Search

Pancreatic lipase related-protein 2 (PLRP2) exhibits remarkable galactolipase and phospholipase A1 activities, which depend greatly on the supramolecular organization of the substrates and the presence of surfactant molecules such as bile salts. The objective of the study was to understand the modulation of the adsorption mechanisms and enzymatic activity of Guinea pig PLRP2 (gPLRP2), by the physical environment of the enzyme and the physical state of its substrate. Langmuir monolayers were used to reproduce homogeneous and heterogeneous photosynthetic model membranes containing galactolipids (GL), and/or phospholipids (PL), and/or phytosterols (pS), presenting uncharged or charged interfaces. The same lipid mixtures were also used to form micrometric liposomes, and their gPLRP2 catalyzed digestion kinetics were investigated in presence or in absence of bile salts (NaTDC) during static in vitro , so called "bulk", digestion. The enzymatic activity of gPLRP2 onto the galactolipid-based monolayers was characterized with an optimum activity at 15 mN/m, in the absence of bile salts. gPLRP2 showed enhanced adsorption onto biomimetic model monolayer containing negatively charged lipids. However, the compositional complexity in the heterogeneous uncharged model systems induced a lag phase before the initiation of lipolysis. In bulk, no enzymatic activity could be demonstrated on GL-based liposomes in the absence of bile salts, probably due to the high lateral pressure of the lipid bilayers. In the presence of NaTDC (4 mM), however, gPLRP2 showed both high galactolipase and moderate phospholipase A1 activities on liposomes, probably due to a decrease in packing and lateral pressure upon NaTDC adsorption, and subsequent disruption of liposomes. [Display omitted] • gPLRP2 showed galactolipase activity onto heterogeneous galactolipid-based monolayers. • Optimal enzymatic activity of gPLRP2 on monolayers was obtained at π∼15 mN/m. • No lipolytic activity of gPLRP2 was detected on liposomes in the absence of NaTDC. • gPLRP2 showed enzymatic activities on liposomes in the presence of NaTDC. [ABSTRACT FROM AUTHOR]

Polyamines are small aliphatic polycations present in all living organisms. In plants, the most abundant polyamines are putrescine (Put), spermidine (Spd) and spermine (Spm). Polyamine levels change in response to different pathogens, including Pseudomonas syringae pv. tomato DC3000 (Pst DC3000). However, the regulation of polyamine metabolism and their specific contributions to defence are not fully understood. Here we report that stimulation of Put biosynthesis by Pst DC3000 is dependent on coronatine (COR) perception and jasmonic acid (JA) signalling, independently of salicylic acid (SA). Conversely, lack of Spm in spermine synthase (spms) mutant stimulated galactolipids and JA biosynthesis, and JA signalling under basal conditions and during Pst DC3000 infection, whereas compromised SA‐pathway activation and defence outputs through SA–JA antagonism. The dampening of SA responses correlated with COR and Pst DC3000‐inducible deregulation of ANAC019 expression and its key SA‐metabolism gene targets. Spm deficiency also led to enhanced disease resistance to the necrotrophic fungal pathogen Botrytis cinerea and stimulated endoplasmic reticulum (ER) stress signalling in response to Pst DC3000. Overall, our findings provide evidence for the integration of polyamine metabolism in JA‐ and SA‐mediated defence responses, as well as the participation of Spm in buffering ER stress during defence. Summary statement: Spermine deficiency stimulates the biosynthesis of galactolipids and jasmonic acid, while it dampens the biosynthesis of salicylic acid, thereby conditioning defence outputs. [ABSTRACT FROM AUTHOR]

Needle abscission in balsam fir has been linked to both cold acclimation and changes in lipid composition. The overall objective of this research is to uncover lipid changes in balsam fir during cold acclimation and link those changes with postharvest abscission. Branches were collected monthly from September to December and were assessed for cold tolerance via membrane leakage and chlorophyll fluorescence changes at −5, −15, −25, −35, and −45 °C. Lipids were extracted and analyzed using mass spectrometry while postharvest needle abscission was determined gravimetrically. Cold tolerance and needle retention each significantly (p < 0.001) improved throughout autumn in balsam fir. There were concurrent increases in DGDG, PC, PG, PE, and PA throughout autumn as well as a decrease in MGDG. Those same lipids were strongly related to cold tolerance, though MGDG had the strongest relationship (R2 = 55.0% and 42.7% from membrane injury and chlorophyll fluorescence, respectively). There was a similar, albeit weaker, relationship between MGDG:DGDG and needle retention (R2 = 24.3%). Generally, a decrease in MGDG:DGDG ratio resulted in better cold tolerance and higher needle retention in balsam fir, possibly due to increased membrane stability. This study confirms the degree of cold acclimation in Nova Scotian balsam fir and presents practical significance to industry by identifying the timing of peak needle retention. It is suggested that MGDG:DGDG might be a beneficial tool for screening balsam fir genotypes with higher needle retention characteristics. [ABSTRACT FROM AUTHOR]

Summary: Plastoglobules (PGs) contiguous with the outer leaflets of thylakoid membranes regulate lipid metabolism, plastid developmental transitions, and responses to environmental stimuli. However, the function of OsFBN7, a PG‐core fibrillin gene in rice, has not been elucidated.Using molecular genetics and physiobiochemical approaches, we observed that OsFBN7 overexpression promoted PG clustering in rice chloroplasts.OsFBN7 interacted with two KAS I enzymes, namely OsKAS Ia and OsKAS Ib, in rice chloroplasts. Lipidomic analysis of chloroplast subcompartments, including PGs in the OsFBN7 overexpression lines, confirmed that levels of diacylglycerol (DAG), a chloroplast lipid precursor and monogalactosyldiacylglycerol (MGDG) and digalactosyldiacylglycerol (DGDG), the main chloroplast membrane lipids, were increased in PGs and chloroplasts. Furthermore, OsFBN7 enhanced the abundances of OsKAS Ia/Ib in planta and their stability under oxidative and heat stresses. In addition, RNA sequencing and real‐time quantitative reverse‐transcription polymerase chain reaction (qRT‐PCR) analyses showed that the expression of the DAG synthetase gene PAP1 and MGDG synthase gene MDG2 was upregulated by OsFBN7.In conclusion, this study proposes a new model in which OsFBN7 binds to OsKAS Ia/Ib in chloroplast and enhances their abundance and stability, thereby regulating the chloroplast and PG membrane lipids involved in the formation of PG clusters. [ABSTRACT FROM AUTHOR]

Lipidomics is a new method in postharvest studies. RNA-seq data on lettuce leaves revealed important changes in gene expression linked to lipid metabolism. Here, we report a global lipid analysis in lettuce leaves over 14 d of storage, in dark cold conditions, with or without an initial light treatment. Our LC-MS analyses of 171 lipid compounds, among which digalactosyldiacylglycerol, triacylglycerol, and phosphatidic acid, revealed variations in response to the postharvest conditions. Our results suggest the reallocation of phospholipid fatty acids to galactolipids through storage time, which may be associated to lettuce capacities to withstand storage without rapid chlorophyll degradation in contrast to other leafy vegetables. The unsaturation level of fatty acids increased over storage time. Intermittent light treatment at the beginning of storage stimulated phosphatidic acid accumulation, concomitantly with upregulation of phospholipase D gene expression. We also observed an increase in triacylglycerol content after light treatment. Usual indicators of membrane damage, electrolyte leakage, and malondialdehyde, were unchanged during storage. Results are discussed in the light of known changes in postharvest life of lettuces and other leafy vegetables. • An extensive lipidomics study of lettuce was performed over cold storage. • An increased unsaturation of the lipids was observed over cold storage. • This was associated with an increased accumulation of most of galactolipids. • Initial light treatment promoted phosphatidic acid and triacylglycerol accumulation. [ABSTRACT FROM AUTHOR]

