The potential of Paeonia lactiflora pall seeds oil as a pure natural cosmetics raw material: In Vitro findings.

BACKGROUND: As a traditional Chinese herbal medicine, Paeonia lactiflora Pall is rich in various active ingredients such as polysaccharides and total flavonoids while having ornamental value. It has potential application value in the development of food and cosmetics. OBJECTIVE: To study the in vitro efficacy of Paeonia lactiflora Pall seeds oil. METHODS: Firstly, the levels of linolenic acid and linoleic acid in Paeonia lactiflora Pall seeds oil were quantified using gas chromatography. The impact of Paeonia lactiflora Pall seeds oil on the proliferation rate of B16F10 cells was assessed through the CCK-8 method, while the melanin content of B16F10 cells was determined using the sodium hydroxide lysis method. The inhibitory effects of Paeonia lactiflora Pall seeds oil on elastase, collagenase and hyaluronidase were evaluated by biochemical techniques in vitro. Lastly, the hen's egg chorioallantoic membrane test (HET-CAM) was conducted to confirm the absence of eye irritation caused by Paeonia lactiflora Pall seeds oil. RESULTS: Paeonia lactiflora Pall seeds oil within a certain volume concentration range (0.5%-4%) had no effect on the proliferation of B16F10 cells. Paeonia lactiflora Pall seeds oil showed significant inhibition of elastase, collagenase and hyaluronidase. Notably, the highest concentration tested, 4% Paeonia lactiflora Pall seed oil, yielded the most pronounced outcomes without causing any irritation. CONCLUSION: A certain concentration of Paeonia lactiflora Pall seeds oil has a significant effect on decreasing the melanin content in B16F10 cells and inhibiting the activities of elastase, collagenase, and hyaluronidase, which can provide a reference for the development of pure natural cosmetics raw materials.

Genome-wide association and RNA-seq analyses reveal a potential gene related to linolenic acid in soybean seeds.

Linolenic acid (LA) has poor oxidative stability since it is a polyunsaturated fatty acid. Soybean oil has a high LA content and thus has poor oxidative stability. To identify candidate genes that affect the linolenic acid (LA) content in soybean seeds, a genome-wide association study (GWAS) was performed with 1,060 soybean cultivars collected in China between 2019-2021 and which LA content was measured using matrix-assisted laser desorption/ionization time-of-flight imaging mass spectrometry (MALDI-TOF IMS). A candidate gene, GmWRI14, encoding an APETALA2 (AP2)-type transcription factor, was detected by GWAS in cultivars from all three study years. Multiple sequence alignments showed that GmWRI14 belongs to the plant WRI1 family. The fatty acid contents of different soybean lines were evaluated in transgenic lines with a copy of GmWRI14, control lines without GmWRI14, and the gmwri14 mutant. MALDI-TOF IMS revealed that GmWRI14 transgenic soybeans had a lower LA content with a significant effect on seed size and shape, whereas gmwri14 mutants had a higher LA content. compared to control. The RNA-seq results showed that GmWRI14 suppresses GmFAD3s (GmFAD3B and GmFAD3C) and GmbZIP54 expression in soybean seeds, leading to decreased LA content. Based on the RNA-seq data, yeast one-hybrid (Y1H) and qRT-PCR were performed to confirm the transcriptional regulation of FAD3s by GmWRI14. Our results suggest that FAD3 is indirectly regulated by GmWRI14, representing a new molecular mechanism of fatty acid biosynthesis, in which GmWRI14 regulates LA content in soybean seeds.

Metabolic profiles in the xylem sap of Brassica juncea exposed to cadmium.

