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Background: Secondary metabolites of several plants, including esculin and digitoxin, which are cardiac glycosides, were previously employed for their therapeutic effects. The current study aims to investigate the functions of the main Na + /K + transport inhibitor digitoxin and the antioxidant esculin for enhancing flax plant growth and production under salinity., Methodology: Flax plants were irrigated with distilled water supplemented with 0.0 and 5000 mg/L salt solution starting from 15 DAS from sowing. Then exogenous treatment with digitoxin and esculin with 50 mg L - 1 and 100 mg L - 1 were used for this work., Results: According to the results of this work, foliar spraying of esculin or digitoxin increased the salinity tolerance of flax plants.The foliar application of either esculin or digitoxin induced an elevation in the contents of photosynthetic pigments, osmolytes including soluble sugar and proline as well as the total phenols in salt-stressed flax plants. Moreover, esculin and digitoxin in particular counteract oxidative stress by increasing the activity of antioxidant enzymes including superoxide dismutase, catalase, peroxidase, phenylalanine ammonia-lyase, and tyrosine ammonia lyase, leading to a decrease in reactive oxygen species and lipid peroxidation levels and electrolyte leakage. The efficiency of esculin and digitoxin to sustain ion homeostasis by inhibiting Na + absorption and increasing potassium, calcium, and phosphorus in flax plants may be the reason for their protective actions towards salinity.As a consequence, esculin and digitoxin increased yield quantity and quality as shown by increases in all investigated yield criteriaas shoot height, root length, their fresh and dry weights as well asseed yield/plant (g), and 1000 seeds weight, especially those that improved the desired oil properties., Conclusion: In conclusion, this study concluded that digitoxin was more effective in inhibiting Na + build-up and increasing flax salinity tolerance, particularly at the high investigated dose as compared to esculin. In this study, we reported the recent findings of exogenousapplication of either digitoxin or esculin glycosides which are new investigated salt alleviators never used before for improving the salt tolerance in flax plants., (© 2024. The Author(s).)

Germination of seeds is known to affect the nutritional composition of cold-pressed oils. This study focused on the effects of germination on the antioxidants and oxidative stability of linseed and sunflower seed oil. As hypothesized, germination led to increased antioxidant activities and tocopherol, chlorophyll and carotenoid content. Analysis revealed a 37.2 ± 3.5-fold and 11.6 ± 1.5-fold increase in polyphenol content in linseed and sunflower seed oil from germinated seeds, respectively. Using LC-HRMS/MS, profiles with up to 69 polyphenolic substances were identified in germinated seed oils for the first time. Germination promoted lipid hydrolysis, as evidenced by NMR, with overall significant decreases in triacylglycerol content leading to increased diacylglycerol and free fatty acid values. Rancimat measurements predicted a 4.10 ± 0.52-fold longer shelf-life for germinated linseed oil. This study successfully demonstrated the potential of germination to develop PUFA-rich oils with enhanced antioxidant capacity and oxidative stability., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Flax ( Linum usitatissimum L.) is an important crop plant with pharmaceutical significance. It is described in pharmacopoeias (the United States Pharmacopeia and the European Pharmacopoeia), which confirms that it (especially the seeds) is a valuable medicinal product. Similar to flax seeds, which accumulate bioactive compounds, flax in vitro cultures are also a rich source of flavonoids, phenolics, lignans and neolignans. In the present study, flax suspension cultures after treatment of the non-pathogenic Fusarium oxysporum strain Fo47 were established and analyzed. The study examined the suitability of Fo47 as an elicitor in flax suspension cultures and provided interesting data on the impact of these endophytic fungi on plant metabolism and physiology. Two flax cultivars (Bukoz and Nike) and two compositions of media for flax callus liquid cultures were tested. Biochemical analysis revealed enhanced levels of secondary metabolites (total flavonoid and total phenolic content) and photosynthetically active pigments in the flax callus cultures after treatment with the non-pathogenic fungal strain F. oxysporum Fo47 when compared to control, untreated cultures. In cultures with the selected, optimized conditions, FTIR analysis was performed and revealed changes in the structural properties of cell wall polymers after elicitation of cultures with F. oxysporum Fo47. The plant cell wall polymers were more strongly bound, and the crystallinity index (Icr) of cellulose was higher than in control, untreated samples. However, lignin and pectin levels were lower in the flax callus liquid cultures treated with the non-pathogenic strain of Fusarium when compared to the untreated control. The potential application of the non-pathogenic strain of F. oxysporum for enhancing the synthesis of desired secondary metabolites in plant tissue cultures is discussed.

Linseed or flaxseed, native to the Indian subcontinent, had undergone domestication, edaphic selection and evolutionary processes that may have resulted in huge genetic variability in Indian genotypes. To understand the hitherto unexplored genetic diversity for sustainable flaxseed production amid challenges of climate fluctuation and identify trait-specific high-yielding genotypes, 2576 unique linseed accessions were comprehensively evaluated for 36 traits for up to six environments representing two major agroecological zones in India. A wide range of variability was recorded for days to initiation of flowering (42.86-114.99), plant height (43.31-122.88 cm), capsules/plant (64.62-375.87), seed size (6.06-14.44 cm 2 ), thousand seed weight (2.80-11.86 g), seed yield (2.93-17.28 g/plant), oil content (30.14-45.96%) and fatty acid profile especially the key constituent omega-3 fatty acid (25.4-65.88%). Most of the traits such as plant height, flowering time, seed yield, seed and capsule size showed a high or moderately high level of variance coupled with high broad sense heritability indicating precise capturing of less heritable quantitative traits. The infraspecific classification of the tested collection revealed the seed/oil type (2498 accessions) as the dominant morphotype over dual-purpose/fiber flax (78 accessions) in the conserved collection. Correlation analysis indicated a significant positive association between flowering time, plant height, days to maturity and oil content. Trait-specific superior genotypes for earliness (50% flowering in < 60 days, maturity in < 122 days), bold seeds with high thousand seed weight (> 11 g), capsules/plant (> 350), oil content (> 45%) and fatty acid composition (> 65% alpha-linolenic acid) were identified to aid genetic improvement of linseed and to broaden the narrow genetic base., (© 2024. The Author(s).)

Sprouting of stored oilseeds due to improper storage can lead to quality defects of cold-pressed oils obtained from them. This study aimed to evaluate the effect of seed sprouting on volatile organic compounds (VOCs), aroma-active compounds, and the content of nonvolatile metabolites in cold-pressed false flax oil obtained from sprouted seeds. In this study, 88 unique VOCs were detected in sprouted oils, whereas only 42 were found in the control oils. The control oils were characterized by a higher abundance of alcohols, while all other groups of compounds were associated with sprouted seeds. The formation of many VOCs was reflected in changes in the nonvolatile precursors. Fifteen aroma-active compounds were identified in sprouted oil, with five compounds playing a significant role (FD ≥ 128) in aroma formation. The presented approach allowed identification of differences caused by seed sprouting, resulting in oils with a much stronger aroma and a richer profile of VOCs due to intensive metabolic changes. The origin of many VOCs can be explained by alterations in the content of nonvolatile metabolites.

