Lipid Composition Of Thylakoid Membranes Research Articles

Changes in intracellular energetic and metabolite states due to increased galactolipid levels in Synechococcus elongatus PCC 7942

The lipid composition of thylakoid membranes is conserved from cyanobacteria to green plants. However, the biosynthetic pathways of galactolipids, the major components of thylakoid membranes, are known to differ substantially between cyanobacteria and green plants. We previously reported on a transformant of the unicellular rod-shaped cyanobacterium Synechococcus elongatus PCC 7942, namely SeGPT, in which the synthesis pathways of the galactolipids monogalactosyldiacylglycerol and digalactosyldiacylglycerol are completely replaced by those of green plants. SeGPT exhibited increased galactolipid content and could grow photoautotrophically, but its growth rate was slower than that of wild-type S. elongatus PCC 7942. In the present study, we investigated pleiotropic effects that occur in SeGPT and determined how its increased lipid content affects cell proliferation. Microscopic observations revealed that cell division and thylakoid membrane development are impaired in SeGPT. Furthermore, physiological analyses indicated that the bioenergetic state of SeGPT is altered toward energy storage, as indicated by increased levels of intracellular ATP and glycogen. We hereby report that we have identified a new promising candidate as a platform for material production by modifying the lipid synthesis system in this way.

Temperature acclimation of net photosynthesis and its underlying component processes in four tropical tree species.

The effect of temperature change on leaf physiology has been extensively studied in temperate trees and to some extent in boreal and tropical tree species. While increased temperature typically stimulates leaf CO2 assimilation and tree growth in high-altitude ecosystems, tropical species are often negatively affected. These trees may operate close to their temperature optima and have a limited thermal acclimation capacity due to low seasonal and historical variation in temperature. To test this hypothesis, we studied the extent to which the temperature sensitivities of leaf photosynthesis and respiration acclimate to growth temperature in four common African tropical tree species. Tree seedlings native to different altitudes and therefore adapted to different growth temperatures were cultivated at three different temperatures in climate-controlled chambers. We estimated the acclimation capacity of the temperature sensitivities of light-saturated net photosynthesis, the maximum rates of Rubisco carboxylation (Vcmax) and thylakoid electron transport (J), and dark respiration. Leaf thylakoid membrane lipid composition, nitrogen content and leaf mass per area were also analyzed. Our results showed that photosynthesis in tropical tree species acclimated to higher growth temperatures, but that this was weakest in the species originating from the coolest climate. The temperature optimum of J acclimated significantly in three species and variation in J was linked to changes in the thylakoid membrane lipid composition. For Vcmax, there was only evidence of significant acclimation of optimal temperature in the lowest elevation species. Respiration acclimated to maintain homeostasis at growth temperature in all four species. Our results suggest that the lowest elevation species is better physiologically adapted to acclimate to high growth temperatures than the highest elevation species, indicating a potential shift in competitive balance and tree community composition to the disadvantage of montane tree species in a warmer world.

Handling the heat - photosynthetic thermal stress in tropical trees.

Warming climate increases the risk for harmful leaf temperatures in terrestrial plants, causing heat stress and loss of productivity. The heat sensitivity may be particularly high in equatorial tropical tree species adapted to a thermally stable climate. Thermal thresholds of the photosynthetic system of sun-exposed leaves were investigated in three tropical montane tree species native to Rwanda with different growth and water use strategies (Harungana montana, Syzygium guineense and Entandrophragma exselsum). Measurements of chlorophyll fluorescence, leaf gas exchange, morphology, chemistry and temperature were made at three common gardens along an elevation/temperature gradient. Heat tolerance acclimated to maximum leaf temperature (Tleaf ) across the species. At the warmest sites, the thermal threshold for normal function of photosystem II was exceeded in the species with the highest Tleaf despite their higher heat tolerance. This was not the case in the species with the highest transpiration rates and lowest Tleaf . The results point to two differently effective strategies for managing thermal stress: tolerance through physiological adjustment of leaf osmolality and thylakoid membrane lipid composition, or avoidance through morphological adaptation and transpiratory cooling. More severe photosynthetic heat stress in low-transpiring montane climax species may result in a competitive disadvantage compared to high-transpiring pioneer species with more efficient leaf cooling.