Metabolic profiles in xylem sap are considered a fundamental mechanism for Cadmium (Cd) detoxification in plants. However, the metabolic mechanism of Brassica juncea xylem sap in response to Cd is still unclear. Here, we investigated the effects on the metabolomics of B. juncea xylem sap treated with Cd at different times by utilizing a nontargeted liquid chromatography-mass spectrometry (LC-MS)-based metabolomics method for further elucidating the response mechanism of Cd exposure. The findings indicated that 48 h and 7 days Cd exposure caused significant differences in metabolic profiles of the B. juncea xylem sap. Those differential metabolites are primarily involved in amino acids, organic acids, lipids, and carbohydrates, and most of them were downregulated, which played essential roles in response to Cd stress. Furthermore, B. juncea xylem sap resisted 48-h Cd exposure via regulation of glycerophospholipid metabolism, carbon metabolism, aminoacyl-tRNA biosynthesis, glyoxylate and dicarboxylate metabolism, linoleic acid metabolism, C5-branched dibasic acid metabolism, alpha-linolenic acid metabolism, cyanoamino acid metabolism, ABC transporters, biosynthesis of amino acids, and pyrimidine metabolism; whereas alpha-linolenic acid metabolism, glycerophospholipid metabolism, photosynthesis, and oxidative phosphorylation were regulated for resisting 7-day Cd exposure.

Effect of Flaxseed Addition on the Quality and Storage Stability of Sesame Paste.

The flaxseed-sesame paste (FSP) was prepared by mixing the heat-treated flaxseed and sesame seeds in different proportions and grinding them in a colloid mill to obtain a FSP. In this study, flaxseed was added to sesame paste (SP) at different addition to assess its effect on the rheological properties, textural properties, and particle size. The effect of flaxseed addition on lipid oxidation and volatile aldehydes and ketones during storage of SP was investigated by accelerated oxidation experiments (63°C, 60 days). Notably, the addition of all different additions of flaxseed increased the linolenic acid content, and also enhanced the hardness, cohesiveness, and viscosity of SP. However, it increased the rate of lipid oxidation in SP during storage, mainly in the form of higher acid value (AV) and malondialdehyde (MDA) content. The content of volatile aldehydes and ketones from lipid oxidation increased significantly with storage time. It was found by using cluster analysis that mixing flaxseed with SP at a ratio of 20 g/100 g had little effect on its storage stability, the sample had a higher overall quality than the addition of 40 g/100 g flaxseed, and its linolenic acid content was 18.7 times higher than that of the SP. Collectively, the results indicated that the addition of flaxseed at an appropriate proportion might be a feasible way to prepare the functional formulated SP.

Difference in the nutritional, in vitro, and functional characteristics of protein and fat isolates of two Indian chia (Salvia hispanica L) seed genotypes with variation in seed coat color.

This study aimed to determine the association between the seed coat color of two chia seed genotypes for their composition, protein content, amino acid, and fatty acid profiles. The optimal pH for protein isolation for both genotypes (BCPI and WCPI) was 10, based on protein purity and solubility. Fatty acid profiling indicated, overall, 18 different fatty acids higher in BCPI10 with linolenic acid domination (∼66%) followed by linoleic acid (∼19%) and oleic acid (∼6%), contributing PUFAs (∼86%). Optimized protein isolates, black (BCPI10) and white (WCPI10) chia, had shown purity, L*-value, solubility, and yields of 90.65%, 75.86%, 77.75%, 11.30%, and 90.00%, 77.83%, 76.07%, 10.69%, respectively. BCPI10 depicted higher EAA (33.19 g/100 g N) and EEA indices (57.676%) compared to WCPI10 (32.14 g/100 g N) and 56.360%, respectively. Amino acid profiling indicated higher, PER, TAA, TEAA, TNEAA, TAAA, TBA, acidic AA values for BCPI10, and higher leucine/isoleucine ratio for WCPI10 having leucine and sulfur amino acids as limiting amino acids. BCPI10 had higher sulfur-containing amino acid contents, as the main contributor to the albumin a water-soluble fraction, leading to its higher in vitro digestibility (71.97%) than WCPI10 (67.70%). Both isolates exhibited good WHC and OHC of 3.18, 2.39 and 3.00, 2.20, respectively. Both protein isolates had similar ∆Td (°C) values with some variation in FTIR spectrum from 1000 cm-1 to 1651 cm-1 having more peak intensity for BCPI10. SDS-PAGE indicated bands at 150 kDa, representing globulin and mild bands at 25-33 kDa for glutelin and albumin. A significant (p < 0.05) variation reported in this study for protein and lipid profiles of both genotype attributes to genetic differences between the seeds. PRACTICAL APPLICATION: Based on the nutritional profile, both chia seed isolates (black and white) are suitable for consumption with an edge for black seed when supplemented with their limiting amino acids. The high values of the functional properties and structural characteristics combined with high nutritional values make the chia protein isolate an excellent source of raw material for various food formulations. Fatty acid profile of the oils from the genotypes showed the presence of high amounts of unsaturated fatty acids, especially the PUFAs with more number of fatty acids in black chia seed. The excellent lipid profile of chia seed oil indicates the benefit of using chia seed oil as a source of essential fatty acids in the human diet for optimal health.