The use of Nuclear Magnetic Resonance spectroscopy for studying lipid digestion in vitro most often consists of quantifying lipolysis products after they have been extracted from the reaction medium using organic solvents. However, the current sensitivity level of NMR spectrometers makes possible to avoid the extraction step and continuously quantify the lipids directly in the reaction medium. We used real-time 1H NMR spectroscopy and guinea pig pancreatic lipase-related protein 2 (GPLRP2) as biocatalyst to monitor in situ the lipolysis of monogalactosyl diacylglycerol (MGDG) in the form of mixed micelles with the bile salt sodium taurodeoxycholate (NaTDC). Residual substrate and lipolysis products (monogalactosyl monoacylglycerol (MGMG); monogalactosylglycerol (MGG) and octanoic acid (OA) were simultaneously quantified throughout the reaction thanks to specific proton resonances. Lipolysis was complete with the release of all MGDG fatty acids. These results were confirmed by thin layer chromatography (TLC) and densitometry after lipid extraction at different reaction times. Using diffusion-ordered NMR spectroscopy (DOSY), we could also estimate the diffusion coefficients of all the reaction compounds and deduce the hydrodynamic radius of the lipid aggregates in which they were present. It was shown that MGDG-NaTDC mixed micelles with an initial hydrodynamic radius r H of 7.3 ± 0.5 nm were changed into smaller micelles of NaTDC-MGDG-MGMG of 2.3 ± 0.5 nm in the course of the lipolysis reaction, and finally into NaTDC-OA mixed micelles (r H of 2.9 ± 0.5 nm) and water soluble MGG. These results provide a better understanding of the digestion of galactolipids by PLRP2, a process that leads to the complete micellar solubilisation of their fatty acids and renders their intestinal absorption possible. [Display omitted] • Digestion of galactolipids, the main plant membrane lipids, by PLRP2. • In situ kinetics of lipolysis monitored by NMR • Quantitative analysis of all lipolysis products formed from model micelles of monogalactosyldiacylglycerol and bile salts. • In situ monitoring of the diffusion coefficients of the lipid substrate and lipolysis products by DOSY-NMR. • Estimation of micelle sizes and their changes upon lipolysis [ABSTRACT FROM AUTHOR]

SUMMARY: Lipids have been observed attached to lumen‐facing surfaces of mature xylem conduits of several plant species, but there has been little research on their functions or effects on water transport, and only one lipidomic study of the xylem apoplast. Therefore, we conducted lipidomic analyses of xylem sap from woody stems of seven plants representing six major angiosperm clades, including basal magnoliids, monocots and eudicots, to characterize and quantify phospholipids, galactolipids and sulfolipids in sap using mass spectrometry. Locations of lipids in vessels of Laurus nobilis were imaged using transmission electron microscopy and confocal microscopy. Xylem sap contained the galactolipids di‐ and monogalactosyldiacylglycerol, as well as all common plant phospholipids, but only traces of sulfolipids, with total lipid concentrations in extracted sap ranging from 0.18 to 0.63 nmol ml−1 across all seven species. Contamination of extracted sap from lipids in cut living cells was found to be negligible. Lipid composition of sap was compared with wood in two species and was largely similar, suggesting that sap lipids, including galactolipids, originate from cell content of living vessels. Seasonal changes in lipid composition of sap were observed for one species. Lipid layers coated all lumen‐facing vessel surfaces of L. nobilis, and lipids were highly concentrated in inter‐vessel pits. The findings suggest that apoplastic, amphiphilic xylem lipids are a universal feature of angiosperms. The findings require a reinterpretation of the cohesion‐tension theory of water transport to account for the effects of apoplastic lipids on dynamic surface tension and hydraulic conductance in xylem. [ABSTRACT FROM AUTHOR]

Across the globe, heat waves are getting more intense and frequent. Diatoms are a major group of microalgae at the base of the marine food webs and an important source of long chain polyunsaturated fatty acids that are transferred through the food web. The present study investigates the possible impacts of temperature increase on lipid classes and expression of genes encoding enzymes related to lipid metabolism in Phaeodactylum tricornutum. The heat wave exposure caused an increase in the relative amounts of plastidial lipids such as the glycolipids monogalactosyldiacylglycerol (MGDG), digalactosyldiacylglycerol (DGDG) and sulphoquinovosyldiacylglycerol (SQDG) in parallel with a decrease in the neutral lipid fraction, which includes triacylglycerols. In agreement, gene expression analyses revealed an up-regulation of a gene encoding one MGDG synthase and down-regulation of a diacylglycerol acyltransferase (DGAT), a key enzyme in triacylglycerol synthesis. Our results show that heat waves not only negatively impact the abundance of unsaturated fatty acids such as eicosapentaenoic acid (20:5n-3, EPA) and hexadecatrienoic acid (16:3n-4) as observed by the decrease in their relative abundance in MGDG and neutral lipids, respectively, but also induce changes in the relative amounts of the diverse membrane lipids as well as the proportion of membrane/storage lipids. The expression study of key genes indicates that some of the aforementioned alterations are regulated at the transcription level whereas others appear to be post-transcriptional. The changes observed in plastidial lipids are related to negative impacts on the photosynthesis. • Heat wave-exposed P. tricornutum cells presented higher galactolipid content. • The MGDG-eicosapentaenoic acid content severely decreased due to heat stress. • Monogalactosyldiacylglycerol (MGDG) synthase was upregulated under heat stress. • A diacylglycerol acyl transferase was downregulated in heat wave-exposed cells. • MGDG and sulfoquinovosyldiacylglycerol content reveals biotechnological potential. [ABSTRACT FROM AUTHOR]

Chloroplast membranes have a high content of the uncharged galactolipids monogalactosyldiacylglycerol (MGDG) and digalactosyldiacylglycerol (DGDG). These galactolipids are essential for the biogenesis of plastids and functioning of the photosynthetic machinery. A monotopic glycosyltransferase, monogalactosyldiacylglycerol synthase synthesizes the bulk of MGDG. It is embedded in the outer leaflet of the inner envelope membrane of chloroplasts. The protein transfers a galactose residue from UDP-galactose to diacylglycerol (DAG); it needs anionic lipids such as phosphatidylglycerol (PG) to be active. The intricacy of the organization and the process of active complex assembly and synthesis have been investigated at the Coarse-Grained and All-Atom of computer simulation levels to cover large spatial and temporal scales. The following self-assembly process and catalytic events can be drawn; (1) in the membrane, in the absence of protein, there is a spontaneous formation of PG clusters to which DAG molecules associate, (2) a reorganization of the clusters occurs in the vicinity of the protein once inserted in the membrane, (3) an accompanying motion of the catalytic domain of the protein brings DAG in the proper position for the formation of the active complex MGD1/UDP-Gal/DAG/PG for which an atomistic model of interaction is proposed. [ABSTRACT FROM AUTHOR]

UDP-glucose epimerases (UGEs) are essential enzymes for catalysing the conversion of UDP-glucose (UDP-Glc) into UDP-galactose (UDP-Gal). Although UDP-Gal has been well studied as the substrate for the biosynthesis of carbohydrates, glycolipids, and glycoproteins, much remains unknown about the biological function of UGEs in plants. In this study, we selected a novel rice fragile culm 24 (Osfc24) mutant and identified it as a nonsense mutation of the FC24/OsUGE2 gene. The Osfc24 mutant shows a brittleness phenotype with significantly altered cell wall composition and disrupted orientation of the cellulose microfibrils. We found significantly reduced accumulation of arabinogalactan proteins in the cell walls of the mutant, which may consequently affect plant growth and cell wall deposition, and be responsible for the altered cellulose microfibril orientation. The mutant exhibits dwarfism and paler leaves with significantly decreased contents of galactolipids and chlorophyll, resulting in defects in plant photosynthesis. Based on our results, we propose a model for how OsUGE2 participates in two distinct metabolic pathways to co-modulate cellulose biosynthesis and cell wall assembly by dynamically providing UDP-Gal and UDP-Glc substrates. [ABSTRACT FROM AUTHOR]

The surface tension of xylem sap has been traditionally assumed to be close to that of the pure water because decreasing surface tension is thought to increase vulnerability to air seeding and embolism. However, xylem sap contains insoluble lipid-based surfactants, which also coat vessel and pit membrane surfaces, where gas bubbles can enter xylem under negative pressure in the process known as air seeding. Because of the insolubility of amphiphilic lipids, the surface tension influencing air seeding in pit pores is not the equilibrium surface tension of extracted bulk sap but the local surface tension at gas–liquid interfaces, which depends dynamically on the local concentration of lipids per surface area. To estimate the dynamic surface tension in lipid layers that line surfaces in the xylem apoplast, we studied the time-dependent and surface area-regulated surface tensions of apoplastic lipids extracted from xylem sap of four woody angiosperm plants using constrained drop surfactometry. Xylem lipids were found to demonstrate potent surface activity, with surface tensions reaching an equilibrium at ~25 mN m-1 and varying between a minimum of 19 mN m-1 and a maximum of 68 mN m-1 when changing the surface area between 50 and 160% around the equilibrium surface area. It is concluded that xylem lipid films in natural conditions most likely range from nonequilibrium metastable conditions of a supersaturated compression state to an undersaturated expansion state, depending on the local surface areas of gas–liquid interfaces. Together with findings that maximum pore constrictions in angiosperm pit membranes are much smaller than previously assumed, low dynamic surface tension in xylem turns out to be entirely compatible with the cohesion–tension and air-seeding theories, as well as with the existence of lipid-coated nanobubbles in xylem sap, and with the range of vulnerabilities to embolism observed in plants. [ABSTRACT FROM AUTHOR]