Betaine lipid and neutral lipid production under nitrogen or phosphorus limitation in the marine microalga Tisochrysis lutea (Haptophyta)

Nitrogen (N) and phosphorus (P) limitations induce neutral lipid accumulation and membrane lipid remodeling in the domesticated oleaginous microalga Tisochrysis lutea. This study compared growth, photosynthetic activity, biochemical and transcriptional responses of T. lutea throughout batch cultures under N or P limitation compared with non-limiting nutrient condition (NP). The results show that, under N limitation, the breakdown and re-use of cellular N-containing compounds contributed to carbohydrates and further triacylglycerol (TAG) accumulation, where the accumulation of docosahexaenoic acid (DHA) in neutral lipids might mainly be derived from the recycling of membrane polar lipids. Conversely, P limitation did not hinder the photosynthetic capacity; a higher efficiency of carbon fixation fueled the allocation of carbon fluxes to the reserves of carbohydrates and neutral lipids. These latter accumulated without massive degradation of essential cellular compounds. Betaine lipids constitute the major compounds of non-plastidial membranes in T. lutea. Given an extremely low constitutive phospholipid level, phospholipids might not be involved in the regulation of P storage. However, transfer of P from non-plastidial to chloroplast membranes might occur, keeping a stable lipid composition of thylakoid membranes and maintaining a relatively high photosynthetic capacity under P deprivation.

Decreased photosynthetic rate under high temperature in wheat is due to lipid desaturation, oxidation, acylation, and damage of organelles

BackgroundHigh temperature is a major abiotic stress that limits wheat (Triticum aestivum L.) productivity. Variation in levels of a wide range of lipids, including stress-related molecular species, oxidative damage, cellular organization and ultrastructural changes were analyzed to provide an integrated view of the factors that underlie decreased photosynthetic rate under high temperature stress. Wheat plants of cultivar Chinese Spring were grown at optimum temperatures (25/15 °C, maximum/minimum) until the onset of the booting stage. Thereafter, plants were exposed to high temperature (35/25 °C) for 16 d.ResultsCompared with optimum temperature, a lower photosynthetic rate was observed at high temperature which is an interplay between thylakoid membrane damage, thylakoid membrane lipid composition, oxidative damage of cell organelle, and stomatal and non-stomatal limitations. Triacylglycerol levels were higher under high temperature stress. Polar lipid fatty acyl unsaturation was lower at high temperature, while triacylglycerol unsaturation was the same at high temperature and optimum temperature. The changes in lipid species indicates increases in activities of desaturating, oxidizing, glycosylating and acylating enzymes under high temperature stress. Cumulative effect of high temperature stress led to generation of reactive oxygen species, cell organelle and membrane damage, and reduced antioxidant enzyme activity, and imbalance between reactive oxygen species and antioxidant defense system.ConclusionsTaken together with recent findings demonstrating that reactive oxygen species are formed from and are removed by thylakoid lipids, the data suggest that reactive oxygen species production, reactive oxygen species removal, and changes in lipid metabolism contribute to decreased photosynthetic rate under high temperature stress.

A Plastid Phosphatidylglycerol Lipase Contributes to the Export of Acyl Groups from Plastids for Seed Oil Biosynthesis.

The lipid composition of thylakoid membranes inside chloroplasts is conserved from leaves to developing embryos. A finely tuned lipid assembly machinery is required to build these membranes during Arabidopsis thaliana development. Contrary to thylakoid lipid biosynthetic enzymes, the functions of most predicted chloroplast lipid-degrading enzymes remain to be elucidated. Here, we explore the biochemistry and physiological function of an Arabidopsis thylakoid membrane-associated lipase, PLASTID LIPASE1 (PLIP1). PLIP1 is a phospholipase A1 In vivo, PLIP1 hydrolyzes polyunsaturated acyl groups from a unique chloroplast-specific phosphatidylglycerol that contains 16:1 Δ3trans as its second acyl group. Thus far, a specific function of this 16:1 Δ3trans -containing phosphatidylglycerol in chloroplasts has remained elusive. The PLIP1 gene is highly expressed in seeds, and plip1 mutant seeds contain less oil and exhibit delayed germination compared with the wild type. Acyl groups released by PLIP1 are exported from the chloroplast, reincorporated into phosphatidylcholine, and ultimately enter seed triacylglycerol. Thus, 16:1 Δ3trans uniquely labels a small but biochemically active plastid phosphatidylglycerol pool in developing Arabidopsis embryos, which is subject to PLIP1 activity, thereby contributing a small fraction of the polyunsaturated fatty acids present in seed oil. We propose that acyl exchange involving thylakoid lipids functions in acyl export from plastids and seed oil biosynthesis.