Characterization and differentiation of pollen lipidomes and proteomes from different intrafloral stamens in heterantherous Senna bicapsularis.

Numerous compounds in pollen can affect the foraging decision-making of bees. Clarification of phytochemical components and identification of key substances for bee foraging preference in pollen are essential steps for apiculture and developing a conservation strategy. Senna bicapsularis, a heterantherous plant that possesses three different stamen types in the same flower, among which bees forage selectively, provides us with an ideal research model for identification of potential substances of bee foraging preference. The lipid and protein compositions of pollen from the anthers of different stamens of S. bicapsularis were investigated and compared. The polyunsaturated fatty acids (PUFAs) and monounsaturated FAs (MUFAs) were highest among lipid molecules in pollen from short (S) stamens than from long (L) and medium (M) stamens. This result is consistent with the FA content measurement, showing the highest FAs and UFAs content in S pollen, especially α-linolenic acid. We inferred that α-linolenic acid might be one of the key substances for bee foraging preference in pollen. Moreover, proteomic analysis showed that several differentially expressed proteins involved in lipid biosynthesis were highly accumulated in S pollen, such as choline kinase 2, 3-oxoacyl-ACP synthase-like protein and choline/ethanolamine phosphotransferase 1, in line with the highest FA content of S pollen. Additionally, DEPs involved in 'starch and sucrose metabolism', 'phenylpropanoid biosynthesis' and 'cyanoamino acid metabolism' were more represented in S compared with L and M pollen. The study suggests that differences in proteomic and lipidomic profiling among the three different stamen types might affect foraging decision-making of bumblebees.

Chemical Characterization of Chinese Perilla Seed Oil.

Physicochemical properties and chemical composition of Chinese perilla seed oil has been characterized in this study. The result showed that both the cold press oil and the solvent extracted oil possessed low acid value and peroxide value. The fatty acid composition result showed that the oil has high content of linolenic acid (C18:3) up to 66.4 g/100 g, followed by linoleic acid (C18:2) of 15.3 g/100 g. The total triacylglycerol (TAG) profiles results showed that the oil contained 20 TAGs including 17 regioisomers, including LnLnLn (35.8 g/100 g), LLnLn (20.2 g/100 g), LLLn (17.7 g/100 g) and PLnLn (14.9 g/100 g) (Ln, linolenic acid; L, linoleic acid; P, palmitic acid). With content of only 0.57 g/100 g oil, the unsaponifiable matters were mainly composed of phytosterols, squalene, tocopherol, alcohols and hydrocarbons. The total phytosterols content was 0.39 g/100 g oil, in which ß-sitosterol has high content of 0.31 g/100 g oil.

Fatty acid profile of milk from Nordestina donkey breed raised on Caatinga pasture.

