Changes in the fatty acid composition of symbiotic dinoflagellates from the hermatypic coral <Emphasis Type="Italic">Echinopora lamellosa</Emphasis> during adaptation to the irradiance level

The composition of fatty acids (FAs) of symbiotic dinoflagellates isolated from the hermatypic coral Echinoporal lamellosa adapted to the irradiance of 95, 30, 8, and 2% PAR was studied. Polar lipids and triacylglycerols (TAG) differed between them in FA composition. Polar lipids were enriched in unsaturated FAs, whereas TAG, in saturated FAs. Light exerted a substantial influence on the FA composition in both polar lipids and TAG. The elevation of irradiance resulted in the accumulation of 16:0 acid in both lipid groups and 16:1(n-7) acid in TAG. It seems likely that de novo synthesis of 16:0 acid occurred actively in the cells of symbiotic dinoflagellates in high light. Since these processes are energy-consuming ones, they utilize excessive energy. When light intensity declined, 18:4(n-3) and 20:5(n-3) acids accumulated in polar lipids, which was accompanied by the increase in the content of chlorophyll a in the cells of zooxanthellae, whereas the levels of 22:6(n-3) and 20:4(n-6) acids reduced. Although the relative content of particular FAs varied substantially in dependence of irradiance, the balance between the sum of saturated and unsaturated FAs changed insignificantly. We concluded that the role of photoadaptation could not be limited only to changes in the degree of lipid unsaturation and membrane fluidity. It is supposed that light-induced changes in the FA composition reflect the interrelation between photosynthesis and FA biosynthesis.     

Fatty acid composition of sprat (<Emphasis Type="Italic">Sprattus sprattus</Emphasis>) and herring (<Emphasis Type="Italic">Clupea harengus</Emphasis>) in the Baltic Sea as potential prey for salmon (<Emphasis Type="Italic">Salmo salar</Emphasis>)

Sprat (Sprattus sprattus) and small herring (Clupea harengus) are the dominant prey fish of Atlantic salmon (Salmo salar) in the Baltic Sea. If the fatty acid (FA) proportions of sprat and herring differ, the dietary history of ascending salmon could be determined from their FA profiles. Therefore, we investigated the FA composition of several age groups of whole sprat and small herring, caught from the three main feeding areas of salmon in autumn and spring. Oleic acid (18:1n-9) was the most prevalent FA in sprat and characteristic of this species. In herring, palmitic acid (16:0) was the most common FA, but herring lipid was characterized by n-6 polyunsaturated FAs, and moreover, by palmitoleic acid (16:1n-7) and vaccenic acid (18:1n-7). Due to the higher lipid content of sprat, the concentrations of all other FAs, excluding these, were higher in sprat than in herring. The concentration of docosahexaenoic acid (DHA, 22:6n-3) increased with an increase in the lipid content and was consequently highest in the youngest specimens, being in young sprat almost double that of young herring, and 2.6 times higher in the sprat biomass than in that of herring. As a result of a decrease in the DHA concentration with age, the ratio thiamine/DHA increased with respect to age in both species, and was lower in sprat than in herring. It is concluded that an abundance of DHA in the diet of salmon most likely increases oxidative stress because of the susceptibility of DHA to peroxidation, and thus decreases thiamine resources of fasting, prespawning salmon. Because the FA composition of sprat and herring differs, and the relative abundancies of prey fish differ between the feeding areas of salmon, the feeding area of ascending salmon can most probably be derived by comparing their FA profiles.     

