `free' pigment dissociated from the enzyme. The role of Pchlide in this dissociation, observed spectroscopically as the `Shibata shift,' is discussed.     

Interaction between photosynthesis and respiration in illuminated leaves

Plants are sessile organisms that often receive excessive amounts of light energy. This excess energy can be exported from the chloroplasts and dissipated by the mitochondrial respiratory chain. The inner membrane of plant mitochondria possesses unique non-phosphorylating pathways, involving alternative oxidase and type II NAD(P)H dehydrogenases. There are accumulating amounts of evidence showing that these energy-wasteful pathways are up-regulated under excess light conditions, suggesting that they play key roles in efficient photosynthesis. Based on recent advances in our understanding about the metabolic interaction between chloroplasts and mitochondria, we discuss the importance of the respiratory chain for stabilizing the photosynthetic system.     

Amyloplast development in etiolated and ethylene-treated pea epicotyls

The amyloplasts found in the apical hook cells of etiolated pea (Pisum sativum L.) epicotyls were randomly distributed. Sedimentation of endodermal amyloplasts in the direction of gravity became apparent in the transition from the hook to the top of the main axis of the epicotyl. Cortical amyloplasts in this region were not, however, sedimented. These patterns of sedimentation could not be related to changes in amyloplast size, and it is proposed that cytoplasmic properties determine amyloplast behaviour.The differentiation of plastids in the hook differed between the amyloplast-containing endodermal cells and the cortical cells, in which amoeboid plastids predominated over amyloplasts. Amyloplasts disappeared from the cortical cells in the main axis of the epicotyl, but in the endodermal cells sedimented amyloplasts were found throughout the upper epicotyl.Etiolated epicotyls induced to grow horizontally by treatment with ethylene had a normal content of amyloplasts, sedimented in the direction of gravity.     

Phytochrome-mediated plastid development in etiolated pea stem apices

The effects of red and far-red light on growth and plastid development in the stem apices of etiolated pea seedlings have been examined. Changes were determined in various growth parameters (DNA, soluble protein and fresh weight) and also in the activities of the plastid-localized enzymes ribulose-1,5-bisphosphate carboxylase, NADP-glyceraldehyde-3-phosphate dehydrogenase and alkaline-1,6-bisphosphatase and the non-photosynthetic (cytoplasmic) enzymes NADP-isocitrate dehydrogenase, enolase and NAD-malate dehydrogenase. Changes in the amounts of Fraction I protein were also measured. Brief daily irradiation with low intensity red light increased growth 5·1–7·6-fold which was correlated with increases of about 3·5-fold in activities of the non-photosynthetic enzymes. The chloroplast enzymes, however, showed much greater increases in activity ranging from 15- to 91-fold. Fraction I protein increased 11·7-fold. These increases approached the levels attained in fully green leaves. All these responses were largely prevented by far-red light indicating that they were mediated by phytochrome. In experiments with red light given at daily intervals there was a lag of 24 hr before the initially very low activity of ribulose-1,5-bisphosphate carboxylase increased. Fraction I protein which was initially present in significant amounts showed a similar lag in its synthesis. However, for 3 days after the initial irradiation, the rate of increase of the enzymic activity was much greater than the rate of net synthesis of Fraction I protein. A single initial red irradiation was as effective as 3 daily irradiations in increasing the activity of ribulose-1,5-bisphosphate carboxylase. A fourth irradiation, however, gave an additional response which exceeded that of the single initial irradiation. It was shown that there was a rapid activation of ribulose-1,5-bisphosphate carboxylase by either continuous white or 3 min of red light. The red light response was slowly reversed in the dark. These results are discussed with particular emphasis on the relation between growth and plastid development in a phytochrome-mediated system.     

Chlorophyll formation and the development of photosynthesis in dark grown seedlings of Picea abies

Abstract Seeds of Picea abies were germinated and grown in either darkness or constant light. The chlorophyll content and photosynthetic carbon dioxide uptake of developing seedlings of different ages was determined. Ten-day-old dark grown seedlings showed an instant ability for photosynthetic carbon dioxide uptake and also formed further chlorophyll most rapidly upon subsequent illumination. These activities progressively diminished when the dark growth period was extended. Light grown seedlings reached a maximum chlorophyll level after 15 days growth, and this preceded maximal photosynthetic development.     