Background: Membrane lipid composition associates closely with membrane stability and fluidity under water stress. In this study, lipidomic analyses based on electrospray ionization mass spectrometry (ESI-MS/MS) were carried out to explore dynamic changes of membrane lipids in term of molecular species caused by PEG (Polyethylene glycol-6000)-induced water stress in wheat seedlings. Results: Among the main phospholipids, phosphatidylcholine (PC), phosphatidylethanolamine (PE), and phosphatidylglycerol (PG) are primary degradation targets, and PC was degraded in the largest degree. Membrane ion leakage dramatically increased later than the significant reduction of these phospholipids, indicating that the loss of membrane integrity lagged behind severe phospholipid degradation. Monogalactosyldiacylglycerol (MGDG) increased firstly and decreased later, while digalactosyldiacylglycerol (DGDG) ratcheted up with stress. DGDG/MGDG increased after stress for 3 days, and unsaturation of DGDG was promoted with stress. Variation trends of galactolipids differed among molecular species. The time when MGDG (34:3), DGDG (34:3) began to decline approached to the time when non-stomatal limitation impaired photosynthesis. While the two predominant molecular species MGDG (36:6) and DGDG (36:6) began to decline later. So we speculated that MGDG (34:3), DGDG (34:3) might be key components in photosynthesis apparatus and participate in photosynthesis directly. While the two predominant molecular species, MGDG (36:6) and DGDG (36:6) might locate in thylakoid lipid bilayer matrix and play roles in stabilizing the membrane. The research provides new insights into the dynamic response of lipid metabolism to PEG-induced water stress. Conclusion: In wheat plants under water stress, the major molecular species of PC, PE and PG were degraded, MGDG and DGDG molecular species had differing degradation time courses. [ABSTRACT FROM AUTHOR]

The chloroplast thylakoid network is a dynamic structure which, through possible rearrangements, plays a crucial role in regulation of photosynthesis. Although the importance of the main components of the thylakoid membrane matrix, galactolipids, in the formation of the network of internal plastid membrane was found before, the structural role of monogalactosyldiacylglycerol (MGDG) and digalactosylidacylglycerol (DGDG) is still largely unknown. We elucidated detailed structural modifications of the thylakoid membrane system in Arabidopsis thaliana MGDG- and DGDG-deficient mutants. An altered MGDG/DGDG ratio was structurally reflected by formation of smaller grana, local changes in grana stacking repeat distance, and significant changes in the spatial organization of the thylakoid network compared with wild-type plants. The decrease of the MGDG level impaired the formation of the typical helical grana structure and resulted in a 'helical-dichotomic' arrangement. DGDG deficiency did not affect spatial grana organization but changed the shape of the thylakoid membrane network in situ from lens like into a flattened shape. Such structural disturbances were accompanied by altered composition of carotenoid and chlorophyll–protein complexes, which eventually led to the decreased photosynthetic efficiency of MGDG- and DGDG-deficient plants. [ABSTRACT FROM AUTHOR]

Highlights • Chlorophyllous fern spores as a new model for the study of desiccation tolerance. • Sustained energy dissipation in the dry state contributes to desiccation tolerance. • Desiccation tolerant spores became rapidly sensitive (72 h) upon imbibition. • Shift from tolerant to sensitive involved profound changes in thylakoids lipid composition. Abstract Fern spores are unicellular structures produced by the sporophyte generation that give rise to the haploid gametophyte. When released from the sporangium, spores are desiccation tolerant (DT) in the royal fern (Osmunda regalis) and contain fully developed chloroplasts. As a consequence, this type of spores is called chlorophyllous spores (CS). Upon transfer to germination conditions, CS initiate a process of imbibition that suppresses DT in 72 h, before the germination starts. In parallel to such change in DT, thylakoids undergo a profound remodelling in composition and function. Firstly, sustained quenching of chlorophyll fluorescence is relaxed, giving rise to photochemically active CS, while lipid composition shifts from that of a resting structure to a metabolically active cell. Basically trigalactolipids decreased in favour of monogalactolipids, with a parallel desaturation of fatty acids. Storage lipids such as triacylglycerol were quickly depleted. These results highlight the importance of the structure of thylakoids lipid as a key to protect membrane integrity during desiccation, together with the saturation of fatty acids and the constitutive chlorophyll quenching to prevent oxidative damage. The CS used here, in which the same cell shifts from DT to sensitive strategy in 72 h, reveal their potential as unicellular models for future studies on DT. [ABSTRACT FROM AUTHOR]

Abstract Diatoms are eukaryotic microalgae that play a pivotal role in biological and geochemical marine cycles. These microorganisms are at the basis of the trophic chain and their lipids are essential components (e.g. eicosapentaenoic acid, EPA) of aquatic food webs. Galactolipids are the primary lipid components of plastid membranes and form the largest lipid family of diatoms. As source of polyunsaturated fatty acids (PUFAs), these compounds are also involved in the synthesis of lipoxygenase (LOX) products such as non-volatile oxylipins and polyunsaturated aldehydes. Here, we report the first identification of two genes, namely PmLAH1 and PaLAH1 , coding for lipolytic enzymes in two diatoms of the genus Pseudo-nitzschia. Functional and modeling studies evidence a patatin-like domain endowed with galactolipase and phospholipase activity at the C-terminus of both proteins. Homologues of Pseudo-nitzschia LAH1 genes were retrieved in other diatom species so far sequenced in agreement with conservation of the functional role of these proteins within the lineage. Highlights • Galactolipids are key components of chloroplast membranes and major lipid class in marine diatoms under natural growth conditions. • By targeting glycolipids and phospholipids, different patatin-like proteins relay a growing number of important biological functions in the development and pathology of photosynthetic organisms. • LAH1 of P. arenysensis and P. multistriata contain the first patatin-like sequence identified in diatoms. The putative genes showed good sequence conservation in the patatin-like PLA (GxSxG) motif and catalytic dyad, as well as high similarity in the domain region (in the protein secondary structure level) with the patatin from Solanum cardiophyllum. • Homologous LAH1 genes are also found in the genomes or transcriptomes of any other diatom so far sequenced. The phylogenetic analysis showed that PmLAH and PaLAH formed a monophyletic group with other secondary red endosymbionts. • In analogy with plants, LAH1 is involved in the biosynthesis of oxylipins and may control the physiological role of these chemical mediators in diatom cells. Analogies with other microalgae and overexpression of LAH1 in P. tricornutum (XP_002180454.1) under N-starvation support also a possible role of this class of enzymes in galactolipid metabolism and lipid remodelling of diatoms. [ABSTRACT FROM AUTHOR]

Abstract Screening of linolipins, i.e. galactolipids containing esterified residues of divinyl ether oxylipins, in the leaves of several higher plants revealed the presence of these complex oxylipins in the meadow buttercup leaves. The rapid accumulation of linolipins occurred in the injured leaves of meadow buttercup, while intact leaves possessed no linolipins. These oxylipins were isolated from the injured leaves, separated and purified by HPLC. The structural analyses of linolipins by UV, mass-spectroscopy and NMR spectroscopy resulted in the identification of eight molecular species. Three of them were identical to linolipins B-D found earlier in the leaves of flax (Linum usitatissimum L.). Other molecular species were identified as 1- O -(ω5 Z)-etherolenoyl-2- O -dinor-(ω5 Z)-etherolenoyl-3- O -β-D-galactopyranosyl- sn -glycerol, 1- O -(ω5 Z)-etherolenoyl-2- O -(7 Z ,10 Z ,13 Z)-hexadecatrienoyl-3- O -β-D-galactopyranosyl- sn -glycerol, 1- O -(ω5 Z)-etherolenoyl-2- O -(7 Z ,10 Z)-hexadecadienoyl-3- O -β-D-galactopyranosyl- sn -glycerol, 1- O -(ω5 Z)-etherolenoyl-2- O -α-linolenoyl-3- O -β-D-galactopyranosyl- sn -glycerol, and 1- O -(ω5 Z)-etherolenoyl-2- O -palmitoyl-3- O -(α-galactopyranosyl-1-6-β-D-galactopyranosyl)- sn -glycerol. The trivial names "linolipins E, F, G, H and I," respectively, have been ascribed to these novel complex oxylipins. Graphical abstract Image 1 Highlights • Leaves of several higher plants were examined for the presence of linolipins. • Five previously unknown linolipins (E-I) were isolated from meadow buttercup leaves. • The structures of linolipins were elucidated by mass spectrometry and NMR. • Damage of meadow buttercup leaves by freezing-thawing induces linolipin synthesis. [ABSTRACT FROM AUTHOR]