Knocking Down of Isoprene Emission Modifies the Lipid Matrix of Thylakoid Membranes and Influences the Chloroplast Ultrastructure in Poplar.

Isoprene is a small lipophilic molecule with important functions in plant protection against abiotic stresses. Here, we studied the lipid composition of thylakoid membranes and chloroplast ultrastructure in isoprene-emitting (IE) and nonisoprene-emitting (NE) poplar (Populus × canescens). We demonstrated that the total amount of monogalactosyldiacylglycerols, digalactosyldiacylglycerols, phospholipids, and fatty acids is reduced in chloroplasts when isoprene biosynthesis is blocked. A significantly lower amount of unsaturated fatty acids, particularly linolenic acid in NE chloroplasts, was associated with the reduced fluidity of thylakoid membranes, which in turn negatively affects photosystem II photochemical efficiency. The low photosystem II photochemical efficiency in NE plants was negatively correlated with nonphotochemical quenching and the energy-dependent component of nonphotochemical quenching. Transmission electron microscopy revealed alterations in the chloroplast ultrastructure in NE compared with IE plants. NE chloroplasts were more rounded and contained fewer grana stacks and longer stroma thylakoids, more plastoglobules, and larger associative zones between chloroplasts and mitochondria. These results strongly support the idea that in IE species, the function of this molecule is closely associated with the structural organization and functioning of plastidic membranes.

LHCII organization and thylakoid lipids affect the sensitivity of the photosynthetic apparatus to high-light treatment.

Pulse-amplitude-modulated (PAM) chlorophyll fluorescence and photosynthetic oxygen evolution were used to investigate the role of the different amount and organization of light-harvesting complexes of photosystem II (LHCII) in four pea species on the susceptibility of the photosynthetic apparatus to high-light treatment. In this work we analyzed the thylakoid membrane lipid composition of the studied pea plants. A relationship between the structural organization of LHCII proteins, the amount of the main lipid classes and the sensitivity of the photosynthetic apparatus to high-light treatment was found. The results reveal that the photosynthetic apparatus, enriched in oligomeric forms of LHCII concomitant with decreased amount of anionic lipids and increased content of the monogalactosyldiacylglycerol (MGDG), is less sensitive to high light. Our data also suggest that the degree of LHCII oligomerization, as well as the lipid composition do not influence the degree of recovery of the PSII photochemistry after excess light exposure.

Alteration in growth and thylakoid membrane lipid composition of Azolla caroliniana under phosphate deficiency

The changes in the fresh biomass accumulation, photosynthetic and anthocyanin pigments, photosystem 2 (PS 2) activity, ultrastructure of chloroplast, total lipids and fatty acid composition of thylakoid membrane were followed in the aquatic fern Azolla caroliniana grown on medium either deficient or supplied with various phosphorus concentrations. The content of photosynthetic pigments and the anthocyanin/chlorophyll ratio increased significantly with increasing PO4 3− concentration. Phosphate deficiency inhibited growth and PS 2 activity and decreased content of total lipids and phospholipids in isolated thylakoids. This was accompanied with a significant increase in the percentage of galalactolipids.

Effect of glycinebetaine on function of thylakoid membranes in wheat flag leaves under drought stress

Two wheat cultivars, HF9703 (drought tolerant) and SN215953 (drought sensitive) were used to examine the effects of glycinebetaine (GB, 100 mM) on lipid composition and function of thylakoid membranes under drought stress. GB application mitigated negative effect of drought on Ca2+-ATPase and Hill reaction activities, chlorophyll content, gas exchange and photosynthesis. These positive effects of GB application maybe, in part, correlated with improving the lipid composition of the thylakoid membranes.