In this research communication we describe the composition of fatty acids (FA) present in the milk of the Nordestina donkey breed, and how they differ during lactation. Milk samples were taken from 24 multiparous lactating Nordestina donkeys that grazed the Caatinga, comprising 5 animals at each of around 30, 60 and 90 d in milk (DIM) and a further 9 animals ranging from 120 to 180 DIM. The milk fat content was analysed by mid infrared spectroscopy and the FA profile by gas chromatography. The milk fat percentage ranged from 0.45 to 0.61%. The main FA found in milk were 16:0 and 18:1c9. These did not differ among DIM classes and comprised 23% and 25% of total FA. Notably, the α-Linolenic acid (18:3 n-3) was the third most abundant FA and differed (P < 0.05) with DIM, being lowest in the 30 and 60 DIM samples (around 10.7% of total FA) and highest in the 60 and 90 DIM classes (around 14.6% of total FA). The low-fat content and the FA profile of the donkey milk gives it potential as a functional ingredient, which could help to preserve the commercial viability of the Nordestina donkey breed.

Profiling and quantitative analysis of underivatized fatty acids in Chlorella vulgaris microalgae by liquid chromatography-high resolution mass spectrometry.

Chlorella vulgaris is a popular microalga used for biofuel production; nevertheless, it possesses a strong cell wall that hinders the extraction of molecules, especially lipids within the cell wall. For tackling this issue, we developed an efficient and cost-effective method for optimal lipid extraction. Microlaga cell disruption by acid hydrolysis was investigated comparing different temperatures and reaction times; after hydrolysis, lipids were extracted with n-hexane. The best recoveries were obtained at 140°C for 90 min. The microalgae were then analyzed by an untargeted approach based on liquid chromatography with high-resolution mass spectrometry, providing the tentative identification of 28 fatty acids. First, a relative quantification on the untargeted data was performed using peak area as a surrogate of analyte abundance. Then, a targeted quantitative method was validated for the tentatively identified fatty acids, in terms of recovery (78-100%), intra- and interday relative standard deviations (<10 and <9%, respectively) and linearity (R2  > 0.98). The most abundant fatty acids were palmitic, palmitoleic, oleic, linoleic, linolenic, and stearic acids.

An alternative solution for α-linolenic acid supplements: in vitro digestive properties of silkworm pupae oil in a pH-stat system.

α-Linolenic acid (ALA) is recognised to have a regulatory effect on cardiovascular diseases. Due to the low bioavailability of linseed oil (LINO), which is the most common ALA supplement, it is necessary to find a replacement for ALA supplements that is more easily accepted by the human body. The content of ALA in silkworm pupae oil (SPO) is 32.60 ± 0.67%, and SPO can be substituted as a dietary lipid to meet the demand of the human body. In the present study, a pH-stat system was used to investigate the release degree of free fatty acids (FFAs) from SPO and construct a first-order kinetic model. Digestion experiments in vitro with different lipids showed that the maximum release FFA levels were SPO > SO (soybean oil) > LO (lard oil) > MSO (mulberry seed oil) > LINO, and the first-order kinetic apparent rate constants were LINO > SPO > LO > SO > MSO. Triacylglycerol (TAG) and fatty acid composition are the decisive factors in determining the level of lipid digestion. Therefore, the maximum level of FFAs released from SPO (84.34 ± 1.37%) was much higher than that of LINO (49.78 ± 0.52%) when the hydrolysis rates were 0.2114 s-1 and 0.2249 s-1, respectively. In addition, the smaller emulsion droplet size (609.24 ± 43.46 nm) and weaker surface charge (-17.93 ± 0.42 mV) also resulted in higher levels of SPO under in vitro digestion conditions. Meanwhile, due to low melting and crystallisation temperature, SPO is quickly absorbed by the human body. Overall, SPO can be used as a new alternative for ALA supplements based on its superior digestive properties.

Optimization of Ultrasonic-assisted Extraction and Fatty Acid Composition of Oil from Paeonia suffruticosa Andr. Seed.