Background: Flax lignan has attracted much attention because of its potential bioactivities. However, the bioavailability of secoisolariciresinol diglucoside (SDG), the main lignan in flaxseed, depends on the bioconversion by the colon bacteria. Lactic acid bacteria (LAB) with β-glucosidase activity has found wide application in preparing bioactive aglycone., Results: LAB strains with good β-glucosidase activity were isolated from fermented tofu. Their bioconversion of flax lignan extract was investigated by resting cell catalysis and microbial fermentation, and the metabolism of SDG by Lactiplantibacillus plantarum C5 following fermentation was characterized by widely targeted metabolomics. Five L. plantarum strains producing β-glucosidase with broad substrate specificity were isolated and identified, and they all can transform SDG into secoisolariciresinol (SECO). L. plantarum C5 resting cell reached a maximum SDG conversion of 49.19 ± 3.75%, and SECO generation of 21.49 ± 1.32% (0.215 ± 0.013 mm) at an SDG substrate concentration of 1 mM and 0.477 ± 0.003 mm SECO was produced at 4 mm within 24 h. Although sixteen flax lignan metabolites were identified following the fermentation of SDG extract by L. plantarum C5, among them, four were produced following the fermentation: SECO, demethyl-SECO, demethyl-dehydroxy-SECO and isolariciresinol. Moreover, seven lignans increased significantly., Conclusion: Fermentation significantly increased the profile and level of flax lignan metabolites, and the resting cell catalysis benefits from higher bioconversion efficiency and more straightforward product separation. Resting cell catalysis and microbial fermentation of flax lignan extract by the isolated β-glucosidase production L. plantarum could be potentially applied in preparing flax lignan ingredients and fermented flaxseed. © 2024 Society of Chemical Industry., (© 2024 Society of Chemical Industry.)

Linseed, also known as flax is an important oilseed crop with many potential uses in paint, textile, food and pharmaceutical industries. Susceptibility to bud fly (Dasyneura lini Barnes) infestation is a serious biotic concern leading to severe yield penalty in linseed. Protease inhibitors (PIs) are potential candidates that activate during the insect-pest attack and modulate the resistance. In the present study, we explored the PI candidates in the linseed genome and a total of 100 LuPI genes were identified and grouped into five distinct subgroups. The analysis of cis-acting elements revealed that almost all LuPI promoters contain several regulatory elementary related to growth and development, hormonal regulation and stress responses. Across the subfamilies of PIs, the specific domains are consistently found conserved in all protein sequences. The tissue-specific in-silico expression pattern via RNA-seq revealed that all the genes were regulated during different stress. The expression through qRT-PCR of 15 genes revealed the significant up-regulation of LuPI-24, LuPI-40, LuPI-49, LuPI-53, and LuPI-63 upon bud fly infestation in resistant genotype EC0099001 and resistant check variety Neela. This study establishes a foundation resource for comprehending the structural, functional, and evolutionary dimensions of protease inhibitors in linseed., (© 2024. The Author(s).)

False flax (Camelina sativa L.), known as camelina, is an ancient oil plant that has gathered renewed interest. In this study, a comprehensive analysis encompassing nontargeted volatilomics and targeted, quantitative metabolomics performed for cold-pressed oil and press cake and was integrated with sensory analysis of cold-pressed camelina oil and the effect of seed roasting was evaluated. Roasting in general resulted in the formation of 22 new volatile organic compounds (VOCs) in oil, while roasting at 140 and 180 °C resulted in the formation of 12 and 124 unique VOCs, respectively. Roasting notably influenced the profile of primary and secondary metabolites in both oil and press cakes, as well as volatilome and aroma of cold-pressed camelina oil. Many VOCs can be attributed to thermal degradation of primary and secondary metabolites. Roasting intensified the flavour of cold-pressed camelina oil, enhancing the perception of notes formed through the Maillard reaction., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Human intestinal microbiota plays a crucial role in converting secoisolariciresinol diglucoside, a lignan found in flaxseed, to enterodiol, which has a range of health benefits: antioxidative, antitumor, and estrogenic/anti-estrogenic effects. Given the high secoisolariciresinol diglucoside content in flaxseed cake, this study investigated the potential of co-fermenting flaxseed cake with fermented soybean product to isolate bacterial strains that effectively convert secoisolariciresinol diglucoside to enterodiol in a controlled environment (in vitro). The co-fermentation process with stinky tofu microbiota significantly altered the lignan, generating 12 intermediate lignan metabolites as identified by targeted metabolomics. One particular promising strain, ZB26, demonstrated an impressive ability to convert secoisolariciresinol diglucoside. It achieved a conversion rate of 87.42 ± 0.33%, with secoisolariciresinol and enterodiol generation rates of 94.22 ± 0.51% and 2.91 ± 0.03%, respectively. Further optimization revealed, under specific conditions (0.5 mM secoisolariciresinol diglucoside, pH 8, 30 °C for 3 days), ZB26 could convert an even higher percentage (97.75 ± 0.05%) of the secoisolariciresinol diglucoside to generate secoisolariciresinol (103.02 ± 0.16%) and enterodiol (3.18 ± 0.31%). These findings suggest that the identified strains ZB26 have promising potential for developing functional foods and ingredients enriched with lignans., Competing Interests: Declaration of competing interest We have no conflicts of interest concerning the work described in this manuscript., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Cadmium is one of the most harmful heavy metals that harm agricultural products. Evaluating microRNAs expression is a new and accurate method to study plant response in various environmental conditions. So this study aimed to evaluate the contribution of two symbiotic fungi in improving flax tolerance in a Cd-polluted soil using microRNAs and their target gene expression. A factorial pot experiment in a completely randomized design was conducted with different levels of Cd (0, 20, and 40 mg kg -1 ) on non-inoculated and inoculated flax with Claroideoglomus etunicatum and Serendipita indica . The results presented that increasing Cd levels caused a constant decline of alkaline phosphatase of soil (from 243 to 210 and 153 μg PNP g -1 h -1 ), respectively, from control (Cd0) to 20 and 40 mg Cd kg -1 . However, the inoculation of flax with fungi significantly enhanced these properties. A negative correlation was observed between the expression level of microRNA 167 and microRNA 398 with their corresponding target genes, auxin response factor 8 and superoxide dismutase zinc/copper 1, respectively. The expression level of both microRNAs and their targets indicated that the inoculation with symbiont fungi could diminish Cd stress and enhance the growth of flax.

The loss of anthocyanin pigments is one of the most common evolutionary transitions in petal color, yet the genetic basis for these changes in flax remains largely unknown. In this study, we used crossing studies, a bulk segregant analysis, genome-wide association studies, a phylogenetic analysis, and transgenic testing to identify genes responsible for the transition from blue to white petals in flax. This study found no correspondence between the petal color and seed color, refuting the conclusion that a locus controlling the seed coat color is associated with the petal color, as reported in previous studies. The locus controlling the petal color was mapped using a BSA-seq analysis based on the F 2 population. However, no significantly associated genomic regions were detected. Our genome-wide association study identified a highly significant QTL ( BP4.1 ) on chromosome 4 associated with flax petal color in the natural population. The combination of a local Manhattan plot and an LD heat map identified LuMYB314 , an R2R3-MYB transcription factor, as a potential gene responsible for the natural variations in petal color in flax. The overexpression of LuMYB314 in both Arabidopsis thaliana and Nicotiana tabacum resulted in anthocyanin deposition, indicating that LuMYB314 is a credible candidate gene for controlling the petal color in flax. Additionally, our study highlights the limitations of the BSA-seq method in low-linkage genomic regions, while also demonstrating the powerful detection capabilities of GWAS based on high-density genomic variation mapping. This study enhances our genetic insight into petal color variations and has potential breeding value for engineering LuMYB314 to develop colored petals, bast fibers, and seeds for multifunctional use in flax.

Kynurenic acid (KYNA) is a bioactive compound exhibiting multiple actions and positive effects on human health due to its antioxidant, anti-inflammatory and neuroprotective properties. KYNA has been found to have a beneficial effect on wound healing and the prevention of scarring. Despite notable progress in the research focused on KYNA observed during the last 10 years, KYNA's presence in flax ( Linum usitatissimum L.) has not been proven to date. In the present study, parts of flax plants were analysed for KYNA synthesis. Moreover, eight different cultivars of flax seeds were tested for the presence of KYNA, resulting in a maximum of 0.432 µg/g FW in the seeds of the cultivar Jan. The level of KYNA was also tested in the stems and roots of two selected flax cultivars: an oily cultivar (Linola) and a fibrous cultivar (Nike). The exposure of plants to the KYNA precursors tryptophan and kynurenine resulted in higher levels of KYNA accumulation in flax shoots and roots. Thus, the obtained results indicate that KYNA might be synthesized in flax. The highest amount of KYNA (295.9 µg/g dry weight [DW]) was detected in flax roots derived from plants grown in tissue cultures supplemented with tryptophan. A spectroscopic analysis of KYNA was performed using the FTIR/ATR method. It was found that, in tested samples, the characteristic KYNA vibration bands overlap with the bands corresponding to the vibrations of biopolymers (especially pectin and cellulose) present in flax plants and fibres.