Talaromyces thermophilus lipase (TTL) was found to hydrolyze monogalactosyl diacylglycerol (MGDG) and digalactosyl diacylglycerol (DGDG) substrates presented in various forms to the enzyme. Different assay techniques were used for each substrate: pHstat with dioctanoyl galactolipid-bile salt mixed micelles, barostat with dilauroyl galactolipid monomolecular films spread at the air-water interface, and UV absorption using a novel MGDG substrate containing α-eleostearic acid as chromophore and coated on microtiter plates. The kinetic properties of TTL were compared to those of the homologous lipase from Thermomyces lanuginosus (TLL), guinea pig pancreatic lipase-related protein 2 and Fusarium solani cutinase. TTL was found to be the most active galactolipase, with a higher activity on micelles than on monomolecular films or surface-coated MGDG. Nevertheless, the UV absorption assay with coated MGDG was highly sensitive and allowed measuring significant activities with about 10 ng of enzymes, against 100 ng to 10 μg with the pHstat. TTL showed longer lag times than TLL for reaching steady state kinetics of hydrolysis with monomolecular films or surface-coated MGDG. These findings and 3D-modelling of TTL based on the known structure of TLL pointed out to two phenylalanine to leucine substitutions in TTL, that could be responsible for its slower adsorption at lipid-water interface. TTL was found to be more active on MGDG than on DGDG using both galactolipid-bile salt mixed micelles and galactolipid monomolecular films. These later experiments suggest that the second galactose on galactolipid polar head impairs the enzyme adsorption on its aggregated substrate. [ABSTRACT FROM AUTHOR]

Drought triggers leaf senescence, leading to a critical decrease in crop yield. The membrane lipid composition generally undergoes major changes during drought. However, little is known about the role of galactolipid remodeling in the drought-induced senescence response in crops. Here, a comparative study of alterations in galactolipid and fatty acid composition during drought and recovery was carried out using two maize cultivars differing in drought-induced leaf senescence. Under drought condition, the monogalactosyldiacylglycerol (MGDG) content was decreased by 18.3% and 25.9% in the cultivar with retarded leaf senescence and the senescent cultivar, respectively, while digalactosyldiacylglycerol (DGDG) was enhanced by 43.2% and 22.9%. Compared with the senescent cultivar, the DGDG content and the DGDG/MGDG ratio were 29.8% and 20.7% higher in the cultivar with retarded leaf senescence, which also maintained stable chloroplast ultrastructure under drought stress. The expressions of key galactolipid biosynthesis genes in both cultivars were up-regulated by the drought stress. In addition, the expression levels were higher in the cultivar with retarded leaf senescence than that in the senescent cultivar. These results suggests that a profound modification of galactolipid composition with high DGDG content and DGDG/MGDG ratio is associated with the alleviation of leaf senescence in response to drought. The regulation of galactolipid remodeling could be a promising strategy for alleviating the leaf senescence and improving drought adaptation in crops. [ABSTRACT FROM AUTHOR]

Monogalactosyldiacylglycerol (MGDG) and digalactosyldiacylglycerol (DGDG) together constitute approximately 80% of chloroplast lipids. Apart from facilitating the photosynthesis light reaction in the thylakoid membrane, these two lipids are important for maintaining chloroplast morphology and for plant survival under abiotic stresses such as phosphate starvation and freezing. Recently it was shown that severe growth retardation phenotypes of the DGDG-deficient mutant dgd1 were due to jasmonate overproduction, linking MGDG and DGDG homeostasis with phytohormone production and suggesting MGDG as a major substrate for jasmonate biosynthesis. Induction of jasmonate synthesis and jasmonic acid (JA) signaling was also observed under conditions of phosphate starvation. We hypothesize that when DGDG is recruited to substitute for phospholipids in extraplastidic membranes during phosphate deficiency, the altered MGDG to DGDG ratio in the chloroplast envelope triggers the conversion of galactolipids into jasmonates. The conversion may contribute to rebalancing the MGDG to DGDG ratio rapidly to maintain chloroplast shape, and jasmonate production can reduce the growth rate and enhance predator deterrence. We also hypothesize that other conditions, such as suppression of dgd1 phenotypes by trigalactosyldiacylglycerol (tgd) mutations, may all be linked to altered jasmonate production, indicating that caution should be exercised when interpreting phenotypes caused by conditions that may alter the MGDG to DGDG ratio at the chloroplast envelope. [ABSTRACT FROM AUTHOR]

Biofortification using agronomic practices can be used to improve the nutritional quality of food crops. Three natural media amendments (dry vermicast, potassium (K)-humate and volcanic minerals) were assessed under greenhouse conditions to determine the effects on the chemical composition of kale ( Brassica oleracea L. var. acephala ‘Ripbor’). The results indicated that K-humate had low pH, while the volcanic minerals had high pH. Plants grown in the dry vermicast amended media had the highest levels of macronutrients and micronutrients except for zinc and iron. However, the glycolipid: phospholipid ratio was lower in kale plants cultivated in dry vermicast compared to plants cultivated in the volcanic minerals or K-humate. Conversely, plants cultivated in the dry vermicast had enhanced levels of polyunsaturated fatty acids. The omega-3 fatty acid content was significantly (P < 0.05) higher in plants cultivated in dry vermicast and potassium humate, while the omega-6 fatty acids were unaffected by media amendments. Dry vermicast was the most effective at increasing plant tissue oleic acid content. The total phenolic content and antioxidant capacity were highest in plants treated with K-humate, but lowest in the dry vermicast treated plants. In conclusion, dry vermicast proved to be the most effective in enhancing the phytochemical composition of kale ‘Ripbor’. These results suggest dry vermicast could be a potential target natural media amendment for biofortifying kale plants during cultivation. [ABSTRACT FROM AUTHOR]

We have identified an enzyme, galactolipase (ckGL), which hydrolyzes the acyl ester bond of galactolipids such as digalactosyldiacylglycerol (DGDG), in the microalga Chlorella kessleri. Following purification of the enzyme to electrophoretic homogeneity from cell-free extract, the maximum activity toward DGDG was observed at pH 6.5 and 37 °C. ckGL was Ca2+-dependent enzyme and displayed an apparent molecular mass of approx. 53 kDa on SDS-PAGE. The substrate specificity was in the order: DGDG (100%) > monogalactosyldiacylglycerol ≈ phosphatidylglycerol (~ 40%) > sulfoquinovosyldiacylglycerol (~ 20%); the enzyme exhibited almost no activity toward glycerides and other phospholipids. Gas chromatography analysis demonstrated that ckGL preferably hydrolyzed the sn-1 acyl ester bond in the substrates. The genomic DNA sequence (5.6 kb) containing the ckGL gene (designated glp1) was determined and the cDNA was cloned. glp1 was composed of 10 introns and 11 exons, and the 1608-bp full-length cDNA encoded a mature ckGL containing 475 amino acids (aa), with a presequence (60 aa) containing a potential chloroplast transit peptide. Recombinant functional ckGL was produced in Escherichia coli. Although the deduced aa sequence of ckGL contained the typical GXSXG motif of serine hydrolases together with conserved histidine and aspartate residues which would form part of the catalytic triad of α/β-hydrolases, ckGL showed no significant overall similarity with known lipases including GLs from Chlamydomonas reinhardtii and Aspergillus japonicus, indicating that ckGL is a novel GL. ckGL, with high specificity for DGDG, could be applicable to food processing as an enzyme capable of improving material textures. [ABSTRACT FROM AUTHOR]

Steroids constitute one of the most important groups of compounds of regulatory properties both in the animal and plant kingdom. In plants, steroids such as brassinosteroids or progesterone, by binding to protein receptors in cell membranes, regulate growth and initiate processes leading to increased tolerance to stress conditions. Due to their structural similarities to sterols, these steroids may also directly interact with cellular membranes. Our aim was to determine the changes of the structural parameters of lipid membranes under the influence of hydrophobic steroid compounds, i.e., 24-epibrassinolide (EBR) and its precursor—24-epicastasterone (ECS) and progesterone (PRO). Lipids were isolated from wheat seedlings with different tolerances to frost, grown at low temperatures (5 °C) for 1.5 and 3 weeks (acclimation process). Control plants were cultured continuously at 20 °C. From galactolipids and phospholipids, the main polar lipid fractions, the monolayers were formed, using a technique of Langmuir trough. EBR and ECS were introduced into monolayers, together with lipids, whereas the PRO was dissolved in the aqueous sub-phase upon which the monolayers were spread. Measurements performed at 25 °C and 10 °C showed a significant action of the tested compounds on the physicochemical properties of the monolayers. EBR and PRO increased the area per lipid molecule in monolayers, resulting in formation of more flexible surface structures while the presence of the ECS induced the opposite effect. The influence of the polarity of lipids and steroids on the interactions in the monolayer was discussed. Lipids extracted from the membranes of wheat with the most tolerance to frost were characterized by the highest fatty acid unsaturation and steroids had a relatively weak effect on the parameters of the structure of their monolayers. [ABSTRACT FROM AUTHOR]