Response surface methodology (RSM) was applied to optimize the effects of extraction parameters including time, power, temperature and liquid-to-solid ratio on peony seed oil yield. Box-Behnken design (BBD) was employed for optimization of extraction parameters in oil yield that extracted assisting by ultrasonic while petroleum ether as solvent. The chemical composition of peony seed oil under optimal condition in ultrasonic-assisted extract method was analyzed by gas chromatography-mass spectrometry (GC-MS). The optimal conditions were that extraction time 45 min, extraction temperature 45°C, extraction power 90 W and liquid-to-solid ratio 7:1, respectively. Under this condition, the extraction yield value was 33.90% which was with 95% confidence level, hence indicated the reliability of RSM in optimizing ultrasonic-assisted extraction of oil from Paeonia suffruticosa Andr. seed. Three unsaturated fatty acid of peony oil such as n-3 α-linolenic acid (39.75%), n-6 linoleic acid (26.32%) and the oleic acid (23.66%), totally more than 89.00% was determined at optimum condition.

Physico-Chemical Analysis and Fatty Acid Profiling of Fenugreek (Trigonella foenum-graecum) Seed Oil Using Different Solvents.

Fenugreek (Trigonella foenum-graecum) a native to Southern Europe, Mediterranean region and Western Asia has been used as a spice all over the world to increase the sensory quality to the food. It is also known for its medicinal properties such as anti-diabetic, anti-carcinogenic, hypocholesterolemic and immunological activities and can also be used as a food stabilizer and emulsifying agent. The ash, protein, moisture and fiber content of defatted fenugreek seed powder obtained were 9%, 23.04%, 3.8%, 25.47% respectively. So, this study is systematically intended to determine the fatty acid composition, to be best among the different solvents used are the ethanol, petroleum ether, acetone and hexane for the extraction of the fenugreek seed oil and to analyze its susceptibility to oxidation. This study was carried out to investigate and examine the results such as acid value, peroxide value, saponification value, iodine value and the physical properties such as the color value and the refractive index of the seed oil. The results stipulate that the oil extracted using the solvent hexane had better quality and yield. Linoleic acid (41.97%) followed by alpha-linolenic acid (29.33%) and cis-9 oleic acid (12.95%) was found as the primary fatty acids present in the oil extracted using hexane. Along with these fatty acids, the PUFA content of hexane oil (71.30%) was also observed to be in a good range. So, on comparing these results with codex standards, it revealed that it can be considered as edible oil with further purifications.

The Impact of Plasma Membrane Lipid Composition on Flagellum-Mediated Adhesion of Enterohemorrhagic Escherichia coli.

Enterohemorrhagic Escherichia coli (EHEC) O157:H7 is a major cause of foodborne gastrointestinal illness. The adhesion of EHEC to host tissues is the first step enabling bacterial colonization. Adhesins such as fimbriae and flagella mediate this process. Here, we studied the interaction of the bacterial flagellum with the host cell's plasma membrane using giant unilamellar vesicles (GUVs) as a biologically relevant model. Cultured cell lines contain many different molecular components, including proteins and glycoproteins. In contrast, with GUVs, we can characterize the bacterial mode of interaction solely with a defined lipid part of the cell membrane. Bacterial adhesion on GUVs was dependent on the presence of the flagellar filament and its motility. By testing different phospholipid head groups, the nature of the fatty acid chains, or the liposome curvature, we found that lipid packing is a key parameter to enable bacterial adhesion. Using HT-29 cells grown in the presence of polyunsaturated fatty acid (α-linolenic acid) or saturated fatty acid (palmitic acid), we found that α-linolenic acid reduced adhesion of wild-type EHEC but not of a nonflagellated mutant. Finally, our results reveal that the presence of flagella is advantageous for the bacteria to bind to lipid rafts. We speculate that polyunsaturated fatty acids prevent flagellar adhesion on membrane bilayers and play a clear role for optimal host colonization. Flagellum-mediated adhesion to plasma membranes has broad implications for host-pathogen interactions.IMPORTANCE Bacterial adhesion is a crucial step to allow bacteria to colonize their hosts, invade tissues, and form biofilm. Enterohemorrhagic Escherichia coli O157:H7 is a human pathogen and the causative agent of diarrhea and hemorrhagic colitis. Here, we use biomimetic membrane models and cell lines to decipher the impact of lipid content of the plasma membrane on enterohemorrhagic E. coli flagellum-mediated adhesion. Our findings provide evidence that polyunsaturated fatty acid (α-linolenic acid) inhibits E. coli flagellar adhesion to the plasma membrane in a mechanism separate from its antimicrobial and anti-inflammatory functions. In addition, we confirm that cholesterol-enriched lipid microdomains, often called lipid rafts, are important in bacterial adhesion. These findings demonstrate that plasma membrane adhesion via bacterial flagella play a significant role for an important human pathogen. This mechanism represents a promising target for the development of novel antiadhesion therapies.

