Mono- and digalactosyldiacylglycerol composition of dinoflagellates. V. The galactolipid profile of Alexandrium tamarense (Dinophyceae) during the course of infection by the parasitic syndinian dinoflagellate Amoebophrya sp.

Amoebophrya is a parasitic, syndinian dinoflagellate genus that must infect another host dinoflagellate in order to reproduce. Work by Park et al. [Mar. Ecol. Prog. Ser., 227: 281–292 (2002)] has led to the hypothesis that Amoebophrya's development within a host cell nucleus disrupts the flow of genetic information involved in plastidial function. The possibility that genetic disruption by this parasite could lead to alterations in plastidial lipid composition during the course of an infection has not yet been elucidated. Our primary objective in this lipidomic study was to examine the chloroplast membrane galactolipid composition of Alexandrium tamarense infected by an Amoebophrya species in order to determine whether infection of A. tamarense causes a phenotypic alteration in the composition of mono- and digalactosyldiacylglycerol (MGDG and DGDG, respectively), two galactolipids that comprise the majority of photosynthetic membranes. Our secondary objective was to determine if non-photosynthetic Amoebophrya sp. either incorporated host cell MGDG and DGDG, and/or itself produced forms of MGDG and DGDG, as has been observed previously in heterotrophic apicomplexan parasites distantly related to Amoebophrya. We found that, despite development of Amoebophrya sp. within the nucleus, the composition of A. tamarense MGDG and DGDG did not change throughout the infection process. The predominant forms of these galactolipids were 18:5/18:4 (sn???1/sn?2) and 20:5/18:4 DGDG, which were present at similar abundances in both an uninfected host and a host late in the infection process just prior to release of Amoebophrya sp. dinospores. Amoebophrya sp. did not possess appreciable amounts of any forms of MGDG and DGDG.     

Mono- and digalactosyldiacylglycerol composition of dinoflagellates. VI. Biochemical and genomic comparison of galactolipid biosynthesis between Chromera velia (Chromerida), a photosynthetic alveolate with red algal plastid ancestry,and the dinoflagellate,Lingulodinium polyedrum

Chromera velia is a recently discovered, photosynthetic, free-living alveolate that is the closest free-living relative to non-photosynthetic apicomplexan parasites. Most plastids, regardless of their origin, have membranes composed chiefly of two galactolipids, mono- and digalactosyldiacylglycerol (MGDG and DGDG, respectively). Because of the hypothesized shared red algal origin between the plastids of C. velia and dinoflagellates, our primary objectives were to examine how growth temperature affects MGDG and DGDG composition via positive-ion electrospray/mass spectrometry (ESI/MS) and positive ion/electrospray/mass spectrometry/mass spectrometry (ESI/MS/MS), and to examine galactolipid biosynthetic genes to determine if shared ancestry translates into shared MGDG and DGDG composition. When growing at 20°C, C. velia produces eicosapentaenoic acid-rich 20:5(n-3)/20:5(n-3) (sn-1/sn-2) MGDG and 20:5(n-3)/20:5(n-3) DGDG as its primary galactolipids, with relative percentage compositions of approximately 35 and 60%, respectively. At 30°C these are lessened by approximately 5 and 8%, respectively, by the corresponding production of 20:5/20:4 forms of these lipids. The presence of 20:5 at the sn-1 position is similar to what has been observed previously in a cluster of peridinin-containing dinoflagellates, but the presence of 20:5(n-3) at the sn-2 position is extremely rare. Thus, the forms of MGDG and DGDG in C. velia displayed similarities and differences to what has been observed in peridinin-containing dinoflagellates, such as Lingulodinium polyedrum, which produces 20:5/18:5 and 20:5/18:4 as the major forms of MGDG and DGDG. We develop conceptual models from the galactolipids observed and galactolipid-relevant gene annotations to explain the presence of polyunsaturated fatty acid-containing MGDG and DGDG in both L. polyedrum and C. velia.     