Thymus is one of the best known genera within the Labiatae (Lamiaceae) family, with more than 200 species and many medicinal and culinary uses. The effects of prolonged drought on lipid profile were investigated in tolerant and sensitive thyme plants (Thymus serpyllum L. and Thymus vulgaris L., respectively). Non-targeted non-polar metabolite profiling was carried out using Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometry with one-month-old plants exposed to drought stress, and their morpho-physiological parameters were also evaluated. Tolerant and sensitive plants exhibited clearly different responses at a physiological level. In addition, different trends for a number of non-polar metabolites were observed when comparing stressed and control samples, for both sensitive and tolerant plants. Sensitive plants showed the highest decrease (55%) in main lipid components such as galactolipids and phospholipids. In tolerant plants, the level of lipids involved in signaling increased, while intensities of those induced by stress (e.g., oxylipins) dramatically decreased (50-60%), in particular with respect to metabolites with m/z values of 519.3331, 521.3488, and 581.3709. Partial least square discriminant analysis separated all the samples into four groups: tolerant watered, tolerant stressed, sensitive watered and sensitive stressed. The combination of lipid profiling and physiological parameters represented a promising tool for investigating the mechanisms of plant response to drought stress at non-polar metabolome level. [ABSTRACT FROM AUTHOR]

Fatty acids and lipids, which are major and essential constituents of all plant cells, not only provide structural integrity and energy for various metabolic processes but can also function as signal transduction mediators. Lipids and fatty acids can act as both intracellular and extracellular signals. In addition, cyclic and acyclic products generated during fatty acid metabolism can also function as important chemical signals. This review summarizes the biosynthesis of fatty acids and lipids and their involvement in pathogen defense. [ABSTRACT FROM AUTHOR]

Grapevine is susceptible to several diseases, being downy mildew, caused by Plasmopara viticola, among the most devastating. Control strategies include the application of pesticides every season through almost all the grapevine developmental stages, jeopardizing viticulture sustainability. Understanding resistance molecular processes is crucial to define alternative strategies and select disease resistance traits. Identifying markers that discriminate between tolerant and susceptible genotypes to P. viticola is an important step. It is shown that lipids and fatty acids (FA) play an important role in grapevine defence against downy mildew and in our previous works we have highlighted that susceptible and resistant grapevine genotypes present contrasting FA profiles even without infection. Here, the constitutive lipid and FA composition of four grapevine genotypes, with contrasting behaviour toward P. viticola was analysed. FA desaturases (FAD) gene expression was also analysed. The results indicate that the susceptible genotypes present higher content of plastidial lipids and unsaturated FA and higher expression levels of FAD genes, whereas tolerant genotypes have higher amounts of neutral lipids and phosphatidic acid as well as saturated FA. In this preliminary study, the concentration of monogalactosyldiacylglycerol and phosphatidylcholine was pinpointed as potential tolerance-associated biomarkers and both linoleic acid and plastidial lipids as susceptibility-associated biomarkers. • Lipids and fatty acid desaturase genes are grapevine tolerance/susceptibility markers to Plasmopara viticola. • Susceptible genotypes have higher plastid lipids, unsaturated fatty acids and fatty acid desaturase expression levels. • Tolerant genotypes have higher neutral lipids, phosphatidic acid and saturated fatty acid amounts. [ABSTRACT FROM AUTHOR]

In this study, we identified a pancreatic lipase-related protein 2 ( PLRP2) gene through searching the transcriptome data of the brown planthopper, Nilaparvata lugens, a monophagous rice pest. PLRP2 mRNAs were first isolated in the human pancreas and play an important role in hydrolysis of galactolipids from the diet. Although homologous PLRP2 genes have been identified in many insect species, their physiological functions remain unknown. The present study for the first time reports the functional role of PLRP2 in an insect species. Differing from mammal PLRP2s, N. lugens PLRP2 was highly expressed in developing oocytes of the ovaries of female adults and little expressed in laid eggs. Suppression of N. lugens PLRP2 expression using RNA interference significantly inhibited egg hatching in rice seedlings, implying that N. lugens PLRP2 was important for oocyte maturation and development. This novel finding will improve our understanding of the reproductive strategies in insects and provides a potential target for future management of crop pests. [ABSTRACT FROM AUTHOR]

The search for nutrient-dense food source is a top-priority in today's food-challenged world of multi-billion population. Food insecurity has become especially critical in developing countries, though potential sources of functional foods are being wasted right before our eyes. Rose hips, the fruits of rose plants ( Rosa sp.) have been discovered to be rich in polyphenols (triterpene acids, flavonoids, proanthocyanidins, catechin), essential fatty acids, galactolipid, folate, vitamin A, C and E, minerals (Ca, Mg, K, S, Si, Se, Mn and Fe), among other bioactive components. The extracts have been proven to possess antioxidant, anti-inflammatory, immunomodulation, anticancer, cardioprotective, antidiabetic, neuroprotective, and antimicrobial properties. The extracts have been validated beneficial against non-alcoholic fatty liver disease, osteoarthritis, rheumatoid arthritis, obesity, cancer, kidney stone, depression, dermal issues, among other pathologies. The mechanisms of therapeutic actions involve intervention in COX-2, iNOS, NF-kappaB, PPAR-γ, p38 MAPK, Bak, Caspase-3, Ca ++ channel blockade pathways. Barring the Rosaceae family-characteristic allergenicity due to LTPs, the rose hips are free of other side effects. This holistic review, based on recent findings is excited to report rose hips as an emerging ‘functional food’ that deserves to be integrated to food platter without delay. [ABSTRACT FROM AUTHOR]

Plants in a temperate climate are often subject to different environmental factors, chilling stress among them, which influence the growth especially during early stages of plant development. Chloroplasts are one of the first organelles affected by the chilling stress. Therefore the proper biogenesis of chloroplasts in early stages of plant growth is crucial for undertaking the photosynthetic activity. In this paper, the analysis of the cotyledon chloroplast biogenesis at different levels of plastid organization was performed in cucumber, one of the most popular chilling sensitive crops. Influence of low temperature on the ultrastructure was manifested by partial recrystallization of the prolamellar body, the formation of elongated grana thylakoids and a change of the prolamellar body structure from the compacted “closed” type to a more loose “open” type. Structural changes are strongly correlated with galactolipid and carotenoid content. Substantial changes in the galactolipid and the carotenoid composition in dark-chilled plants, especially a decrease of the monogalactosyldiacylglycerol to digalactosyldiacylglycerol ratio (MGDG/DGDG) and an increased level of lutein, responsible for a decrease in membrane fluidity, were registered together with a slower adaptation to higher light intensity and an increased level of non-photochemical reactions. Changes in the grana thylakoid fluidity, of their structure and photosynthetic efficiency in developing chloroplasts of dark-chilled plants, without significant changes in the PSI/PSII ratio, could distort the balance of photosystem rearrangements and be one of the reasons of cucumber sensitivity to chilling. [ABSTRACT FROM AUTHOR]

In this work biomimetic monolayers of a MGDG, monogalactosyldiacylglycerol, and DGDG, digalactosyldiacylglycerol mixture (MD), in a ratio close to that of the thylakoid membranes of oxygenic photosynthetic organisms, have been prepared. The lipid mixture incorporates plastoquinone-9 (PQ), that is the electron and proton shuttle of the photosynthetic reaction centres. The MD:PQ mixtures have been firstly studied using surface pressure-area isotherms. Langmuir-Blodgett (LB) films of those mixtures have been transferred onto a substrate forming a monolayer that mimics one of the bilayer sides of the thylakoid membranes. These monolayers have been characterized topographically and electrochemically. The results show the influence of PQ in the MD matrix and its partial expulsion when increasing the surface pressure, obtaining two main PQ positions in the MD matrix. The calculated apparent electron transfer rate constants indicate a different kinetic control for the reduction and the oxidation of the PQ/PQH 2 couple, being k Rapp (I) = 0.7 · 10 − 6 s − 1 , k Rapp (II) = 2.2 · 10 − 9 s − 1 , k Oapp (I) = 7.4 · 10 − 4 s − 1 and k Oapp (II) = 5.2 · 10 − 5 s − 1 , respectively. The comparison of the different galactolipid:PQ systems that our group has studied is also presented, concluding that the PQ position in the galactolipid matrix can be tuned according to several controlled variables. [ABSTRACT FROM AUTHOR]