Revision of Coelastrella (Scenedesmaceae, Chlorophyta) and first register of this green coccoid microalga for continental Norway.

A terrestrial green microalga was isolated at Ås, in Akershus County, Norway. The strain corresponded to a coccoid chlorophyte. Morphological characteristics by light and electron microscopy, in conjunction with DNA amplification and sequencing of the 18 s rDNA gene and ITS sequences, were used to identify the microalgae. The characteristics agree with those of the genus Coelastrella defined by Chodat, and formed a sister group with the recently described C. thermophila var. globulina. Coelastrella is a relatively small numbered genus that has not been observed in continental Norway before; there are no previous cultures available in collections of Norwegian strains. Gas chromatography analyses of the FAME-derivatives showed a high percentage of polyunsaturated fatty acids (44-45%) especially linolenic acid (C18:3n3; 30-34%). After the stationary phase, the cultures were able to accumulate several carotenoids as neoxanthin, pheophytin a, astaxanthin, canthaxanthin, lutein, and violaxanthin. Due to the scarcity of visual characters suitable for diagnostic purposes and the lack of DNA sequence information, there is a high possibility that species of this genus have been neglected in local environmental studies, even though it showed interesting properties for algal biotechnology.

Structure determination of conjugated linoleic and linolenic acids.

Conjugated linoleic and linolenic acids (CLA and CLnA) can be found in dairy, ruminant meat and oilseeds, these types of unsaturated fatty acids consist of various positional and geometrical isomers, and have demonstrated health-promoting potential for human beings. Extensive reviews have reported the physiological effects of CLA, CLnA, while little is known regarding their isomer-specific effects. However, the isomers are difficult to identify, owing to (i) the similar retention time in common chromatographic methods; and (ii) the isomers are highly sensitive to high temperature, pH changes, and oxidation. The uncertainties in molecular structure have hindered investigations on the physiological effects of CLA and CLnA. Therefore, this review presents a summary of the currently available technologies for the structural determination of CLA and CLnA, including the presence confirmation, double bond position determination, and the potential stereo-isomer determination. Special focus has been projected to the novel techniques for structure determination of CLA and CLnA. Some possible future directions are also proposed.

A comparison of fatty acid and sensory profiles of raw and roasted pecan cultivars.