A study was conducted to evaluate standardized ileal digestibility (SID) of amino acids (AA) and N-corrected apparent metabolizable energy (AMEn) values of cold pressed flaxseed meal (CPFM) for broilers. One hundred and twenty broiler chicks were divided into 20 groups of 6 birds/group and fed 2 diets in a completely randomized design (10 groups/diet) from 14 to 21 d of age. The diets were cornstarch-based containing CPFM or conventional soybean meal (SBM; reference feedstuff) as the sole protein source. A N-free diet fed in another study conducted in the same facility and at the same time that the current study was conducted was used to estimate basal endogenous AA losses, and to calculate gross energy retention by difference method. The CPFM had greater neutral detergent fiber and ether extract contents (21.40 vs. 8.18% and 20.4 vs. 2.47% as is, respectively), but lower CP (20.47 vs. 48.28% as is) than SBM. The SID values of all indispensable AA (except for Leu, Phe and Trp) for CPFM were lower (P < 0.05) than those for SBM. The apparent retention of gross energy (65.95 vs. 44.24%) and N (55.53 vs. 9.79%), and AMEn (2,699 vs. 2,491 kcal/kg) for CPFM were lower (P < 0.05) than those for SBM. In conclusion, CPFM can serve as alternative oilseed co-product feedstuff for poultry. However, the CPFM has lower SID of AA and AMEn values than SBM likely due to the greater fiber content in the former than in the latter., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Lignan, a beneficial constituent of Flaxseed (Linum usitatissimum L.) showed great interest in researchers because of its multiple functional properties. Nonetheless, a challenge arises due to the glycosidic structure of lignans, which the gut epithelium cannot readily absorb. Therefore, we screened 18 strains of Lactiplantibacillus plantarum, Lacticaseibacillus casei, Lactobacillus acidophilus, Lacticaseibacillus rhamnosus, Pediococcus pentosaceus, Pediococcus acidilactici, and Enterococcus durans to remove glycosides from flaxseed lignan extract enzymatically. Among our findings, Lactiplantibacillus plantarum SCB0151 showed the highest activity of β-glucosidase (8.91 ± 0.04 U/mL) and higher transformed efficiency of Secoisolariciresinol (SECO) (8.21 ± 0.13%). The conversion rate of Secoisolariciresinol diglucoside (SDG) and the generation rate of SECO was 58.30 ± 3.78% and 32.13 ± 2.78%, respectively, under the optimized conditions. According to the LC-HRMSMS analysis, SECO (68.55 ± 6.57 µM), Ferulic acid (FA) (32.12 ± 2.50 µM), and Coumaric acid (CA) (79.60 ± 6.21 µM) were identified in the biotransformation products (TP) of flaxseed lignan extract. Results revealed that the TP exhibited a more pronounced anti-inflammatory effect than the flaxseed lignan extract. SECO, FA, and CA demonstrated a more inhibitory effect on NO than that of SDG. The expression of iNOS and COX-2 was significantly suppressed by TP treatment in LPS-induced Raw264.7 cells. The secretion of IL-6, IL-2, and IL-1β decreased by 87.09 ± 0.99%, 45.40 ± 0.87%, and 53.18 ± 0.83%, respectively, at 60 µg/mL of TP treatment. Given these data, the bioavailability of flaxseed lignan extract and its anti-inflammatory effect were significantly enhanced by Lactiplantibacillus plantarum SCB0151, which provided a novel approach to commercializing flaxseed lignan extract for functional food., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

The present study demonstrates the in vivo soft tissue regenerative potential of flax seed mucilage (FSM) reinforced collagen aerogels in Wistar rats. The physiochemical, mechanical, and thermal properties were significantly improved upon the incorporation of flax mucilage into collagen when compared to the native collagen scaffold. In addition, the functional group of flax mucilage notably contributed to a better anti-oxidative potential than the control collagen. The flax mucilage-reinforced collagen at 4 mg ml -1 concentration showed a 2-fold increase in porosity compared to native collagen. The tensile strength of native collagen, 2 mg ml -1 , and 4 mg ml -1 FSM reinforced collagen was 5.22 MPa, 9.76 MPa, and 11.16 MPa, respectively, which indicated that 2 mg ml -1 and 4 mg ml -1 FSM showed an 87% and 113% percentage increase respectively in tensile strength compared to the native collagen control. FSM-reinforced biomatrix showed 97% wound closure on day 15 post-wounding, indicating faster healing than controls, where complete healing occurred only on day 21. The mechanical properties of skin treated with FSM-reinforced collagen scaffold post-healing were considerably better than native collagen. The histological and immunohistochemistry analysis also showed complete restoration of wounded tissue like intact normal skin. The findings paved the way for the development of collagen-polysaccharide mucilage wound dressing materials and their further application in skin tissue engineering., (© 2024 IOP Publishing Ltd.)

Polycystic ovarian syndrome (PCOS) is an endocrinological disorder aroused due to hormonal disturbances. It is characterized by anovulation due to an excess of androgen and estrogen hormones, thus leading to the formation of multiple cysts, imposing life-threatening conditions. This manuscript aimed to introduce a natural estrogen receptor (ESR) inhibitors that can provide protection against PCOS. The computational analysis of Linum usitatissimum seeds  compounds against ESR alpha receptor was performed, and the binding affinities of the ligand compounds and receptor proteins were scrutinized. Nine lignin compounds were docked, and the results were compared with that of reference estrogen receptor inhibitors, clomiphene, and tamoxifen. The binding affinity scores for pinoresinol, lariciresinol, secoisolariciresinol, and matairesinol were -10.67, -10.66, -10.91, and -10.60 kcal mol -1 , respectively. These were comparable to the binding affinity score of reference compounds -11.406 kcal mol -1 for clomiphene and -10.666 kcal mol -1 for tamoxifen. Prime MM-GBSA studies showcased that Linum usitatissimum seeds compounds exhibit significant efficacy and efficiency towards receptor protein. Moreover, MD-simulation studies were performed and the results depict that the lignin compounds form stable complexes at 300 K throughout the simulation time. For further clarity, in-vitro experiments were carried out. The results exhibit the decline in cell proliferation in a concentration-dependent manner by extract 1 (ethyl acetate) EX1 and extract 2 (petroleum ether) EX2. Hence, providing evidence regarding the anti-estrogenic activity of the sample extracts. Collectively, these results showed that flax seed can reduce the levels of estrogen, which can induce ovulation and prevent cyst formation, and ultimately can provide protection against PCOS., (© 2023 John Wiley & Sons Ltd.)

Nitrogen (N), the most important macro-nutrient for plant growth and development, is a key factor that determines crop yield. Yet its excessive applications pollute the environment and are expensive. Hence, studying nitrogen use efficiency (NUE) in crops is fundamental for sustainable agriculture. Here, an association panel consisting of 123 flax accessions was evaluated for 21 NUE-related traits at the seedling stage under optimum N (N+) and N deficiency (N-) treatments to dissect the genetic architecture of NUE-related traits using a multi-omics approach integrating genome-wide association studies (GWAS), transcriptome analysis and genomic selection (GS). Root traits exhibited significant and positive correlations with NUE under N- conditions (r = 0.33 to 0.43, p < 0.05). A total of 359 QTLs were identified, accounting for 0.11% to 23.1% of the phenotypic variation in NUE-related traits. Transcriptomic analysis identified 1034 differentially expressed genes (DEGs) under contrasting N conditions. DEGs involved in N metabolism, root development, amino acid transport and catabolism and others, were found near the QTLs. GS models to predict NUE stress tolerance index (NUE&#95;STI) trait were tested using a random genome-wide SNP dataset and a GWAS-derived QTLs dataset. The latter produced superior prediction accuracy (r = 0.62 to 0.79) compared to the genome-wide SNP marker dataset (r = 0.11) for NUE&#95;STI. Our results provide insights into the QTL architecture of NUE-related traits, identify candidate genes for further studies, and propose genomic breeding tools to achieve superior NUE in flax under low N input., Competing Interests: The authors declare no conflict of interest.