Galactolipids [monogalactosyldiacylglycerol (MGDG) and digalactosyldiacylglycerol (DGDG)] are the hallmark lipids of photosynthetic membranes. The galactolipid synthases MGD1 and DGD1 catalyze consecutive galactosyltransfer reactions but localize to the inner and outer chloroplast envelopes, respectively, necessitating intermembrane lipid transfer. Here we show that the N-terminal sequence of DGD1 (NDGD1) is required for galactolipid transfer between the envelopes. Different diglycosyllipid synthases (DGD1, DGD2, and Chloroflexus glucosyltransferase) were introduced into the dgd1-1 mutant of Arabidopsis in fusion with N-terminal extensions (NDGD1 and NDGD2) targeting to the outer envelope. Reconstruction of DGDG synthesis in the outer envelope membrane was observed only with diglycosyllipid synthase fusion proteins carrying NDGD1, indicating that NDGD1 enables galactolipid translocation between envelopes. NDGD1 binds to phosphatidic acid (PA) in membranes and mediates PA-dependent membrane fusion in vitro. These findings provide a mechanism for the sorting and selective channeling of lipid precursors between the galactolipid pools of the two envelope membranes. [ABSTRACT FROM AUTHOR]

Arabidopsis produces galactolipids containing esters of 12-oxo-phytodienoic acid (OPDA) and dinor-12-oxo-phytodienoic acid (dnOPDA). These lipids are referred to as arabidopsides and accumulate in response to abiotic and biotic stress. We explored the natural genetic variation found in 14 different Arabidopsis accessions to identify genes involved in the formation of arabidopsides. The accession C24 was identified as a poor accumulator of arabidopsides whereas the commonly used accession Col-0 was found to accumulate comparably large amounts of arabidopsides in response to tissue damage. A quantitative trait loci analysis of an F2 population created from a cross between C24 and Col-0 located a region on chromosome four strongly linked to the capacity to form arabidopsides. Expression analysis of HYDROPEROXIDE LYASE 1 (HPL1) showed large differences in transcript abundance between accessions. Transformation of Col-0 plants with the C24 HPL1 allele under transcriptional regulation of the 35S promoter revealed a strong negative correlation between HPL1 expression and arabidopside accumulation after tissue damage, thereby strengthening the view that HPL1 competes with ALLENE OXIDE SYNTHASE (AOS) for lipid-bound hydroperoxide fatty acids. We further show that the last step in the synthesis of galactolipid-bound OPDA and dnOPDA from unstable allene oxides is exclusively enzyme-catalyzed and not the result of spontaneous cyclization. Thus, the results presented here together with previous studies suggest that all steps in arabidopside biosynthesis are enzyme-dependent and apparently all reactions can take place with substrates being esterified to galactolipids. [ABSTRACT FROM AUTHOR]

Ribosomal protein S6 kinase (S6K) functions as a key component in the target of rapamycin (TOR) pathway involved in multiple processes in eukaryotes. The role and regulation of TOR-S6K in lipid metabolism remained unknown in plants. Here we provide genetic and pharmacological evidence that TOR-Raptor2-S6K1 is important for thylakoid galactolipid biosynthesis and thylakoid grana modeling in rice ( Oryza sativa L.). Genetic suppression of S6K1 caused pale yellow-green leaves, defective thylakoid grana architecture. S6K1 directly interacts with Raptor2, a core component in TOR signaling, and S6K1 activity is regulated by Raptor2 and TOR. Plants with suppressed Raptor2 expression or reduced TOR activity by inhibitors mimicked the S6K1-deficient phenotype. A significant reduction in galactolipid content was found in the s6k1 , raptor2 mutant or TOR-inhibited plants, which was accompanied by decreased transcript levels of the set of genes such as lipid phosphate phosphatase α5 ( LPPα5 ), MGDG synthase 1 ( MGD1 ), and DGDG synthase 1 ( DGD1 ) involved in galactolipid synthesis, compared to the control plants. Moreover, loss of LPPα5 exhibited a similar phenotype with pale yellow–green leaves. These results suggest that TOR-Raptor2-S6K1 is important for modulating thylakoid membrane lipid biosynthesis, homeostasis, thus enhancing thylakoid grana architecture and normal photosynthesis ability in rice. [ABSTRACT FROM AUTHOR]

Membrane lipids contain trienoic fatty acids which enhance fluidity, an essential feature for optimal membrane performance. However highly unsaturated molecules are prone to lipoperoxidation processes. In V. unguiculata, linolenic acid (C18:3) accounts for more than 90% of the total fatty acids of galactolipids. High C18:3 contents causes galactolipids molecules, especially monogalactosyl-diacylglycerol (MGDG), to become more susceptible to drought-induced peroxidative or enzymatic degradation. In this work 8 Vigna genotypes differing in drought resistance in the field were compared to search for an eventual correlation between genotypes ability to resist to water stress and some characteristics of their membrane lipids in well-watered plants. Plants were grown in a greenhouse with semi-controlled environmental conditions, and kept fully hydrated. Leaves were collected from one month old plants to perform electrolyte leakage test under PEG-induced dehydration and lipid analysis. Results showed that both a reduced amount of total lipids, which reflects a small amount of membranes, and a reduced amount of C18:3 in MGDG, which indicates a low membrane fluidity, were strongly correlated with a high membrane resistance to dehydration, and therefore seem to be useful screening tools for drought-tolerant genotypes. Taking into account what is known concerning drought resistance of the genotypes under study, it was also concluded that the degree of membrane resistance to osmotic stress could be extrapolated to drought resistance in the field. [ABSTRACT FROM AUTHOR]

The rapid and preferential adsorption of a gastric lipase recombinant dog gastric lipase (rDGL) in heterogeneous films of phospholipids and triacylglycerols has previously been unveiled using Langmuir films analyzed by tensiometry, ellipsometry and Langmuir-Blodgett transfer coupled to atomic force microscopy. Here we invest the adsorption behavior of rDGL in heterogeneous galactolipid and mixed galactolipid-phospholipid or galactolipid-phospholipid-phytosterol films representative of plant membrane. Again rDGL, preferentially got adsorbed at the expanded lipid phases of the films underlining the genericity of such adsorption behavior. The addition of phytosterols to these mixtures resulted in the creation of defects, favoring the adsorption of rDGL at the fluid phases, but also improving the adsorption capacities of the lipase at the phase boundaries and towards the defects in the condensed phase. rDGL, like all gastric lipases, does not show any activity on galactolipids and phospholipids but its adsorption impacts their lateral organization and may change the adsorption and activity of other lipolytic enzymes in the course of digestion. [Display omitted] • rDGL binds to the fluid phase of homogeneous galactolipid monolayer. • The orientation of galactolipid polar heads limits the adsorption capacity of rDGL. • rDGL was excluded from condensed phase domains in mixed heterogenous galactolipid-phospholipid monolayer. • Defects created in the condensed phase by addition of phytosterol favor rDGL adsorption. [ABSTRACT FROM AUTHOR]

Silicon, the second abundant element in the crust, is beneficial for plant growth, mechanical strength, and stress responses. Here we show that manipulation of the non-specific phospholipase C1, NPC1, alters silicon content in nodes and husks of rice ( Oryza sativa). Silicon content in NPC1-overexpressing ( OE) plants was decreased in nodes but increased in husks compared to wild-type, whereas RNAi suppression of NPC1 resulted in the opposite changes to those of NPC1- OE plants. NPC1 from rice hydrolyzed phospholipids and galactolipids to generate diacylglycerol that can be phosphorylated to phosphatidic acid. Phosphatidic acid interacts with Lsi6, a silicon transporter that is expressed at the highest level in nodes. In addition, the node cells of NPC1- OE plants have lower contents of cellulose and hemicellulose, and thinner sclerenchyma and vascular bundle fibre cells than wild-type plants; whereas NPC1- RNAi plants displayed the opposite changes. These data indicate that NPC1 modulates silicon distribution and secondary cell wall deposition in nodes and grains, affecting mechanical strength and seed shattering. [ABSTRACT FROM AUTHOR]

The protective ability of α-tocopherol (TOH) and gallic acid (GA) acting simultaneously at the moment of oxidizer application was evaluated by determination of galactolipid layers' oxidation degree. Addition of GA resulted in a significant decrease of ozone-derived radicals shifting the threshold of lipid sensitivity by an amount approximately corresponding to the GA intake in bulk reaction with ozone. TOH presence in lipid layers results in a change of the role of GA which additionally may be involved in the reduction of tocopheroxyl radical formed during oxidation. This leads to a decrease in effectiveness of GA in diminishing the amount of ozone radicals. Such an effect was not observed for mixed layers containing galactolipid and pre-oxidized tocopherol where the ozone threshold level was associated with a stoichiometry of GA + O3 reaction. It was concluded that probably subsequent transformations of tocopheroxyl radical to less reactive forms prevent its reaction with GA the entire quantity of which is used for radicals scavenging. This result shows the role of time parameter in systems where substrates are engaged in various reactions taking place simultaneously. The inactivation of 1,1-diphenyl-2-picrylhydrazyl radical by studied antioxidants in homogeneous system confirmed observations made on the basis of lipid layer properties indicating their antagonistic action (at least at studied conditions). Formation of layers in post-oxidation situation did not depend whether tocopherol was oxidized during oxidation of lipid/tocopherol mixture or was introduced as pre-oxidized. This may be interpreted as indication that products of tocopherol oxidation may stabilize lipid layers. [ABSTRACT FROM AUTHOR]