High level of tetracosapolyenoic fatty acids in the cold-water mollusk <Emphasis Type="Italic">Tochuina</Emphasis><Emphasis Type="Italic">tetraquetra</Emphasis> is a result of the nudibranch feeding on soft corals

Fatty acid (FA) markers are used to trace predator–prey relationships in the marine environment. Soft corals contain tetracosapolyenoic acids (TPA), namely 24:5n-6 and 24:6n-3, which are considered as octocoral FA markers. The nudibranch mollusks are known to feed on soft corals. To check whether TPA are transferred from soft corals to nudibranch mollusks during their nutrition, we determined and compared FA compositions of total lipids of Tochuina tetraquetra and soft corals (Acanella sp., Anthomastus rylovi, Gersemia fruticosa, and Paragorgia arborea) collected together near the Kuril Islands (the depths of 80–550 m). FA compositions of T. tetraquetra, Acanella sp. and A. rylovi were described for the first time. In Acanella sp., G. fruticosa, A. rylovi, and P. arborea, the average contents of TPA were 6.5, 13.4, 5.9, and 12.0 % of total FAs, whereas the 24:5n-6/24:6n-3 ratio was 1.0, 1.4, 5.4, and 2.6, respectively. The high level of TPA (21.7 %) found in T. tetraquetra indicates that 24:5n-6 and 24:6n-3 are transferred from soft corals to the mollusks during their feeding and accumulated in mollusk tissues. The most possible feed source of the mollusk is suggested to be G. fruticosa and/or Acanella sp., because the 24:5n-6/24:6n-3 ratios in T. tetraquetra and these soft corals were similar. Thus, the TPA could be used as FA markers to estimate a proportion of soft corals in feeding of cold-water nudibranch mollusks.     

The composition of fatty acids in the endosperm and embryo lipids of <Emphasis Type="Italic">Pinus sibirica</Emphasis> and <Emphasis Type="Italic">P. sylvestris</Emphasis> seeds

The fatty acid (FA) composition of storage lipids in the seed endosperms and embryos of two pine species, Pinus sibirica and P. sylvestris, and possible biosynthetic pathways of these acids were studied by the GLC method. Linoleic acid predominated in the embryo and endosperm lipids of both P. sibirica (43.5 and 42.6%) and P. sylvestris (44.8 and 46.8%); this was evidently determined by a high expression of the gene encoding stearoyl-Δ9 acyl-lipid desaturase and the fad2 gene encoding microsomal ω6 acyl-lipid desaturase. P. sibirica lipids of the embryo and endosperm contained more oleic acid (22.0 and 24.0%, respectively) than corresponding P. sylvestris lipids (18.7 and 14%). Storage lipids of conifer seeds contain Δ5-unsaturated FAs: taxoleic (18:2Δ5,9), ephedrenic (18:2Δ5,11), pinoleenic (18:3Δ5,9,12), skiadonic (18:3Δ5,11, 14), and coniferonic (18:4Δ5,9,12,15). In the endosperm and embryos of P. sylvestris, the content of pinolenic acid was higher (22.1 and 19.6%) than in P. sibirica seeds (19.1 and 18.6%).     

Fatty acid composition of lipids from the vacuolar membranes of the roots of root vegetables

The fatty acid (FA) composition of vacuolar membrane lipids from the storage tissues of parsnip (Pastinaca sativa L.), parsley (Petroselinum crispum L.), and carrot (Daucus carota L.) was studied by gasliquid chromatography, and possible pathways of the biosynthesis of these acids are considered. A high level of unsaturated FAs (up to 78% of the total FA amount) was characteristic of these membrane lipids with the predominance of linoleic acid, which content in vacuolar lipids of parsnip, parsley, and carrot was 53.5, 55.1, and 54.4%, respectively. A high content of hexadienoic acid (C16:2ω6) was characteristic of the vacuolar lipids of parsnip and parsley (8.0 and 4.6%. respectively); the content of α-linolenic acid in the vacuolar lipids of these plants was 4.4–7.3%. Palmitic acid predominated among the saturated FAs (18.0–20.4%).     