Flax is an important cash crop globally with a variety of commercial uses. It has been widely used for fiber, oil, nutrition, feed and in composite materials. Growth regulatory factor (GRF) is a transcription factor family unique to plants, and is involved in regulating many processes of growth and development. Bioinformatics analysis of the GRF family in flax predicted 17 LuGRF genes, which all contained the characteristic QLQ and WRC domains. Equally, 15 of 17 LuGRFs (88%) are predicted to be regulated by lus-miR396 miRNA. Phylogenetic analysis of GRFs from flax and several other well-characterized species defined five clades; LuGRF genes were found in four clades. Most LuGRF gene promoters contained cis-regulatory elements known to be responsive to hormones and stress. The chromosomal locations and collinearity of LuGRF genes were also analyzed. The three-dimensional structure of LuGRF proteins was predicted using homology modeling. The transcript expression data indicated that most LuGRF family members were highly expressed in flax fruit and embryos, whereas LuGRF3 , LuGRF12 and LuGRF16 were enriched in response to salt stress. Real-time quantitative fluorescent PCR (qRT-PCR) showed that both LuGRF1 and LuGRF11 were up-regulated under ABA and MeJA stimuli, indicating that these genes were involved in defense. LuGRF1 was demonstrated to be localized to the nucleus as expected for a transcription factor. These results provide a basis for further exploration of the molecular mechanism of LuGRF gene function and obtaining improved flax breeding lines.

One of the most intriguing areas of research and innovation in the animal production and food sector recently has been designed-enriched products. These items are regarded as functional foods because they feature components that have advantageous physiological impacts on human health. In the production of poultry, designed eggs constitute a significant category of functional foods. The present study hypothesized that adding different kinds of oils to quail diets will help produce designer eggs rich in omega-3 and 6 fatty acids in addition to enhancing productive performance. So, this study examined how linseed (flaxseed) and canola oils with various levels can affect lipid metabolism, immune function, and the amount of n-3 polyunsaturated fatty acids (n-3 PUFA) in Japanese quail eggs. This work was conducted using 3 different vegetable oils (sunflower, linseed, and canola oils) and 3 different antioxidant supplements (0, 250 mg vitamin E/kg feed, and 1,000 mg ginger/kg feed) in a 3 × 3 factorial experiment. When linseed or canola oil was added to the diet, the number of fatty acids in the egg yolks of Japanese quail layers fell by (12.7 and 18.9%) and (41.4 and 24.6%), respectively. The amounts of saturated and monounsaturated fatty acids in total eggs fell by 21.9 and 14.6% and 24.5 and 15.8%, respectively, at 20 wk of age. However, when linseed and canola oil were added to the diet, the sum n-3 PUFA content in the egg yolk of Japanese quail-laying hens was noticeably raised at 15 and 20 wk of age. At 15 and 20 wk of age, the same groups' total n-6 PUFA content considerably increased compared to the group that did not receive flaxseed. In conclusion, during the laying period of Japanese quail, linseed oil, canola oil, vitamin E, or ginger positively affected productivity, blood hematology, constituents, resistance, lipid digestion system, and antioxidative properties in serum and egg yolk., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

FAD (fatty acid desaturase) and SAD (stearoyl-ACP desaturase) genes play key roles in the synthesis of fatty acids (FA) and determination of oil composition in flax ( Linum usitatissimum L.). We searched for FAD and SAD genes in the most widely used flax genome of the variety CDC Bethune and three available long-read assembled flax genomes-YY5, 3896, and Atlant. We identified fifteen FAD2 , six FAD3 , and four SAD genes. Of all the identified genes, 24 were present in duplicated pairs. In most cases, two genes from a pair differed by a significant number of gene-specific SNPs (single nucleotide polymorphisms) or even InDels (insertions/deletions), except for FAD2a-1 and FAD2a-2 , where only seven SNPs distinguished these genes. Errors were detected in the FAD2a-1 , FAD2a-2 , FAD3c-1 , and FAD3d-2 sequences in the CDC Bethune genome assembly but not in the long-read genome assemblies. Expression analysis of the available transcriptomic data for different flax organs/tissues revealed that FAD2a-1 , FAD2a-2 , FAD3a , FAD3b , SAD3-1 , and SAD3-2 were specifically expressed in embryos/seeds/capsules and could play a crucial role in the synthesis of FA in flax seeds. In contrast, FAD2b-1 , FAD2b-2 , SAD2-1 , and SAD2-2 were highly expressed in all analyzed organs/tissues and could be involved in FA synthesis in whole flax plants. FAD2c-2 , FAD2d-1 , FAD3c-1 , FAD3c-2 , FAD3d-1 , FAD3d-2 , SAD3-1 , and SAD3-2 showed differential expression under stress conditions- Fusarium oxysporum infection and drought. The obtained results are essential for research on molecular mechanisms of fatty acid synthesis, FAD and SAD editing, and marker-assisted and genomic selection for breeding flax varieties with a determined fatty acid composition of oil.

In this study, 16S rRNA sequencing and GC-MS (gas chromatography-mass spectrometry) techniques were employed to examine the relationship between bacterial succession and metabolite alterations during the dew retting process of flax. The results indicated that the addition of compound microbial agents may affect the production and transformation of metabolites by re-establishing bacterial communities and promoting the degradation of pectic substances and the release of metabolites, and the best retting effect was achieved under the combined addition (BA). In addition, Chryseobacterium, Bacillus, and Pseudoonas were closely associated with the production of fatty acids and alcohols; the addition of compound microbial agents increased the content of critical metabolites while decreasing the environmental pollutant bis(2-ethylhexyl) phthalate. In summary, the addition of compound microbial agents can positively regulate the retting process of flax, shorten the retting cycle, improve the quality of flax fibre, and reduce the pollution of the environment., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier Ltd.)