Glycoengineering of Cyanobacterial Thylakoid Membranes for Future Studies on the Role of Glycolipids in Photosynthesis

The lipid composition of thylakoid membranes is conserved from cyanobacteria to angiosperms. The predominating components are monogalactosyl- and digalactosyldiacylglycerol. In cyanobacteria, thylakoid membrane biosynthesis starts with the formation of monoglucosyldiacylglycerol which is C4-epimerized to the corresponding galactolipid, whereas in plastids monogalactosyldiacylglycerol is formed at the beginning. This suggests that galactolipids have specific functions in thylakoids. We wanted to investigate whether galactolipids can be replaced by glycosyldiacylglycerols with headgroups differing in their epimeric and anomeric details as well as the attachment point of the terminal hexose in diglycosyldiacylglycerols. For this purpose putative glycosyltransferase sequences were identified in databases to be used for functional expression in various host organisms. From 18 newly identified sequences, four turned out to encode glycosyltransferases catalyzing final steps in glycolipid biosynthesis: two alpha-glucosyltransferases, one beta-galactosyltransferase and one beta-glucosyltransferase. Their functional annotation was based on detailed structural characterization of the new glycolipids formed in the transformant hosts as well as on in vitro enzymatic assays. The expression of alpha-glucosyltransferases in the cyanobacterium Synechococcus resulted in the accumulation of the new alpha-galactosyldiacylglycerol which is ascribed to epimerization of the corresponding glucolipid. The expression of the beta-glucosyltransferase led to a high proportion of new beta-glucosyl-(1-->6)-beta-galactosyldiacylglycerol almost entirely replacing the native digalactosyldiacylglycerol. These results demonstrate that modifications of the glycolipid pattern in thylakoids are possible.

Membrane lipids of symbiotic algae are diagnostic of sensitivity to thermal bleaching in corals.

Over the past three decades, massive bleaching events of zooxanthellate corals have been documented across the range of global distribution. Although the phenomenon is correlated with relatively small increases in sea-surface temperature and enhanced light intensity, the underlying physiological mechanism remains unknown. In this article we demonstrate that thylakoid membrane lipid composition is a key determinate of thermal-stress sensitivity in symbiotic algae of cnidarians. Analyses of thylakoid membranes reveal that the critical threshold temperature separating thermally tolerant from sensitive species of zooxanthellae is determined by the saturation of the lipids. The lipid composition is potentially diagnostic of the differential nature of thermally induced bleaching found in scleractinian corals. Measurements of variable chlorophyll fluorescence kinetic transients indicate that thermally damaged membranes are energetically uncoupled but remain capable of splitting water. Consequently, a fraction of the photosynthetically produced oxygen is reduced by photosystem I through the Mehler reaction to form reactive oxygen species, which rapidly accumulate at high irradiance levels and trigger death and expulsion of the endosymbiotic algae. Differential sensitivity to thermal stress among the various species of Symbiodinium seems to be distributed across all clades. A clocked molecular phylogenetic analysis suggests that the evolutionary history of symbiotic algae in cnidarians selected for a reduced tolerance to elevated temperatures in the latter portion of the Cenozoic.

Effects of phosphate deficiency on the lipid composition in cucumber thylakoid membranes and PSII particles

Phosphatidylglycerol (PG) is the sole phospholipid in thylakoid membranes. Phosphate deficiency clearly affected the thylakoid membrane lipid composition of cucumber leaves. Phosphate deficiency induced the decrease in PG content and the concomitant increases in the content of digalactosyl diacylglycerol (DGDG) and sulfoquinovosyl diacylglycerol (SQDG) in cucumber leaves. Although phosphate deficiency led to the 44 mol% decrease of thylakoid PG, the chlorophyll fluorescence coefficients and photosystem II (PSII) oxygen-evolving activity under phosphate deficiency showed virtually no difference compared with those under phosphate sufficiency. It was suggested that phosphate deficiency had no clear effect on PSII function. Besides, PG still maintained in a high level in PSII under phosphate-deficient conditions. These results showed that phosphate deficiency could result in PG decrease in some thylakoid membrane domains but had little effect on the content of PG existed in PSII membrane domains of thylakoid. Maybe, there were two types of PG in thylakoid membrane. Some PG molecules only played a role in the structure of thylakoid membrane and other lipid molecules such as SQDG can substitute for them under phosphate deficiency while some PG molecules may play an important role in maintaining the structure and function of PSII and other lipid molecules cannot substitute for them.

Effects of slow clinorotation on lipid contents and proton permeability of thylakoid membranes of pea chloroplasts.