Fatty-Acid and Stable-Isotope Compositions in Shallow-Water Bivalve Mollusks and their Food

We conducted a comparative analysis of the fatty acid (FA) composition and the ratios of stable isotopes of carbon (δ¹³C) and nitrogen (δ¹⁵N) in soft tissues of ten species of bivalve mollusks collected simultaneously on adjacent biotopes in shallow Vostok Bay (the Sea of Japan). Comparison of the FA composition of the lipids of digestive gland and all soft tissues showed that the percentages of C₁₆ and C₁₈ marker FAs were greater in the digestive gland and the levels of marker C₂₀ and C₂₂ FAs were, in most cases, higher in soft tissues. According to the results of cluster analysis and principal component analysis, four groups of samples were identified with a similarity of the FA composition of more than 80% within groups. The carbon stableisotope ratios varied within very wide limits in the studied species of bivalves; the range of δ¹³C variations was 8.1‰. The range of δ15N variations was much smaller, 2.5‰. Two pairs of species of mollusks (Saxidomus purpurata–Protothaca euglypta and P. jedoensis–Diplodonta semiasperoides) did not differ in the values of both δ¹⁵N and δ¹³C, the remaining species differed in at least one of these parameters. The greatest similarity of the FA composition and stable-isotope ratios was found in species that inhabit similar substrates, except Macoma irus and D. semiasperoides. Particularly marked differences in the FA composition and stable-isotope ratios were found between a filter-/surface deposit-feeder M. irus and filter-feeders Arca boucardi and Mytilus coruscus that live next to this species.     

Salinity tolerance of four freshwater microalgal species and the effects of salinity and nutrient limitation on biochemical profiles

Microalgae are ideal candidates for bioremediation and biotechnological applications. However, salinity and nutrient resource availability vary seasonally and between cultivation sites, potentially impacting on biomass productivity. The aim of this study was to screen pollutant-tolerant freshwater microalgae (Desmodesmus armatus, Mesotaenium sp., Scenedesmus quadricauda and Tetraedron sp.), isolated from Tarong power station ash-dam water, for their tolerance to cultivation at a range of salinities. To determine if biochemical composition could be manipulated, the effects of 4-day nutrient limitation were also determined. Microalgae were cultured at 2, 8, 11 and 18 ppt salinity, and nutrient uptake was monitored daily. Growth, total lipid, fatty acid (FA), and amino acid contents were quantified in biomass harvested while nutrient-replete and, after 4 days, nutrient-deplete. D. armatus showed the highest salinity tolerance actively growing in up to 18 ppt while Mesotaenium sp. was the least halotolerant with decreasing growth rates from 11 ppt. However, Mesotaenium sp. at 2 and 8 ppt had the highest biomass productivity and nutrient requirements of the four species, making it ideal for nutrient remediation of eutrophic freshwater effluents. Salinity and nutrient status had minimal influence on total lipid and FA contents in D. armatus and Mesotaenium sp., while nutrient depletion induced an increase of total lipid and FAs in S. quadricauda and Tetraedron sp., which was further increased with increasing salinity. As none of the growth conditions affected amino acid profiles of the species, these findings provide a basis for species selection based on site-specific salinity conditions and nutrient resource availability.     

Assessing biodegradation of furfuryl alcohol using <Emphasis Type="Italic">Pseudomonas putida</Emphasis> MTCC 1194 and <Emphasis Type="Italic">Pseudomonas aeruginosa</Emphasis> MTCC 1034 and its kinetics

The biodegradation of furfuryl alcohol (FA) in shake flask experiments using a pure culture of Pseudomonas putida (MTCC 1194) and Pseudomonas aeruginosa (MTCC 1034) was studied at 30 °C and pH 7.0. Experiments were performed at different FA concentrations ranging from 50 to 500 mg/l. Before carrying out the biodegradation studies, the bacterial strains were acclimatized to the concentration of 500 mg/l of FA by gradually raising 100 mg/l of FA in each step. The well acclimatized culture of P. putida and P. aeruginosa degraded about 80 and 66% of 50 mg/l FA, respectively. At higher concentration of FA, the percentage of FA degradation decreased. The purpose of this study was to determine the kinetics of biodegradation of FA by measuring biomass growth rates and concentration of FA as a function of time. Substrate inhibition was calculated from experimental growth parameters using the Haldane equation. Data for P. putida were determined as µ _max ?=?0.23 h^?1, K _s ?=?23.93 mg/l and K _i ?=?217.1 mg/l and for P. aeruginosa were determined as µ _max ?=?0.13 h^?1, K _s ?=?21.3 mg/l and K _i ?=?284.9 mg/l. The experimental data were fitted in Haldane, Aiba and Edwards inhibition models.     