Flaxseed is a rich source of α-linolenic acid (ALA, 18:3 n-3) and can be used to enrich chicken tissues with n-3 fatty acids (FA). However, antinutritional factors in flaxseed compromise the live performance of birds coupled with increased oxidative stress. Chromium (Cr) is a trace element with antioxidant properties. It is hypothesized that Cr supplementation will affect the hepatic total lipid profile, phospholipid n-3 and n-6 FA molecular species, lipid oxidation products, and transcription of genes associated with lipid metabolism in broiler chickens fed flaxseed. Ninety (n = 90), day-old Cornish cross chicks were fed a corn-soybean meal-based diet containing 0% flaxseed (CTR), 10% flaxseed (FLAX), and FLAX + 0.05% organic Cr (FLAXCr) for 42 d. The chicks were kept in 18 pens with 5 chicks per pen. For all response variables, the effect of dietary treatments were compared separately using SAS 9.4. P values were considered significant at ≤0.05. Total lipids, saturated FA, long-chain (≥20C) n-6 FA were reduced while total n-3 FA and long-chain n-3 FA were higher in the liver of FLAX and FLAXCr than CTR (P < 0.05). Hepatic phosphatidylcholine (PC) and phosphatidylethnolamine (PE) n-3 species (36:5, 38:6) were higher in FLAX and FLAXCr compared to CTR (P < 0.05). On the contrary, n-6 species in PC (36:4, 38:4) and PE (38:4) were lower in FLAX and FLAXCr compared to CTR (P < 0.05). Addition of Cr to a flaxseed-containing diet led to an increase in PE 36:4 (P < 0.05). A decrease in the transcription of ELOVL6 gene involved in de novo lipid synthesis was observed in FLAXCr (P = 0.01). An increase in the transcription of genes involved in FA oxidation (ACAA2, ACOX1) was observed in FLAX compared to FLAXCr (P = 0. 05; P = 0.02). A trend for a decrease in the transcription of FADS2 and HMGCS1 was observed in FLAXCr than CTR and FLAX (P = 0.06; 0.08). Transcription of other genes involved in de novo lipid synthesis (FASN, PPARA), FA oxidation (CPT1A, CPT2, ACAA1), and oxidative stress response (GPX1, NQO11, GSTA2, SLC40A1, NFE2L2) were not affected by the diets (P > 0.05). Lipid peroxidation products measured as thiobarbituric acid reactive substances (TBARS) in liver was reduced in FLAXCr than CTR (P < 0.05) and was not different from FLAX (P > 0.05). Serum cholesterol and aspartic aminotransferase were reduced in FLAX and FLAXCr compared to CTR (P < 0.05). The serum glucose level was decreased in FLAX compared to CTR (P < 0.05) and a trend in decrease was noticed in FLAXCr vs. CTR (P = 0.10). Serum TBARS were higher in CTR and FLAXCr compared to FLAX (P < 0.05). In conclusion, flaxseed supplementation enhances total and long-chain n-3 FA while reducing total lipids, saturated, and n-6 FA in the liver. Supplementing Cr along with flaxseed increased n-6 FA species in the hepatic PE and decreased the transcription of genes involved in FA oxidation and lipid synthesis., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

Objectives: A healthy lifestyle, including a healthy diet has been associated with an improvement in cardiovascular risk factors. The aim of the present study was to assess the effect of olive oil and flaxseed consumption as part of a healthy diet on endothelial function, plasma inflammatory factors, and lipid profile in patients with coronary heart disease (CHD)., Method: This randomized nonblinded trial was performed on CHD patients. In the control group, participants received general heart-healthy dietary recommendations while in the intervention group, in addition to these recommendations, the participants consumed 25 ml of olive oil and 30 g of flaxseeds daily for 3 months. At baseline and after 3 months, changes in brachial flow-mediated dilation (FMD), plasma asymmetric dimethyl arginine, interleukin-6 (IL-6), IL-10, high-sensitivity C-reactive protein (hs-CRP), tumor necrosis factor-α (TNF-α), monocyte chemoattractant protein-1 (MCP-1), and lipids and lipoproteins were measured., Results: A total of 50 patients finished the trial ( n  = 24 in the intervention and n  = 26 in the control groups). Compared to the control group, consumption of flaxseed and olive oil significantly improved brachial artery FMD%, and reduced plasma IL-6, TNF-α, MCP-1, total cholesterol, and tended to reduce hs-CRP, and non-HDL-cholesterol but the concentration of other study indices were not different between the two groups., Conclusion: Inclusion of olive oil and flaxseed in the diet of CHD patients may contribute to secondary prevention by improving endothelial function and plasma inflammatory factors., (Copyright © 2023 Wolters Kluwer Health, Inc. All rights reserved.)

Flaxseed (Linum usitatissimum L) is an ancient perennial plant species regarded as a multipurpose plant owing to its richness in omega-3 polyunsaturated fatty acids (PUFA) including α-linolenic acid (ALA). The extensive biochemical analysis of flaxseed resulted in the identification of its bioactive, i.e., lignans with potential application in the improvement of human health. Flaxseed oil, fibers, and lignans exert potential health benefits including reduction of cardiovascular disease, atherosclerosis, diabetes, cancer, arthritis, osteoporosis, and autoimmune and neurological disorders that have led to the diversification of flaxseed plant applications. This comprehensive review focuses on flaxseed oil as the major product of flaxseed with emphasis on the interrelationship between its chemical composition and biological effects. Effects reviewed include antioxidant, anti-inflammatory, antimicrobial, anticancer, antiulcer, anti-osteoporotic, cardioprotective, metabolic, and neuroprotective. This study provides an overview of flaxseed oil effects with the reported action mechanisms related to its phytochemical composition and in comparison, to other PUFA-rich oils. This study presents the most updated and comprehensive review summarizing flaxseed oil's health benefits for the treatment of various diseases., (© 2023. The Author(s).)

Auxin response factors (ARFs) are critical components of the auxin signaling pathway, and are involved in diverse plant biological processes. However, ARF genes have not been investigated in flax ( Linum usitatissimum L.), an important oilseed and fiber crop. In this study, we comprehensively analyzed the ARF gene family and identified 33 LuARF genes unevenly distributed on the 13 chromosomes of Longya-10, an oil-use flax variety. Detailed analysis revealed wide variation among the ARF family members and predicted nuclear localization for all proteins. Nineteen LuARFs contained a complete ARF structure, including DBD, MR, and CTD, whereas the other fourteen lacked the CTD. Phylogenetic analysis grouped the LuARFs into four (I-V) clades. Combined with sequence analysis, the LuARFs from the same clade showed structural conservation, implying functional redundancy. Duplication analysis identified twenty-seven whole-genome-duplicated LuARF genes and four tandem-duplicated LuARF genes. These duplicated gene pairs' K a /K s ratios suggested a strong purifying selection pressure on the LuARF genes. Collinearity analysis revealed that about half of the LuARF genes had homologs in other species, indicating a relatively conserved nature of the ARFs . The promoter analysis identified numerous hormone- and stress-related elements, and the qRT-PCR experiment revealed that all LuARF genes were responsive to phytohormone (IAA, GA3, and NAA) and stress (PEG, NaCl, cold, and heat) treatments. Finally, expression profiling of LuARF genes in different tissues by qRT-PCR indicated their specific functions in stem or capsule growth. Thus, our findings suggest the potential functions of LuARFs in flax growth and response to an exogenous stimulus, providing a basis for further functional studies on these genes.

This paper describes the effects of flaxseed (Linum usitatissimum) oil (FSO) as a feed additive on growth performance, oxidative stress, immunity, and disease resistance in rainbow trout (Oncorhynchus mykiss). Eight-hundred-and-forty rainbow trout individuals (mean weight: 25.66 ± 1.33 g) were fed with different doses of FSO (0.5, 1, and 1.5%) ad libitum two times a day for 9 weeks. At the end of the feeding, growth performance was evaluated and the fish were challenged with two different bacteria (Yersinia ruckeri and Aeromonas hydrophila). At the end of the 3rd, 6th, and 9th weeks, blood and tissue samples were taken from 9 fish per treatment to evaluate innate immune response, cytokine gene expression levels, antioxidant enzyme activities and lipid peroxidation levels, and digestive enzyme activities. Determination of haematological parameters and histological examination was also carried out to evaluate the general health status of the fish. Results showed that the final weight and specific growth rate of FSO-supplemented fish increased significantly (p < 0.05). FSO-supplemented fish showed higher resistance to Y. ruckeri than the control group (p < 0.05). However, survival rates of all groups in A. hydrophila challenge test were similar (p > 0.05). Among the investigated innate immune response parameters, the potential killing activity of phagocytes, myeloperoxidase activity, and lysozyme activity increased in the FSO-supplemented groups (p < 0.05). Almost all cytokine gene expression levels in the experimental groups up-regulated especially after 9 weeks of feeding in the head kidney and intestine (p < 0.05). Similarly, superoxide dismutase and catalase activities were found to be significantly higher in the FSO group than in the control (p < 0.05) whereas, the lipid peroxidation levels drastically declined as a result of the FSO supplementation (p < 0.05). These results suggest that FSO can improve growth, enhance immune response, and lower oxidative damage in rainbow trout when supplemented at the rates of 0.5-1.5% for 9 weeks., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Background: Oilseeds such as linseed, canola and sunflower contain unsaturated fatty acids that play important functions in the body. The aim of this study was to assess the effects of different levels of processing linseed on growth performance, nutrient digestibility, blood parameters and ruminate behaviour of lambs., Methods: Fifty-six Moghani male lambs (3 months of age, initial average body weight = 28 ± 1.2 kg) were allocated to seven experimental diets in randomized design (eight lambs per each treatment). The experimental diets were as follows: (1) control diet (without linseed), (2) 5% raw linseed, (3) 10% raw linseed, (4) 5% micronized linseed, (5) 10% micronized linseed, (6) 5% extruded linseed and (7) 10% extruded linseed. Lambs were fed ad libitum a basal diet as total mixed ration consisting of 25% concentrate and 75% hay., Results: The results showed that linseed level and processing method had no significant effect on dry matter intake. Average daily gain, final body weight and feed conversion ratio (FCR) in lambs were affected by experimental diets. The use of 10% micronized linseed and 10% of extruded linseed in the lambs' diet improved dry matter and crude protein digestibility significantly (p < 0.001). Blood glucose concentration observed for lambs fed 10% of micronized or extruded linseed (LS) was not different from that observed in other groups, only from the values shown by lambs fed diets 1 (control) and 2 (5% raw LS). The lowest cholesterol and the highest blood urea nitrogen concentrations were related to lambs fed the control diet (p < 0.001). Feeding processed linseed relative to control diet had no effect on feeding behaviour in lambs., Conclusion: Results of this research showed that the use of extruded and micronized linseed at the level of 10% can improve FCR, nutrient digestibility, and blood parameters., (© 2023 The Authors. Veterinary Medicine and Science published by John Wiley & Sons Ltd.)