Methionine metabolism and ethylene formation in etiolated pea stem sections

Stem sections of etiolated pea seedlings (Pisum sativum L. cv. Alaska) were incubated overnight on tracer amounts of l-[U-(14)C]methionine and, on the following morning, on 0.1 millimolar indoleacetic acid to induce ethylene formation. Following the overnight incubation, over 70% of the radioactivity in the soluble fraction was shown to be associated with S-methylmethionine (SMM). The specific radioactivity of the ethylene evolved closely paralleled that of carbon atoms 3 and 4 of methionine extracted from the tissue and was always higher than that determined for carbon atoms 3 and 4 of extracted SMM.Overnight incubation of pea stem sections on 1 millimolar methionine enhanced indoleacetic acid-induced ethylene formation by 5 to 10%. Under the same conditions, 1 millimolar homocysteine thiolactone increased ethylene synthesis by 20 to 25%, while SMM within a concentration range of 0.1 to 10 millimolar did not influence ethylene production. When unlabeled methionine or homocysteine thiolactone was applied to stem sections which had been incubated overnight in l-[U-(14)C]methionine, the specific radioactivity of the ethylene evolved was considerably lowered. Application of unlabeled SMM reduced the specific radioactivity of ethylene only slightly.     

Haem and chlorophyll formation in etiolated and greening leaves of barley

The amounts of protochlorophyllide (P650) and protohaem were measured in ageing dark-grown barley leaves. Maximum amounts of P650 and protohaem were found in 6- to 8-day-old material after which P650 declined rapidly and protohaem more slowly. In leaves exposed to light maximum chlorophyll was produced in 6-day-old material with progressively less the older the leaves. Haem concentrations increased in seedlings of all ages exposed to light. A lag phase was observed for both chlorophyll and haem formation in leaves given a light treatment. Haem, however, showed a slight yet sig nificant decline as chlorophyll production commenced. The results indicate that chlorophyll and haem synthesis share a common pool of δ-aminolae vulinic acid (ALA). At a certain stage of development, the magnesium porphyrin pathway diverts precursors away from haem synthesis. It is only when the ALA synthesising system is well developed that the production of ALA can satisfy pathways to both haem and chlorophyll. The observed changes in haem under certain conditions suggest that, as in animal systems, haem levels may regulate porphyrin formation (chlorophylls) by controlling the supply of ALA.     

Quenching of the fluorescence emitted by P695-682 at room temperature in etiolated illuminated leaves

Chlorophyll(ide) fluorescence emission decreases at room temperature during completion of protochlorophyll(ide) reduction. The process responsible for this quenching is parallel to the P_688-676 P_695-682 transition. It proceeds equally well in darkness and in the light. It consists in a decrease of the fluorescence yield of chlorophyll(ide) in P_695-682. Apparently, room temperature P_695-682 fluorescence is regulated by a conjunction of factors such as energy transfers and photobiochemical activities.Abbreviations NADP nicotinamide-adenine dinucleotide phosphate - CPI chlorophyll-protein-complex I - CPII chlorophyll-protein-complex II Aspirant du Fond National de la Recherche Scientifique, Belgium     

Local weak light induces the improvement of photosynthesis in adjacent illuminated leaves in maize seedlings