Growth light level and photon absorption by cells of Chlamydomonas rheinhardii,Dunaliella tertiolecta (Chlorophyceae,Volvocales), Scenedesmus obliquus (Chlorophyceae,Chlorococcales) and Euglena viridis (Euglenophyceae,Euglenales)

Reductions in the growth light level (40 to 6 μmol m^-2 s^-1) resulted in increases in chlorophyll and protein per cell for all of the species examined. Only Dunaliella tertiolecta exhibited a reduction in chlorophyll a:b ratio with decreases in the photon flux density. However, the specific absorption coefficient (ā? _i ) normalized to chlorphyll a (ā? _a remained invariant for all of the microalgae studied. Constant values for the specific absorption coefficient normalized to the total pigment content (ā? _a+b ) were also found for the species Chlamydomonas rheinhardii, Euglena viridis and Scenedesmus obliquus. In contrast ā? _a+b for D. tertiolecta decreased with a reduction in light level due to an increase in the proportion of chlorophyll b. Differences in ā? _i were related to cell size and pigment content and possible reasons for the constancy of ā? _a discussed. Increases in the absorption cross sections (¯sQ _a ) were also found at reduced light levels due to an increase in the absorptance per cell (α_cell). The lower α_cell for D. tertiolecta, compared with C. rheinhardii was exactly compensated for by a larger light-capturing area. Although the increase in α_cell does not compensate for the reduction in the incident light level, it does reduce this range by half on an absorbed light basis.     

Mono‐ and digalactosyldiacylglycerol composition of the marennine‐producing diatom,Haslea ostrearia: Comparison to a selection of pennate and centric diatoms

Diatoms are one of the largest groups of primary producers in the oceans, yet despite their environmental importance little is known about their plastidial lipid biochemistry. It has been previously reported that Skeletonema species contain primarily C₁₆/C₁₆ and C₂₀/C₁₆ forms of mono‐ and digalactosyldiacylglycerol (MGDG and DGDG, respectively). Likewise, it was also reported that Phaeodactylum tricornutum contains primarily C₁₆/C₁₆ and C₂₀/C₂₀ forms of MGDG and DGDG. We seek to relate their studies to other diatoms, both in the centrics and pennates, with particular focus on the marennine‐producing pennate diatom, Haslea ostrearia. To this end, the composition and positional distribution of fatty acids of MGDG and DGDG were examined using positive‐ion electrospray ionization/mass spectrometry (ESI/MS). Two centric diatoms, Skeletonema marinoi and Thalassiosira weissflogii, and the pennate diatom, P. tricornutum, contained primarily C₂₀/C₁₆ (sn‐1/sn‐2) and C₁₈/C₁₆ forms of MGDG and DGDG. The other pennate diatoms, H. ostrearia and Navicula perminuta, contained primarily C₁₈/C₁₆ or C₁₈/C₁₈ forms of MGDG and DGDG, indicating a previously unrecognized fatty acid diversity in diatom MGDG and DGDG.     

Galactolipid composition of the star-shaped dinoflagellate Asterodinium gracile (Kareniaceae): presence of hexadecatetraenoic acid (16:4(n-3))-containing monogalactosyldiacylglycerol as the predominant galactolipid and chemotaxonomic closeness to Karenia mikimotoi as the only other known Kareniacean producer of this fatty acid

Asterodinium gracile is a morphologically distinct, star-shaped member of the Kareniaceae with, like canonical Kareniaceae, a tertiary plastid of haptophyte origin. However, A. gracile's complement of carotenoid photosynthetic pigments has been shown to be chemotaxonomically atypical in that it possesses much less fucoxanthin when compared to that of other, canonical Kareniaceae in the genera Karenia, Karlodinium, and Takayama, also with a tertiary plastid of haptophyte origin. To date, Karenia mikimotoi, Karenia papilionacea, and Karenia selliformis are the only canonical Kareniaceae that have been shown to have a chemotaxonomically atypical carotenoid pigment composition in that they possess a gyroxanthin diester-like carotenoid not observed in other species of Karenia, Karlodinium, or Takayama (recognizing that Karenia, in general, produces fucoxanthin derivatives not observed in Karlodinium or Takayama). As a photosynthetic organism, K. mikimotoi has been shown to resemble Karenia brevis such that both species possess the chloroplast-associated galactolipids mono- and digalactosyldiacylglycerol (MGDG and DGDG, respectively) enriched with octadecapentaenoic acid (18:5(n-3)) in the sn-1 position, and hexadecenoic acid (16:0) and tetradecanoic acid (14:0) at the sn-2 position. However, K. mikimotoi is chemotaxonomically atypical beyond its carotenoid composition in that it possesses MGDG and DGDG with hexadecatetraenoic acid (16:4(n-3)), which has not been observed in any other members of the Kareniaceae, in the sn-2 position as major galactolipids. The goal of this study was to characterize the galactolipids of A. gracile with the hypothesis that they would also be atypical when compared to other canonical Kareniaceae because of A. gracile's atypical carotenoid pigment composition. To this end, we report that like K. brevis and K. mikimotoi, A. gracile produces MGDG and DGDG enriched in 18:5(n-3) at the sn-1 position and C₁₄ fatty acids, such as 14:0, at the sn-2 position, and like K. mikimotoi, it produces 18:5(n-3)/16:4(n-3) MGDG, yet here as its most abundant galactolipid.     