Intra-and interspecific variability of fatty acid composition of soft corals

The intra-and interspecific variability of fatty acid (FA) composition of soft corals was examined in the tropical alcyonarian Sarcophyton sp., tropical gorgonian Euplexaura erecta, and boreal alcyonarian Gersemia rubiformis. Characteristic significant differences in the FA composition were found between these species belonging to different taxonomic groups and habitats. We assume that the FA groups 14: 0 + 16: 0 + 18: 3n-6, 16: 2 + 20: 4n-6 + 24: 5n-6, and 18: 1n-7 + 20: 1n-7 + 20: 5n-3 + 24: 6n-3 are characteristic of Sarcophyton sp., E. erecta, and G. rubiformis respectively. There were no significant differences (p > 0.05) between the three soft coral species in the content of oleic, linoleic, and docosahexaenoic acids. The relative content of more than 45% of individual FA did not significantly differ (p > 0.05) between the pairs of species, i.e., intraspecific variations exceeded interspecific ones. The reasons for intraspecific variability of soft coral FA composition are discussed. Control of this variability is needed when using individual FA as chemotaxonomic and food markers.     

Distribution of unusual fatty acids in the mesocarp triacylglycerols of maturing sea buckthorn fruits

The pattern of distribution of X-FAs among individual positional species of the X-triacylglycerols (X-TAGs) in the sea buckthorn (Hippophaë rhamnoides L.) mesocarp oil by the 80th, 90th, and 105th day after pollination (DAP) was investigated. The X-FAs comprised oleic acid (O) as well as unusual FAs, viz. hexadecenoic (H), palmitolinoleic (Pl), and cis-vaccenic acid (V). During mesocarp growth, the number of X-TAG species decreased dramatically as a result of their metabolism, but their proportion in the total TAGs remained constant. H-TAGs predominating in the oil were similar to total TAGs in their composition and the pattern of their dynamics during maturation. As regards minor Pl-TAGs, there was a tendency for an increase in the level of species including palmitolinoleic acid in the middle (sn-2) position of their molecules. Throughout the entire process of fruit maturation, oleic acid was by 98.0–98.8% concentrated in the sn-2 position of O-TAGs. At the same time, cis-vaccenic acid, a Δ11-positional isomer of oleic acid, was predominantly incorporated in the side positions of V-TAGs, and its 98.1% concentration in these positions was achieved only by the 105th DAP. A virtually absolute selectivity of the distribution of oleic and cis-vaccenic acids in TAGs was found here for the first time. Discussed are the possible physicochemical and biochemical factors of a highly selective incorporation of these FAs in the sea buckthorn TAGs, the pathways of enzymatic biosynthesis of O- and V-TAGs, the metabolic role of discrimination of incorporation of some FA species in the glycerol residue of glycerolipids, and the causes for the disappearance of some TAG species during maturation.     

The role of acyl-CoA thioesterase ACOT8I in mediating intracellular lipid metabolism in oleaginous fungus <Emphasis Type="Italic">Mortierella alpina</Emphasis>

Thioesterases (TEs) play an essential role in the metabolism of fatty acids (FAs). To explore the role of TEs in mediating intracellular lipid metabolism in the oleaginous fungus Mortierella alpina, the acyl-CoA thioesterase ACOT8I was overexpressed. The contents of total fatty acids (TFAs) were the same in the recombinant strains as in the wild-type M. alpina, whilst the production of free fatty acids (FFAs) was enhanced from about 0.9% (wild-type) to 2.8% (recombinant), a roughly threefold increase. Linoleic acid content in FFA form constituted about 9% of the TFAs in the FFA fraction in the recombinant strains but only about 1.3% in the wild-type M. alpina. The gamma-linolenic acid and arachidonic acid contents in FFA form accounted for about 4 and 25%, respectively, of the TFAs in the FFA fraction in the recombinant strains, whilst neither of them in FFA form were detected in the wild-type M. alpina. Overexpression of the TE ACOT8I in the oleaginous fungus M. alpina reinforced the flux from acyl-CoAs to FFAs, improved the production of FFAs and tailored the FA profiles of the lipid species.     