To copy with highly heterogeneous light environment, plants can regulate photosynthesis locally and systemically, thus, maximizing the photosynthesis of individual plants. Therefore, we speculated that local weak light may induce the improvement of photosynthesis in adjacent illuminated leaves in plants. In order to test this hypothesis, maize seedlings were partially shaded, and gas exchange, chlorophyll a fluorescence and biochemical analysis were carefully assessed. It was shown that local shading exacerbated the declines in the photosynthetic rates, chlorophyll contents, electron transport and carbon assimilation‐related enzyme activities in shaded leaves as plants growth progressed. While, the decreases of these parameters in adjacent illuminated leaves of shaded plants were considerably alleviated compared to the corresponding leaves of control plants. Obviously, the photosynthesis in adjacent illuminated leaves in shaded plants was improved by local shading, and the improvement in adjacent lower leaves was larger than that in adjacent upper ones. As growth progressed, local shading induced higher abscisic acid contents in shaded leaves, but it alleviated the increase in the abscisic acid contents in adjacent leaves in shaded plants. Moreover, the difference in sugar content between shaded leaves and adjacent illuminated ones was gradually increased. Consequently, local weak light suppressed the photosynthesis in shaded leaves, while it markedly improved the photosynthesis of adjacent illuminated ones. Sugar gradient between shaded leaves and adjacent illuminated ones might play a key role in photosynthetic regulation of adjacent illuminated leaves.     

Alterations in photosynthesis and pigment distributions in pea leaves following UV-B exposure

We compared photosynthetic and UV-B-absorbing pigment concentrations, gas-exchange rates and photosystem II (PSII) electron transport rates in leaves of pea (Pisum sativum mutant Argenteum) grown without UV-B or under an enhanced UV-B treatment (18 kJ m^?2 biologically effective daily dose) in a greenhouse. We also compared the distribution of chlorophyll by depth within leaves of each treatment by using image analysis of chlorophyll autofluorescence. Ultraviolet-B treatment elicited putative protective responses such as an 80% increase in UV-B-absorbing compound concentrations (leaf-area basis), and a slight increase in mesophyll thickness (178 in controls compared to 191 μm in UV-B-treated leaves). However, photosynthetic rates of UV-B-treated leaves were only 80% of those of controls. This was paralleled by reductions in leaf conductance to water vapor (50% of controls) and intercellular CO₂ concentrations, suggesting that stomatal limitations were at least partly responsible for lower photosynthetic rates under the UV-B treatment. Total chlorophyll concentrations (leaf-area basis) in UV-B-treated leaves were only 70% of controls, and there was a shift in the relative distribution of chlorophyll with depth in UV-B-treated leaves. In control leaves chlorophyll concentrations were highest near the adaxial surface of the upper palisade, dropped with depth and then increased slightly in the bottom of the spongy mesophyll nearest the abaxial surface. In contrast, in UV-B-treated leaves chlorophyll concentrations were lowest at the adaxial surface of the upper palisade and increased with depth through the leaf. The most notable treatment difference in chlorophyll concentrations was in the upper palisade near the adaxial surface of leaves, where we estimate that chlorophyll concentrations in each 1-μm-thick paradermal layer were about 50% lower in UV-B-treated leaves than in controls. We found reduced electron transport capacity in UV-B-treated leaves, based on lower maximum fluorescence (F_m), variable to maximum fluorescence ratios (F,/F_m) and quantum yield of PSII electron transport (Y). However, the above were assessed from fluorometer measurements on the adaxial leaf surface and may reflect the markedly lower chlorophyll concentrations in the upper palisade of UV-B-treated leaves.     

Chlorophyll a and b biosynthesis in the dark in etiolated leaves infiltrated by exogenous chlorophyllide a]

Exogenous chlorophyllide a was introduced into etiolated rye leaves by the vacuum-infiltration technique. Appearance and accumulation of chlorophylls a and b within the leaves are observed during continued darkening, protochlorophyllide photoreduction being avoided. The pigments are identified by the solubility in petroleum ether, paper chromatograms, the fluorescence maxima, the peculiarities of exciting light 430 and 460 nm effects on fluorescence intensity, the specific interaction with hydrochloric hydroxylamine. The conclusion is made that before illumination etioplasts already contain enzyme systems and substrates which provide esterification of chlorophyllide a to chlorophyll a and conversion of chlorophyll a into chlorophyll b.     