Lipid composition of the pea aphid,Acyrthosiphon pisum (Harris) (homoptera: Aphididae), reared on host plant and on artificial media

Polar and neutral lipids and their constitutive fatty acids were quantified in the pea aphid, Acyrthosiphon pisum (Harris), grown on host plant or on a lipid free artificial diet. The results were compared to determine if lipids were involved in the suitability of the diet for continuous rearing of this A. pisum biotype. For apterous adults grown on plants, the lipids were characterized by a low amount of neutral lipids (2.5% weight/fresh weight) almost entirely (96.4%) composed of hexanoyl and sorboyl dimyristin. These storage lipids were higher in the alatae (3.8%), probably correlated with potential flight activity. The phospholipid amounts were identical in these two morphs (1.3–1.4% weight/fresh weight), comprised mainly of phosphatidylethanolamines (52%) and phosphatidylcholines (40.6%). These phospholipids contained a still unidentified fatty acid, with a retention time close to that of linolenic acid and synthesized by the aphid or its bacterial symbionts (not found in plants). The apterous adult aphids reared on an artificial diet showed an accumulation of neutral lipids (8.9% for the first generation); this increase was shown to be slightly greater for the hexanoyl and sorboyl triglycerides. In contrast, the phospholipids decreased in aphids reared on an artificial diet (1.1% and 0.9%, respectively, for first and second generation), correlated with a phospholipid fatty acid profile significantly deficient in C18:3 and in the unidentified peculiar fatty acid. These phospholipids are essential components of biological membranes and a diet-driven deficient synthesis in some of their components may result in the observed symptoms. © 1992 Wiley-Liss, Inc.     

Formation of biomass,total protein,chlorophylls, lipids and fatty acids in green and blue-green algae during one growth phase.

Batch cultures (8–32 l.) of Chlorella vulgaris and Scenedesmus obliquus and of Anacystis nidulans and Microcystis aeruginosa were grown in media containing 0.001 % KNO₃ and at several stages in growth sampled for biomass, total protein, chlorophylls, lipids and fatty acids. With increasing time and decreasing nitrogen concentrations, the biomass of all of the algae increased, whereas the total protein and chlorophyll content dropped. Green and blue-green algae, however, behaved differently in their lipid metabolism. In the green algae the total lipid and fatty acid content as well as the composition of these compounds changed considerably during one growth phase and was dependent on the nitrogen concentration in the media at any given day of growth. More specifically, during the initial stages of growth the green algae produced larger amounts of polar lipids and polyunsaturated C₁₆ and C₁₈ fatty acids. Towards the end of growth, however, these patterns changed in that the main lipids of the green algae were neutral with mainly saturated fatty acids (mostly 18:1 and 16:0). Such changes did not occur in the blue-green algae. These differences between prokaryotic and eukaryotic algae can possibly be explained by the ‘endosymbiont theory’.     

Correlations between the thermal stability of chloroplast (thylakoid) membranes and the composition and fluidity of their polar lipids upon acclimation of the higher plant,Nerium oleander,to growth temperature

The thermal stability of excitation transfer from pigment proteins to the Photosystem II reaction center of Nerium oleander adjusts by 10 Celsius degrees when cloned plants grown at 20°C/15°C, day/night growth temperatures are shifted to 45°C/32°C growth temperature or vice versa. Concomitant with this adjustment is a decrease in the fluidity of thylakoid membrane polar lipids as determined by spin labeling. The results are consistent with the hypothesis that there is a limiting maximum fluidity compatible with maintenance of native membrane structure and function. This limiting fluidity was about the same as for a number of other species which exhibit a range of thermal stabilities. Inversely correlated shifts in lipid fluidity and thermal stability occurred during the time course of acclimation of N. oleander to new growth temperatures. Thus, the temperature at which the limiting fluidity was reached changed during acclimation while the limiting fluidity remained constant. Although the relative proportion of the major classes of membrane polar lipids remained constant during adjustments in fluidity, large changes occured in the abundance of specific fatty acids. These changes were different for the phospho- and galacto-lipids suggesting that the fatty acid composition of these two lipid classes is regulated by different mechanisms. Comparisons between membrane lipid fluidity and fatty acid composition indicate that fluidity is not a simple linear function of fatty acid composition.     