Studying how photosynthetic cells modify membrane lipids in response to heat stress is important to understand how plants and microalgae adapt to daily fluctuations in temperature and to investigate new lipid pathways. Here, we investigate changes occurring in lipid molecular species and lipid metabolism genes during early response to heat stress in the model photosynthetic microorganism Chlamydomonas reinhardtii. Lipid molecular species analyses revealed that, after 60 min at 42 °C, a strong decrease in specific polyunsaturated membrane lipids was observed together with an increase in polyunsaturated triacylglycerols (TAGs) and diacylglycerols (DAGs). The fact that decrease in the major chloroplastic monogalactosyldiacylglycerol sn1-18:3/ sn2-16:4 was mirrored by an accumulation of DAG sn1-18:3/ sn2-16:4 and TAG sn1-18:3/ sn2-16:4/ sn3-18:3 indicated that newly accumulated TAGs were formed via direct conversion of monogalactosyldiacylglycerols to DAGs then TAGs. Lipidomic analyses showed that the third fatty acid of a TAG likely originated from a phosphatidylethanolamine or a diacylglyceryl- O-4′-(N,N,N,-trimethyl)-homoserine betaine lipid species. Candidate genes for this TAG synthesis pathway were provided through comparative transcriptomic analysis and included a phospholipase A2 homolog and the DAG acyltransferase DGTT1. This study gives insights into the molecular events underlying changes in membrane lipids during heat stress and reveals an alternative route for TAG synthesis. [ABSTRACT FROM AUTHOR]

Background: Glycerolipids are the principal constituent of cellular membranes; remodelling of glycerolipids plays important roles in temperature adaptation in plants. Temperate plants can endure freezing stress, but even chilling at above-zero temperatures can induce death in tropical species. However, little is known about the differences in glycerolipid response to low temperatures between chilling-sensitive and freezing-tolerant plants. Using ESI-MS/ MS-based lipidomic analysis, we compared the glycerolipidome of chilling (4 and 10 °C)-treated rice with that of freezing (-6 and -12 °C)-treated Arabidopsis, both immediately after these low-temperature treatments and after a subsequent recovery culture period. Results: Arabidopsis is a 16:3 plant that harbours both eukaryotic and prokaryotic-type lipid synthesis pathways, while rice is an 18:3 plant that harbours only the eukaryotic lipid synthesis pathway. Arabidopsis contains higher levels of galactolipids than rice and has a higher double bond index (DBI). Arabidopsis contains lower levels of high melting point phosphatidylglycerol (PG) molecules and has a lower average acyl chain length (ACL). Marked phospholipid degradation occurred during the recovery culture period of non-lethal chilling treated rice, but did not occur in non-lethal freezing treated Arabidopsis. Glycerolipids with larger head groups were synthesized more in Arabidopsis than in rice at sub-lethal low-temperatures. Levels of phosphatidic acid (PA) and phosphatidylinositol (PI) rose in both plants after low-temperature treatment. The DBI and ACL of total lipids did not change during low-temperature treatment. Conclusions: A higher DBI and a lower ACL could make the membranes of Arabidopsis more fluid at low temperatures. The ability to synthesize glycerolipids containing a larger head group may correlate with lowtemperature tolerance. The low-temperature-induced increase of PA may play a dual role in plant responses to low temperatures: as a lipid signal that initiates tolerance responses, and as a structural molecule that, on extensive in large accumulation, could damage the integrity of membranes. Changes in ACL and DBI are responses of plants to long-term low temperature. [ABSTRACT FROM AUTHOR]

Monogalactosyldiacylglycerol ( MGDG) and digalactosyldiacylglycerol ( DGDG) are the major lipid components of photosynthetic membranes, and hence the most abundant lipids in the biosphere. They are essential for assembly and function of the photosynthetic apparatus. In Arabidopsis, the first step of galactolipid synthesis is catalyzed by MGDG synthase 1 ( MGD1), which transfers a galactosyl residue from UDP-galactose to diacylglycerol ( DAG). MGD1 is a monotopic protein that is embedded in the inner envelope membrane of chloroplasts. Once produced, MGDG is transferred to the outer envelope membrane, where DGDG synthesis occurs, and to thylakoids. Here we present two crystal structures of MGD1: one unliganded and one complexed with UDP. MGD1 has a long and flexible region (approximately 50 amino acids) that is required for DAG binding. The structures reveal critical features of the MGD1 catalytic mechanism and its membrane binding mode, tested on biomimetic Langmuir monolayers, giving insights into chloroplast membrane biogenesis. The structural plasticity of MGD1, ensuring very rapid capture and utilization of DAG, and its interaction with anionic lipids, possibly driving the construction of lipoproteic clusters, are consistent with the role of this enzyme, not only in expansion of the inner envelope membrane, but also in supplying MGDG to the outer envelope and nascent thylakoid membranes. [ABSTRACT FROM AUTHOR]

Abstract.: The relevance of biomimetic membranes using galactolipids has not been expressed in any extensive experimental study of these lipids. Thus, on the one hand, we present an in-depth article about the presence and role of monogalactosyldiacylglycerol (MGDG) and digalactosyldiacylglycerol (DGDG) in thylakoid membranes, their physical states and their applications. On the other hand, we use the Langmuir and Langmuir-Blodgett (LB) techniques to prepare biomimetic monolayers of saturated galactolipids MGDG, DGDG and MGDG:DGDG 2:1 mixture (MD)--biological ratio--. These monolayers are studied using surface pressure-area isotherms and their data are processed to enlighten their physical states and mixing behaviour. These monolayers, once transferred to a solid substrate at several surface pressures are topographically studied on mica using atomic force microscopy (AFM) and using cyclic voltammetry for studying the electrochemical behaviour of the monolayers once transferred to indium-tin oxide (ITO), which has good optical and electrical properties. Moreover, MD presents other differences in comparison with its pure components that are explained by the presence of different kinds of galactosyl headgroups that restrict the optimal orientation of the MGDG headgroups. Graphical abstract: [ABSTRACT FROM AUTHOR]

Sulfatide (3- O-sulfogalactosylceramide, SM4s) was isolated by Thudichum from the human brain in 1884. Together with galactosylceramide, its direct metabolic precursor in the biosynthetic pathway, sulfatide is highly enriched in myelin in the central and peripheral nervous system, and it has been implicated in several aspects of the biology of myelin-forming cells. Studies obtained using galactolipid-deficient mice strongly support the notion that sulfatide plays critical roles in the correct structure and function of myelin membrane. A number of papers are suggesting that these roles are mediated by a specific function of sulfatide in the lateral organization of myelin membrane, thus affecting the sorting, lateral assembly, membrane dynamics and also the function of specific myelin proteins in different substructures of the myelin sheath. The consequences of altered sulfatide metabolism and sulfatide-mediated myelin organization with respect to myelin diseases are still poorly understood, but it's very likely that sulfatide might represent not only a critical player in the pathogenesis of several diseases, including multiple sclerosis and Alzheimer's disease, but also a potentially promising therapeutic target. [ABSTRACT FROM AUTHOR]

The lipid phase of the thylakoid membrane is mainly composed of the galactolipids mono- and digalactosyl diacylglycerol (MGDG and DGDG, respectively). It has been known since the late 1960s that MGDG can be acylated with a third fatty acid to the galactose head group (acyl-MGDG) in plant leaf homogenates. In certain brassicaceous plants like Arabidopsis thaliana, the acyl-MGDG frequently incorporates oxidized fatty acids in the form of the jasmonic acid precursor 12-oxo-phytodienoic acid (OPDA). In the present study we further investigated the distribution of acylated and OPDA-containing galactolipids in the plant kingdom. While acyl-MGDG was found to be ubiquitous in green tissue of plants ranging from non-vascular plants to angiosperms, OPDA-containing galactolipids were only present in plants from a few genera. A candidate protein responsible for the acyl transfer was identified in Avena sativa (oat) leaf tissue using biochemical fractionation and proteomics. Knockout of the orthologous gene in A. thaliana resulted in an almost total elimination of the ability to form both non-oxidized and OPDA-containing acyl-MGDG. In addition, heterologous expression of the A. thaliana gene in E. coli demonstrated that the protein catalyzed acylation of MGDG. We thus demonstrate that a phylogenetically conserved enzyme is responsible for the accumulation of acyl-MGDG in A. thaliana. The activity of this enzyme in vivo is strongly enhanced by freezing damage and the hypersensitive response. [ABSTRACT FROM AUTHOR]