Prokaryotic photosynthesis and phototrophy illuminated

Genome sequencing projects are revealing new information about the distribution and evolution of photosynthesis and phototrophy. Although coverage of the five phyla containing photosynthetic prokaryotes (Chlorobi, Chloroflexi, Cyanobacteria, Proteobacteria and Firmicutes) is limited and uneven, genome sequences are (or soon will be) available for >100 strains from these phyla. Present knowledge of photosynthesis is almost exclusively based on data derived from cultivated species but metagenomic studies can reveal new organisms with novel combinations of photosynthetic and phototrophic components that have not yet been described. Metagenomics has already shown how the relatively simple phototrophy based upon rhodopsins has spread laterally throughout Archaea, Bacteria and eukaryotes. In this review, we present examples that reflect recent advances in phototroph biology as a result of insights from genome and metagenome sequencing.     

Orientation of cell growth in the etiolated pea stem

Summary Both ethylene and IAA induce swelling in the sub-apical region of etiolated pea plants. The modified cells of these two types of swellings differ both morphologically and in their enzyme composition. In ethylene the cell walls become thickened within 24 h and the level of peroxidase is enhanced; ethylene does not affect cellulase levels. IAA induced swellings are not accompanied by early thickening of cell walls or enhanced peroxidase activity, but IAA greatly increases the level of cellulase. It is proposed that the retardation of extension growth by ethylene treatment results from the deposition of longitudinal microfibrils in the walls and that cross linking bonds in the polysaccharide matrix prevent their separation. Lateral expansion can occur, however, in the presence of auxin-induced cellulase which breaks or prevents the formation of these bonds.     

Ethylene formation from 1-aminocyclopropane-1-carboxylic acid in homogenates of etiolated pea seedlings

Homogenates of etiolated pea (Pisum sativum L.) shoots formed ethylene upon incubation with 1-aminocyclopropane-1-carboxylic acid (ACC). In-vitro ethylene formation was not dependent upon prior treatment of the tissue with indole-3-acetic acid. When homogenates were passed through a Sephadex column, the excluded, high-molecular-weight fraction lost much of its ethylene-synthesizing capacity. This activity was largely restored when a heat-stable, low-molecular-weight factor, which was retarded on the Sephadex column, was added back to the high-molecular-weight fraction. The ethylene-synthesizing system appeared to be associated, at least in part, with the particulate fraction of the pea homogenate. Like ethylene synthesis in vivo, cell-free ethylene formation from ACC was oxygen dependent and inhibited by ethylenediamine tetraacetic acid, n-propyl gallate, cyanide, azide, CoCl₃, and incubation at 40°C. It was also inhibited by catalase. In-vitro ethylene synthesis could only be saturated at very high ACC concentrations, if at all. Ethylene production in pea homogenates, and perhaps also in intact tissue, may be the result of the action of an enzyme that needs a heat-stable cofactor and has a very low affinity for its substrate, ACC, or it may be the result of a chemical reaction between ACC and the product of an enzyme reaction. Homogenates of etiolated pea shoots also formed ethylene with 2-keto-4-mercaptomethyl butyrate (KMB) as substrate. However, the mechanism by which KMB is converted to ethylene appears to be different from that by which ACC is converted.Abbreviations ACC 1-aminocyclopropane-1-carboxylic acid - IAA indole-3-acetic acid - KMB 2-keto-4-mercaptomethyl butyrate - SAM S-adenosylmethionine     

Genotype-specific soluble auxin-binding in etiolated pea epicotyls

Using different independent procedures for assaying soluble auxin-binding in etiolated pea epicotyls, wo could prove the reliability of the (XH₄)₂SO₄-pelleting assay both for crude cytosols as well as for specific protein fractions obtained after chromatofocusing. Three distinct genotypes (two parent lines, one tall recombinant) investigated so far exhibit characteristic differences with respect to soluble auxin-binding kinetics in their cytosols.     

Superoxide dismutase in wounded etiolated pea seedlings

The activities of three related enzymes, superoxide dismutase, peroxidase and catalase, were followed in decapitated (wounded) etiolated pea stem tissue. Of the three enzyme activities, only that of superoxide dismutase showed a significant loss and recovery pattern within 1 hr. The change in enzymatic activity appears to result from protein loss and recovery.