Structural organization of fatty acid desaturase loci in linseed lines with contrasting linolenic acid contents

Flax (Linum usitatissimum L.), the richest crop source of omega-3 fatty acids (FAs), is a diploid plant with an estimated genome size of ~370 Mb and is well suited for studying genomic organization of agronomically important traits. In this study, 12 bacterial artificial chromosome clones harbouring the six FA desaturase loci sad1, sad2, fad2a, fad2b, fad3a and fad3b from the conventional variety CDC Bethune and the high linolenic acid line M5791 were sequenced, analysed and compared to determine the structural organization of these loci and to gain insights into the genetic mechanisms underlying FA composition in flax. With one gene every 3.2–4.6 kb, the desaturase loci have a higher gene density than the genome’s average of one gene per 7.8–8.2 kb. The gene order and orientation across the two genotypes were generally conserved with the exception of the sad1 locus that was predicted to have additional genes in CDC Bethune. High sequence conservation in both genic and intergenic regions of the sad and fad2b loci contrasted with the significant level of variation of the fad2a and fad3 loci, with SNPs being the most frequently observed mutation type. The fad2a locus had 297 SNPs and 36 indels over ~95 kb contrasting with the fad2b locus that had a mere seven SNPs and four indels in ~110 kb. Annotation of the gene-rich loci revealed other genes of known role in lipid or carbohydrate metabolic/catabolic pathways. The organization of the fad2b locus was particularly complex with seven copies of the fad2b gene in both genotypes. The presence of Gypsy, Copia, MITE, Mutator, hAT and other novel repeat elements at the desaturase loci was similar to that of the whole genome. This structural genomic analysis provided some insights into the genomic organization and composition of the main desaturase loci of linseed and of their complex evolution through both tandem and whole genome duplications.     

Fatty Acid Composition of Lipids from Leaves and Strobila of Cycas Revoluta (<Emphasis Type="Italic">Cycas revoluta</Emphasis>)

Fatty acid (FA) composition of lipids from leaves and differentiated fleshy strobila tissues and sporangia with spores of Cycas (Cycas revoluta Thumb.) after their step quantitative extraction from plant material was investigated. Quantitative content and qualitative composition of FAs of extractable and nonextractable leaf lipids were determined. It was established that flesh lipids of sporophylls are characterized by a high saturation level and contain a considerable proportion of saturated FAs with the usual chain length (C₁₂–C₁₈, 53–57%). At the same time, total amount of etherified FAs with a very long chain in lipids not extractable by the method of Zhukov and Vereshchagin exceeds several times that found in extractable lipids (~15 and ~4%, respectively). Neutral lipids of Cycas spores were represented by triacylglycerols, the lower-alkyl esters of FAs, free FAs, and sterol esters.     

Fatty acid composition and <Emphasis Type="Italic">PlFADs</Emphasis> expression related to α-linolenic acid biosynthesis in herbaceous peony (<Emphasis Type="Italic">Paeonia lactiflora</Emphasis> Pall.)

As a new α-linolenic acid (ALA) resource, there has been little known about the relationship between expression levels of fatty acid desaturase (FAD) genes during seed development and fatty acid (FA) composition in herbaceous peony (Paeonia lactiflora Pall.). In this study, oil content and FA composition of nine cultivars were measured at four different stages during seed development. Moreover, five genes including PlFAD2-1, PlFAD2-2, PlFAD3-2, PlFAD6 and PlFAD7 related to the ALA biosynthesis were isolated. Furthermore, the relative expression levels of these genes in seeds were investigated in two cultivars, that is, ‘PL1’ and ‘PL2’ with higher and lower ALA content, respectively. The results showed that oil content was from 17.03 ± 0.15 to 24.51 ± 0.15%; 15 kinds of FA were detected. However, the relative content of ALA was decreased during seed development. Although, the genes PlFAD2-1, PlFAD2-2, PlFAD3-2, PlFAD6 and PlFAD7 were all expressed during the seed development, the expression levels varied significantly. Most of the genes were expressed strongly in the early stage but weakly in the late stage except PlFAD6. Moreover, expression level of gene PlFAD2-2 was the highest while that of PlFAD7 was lowest at S1. In addition, the relative expression level of genes in ‘PL1’ (high ALA content) was higher than those in ‘PL2’ (low ALA content) during the early stage of the seed development apart from gene PlFAD6. This study provides a reliable theoretical basis for improving the contents of ALA by means of genetic engineering in the herbaceous peony seed oil.     