Linum album is a well-known rich source of anticancer compounds, i.e., podophyllotoxin (PTOX) and other lignans. These compounds play an important role in the plant's defensive system. The RNA-Seq data of flax (L. usitatissimum) were analyzed under various biotic and abiotic stresses to comprehend better the importance of lignans in plant defense responses. Then, the association between the lignan contents and some related gene expressions was experimented with HPLC and qRT-PCR, respectively. Transcriptomic profiling showed a specific expression pattern in different organs, and just the commonly regulated gene EP3 was detected with a significant increase under all stresses. The in silico analysis of the PTOX biosynthesis pathway identified a list of genes, including laccase (LAC11), lactoperoxidase (POD), 4-coumarate-CoA ligase (4CL), and secoisolariciresinol dehydrogenase (SDH). These genes increased significantly under individual stresses. The HPLC analysis showed that the measured lignan contents generally increased under stress. In contrast, a quantitative expression of the genes involved in this pathway using qRT-PCR showed a different pattern that seems to contribute to regulating PTOX content in response to stress. Identified modifications of critical genes related to PTOX biosynthesis in response to multiple stresses can provide a baseline for improving PTOX content in L. album., (© 2023. The Author(s).)

Flax is a flowering plant cultivated for its oil and contains various unsaturated fatty acids. Linseed oil is known as the "deep-sea fish oil" of plants, and is beneficial to brain and blood lipids, among other positive effects. Long non-coding RNAs (lncRNAs) play an important role in plant growth and development. There are not many studies assessing how lncRNAs are related to the fatty acid synthesis of flax. The relative oil contents of the seeds of the variety Heiya NO.14 (for fiber) and the variety Macbeth (for oil) were determined at 5 day, 10 day, 20 day, and 30 day after flowering. We found that 10-20 day is an important period for ALA accumulation in the Macbeth variety. The strand-specific transcriptome data were analyzed at these four time points, and a series of lncRNAs related to flax seed development were screened. A competing endogenous RNA (ceRNA) network was constructed and the accuracy of the network was verified using qRT-PCR. MSTRG.20631.1 could act with miR156 on the same target, squamosa promoter-binding-like protein (SPL), to influence fatty acid biosynthesis through a gluconeogenesis-related pathway during flax seed development. This study provides a theoretical basis for future studies assessing the potential functions of lncRNAs during seed development., Competing Interests: The authors declare no conflict of interest.

Background: Phytic acid is an antinutritional compound that reduces the bioavailability of minerals and proteins. Flaxseed is a rich source of unsaturated fatty acids and antioxidants. In this study, flaxseed flour was dephytinized by phytase enzyme and fermentation. Untreated and dephytinized flaxseed flours were used in noodle production at 0%, 10%, 20% and 30% levels to improve nutritional quality., Results: The addition of flaxseed flours did not show an adverse effect on the cooking loss values of noodles (4.22-4.85%). Firmness values of noodles containing 10% flaxseed flour were close to the control (0% flaxseed flour). The ash, protein and fat levels of noodles were enhanced with the addition of flaxseed flours. The use of 30% phytase-treated flaxseed flour in noodles elicited lower phytic acid concentration (1.35 g kg -1 ) than the control (1.52 g kg -1 ). The greatest total phenolic contents in noodles were obtained with the inclusion of fermented flour. Besides that, noodles made from flaxseed flours exhibited stronger antioxidant activity than the control. The overall acceptability scores of noodles containing 10% and 20% dephytinized flours were close to the control., Conclusion: The findings demonstrated that the inclusion of dephytinized flaxseed flours up to 20% can allow the production of functional noodles with acceptable quality. Moreover, phytase enzyme treatment method revealed lower phytic acid content than fermentation method in noodles and better antioxidant properties than untreated noodles. © 2022 Society of Chemical Industry., (© 2022 Society of Chemical Industry.)

Melatonin is a pleiotropic molecule that can influence various aspects of plant performance. Recent studies have exhibited that it mediates plant defensive responses, probably through managing redox homeostasis. We tried to track the regulatory effects of melatonin on the antioxidant machinery of Linum album cell culture. To this, different concentrations of melatonin were applied, and the oxidative status of cells was investigated by measuring the levels of oxidative molecules and antioxidant agents. The results showed that H 2 O 2 content did not change at the low melatonin levels, while it increased at the high concentrations. It can be correlated with the low melatonin dosages capacity to remove excessive amounts of H 2 O 2 , while the high melatonin dosages exhibit toxicity effects. In contrast, the NO enhancement occurred at 50 μM melatonin, proposing its role in triggering melatonin-induced defensive responses. The MDA results stated that NO led to oxidative stress in melatonin-treated cells at 50 μM melatonin. Antioxidant enzyme POD was activated by melatonin treatment, while SOD enzyme behaved reversely which can explain the changes in the H 2 O 2 level. In addition, the analysis of the phenolics profile showed that the contents of phenolic acids, flavonoids, and lignans enhanced following an increase in PAL enzyme activity. The increased level of phenolic hormone SA can indicate that melatonin affects the defensive responses in L. album cells through a SA-dependent pathway. In general, it seems that melatonin, by modulating NO and SA levels, can induce the activity of antioxidant enzymes and the production of phenolics, especially lignans, in L. album cells., (© 2023. The Author(s).)

In this study, the combined addition of Bacillus licheniformis HDYM-04 and Bacillus subtilis ZC-01 to flax degradation increased the degradation rates of pectin (74.7 %) and pectinic acid (59.3 %) and increased the maximum activities of pectinase (610.66 ± 7.03 U/mL) and mannanase (656.97 ± 13.16 U/mL). 16S rRNA sequencing showed that the added bacterial agent (Bacillus) was the dominant bacterium, and its addition increased the relative abundance (RA) of Firmicutes and decreased the RA of Bacteroidetes. The core bacterial community linked to degradation (Firmicutes) was determined by RDA. Network analysis showed that the number of bacteria related to pectin and hemicellulose degradation increased with the addition of the bacteria combination. SEM analysis showed that Bacillus was positively correlated with the degradation of pectic substances. These results provide new ideas for improving the utilization of agricultural waste resources and promoting sustainable development in modern agriculture., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022. Published by Elsevier Ltd.)