Chemical compositions and physical states of chloroplast lipids related to atrazine resistance in Conyza canadensis L.

A comparison of the chemical composition and physical states of chloroplast lipids, of atrazine-resistant (R) and sensitive (S) biotypes of Conyza canadensis L. (horseweed), in the rosetta stage showed: (1) the R biotype contains lower amounts of polar lipids in its thylakoids, as expressed on a chlorophyll basis, than the S biotype. (2) The chloroplasts of the R biotype have higher contents of monogalactosyl diacylglycerol (MGDG) and lower contents of digalactosyl diacylglycerol (DGDG) and phosphatidylglycerol (PG), than those of the S biotype. (3) The chloroplast total lipids exhibit a higher degree of unsaturation in the R biotype. This is due to a higher level of linolenic acid, and a lower level of palmitic acid in the glycolipids. The fatty acid compositions of the phospholipids, except that of PG, do not differ significantly. (4) The lipid matrix of the thylakoid membranes of the R biotype is more fluid than that of the S biotype, as measured by the fluorescence polarization technique. The results are discussed in terms of whether these differences are responsible for the herbicide resistance.     

LYCAT, a homologue of C. elegans acl-8, acl-9, and acl-10, determines the fatty acid composition of phosphatidylinositol in mice

Mammalian phosphatidylinositol (PI) has a unique fatty acid composition in that 1-stearoyl-2-arachidonoyl species is predominant. This fatty acid composition is formed through fatty acid remodeling by sequential deacylation and reacylation. We recently identified three Caenorhabditis elegans acyltransferases (ACL-8, ACL-9, and ACL-10) that incorporate stearic acid into the sn-1 position of PI. Mammalian LYCAT, which is the closest homolog of ACL-8, ACL-9, and ACL-10, was originally identified as a lysocardiolipin acyltransferase by an in vitro assay and was subsequently reported to possess acyltransferase activity toward various anionic lysophospholipids. However, the in vivo role of mammalian LYCAT in phospholipid fatty acid metabolism has not been well elucidated. In this study, we generated LYCAT-deficient mice and demonstrated that LYCAT determined the fatty acid composition of PI in vivo. LYCAT-deficient mice were outwardly healthy and fertile. In the mice, stearoyl-CoA acyltransferase activity toward the sn-1 position of PI was reduced, and the fatty acid composition of PI, but not those of other major phospholipids, was altered. Furthermore, expression of mouse LYCAT rescued the phenotype of C. elegans acl-8 acl-9 acl-10 triple mutants. Our data indicate that LYCAT is a determinant of PI molecular species and its function is conserved in C. elegans and mammals.     

Are Sulfurous Soil Amendments (S0, Fe(II)SO4, Fe(III)SO4) an Effective Tool in the Restoration of Heathland and Acidic Grassland after Four Decades of Rock Phosphate Fertilization?

This paper deals with the complex issue of reversing long‐term improvements of fertility in soils derived from heathlands and acidic grasslands using sulfur‐based amendments. The experiment was conducted on a former heathland and acid grassland in the U.K. that was heavily fertilized and limed with rock phosphate, chalk, and marl. The experimental work had three aims. First, to determine whether sulfurous soil amendments are able to lower pH to a level suitable for heathland and acidic grassland re‐creation (approximately 3 pH units). Second, to determine what effect the soil amendments have on the available pool of some basic cations and some potentially toxic acidic cations that may affect the plant community. Third, to determine whether the addition of Fe to the soil system would sequester PO₄^? ions that might be liberated from rock phosphate by the experimental treatments. The application of S⁰ and Fe^(II)SO₄^? to the soil was able to reduce pH. However, only the highest S⁰ treatment (2,000 kg/ha S) lowered pH sufficiently for heathland restoration purposes but effectively so. Where pH was lowered, basic cations were lost from the exchangeable pool and replaced by acidic cations. Where Fe was added to the soil, there was no evidence of PO₄^? sequestration from soil test data (Olsen P), but sequestration was apparent because of lower foliar P in the grass sward. The ability of the forb Rumex acetosella to apparently detoxify Al³⁺, prevalent in acidified soils, appeared to give it a competitive advantage over other less tolerant species. We would anticipate further changes in plant community structure through time, driven by Al³⁺ toxicity, leading to the competitive exclusion of less tolerant species. This, we suggest, is a key abiotic driver in the restoration of biotic (acidic plant) communities.     