Lipid fatty acid composition of potato plants transformed with the Δ12-desaturase gene from cyanobacterium

Potato (Solanum tuberosum L.) plants were transformed with the desA gene encoding Δ12 acyl-lipid desaturase in the cyanobacterium Synechocystis sp. PCC 6803. To evaluate the efficiency of this gene expression in the plant, its sequence was translationally fused with the sequence of the reporter gene encoding thermostable lichenase. A comparison of native and hybrid gene expression showed that lichenase retained its activity and thermostability within the hybrid protein, whereas desaturase retained its capability of inserting the double bond in fatty acid (FA) chains and, thus, to modify their composition in membrane lipids. In most transformed plants, shoots contained higher amounts of polyunsaturated FAs, linoleic and linolenic (by 39–73 and 12–41%, respectively). The total absolute content of unsaturated FAs was also higher in transformants by 20–42% as compared to wild-type plants. When transformed plants were severely cooled (to ?7°C), the rate of their membrane lipid peroxidation was not enhanced, whereas in wild-type plants, it increased substantially (by 25%) under such conditions. These results could indicate a higher tolerance of transformed plants to low temperatures and the oxidative stress induced by hypothermia.     

Optimization of <Emphasis Type="Italic">Prochlorothrix hollandica</Emphasis> cyanobacteria culturing for obtaining myristoleic acid

Plankton filament cyanobacteria Prochlorothrix hollandica is characterized by a very high content of C₁₄ and C₁₆ fatty acids (FA) in the lipid membranes. Depending on culturing conditions of the cyanobacteria, total concentrations of myristic and myristoleic acids can reach 35% and those of palmitic and palmitoleic acids can reach 60% of all esterified FA cells. In P. hollandica, a variety of monounsaturated FA is represented by myristoleic and palmitic acids, and by hexadecenoic (C_16:1) acid with olefin bond of cis-configuration, located in the Δ4 position. The process of intensive culturing for P. hollandica cells to yield a maximal biomass in order to isolate the pure drug of myristoleic acid derivative has been optimized. The use of a threestage purification gives 30 mg of chromatographically pure myristoleic acid methyl ester from 17 g of P. hollandica raw biomass (dry mass is 3 g), which is 1% of dry cell mass.     

Substrate specificity of acyl-lipid Δ9-desaturase from <Emphasis Type="Italic">Prochlorothrix hollandica</Emphasis> cyanobacterium producing myristoleic acid

Chlorophyll b-containing cyanobacterium Prochlorothrix hollandica is characterized by a high content of esterified fatty acids (FA) with 14 and 16 carbon atoms in the membrane lipids. Depending on the conditions of cultivation, the relative amount of myristic (C_14:0) and myristoleic (C_14:1) acids can reach 35%, and palmitic (С_16:0) and palmitoleic (С_16:1) acids can reach 60% of the sum of all fatty acids in cells. Monounsaturated FAs are represented by C_14:1, and C_16:1 with an olefinic bond presumably located in the Δ⁹ position. We cloned the gene of acyl-lipid Δ9-desaturase, desC1, from Prochlorothrix hollandica and characterized its specificity to the length of the substrate using the heterologous expression in Escherichia coli cells adding C_14:0 or stearic (C_18:0) acids as exogenous substrates. The results show that DesC1 Δ⁹ desaturase generates olefinic bonds in the FAs with a length of 14 to 18 carbon atoms with an approximately equal efficiency. This indicates that the length of the FA chain in P. hollandica is determined by the activity of the FA synthase, and the chain is desaturated at the Δ⁹ position nonspecifically relatively to its length.