Liver fibrosis is a major pathological feature of chronic liver disease and effective therapies are limited at present. The present study focuses on the hepatoprotective potential of L . corymbulosum against carbon tetrachloride (CCl 4 )-induced liver damage in rats. Analysis of Linum corymbulosum methanol extract (LCM) using high-performance liquid chromatography (HPLC) revealed the presence of rutin, apigenin, catechin, caffeic acid and myricetin. CCl 4 administration lowered ( p < 0.01) the activities of antioxidant enzymes and reduced glutathione (GSH) content as well as soluble proteins, whereas the concentration of H 2 O 2 , nitrite and thiobarbituric acid reactive substances was higher in hepatic samples. In serum, the level of hepatic markers and total bilirubin was elevated followed by CCl 4 administration. The expression of glucose-regulated protein (GRP78), x-box binding protein-1 total (XBP-1 t), x-box binding protein-1 spliced (XBP-1 s), x-box binding protein-1 unspliced (XBP-1 u) and glutamate-cysteine ligase catalytic subunit (GCLC) was enhanced in CCl 4 -administered rats. Similarly, the expression of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and monocyte chemo attractant protein-1 (MCP-1) was strongly increased with CCl 4 administration to rats. Co-administration of LCM along with CCl 4 to rats lowered ( p < 0.05) the expression of the above genes. Histopathology of the liver showed hepatocyte injury, leukocyte infiltration and damaged central lobules in CCl 4 -treated rats. However, LCM administration to CCl 4 -intoxicated rats restored the altered parameters towards the levels of control rats. These outcomes indicate the existence of antioxidant and anti-inflammatory constituents in the methanol extract of L. corymbulosum .

Metal retention in wastewater fertigated crops poses a potential hazard to food chain. Current work demonstrates the bioremediation and growth-promoting potential of Acinetobacter schindleri SR-5-1 by using nitrogen-fixing (pea) and non-nitrogen fixing (linseed) plants under cadmium (Cd) and wastewater irrigation regimes. Both plants were grown at 250 or 500 CdCl 2 and 75 or 100% wastewater, each separately with and without A. schindleri SR-5-1 inoculation. The results revealed that Cd and wastewater significantly decreased growth, biomass, antioxidants, and nutrient acquisition through increased malondialdehyde, H 2 O 2 , and Cd accumulation. However, application of A. schindleri SR-5-1 significantly promoted morpho-physio-biochemical attributes while diminishing MDA and H 2 O 2 under applied Cd and wastewater stress levels in both pea and linseed. Further, PGPR inoculation positively influenced pea and linseed seedlings through a substantial decline in Cd accumulation in roots/shoots and retained the optimal level of essential nutrients. It was inferred that both pea and linseed, with A. schindleri SR-5-1 application, exhibited higher growth and metabolism under Cd and wastewater stress but substantial tolerance was acquired under wastewater stress. Studied plants exhibited tolerance in order of 75% WW ≥ 250 µM Cd ≥ 100%WW ≥ 500 µM Cd treatment under A. schindleri inoculation. Current findings revealed the potential of A. schindleri to be exploited both for bioremediation and bio-fertilization under Cd, and wastewater-polluted regimes to reduce metal contamination of edible plants. It was suggested that with inoculation of A. schindleri SR-5-1, 75% WW dilution can be applied for irrigation of both nitrogen-fixing and non-nitrogen-fixing crops., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Linseed as a common oil crop that has been implicated in regulating the meat quality of animals. The aim of this investigation was to establish the effects of dietary linseed supplementation on carcass traits, meat quality, muscle fiber characteristics, volatile compounds and antioxidant capacity of sheep. Twenty-four Sunit sheep were distributed into 2 groups with different diet: control (CO) fed a standard diet and linseed (LS, 8% of extruded linseed). The results showed that linseed not only increased M. longissimus thoracis (LT) increased the loin-eye area but also decreased pH24h, L* and shear force. Meanwhile, linseed increased mRNA expression of MyHCI and MyHCIIx and decreased in cross-sectional area (CSA) and muscle fiber diameter. In addition, linseed altered the composition of meat volatile flavor compounds, such as Z-10-Pentadecen-1-ol, pentanal, 2-Octenal, (E)-, decanal, butane, and 2-heptanone. Moreover, linseed increased total antioxidative capacity (T-AOC) activity, the mRNA expression of glutathione peroxidase (GSH-Px) and catalase (CAT), decreased malondialdehyde (MDA) content. Overall, the results suggest that linseed is an effective feed additive in improving meat quality and flavor. The underlying mechanisms for its effectiveness may be partly due to a change in muscle fiber characteristics and antioxidant capacity., (© 2023 Japanese Society of Animal Science.)

Hyperlipidemia, high levels of blood lipids including cholesterol and triglycerides, is a major risk factor for cardiovascular disease. Traditional treatments of hyperlipidemia often include lifestyle changes and pharmacotherapy. Recently, flaxseed has been approved as a nutrient that lowers blood lipids. Several metabolites of flaxseed lignan secoisolariciresinol diglucoside (SDG), have been identified that reduce blood lipids. SDG is present in flaxseed hull as an ester-linked copolymer with 3-hydroxy-3-methylglutaric acid (HMGA). However, purification processes involved in hydrolysis of the copolymer and enriching SDG are often expensive. The natural copolymer of SDG with HMGA (SDG polymer) is a source of bioactive compounds useful in prophylaxis of hypercholesterolemia. After consumption of the lignan copolymer, SDG and HMGA are released in the stomach and small intestines. SDG is metabolized to secoisolariciresinol, enterolactone and enterodiol, the bioactive forms of mammalian lignans. These metabolites are then distributed throughout the body where they accumulate in the liver, kidney, skin, other tissues, and organs. Successively, these metabolites reduce blood lipids including cholesterol, triglycerides, low density lipoprotein cholesterol, and lipid peroxidation products. In this review, the metabolism and efficacies of flaxseed-derived enriched SDG and SDG polymer will be discussed.

Flax (Linum usitatissimum) is an important oil crop in arid and semi-arid regions of North and Northwest China, and its seeds are rich in nutritious storage reserves, such as polyunsaturated fatty acids (FAs) and proteins. However, the regulatory networks that control the accumulation of seed storage reserves in flax are still largely unknown. In this study, we found that LuABI3-1 and LuABI3-2 homologs from the flax cultivar 'Longya 10' play important roles in regulating the accumulation of seed storage reserves in Arabidopsis thaliana. The results of subcellular localization and transcriptional activity assays showed that both LuABI3-1 and LuABI3-2 function as transcription factors. Overexpression of either LuABI3-1 or LuABI3-2 resulted in the significant increase in the contents of total seed FAs and storage proteins, but did not alter other key agronomic traits in A. thaliana. Accordingly, the expression of key genes involved in the biosynthesis of FAs and storage proteins was also greatly up-regulated in the developing seeds of LuABI3-1-overexpression lines. Additionally, both LuABI3-1 and LuABI3-2 enhanced the tolerance to the high salt and mannitol stresses during seed germination and seedling establishment in A. thaliana. These results increase our understanding of the LuABI3 regulatory functions and provide promising targets for genetic manipulation of L. usitatissimum to innovate the germplasm resources and cultivate high yield and quality varieties., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022. Published by Elsevier GmbH.)

Linosorbs (Los) are cyclic peptides from flaxseed oil composed of the LO mixture (LOMIX). The activity of LO has been reported as being anti-cancer and anti-inflammatory. However, the study of skin protection has still not proceeded. In particular, there are poorly understood mechanisms of melanogenesis to LO. Therefore, we investigated the anti-melanogenesis effects of LOMIX and LO, and its activity was examined in mouse melanoma cell lines. The treatment of LOMIX (50 and 100 μg/mL) and LO (6.25-50 μM) suppressed melanin secretion and synthesis, which were 3-fold increased, in a dose-dependent manner, up to 95%. In particular, [1-9-NαC]-linusorb B3 (LO1) and [1-9-NαC]-linusorb B2 (LO2) treatment (12.5 and 25 μM) highly suppressed the synthesis of melanin in B16F10 cell lines up to 90%, without toxicity. LOMIX and LOs decreased the 2- or 3-fold increased mRNA levels, including the microphthalmia-associated transcription factor (MITF), Tyrosinase, tyrosinase-related protein 1 (TYRP1), and tyrosinase-related protein 2 (TYRP2) at the highest concentration (25 μM). Moreover, the treatment of 25 μM LO1 and LO2 inhibited the expression of MITF and phosphorylation of upper regulatory proteins such as CREB and PKA. Taken together, these results suggested that LOMIX and its individual LO could inhibit melanin synthesis via downregulating the CREB-dependent signaling pathways, and it could be used for novel therapeutic materials in hyperpigmentation.