Photochemical characteristics and lipid composition of thylakoid membranes from 12 day-old pea leaves that were exposed to slow clino-rotation were examined and compared with a vertical control. Proton permeability of thylakoid membranes was estimated from light-induced proton uptake (delta H+) and post-illumination proton efflux in chloroplast suspensions. The delta pH magnitude was calculated from the level of light-induced quenching of 9-aminoacridine fluorescence. Proton permeability of thylakoid membranes increased during exposure to clino-rotation. When subsequently transferred to darkness, proton efflux increased almost 2-fold in clinorotated leaves. The results were compared with data on pigment and polar lipid composition of photosynthetic membranes in clino-rotated and control plants. It was concluded that both the increase of proton permeability and the decrease of polar lipid content in chloroplasts were induced by clino-rotation.

Lipid composition of thylakoid membranes from leaves and regreened spathes of Zantedeschia aethiopica

The glycerolipid and fatty acid composition of thylakoid membranes from two photosynthetic organs, the leaf and regreened spathe, of Zantedeschia aethiopica was studied. In regreened spathes, there was a decrease in the monogalactosyl- and digalactosyldiacylglycerols, and an increase in the contents of the sulpholipid, sulphoquinovosyldiacylglycerol and in the phospholipids phosphatidylglycerol and phosphatidylcholine, when compared with those isolated from the leaf. The major difference detected in fatty acid composition of both photosynthetic structures was the absence of the unusual fatty acid, Δ 3- trans-hexadecenoic, from phosphatidylglycerol in regreened spathe thylakoid membranes. In spite of these differences, chloroplasts from regreened spathes exhibited a typical organization, with grana and stromal regions. The results obtained suggest, that in Z. aethiopica, there is no evident correlation between the presence of Δ 3- trans-hexadecenoic acid in thylakoid membranes and thylakoid-stacking. The absence of this fatty acid in the regreened spathe is discussed in terms of chloroplast function.

Organization of the photosynthetic apparatus from triazine-resistant and -susceptible biotypes of several plant species

To specify the relations between triazine resistance and modification of the photosynthetic apparatus, the organization of plastids from herbicide-resistant and -susceptible biotypes of Chenopodium album L., Amaranthus retroflexus L., Solanum nigrum L., Brassica campestris L., and Brassica napus L. has been compared. The plastids from resistant biotypes exhibit a higher degree of stacking owing to a paucity of stroma lamellae and irrespective of the granum size distribution. Their enrichment in chlorophyll b and xanthophylls results from a higher content in light-harvesting chlorophyll a/b – proteins (LHCP), especially of the oligomer LHCP1. The resulting enlargement of photosystem II antennae is correlated to the fluorescence properties of isolated plastids. The most important difference in the lipid composition of thylakoid membranes is the increase of the Δ3-trans-hexadecenoic acid proportion in phosphatidylglycerol from resistant plastids, which is thought to participate in the formation of the oligomeric structure of LHCP1. The slight increase of the unsaturation of lipids, especially of mono-galactosyldiacylglycerol, from resistant plastids, is but a consequence of their higher proportion of appressed membranes. These alterations in lipid composition and ultrastructure of the chloroplasts resemble those occurring in plants grown under low irradiance. This adaptative reorganization of thylakoid components seems to be a compensatory mechanism allowing the overcoming of limitations in the QA – QB electron transfer and, thus, its occurrence and extent depend on the light and temperature conditions of growth.

Analysis of temperature-jump chlorophyll fluorescence induction in plants

A newly observed general chlorophyll fluorescence induction effect in plants is described. Fluorescence yield can rise through as many as four different phases (α, β, γ, δ) in the dark, when intact cells or leaves are rapidly heated (within approx. 2.5 s) from 20 to 40–50 °C. An analysis of this temperature-jump fluorescence induction in Scenedesmus obliquus leads to the following: 1. 1. Phase α is due to removal of S-quenching and appears to be related to heat deactivation of the water-splitting enzyme system. With prolonged heating, irreversibility of α upon recooling reflects irreversible damage to the water-splitting enzyme system. 2. 2. β is independent of the S-states and of the redox state of primary System II acceptor Q. It is suggested that β parallels functional separation of Q from the System II trapping centre. This effect is highly reversible. 3. 3. γ and δ reflect reduction of primary System II acceptor Q by a heat-induced endogenous reductant, which is probably identical to hydrogenase. Critical temperatures for pronounced α and β phases differ markedly in different plants. Possible correlations between temperature-jump fluorescence induction, thylakoid membrane lipid composition, lipid phase transition and lipid-protein interactions are discussed.