Apoptosis of human carcinoma cells in the presence of potential anti-cancer drugs: III. Treatment of Colo-205 and SKBR3 cells with: cis-platin, Tamoxifen, Melphalan, Betulinic acid, L-PDMP, L-PPMP, and GD3 ganglioside

Breast cancer is the most common type of cancer, predominantly among women over 20, whereas colo-rectal cancer occurs in both men and women over the age of 50. Chemotherapy of both cancers affect rapidly growing normal as well as cancer cells. Cancer cells are non-apoptotic. Seven anti-cancer agents (cis-platin, Tamoxifen, Melphalan, Betulinic acid, D-PDMP, L-PPMP, and GD3) have been tested with human breast (SKBR3) and colon (Colo-205) carcinoma cells for their apoptotic effect and found to be positive by several assay systems. Colo-205 cells were obtained from ATCC, and the SKBR3 cells were a gift from the Cleveland Clinic. All of these six agents killed those two cell lines in a dose-dependent manner. In the early apoptotic stage (6 h), these cells showed only a flopping of phosphatidylserine on the outer lamella of the plasma membranes as evidenced by the binding of a novel fluorescent dye PSS-380. After 24 h of the treatment, those apoptotic cells showed damage of the plasma as well as the nuclear membrane as evidenced by binding of propidium iodide to the nuclear DNA. DNA laddering assay viewed further breakdown of DNA by 1% agarose gel electrophoresis analysis. It is concluded that during apoptosis the signaling by Mitochondrial Signaling Pathway (MSP) is stimulated by some of these agents. Caspase 3 was activated with the concomitant appearance of its p17 polypeptide as viewed by Westernblot analyses. Incorporation of radioactivity from [U-¹⁴C]-L-serine in total sphingolipid mixture was observed between 2 and 4 micromolar concentrations of most of the agents except cis-platin. However, apoptosis in carcinoma cells in the presence of cis-platin is induced by a caspase 3 activation pathway without any increase in synthesis of ceramide. Published in 2004..     

Lipid extracts from different algal species:1H and13C-NMR spectroscopic studies as a new tool to screen differences in the composition of fatty acids,sterols and carotenoids

One- and two-dimensional¹H- and¹³C-NMR spectra of lipid extracts fromUlva rigida, Gracilaria longa, Fucus virsoides andCodium tomentosum collected in the northern Adriatic Sea allowed screening of the content of fatty acid chains, carotenoids, free and acylated cholesterol and chlorophylls. The carotenoid-to-polyunsaturated fatty acid molar ratio was taken as a comparison parameter in samples ofUlva rigida collected in differentloci and seasons; the value was markedly higher in samples from the Lagoon of Venice than from marine coastal waters. The total cholesterol concentration was evaluated by¹H-NMR spectroscopy and similar values were found for all species. Two-dimensional heterocorrelated NMR spectroscopy was shown to give characteristic fingerprints of the lipid extracts from algal samples as regards the content in chlorophylls, unsaturated fatty acids and carotenoids.author for correspondence     

Steric effects on the substitution of pentaamine-chromium(III) and -rhodium(III) complexes. Anation reaction rate constants as an indicative of the dissociative shift of the mechanism on crowding the [M(RNH2)5H2O] (M = Rh, R = H, Me, Et, Pr; M = Cr, R = H, Me, Pr) complexes

The kinetics of the anation reactions of [M(RNH₂)₅H₂O]³⁺ (M = Rh, R = H, Me, Et, Pr; M = Cr, R = H, Me, Pr) with several ligands (H₃PO₄/H₂PO₄⁻, H₃PO₃/H₂PO₃⁻, CF₃COO⁻, Br⁻, Cl⁻, SCN⁻) have been studied at different temperatures and acidities at I = 1.0 M (LiClO₄. Results obtained for the anation rate constants and thermal activation parameters are compared with the previously published data for R = H, in order to establish the effects of the amine substituents in the reaction mechanism proposed for the substitution reactions of these complexes. The results obtained are interpreted on the basis of a mechanism where the bond formation process is more important in the substitution on M = Cr complexes than in that of the M = Rh complexes, as already pointed out for the published ΔΛ^≠ values for the water exchange on these systems. A simple Langford-Gray classification becomes inadequate to describe these situations where the increase of the steric demand of the amine substituents shifta the I_a-I_d classification to the I_d side, although no dramatic changes in the reaction mechanism are found. It is concluded that a More O'Ferall ‘continuous’ type of approach to the mechanism classification of the substitution reactions is much more useful in this case.