Five fatty acids comprise the bulk of the lipid content in pecans: palmitic acid, stearic acid, oleic acid, linoleic acid, and linolenic acid. Understanding the profiles of these fatty acids and how they relate to sensory characteristics may offer an explanation for flavor and flavor defects that may exist in certain cultivars of pecans. The objective of this study was to examine and compare fatty acid profiles of three cultivars of pecans (Major, Lakota, and Chetopa), over two crop years, under raw and roasted preparation methods, and understand the fatty acids association with sensory attributes. Percentages of palmitic, stearic, oleic, linoleic, and linolenic acids to total fatty acid content were determined using gas chromatography, and sensory profiles were generated using descriptive sensory analysis. Similar trends were seen across samples, with oleic acid comprising the majority of the total fatty acids and linolenic acid comprising the smallest percentage. There were significant differences in fatty acid content among cultivars and between pecans in the first and second crop year. Few associations were found between the fatty acids and sensory attributes, which suggest that combinations of the fatty acids contribute to certain pleasant or undesirable flavor attributes in the pecans. Subtle differences in fatty acid composition may lead to variation in flavor and flavor intensity or draw attention to or from certain attributes during consumption. Differences in crop year indicated that fatty acid content and therefore flavor are variable year to year. PRACTICAL APPLICATION: This study will help understand how fatty acid content of pecans varies from year to year. This should be taken into account when manufacturing products with pecans as the nutritional content of the product may change as the result.

Marker assisted selection of new high oleic and low linolenic winter oilseed rape (Brassica napus L.) inbred lines revealing good agricultural value.

Development of oilseed rape (Brassica napus L.) breeding lines producing oil characterized by high oleic and low linolenic acid content is an important goal of rapeseed breeding programs worldwide. Such kind of oil is ideal for deep frying and can also be used as a raw material for biodiesel production. By performing chemical mutagenesis using ethyl methanesulfonate, we obtained mutant winter rapeseed breeding lines that can produce oil with a high content of oleic acid (C18:1, more than 75%) and a low content of linolenic acid (C18:3, less than 3%). However, the mutant lines revealed low agricultural value as they were characterized by low seed yield, low wintering, and high content of glucosinolates in seed meal. The aim of this work was to improve the mutant lines and develop high-oleic and low-linolenic recombinants exhibiting both good oil quality and high agronomic value. The plant materials used in this study included high-oleic and low-linolenic mutant breeding lines and high-yielding domestic canola-type breeding lines of good agricultural value with high oleic acid content and extremely low glucosinolates content. Field trials were conducted in four environments, in a randomized complete block design. Phenotyping was performed for wintering, yield of seed and oil, and seed quality traits. Genotype × environment interaction was investigated with respect to the content of C18:1 and C18:3 acids in seed oil. Genotyping was done for the selection of homozygous high oleic and low linolenic lines using allele-specific CAPS markers and SNaPshot assay, respectively. Finally, new high oleic and low linolenic winter rapeseed recombinant lines were obtained for use as a starting material for the development of new varieties that may be of high value on the oil crop market.

Esterification and modification of [1-14C] n-3 and n-6 polyunsaturated fatty acids in pikeperch (Sander lucioperca) larvae reared under linoleic or α-linolenic acid-based diets and variable environmental salinities.

To elucidate the in vivo endogenous ability of pikeperch (Sander lucioperca) larvae to deacylate and reacylate phospholipids and to elongate and desaturate PUFAs, 20 days post hatch (DPH) fish were incubated with either [1-14C]20:4n-6 bound to PC and PE, or with free [1-14C]-labelled fatty acids (18:2n-6, 18:3n-3, 20:4n-6, 20:5n-3 and 22:6n-3). The modulation capacity of both low LC-PUFAs but high 18C PUFAs precursors dietary supply and increasing salinity on larval fatty acid metabolic pathways was also investigated. [1-14C]DHA was incorporated into larval tissues to a lower extent than [1-14C]ARA or [1-14C] EPA. [1-14C]ARA was significantly less abundant in larval tissues when provided bound to PE than when esterified into PC, indicating that PC is a better phospholipid source to provide LC-PUFA to pikeperch larvae. Radioactivity was mainly recovered into phospholipids, especially that of the three LC-PUFAs ARA, EPA and DHA. All substrates were primarily incorporated into PC except [1-14C]ARA which significantly did into PI. Both [1-14C]EPA and [1-14C]DHA showed a similar esterification pattern into lipid classes: PC > PE > PI > TAG, with [1-14C]DHA presenting the highest esterification into PE of all radiolabelled compounds (26.3% vs 3.6-14.2%). Although higher rearing salinities tended to increase ∆6 desaturase activity, no radioactivity from [1-14C]18:2n-6 or [1-14C]18:3n-3 was detected in ARA or EPA, proving a deficiency of Δ5 activity and the inability of pikeperch to biosynthesize DHA. This work provides novel information on the lipid metabolism of pikeperch at early development necessary for the design of live prey enrichment protocols and dietary formulations adapted to larval metabolic capabilities.