Several classes of glycerolipids were isolated from the total lipids of the algae Saccharina cichorioides, Eualaria fistulosa, Fucus evanescens, Sargassum pallidum, Silvetia babingtonii (Ochrophyta, Phaeophyceae), Tichocarpus crinitus, and Neorhodomela larix (Rhodophyta, Florideophyceae). The structures of these lipids were examined by nuclear magnetic resonance (NMR) spectroscopy, including 1D (¹H and 13C) and 2D (COSY, HSQC and HMBC) experiments. All of the investigated algae included common galactolipids and sulfonoglycolipids as the major glycolipids. Minor glycolipids isolated from S. cichorioides, T. crinitus, and N. laris were identified as lyso-galactolipids with a polar group consisted of the galactose. Comparison of the ¹H NMR data of minor nonpolar lipids isolated from the extracts of the brown algae S. pallidum and F. evanescens with the ¹H NMR data of other lipids allowed them to be identified as diacylglycerols. The structures of betaine lipids isolated from brown algae were confirmed by NMR for the first time. The fatty acid compositions of the isolated lipids were determined by gas chromatography-mass spectrometry. [ABSTRACT FROM AUTHOR]

Pancreatic triglyceride lipase (PNLIP) is essential for dietary fat digestion in children and adults, whereas a homolog, pancreatic lipase-related protein 2 (PNLIPRP2), is critical in newborns. The two lipases are structurally similar, yet they have different substrate specificities. PNLIP only cleaves neutral fats. PNLIPRP2 cleaves neutral and polar fats. To test the hypothesis that the differences in activity between PNLIP and PNLIPRP2 are governed by surface loops around the active site, we created multiple chimeras of both lipases by exchanging the surface loops singly or in combination. The chimeras were expressed, purified, and tested for activity against various substrates. The structural determinants of PNLIPRP2 galactolipase activity were contained in the N-terminal domain. Of the surface loops tested, the lid domain and the β5-loop influenced activity against triglycerides and galactolipids. Any chimera on PNLIP with the PNLIPRP2 lid domain or β5-loop had decreased triglyceride lipase activity similar to that of PNLIPRP2. The corresponding chimeras of PNLIPRP2 did not increase activity against neutral lipids. Galactolipase activity was abolished by the PNLIPβ5-loop and decreased by the PNLIP lid domain. The source of the β9-loop had minimal effect on activity. We conclude that the lid domain and β5-loop contribute to substrate specificity but do not completely account for the differing activities of PNLIP and PNLIPRP2. Other regions in the N-terminal domain must contribute to the galactolipase activity of PNLIPRP2 through direct interactions with the substrate or by altering the conformation of the residues surrounding the hydrophilic cavity in PNLIPRP2. [ABSTRACT FROM AUTHOR]

The plant hormone abscisic acid (ABA) controls growth and development and regulates plant water status through an established signaling pathway. In the presence of ABA, pyrabactin resistance/regulatory component of ABA receptor proteins inhibit type 2C protein phosphatases (PP2Cs). This, in turn, enables the activation of Sucrose Nonfermenting1-Related Protein Kinases2 (SnRK2). Open Stomata1 (OST1)/SnRK2.6/SRK2E is a major SnRK2-type protein kinase responsible for mediating ABA responses. Arabidopsis (Arabidopsis thaliana) expressing an epitope-tagged OST1 in the recessive ost1-3 mutant background was used for the copurification and identification of OST1-interacting proteins after osmotic stress and ABA treatments. These analyses, which were confirmed using bimolecular fluorescence complementation and coimmunoprecipitation, unexpectedly revealed homo- and heteromerization of OST1 with SnRK2.2, SnRK2.3, OST1, and SnRK2.8. Furthermore, several OST1-complexed proteins were identified as type 2A protein phosphatase (PP2A) subunits and as proteins involved in lipid and galactolipid metabolism. More detailed analyses suggested an interaction network between ABA-activated SnRK2-type protein kinases and several PP2A-type protein phosphatase regulatory subunits. pp2a double mutants exhibited a reduced sensitivity to ABA during seed germination and stomatal closure and an enhanced ABA sensitivity in root growth regulation. These analyses add PP2A-type protein phosphatases as another class of protein phosphatases to the interaction network of SnRK2-type protein kinases. [ABSTRACT FROM AUTHOR]

Mono- and digalactosyldiacylglycerol (MGDG and DGDG, respectively) are important galactolipids that comprise photosynthetic membranes in almost all photosynthetic organisms. Intact forms of MGDG and DGDG of Euglena gracilis and Lepocinclis acus, two example euglenids with secondary plastids of green algal origin, were elucidated with fatty acid regiochemistry via positive-ion electrospray ionization/mass spectrometry at two growth temperatures. At 20°C, E. gracilis and L. acus produced predominantly 18:3/16:4 (sn-1/sn-2) MGDG, whereas at 30°C this was supplanted by 18:2/16:2 MGDG. At both temperatures were also observed a variety of other MGDG and DGDG forms, including C20 fatty acidcontaining forms not expected in a green algal-derived plastid. In addition to providing structural details of MGDG and DGDG not available in past studies, these results suggest a previously unknown relationship between these two organisms and the red algae. This study also illustrates that temperature modulation of galactolipids occurs via modification of unsaturation of both the sn-1 and sn-2 fatty acids; this is fundamentally different from previously published studies from our laboratory on other algal classes. [ABSTRACT FROM AUTHOR]

The activity of bc1 complex kinase (ABC1K) family is a large group of atypical protein kinases found in prokaryotes and eukaryotes. In bacteria and mitochondria, ABC1K kinases are necessary for the synthesis of coenzyme Q and are therefore involved in the respiratory pathway. In chloroplasts, they are involved in prenylquinone synthesis and stress responses, but their functional role remains unclear. Plants can respond to biotic and abiotic stress by modifying membrane fluidity in order to create a suitable environment for the activity of integral membrane proteins. Therefore, this work was focused on the analysis of the effect of ABC1K7 and ABC1K8 on the production of polar lipids and their accumulation in Arabidopsis thaliana leaves. A comparison of abc1k7 and abc1k8 single mutants and the abc1k7/abc1k8 double mutant with wild-type plants and transgenic lines overexpressing ABC1K7 and ABC1K8 was performed using untargeted lipidomic analysis based on liquid chromatography coupled to mass spectrometry. Multivariate data analysis identified sets of chloroplast lipids representing the different genotypes. The abc1k7 and abc1k8 single mutants produced lower levels of the highly unsaturated lipid digalactosyldiacylglycerol (DGDG) than wild-type plants and also different forms of monogalactosyldiacylglycerol (MGDG) and kaempferol. The abc1k8 mutant also produced higher levels of oxylipin-conjugated DGDG and sinapates. The double mutant produced even higher levels of oxylipin- conjugated MGDG and DGDG. These results show that ABC1K7 and ABC1K8 influence chloroplast lipid synthesis or accumulation and modulate chloroplast membrane composition in response to stress. [ABSTRACT FROM AUTHOR]

The thylakoid membrane is mainly composed of non-common lipids, so called galactolipids. Despite the importance of these lipids for the function of the photosynthetic reaction centers, the molecular organization of these membranes is largely unexplored. Here we use multiscale molecular dynamics simulations to characterize the thylakoid membrane of both cyanobacteria and higher plants. We consider mixtures of up to five different galactolipids plus phosphatidylglycerol to represent these complex membranes. We find that the different lipids generally mix well, although nanoscale heterogeneities are observed especially in case of the plant membrane. The fluidity of the cyanobacterial membrane is markedly reduced compared to the plant membrane, even considering elevated temperatures at which thermophilic cyanobacteria are found. We also find that the plant membrane more readily undergoes a phase transformation to an inverted hexagonal phase. We furthermore characterized the conformation and dynamics of the cofactors plastoquinone and plastoquinol, revealing of the fast flip-flop rates for the non-reduced form. Together, our results provide a molecular view on the dynamical organization of the thylakoid membrane. [ABSTRACT FROM AUTHOR]

This study describes an interesting observation that the mixture of anomeric galactolipids has synergistic effects on the growth inhibition of human multiple myeloma (MM) cells. We determine that the equivalent mixture of a pair of α- and β-galactolipids with a 14-carbon lipid chain can cause stronger poly ADP-ribose polymerase cleavage and DNA damage, producing more late apoptotic MM cells, than either anomer alone. [ABSTRACT FROM AUTHOR]