Modifications of fatty acid profile through targeted mutation at BnaFAD2 gene with CRISPR/Cas9-mediated gene editing in Brassica napus.

KEY MESSAGE: Genomic editing with CRISPR/Cas9 system can simultaneously modify multiple copies of theBnaFAD2 gene to develop novel variations in fatty acids profiles in polyploidy rapeseed. Fatty acid composition affects edible and processing quality of vegetable oil and has been one of the primary targets for genetic modification in oilseed crops including rapeseed (Brassica napus). Fatty acid desaturase 2 gene, FAD2, is a key player that affects three major fatty acids, namely oleic, linoleic and linolenic acid, in oilseed plants. Previously, we showed that there are four copies of BnaFAD2 in allotetraploid rapeseed. In this study, we further established spatiotemporal expression pattern of each copy of BnaFAD2 using published RNA-seq data. Genomic editing technology based on CRISPR/Cas9 system was used to mutate all the copies of BnaFAD2 to create novel allelic variations in oleic acid and other fatty acid levels. A number of mutants at two targeting sites were identified, and the phenotypic variation in the mutants was systematically evaluated. The oleic acid content in the seed of the mutants increased significantly with the highest exceeding 80% compared with wild type of 66.43%, while linoleic and linolenic acid contents decreased accordingly. Mutations on BnaFAD2.A5 caused more dramatic changes of fatty acid profile than the mutations on BnaFAD2.C5 alleles that were identified with gene editing technique for the first time. Moreover, combining different mutated alleles of BnaFAD2 can even broaden the variation more dramatically. It was found that effects of different mutation types at BnaFAD2 alleles on oleic levels varied, indicating a possibility to manipulate fatty acid levels by precise mutation at specific region of a gene.

Spleen Oxylipin and Polyunsaturated Fatty Acid Profiles are Altered by Dietary Source of Polyunsaturated Fatty Acid and by Sex.

As the largest secondary lymphoid organ, the spleen plays an important role in immune responses. The role of arachidonic acid (ARA) and its 20-carbon eicosanoids in modulating immune function has long been of interest. However, recent advances have enabled the identification of numerous other n-6 and n-3 polyunsaturated fatty acid (PUFA)-derived oxylipins. Here, we investigate the effects of diet and sex on the spleen nonesterified oxylipin profiles and phospholipid and neutral lipid PUFA composition in Sprague-Dawley rats supplemented with oils rich in α-linolenic acid (ALA), eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), or linoleic acid. Dietary ALA, EPA, and DHA resulted in lower levels of ARA and ARA oxylipins. Oxylipins derived from other n-6 PUFA were also reduced despite no or opposite effect on their PUFA levels. Each diet also resulted in higher levels of oxylipins almost exclusively derived from the supplemented PUFA, despite PUFA in the same biosynthetic pathway also often being increased. Further, while oxylipin differences often reflected changes to phospholipid PUFA, there were instances where they corresponded more closely to changes in neutral lipid PUFA. With respect to sex effects, >50% of lipoxygenase ARA-derived oxylipins were higher in males in at least one diet group, while multiple DHA oxylipins were lower in males only in rats provided the DHA diet. This fundamental description of oxylipin composition in the spleen, including the influence of diet and sex and the relationship to PUFA composition, will help inform future studies examining the functions of these oxylipins under physiological and pathological conditions.