The imbalance in polyunsaturated fatty acid (PUFA) composition in human food is ubiquitous and closely related to obesity and cardiovascular diseases. The development of n-3 PUFA-enriched poultry products is of great significance for optimizing fatty acid composition. This study aimed to improve our understanding of the effects of dietary linseed oil on hepatic metabolism using untargeted metabolomics and 4D label-free proteome analysis. A total of 91 metabolites and 63 proteins showed differences in abundance in duck livers between the high linseed oil and control groups. Pathway analysis revealed that the biosynthesis of unsaturated fatty acids, linoleic acid, glycerophospholipid, and pyrimidine metabolisms were significantly enriched in ducks fed with linseed oil. Meanwhile, dietary linseed oil changed liver fatty acid composition, which was reflected in the increase in the abundance of downstream metabolites, such as α-linolenic acid (ALA; 18:3n-3) as a substrate, including n-3 PUFA and its related glycerophospholipids, and a decrease in downstream n-6 PUFA synthesis using linoleic acid (LA; 18:2n-6) as a substrate. Moreover, the anabolism of PUFA in duck livers showed substrate-dependent effects, and the expression of related proteins in the process of fatty acid anabolism, such as FADS2, LPIN2, and PLA2G4A, were significantly regulated by linseed oil. Collectively, our work highlights the ALA substrate dependence during n-3 PUFA synthesis in duck livers. The present study expands our knowledge of the process products of PUFA metabolism and provides some potential biomarkers for liver health.

This study was carried out to determine the effect of dietary flaxseed meal (FSM) supplemented with dried tomato pomace (DTP) and dried grape pomace (DGP) on performance, egg quality, biochemical parameters traits and antioxidant status of laying hens. Birds (1825 ± 87 g of body weight) were divided into 12 dietary groups with six replicates per group (eight birds per replicate), under a completely randomized design with factorial arrangement 2 × 3 × 2 consisted of two levels of DTP (0 and 15%), three FSM levels (0, 4 and 8%) and two levels of DGP (0 and 5%). As a result of this study, there were no significant differences in egg production and weight as well in feed conversion ratio (FCR) among treatments ( p  > 0.05). Feeding of DGP reduced significantly feed intake and egg mass when compared to control group ( p  < 0.05). There was no effect ( p  > 0.05) of dietary treatment on shell thickness and strength, shape index, Haugh unit and egg specific gravity. Hens consuming 15% DTP and 5% DGP revealed a significantly higher yolk color compared to the other dietary treatments ( p  < 0.05). Moreover, there was no difference among dietary treatments in terms of serum low-density lipoprotein cholesterol (LDL) and high-density lipoprotein cholesterol (HDL) cholesterol, atherogenic index, triglycerides, total cholesterol levels ( p  > 0.05). Serum antioxidant parameters as glutathione peroxidase (GSH-Px) activity, malondialdehyde (MDA), total superoxide dismutase (TSOD) and total antioxidant capacity (T-AOC) were not influenced by treatments ( p  > 0.05). Based on findings, FSM and DTP supplements did not significantly impact most of hens' performance indicators and egg quality parameters, whereas significant improvements were observed by feeding of 15% DTP and 5% DGP on egg traits, in particular on egg-yolk color that plays a key-role in consumer's choice. However, the supplementation of FSM and DTP or DGP even in laying hen diet is still controversial and further research is needed.

In this paper, we explore potential genetic factors in control of flax phenotypes associated with fiber by mining a collection of 306 flax accessions from the Federal Research Centre of the Bast Fiber Crops, Torzhok, Russia. In total, 11 traits were assessed in the course of 3 successive years. A genome-wide association study was performed for each phenotype independently using six different single-locus models implemented in the GAPIT3 R package. Moreover, we applied a multivariate linear mixed model implemented in the GEMMA package to account for trait correlations and potential pleiotropic effects of polymorphisms. The analyses revealed a number of genomic variants associated with different fiber traits, implying the complex and polygenic control. All stable variants demonstrate a statistically significant allelic effect across all 3 years of the experiment. We tested the validity of the predicted variants using gene expression data available for the flax fiber studies. The results shed new light on the processes and pathways associated with the complex fiber traits, while the pinpointed candidate genes may be further used for marker-assisted selection.

Soil salinity constrains agricultural development in arid regions. Flax is an economically important crop in many countries, and screening or breeding salinity-resistant flax cultivars is necessary. Based on the previous screening of flaxseed cultivars C71 (salt-sensitive) and C116 (salt-tolerant) as test materials, flax seedlings stressed with different concentrations of NaCl (0, 100, 150, 200, and 250 mmol/L) for 21 days were used to investigate the effects of salt stress on the growth characteristics, osmotic regulators, and antioxidant capacity of these flax seedlings and to reveal the adaptive responses of flax seedlings to salt stress. The results showed that plant height and root length of flax were inhibited, with C116 showing lower growth than C71. The concentrations of osmotic adjustment substances such as soluble sugars, soluble proteins, and proline were higher in the resistant material, C116, than in the sensitive material, C71, under different concentrations of salt stress. Consistently, C116 showed a better rapid scavenging ability for reactive oxygen species (ROS) and maintained higher activities of antioxidant enzymes to balance salt injury stress by inhibiting growth under salt stress. A transcriptome analysis of flax revealed that genes related to defense and senescence were significantly upregulated, and genes related to the growth and development processes were significantly downregulated under salt stress. Our results indicated that one of the important adaptations to tolerance to high salt stress is complex physiological remediation by rapidly promoting transcriptional regulation in flax.

Intrusive growth is a type of growth in which a cell exceeds the growth rate of its neighbours and intrudes between them, reaching a much greater length. This process provides plant fibres with their exceptional length. Fibres are the most abundant cell type in the mechanical tissues of plants. At the same time, the plant fibres are of fundamental importance for the production of textiles, paper, biocomposites, etc. Here we describe a mutant of flax (reduced fibre 1, rdf) in which intrusive growth of fibres is impaired in both phloem and xylem. In addition to the intrinsic differences in fibre length, the mutant is characterized by a constitutive gravitropic response, mechanical aberrations at the macro- and nanolevels, disruption of the cambium and uneven transition of xylem cells to secondary cell wall formation. Gelatinous cell walls in both phloem and xylem of mutant plants have disturbed structure and reduced elasticity. The existence of this mutant-control pair offers both prospects for finding the molecular players involved in triggering intrusive growth, cell wall thickening and for understanding the principles of plant mechanical tissue functioning., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Omega-3 fatty acids support cardiometabolic health and reduce chronic low-grade inflammation. These fatty acids may impart their health benefits partly by modulating the endocannabinoidome and the gut microbiome, both of which are key regulators of metabolism and the inflammatory response. Whole hemp seeds ( Cannabis sativa ) are of exceptional nutritional value, being rich in omega-3 fatty acids. We assessed the effects of dietary substitution (equivalent to about 2 tablespoons of seeds a day for humans) of whole hemp seeds in comparison with whole linseeds in a diet-induced obesity mouse model and determined their effects on obesity and the gut microbiome-endocannabinoidome axis. We show that whole hemp seed substitution did not affect weigh gain, adiposity, or food intake, whereas linseed substitution did, in association with higher fasting glucose levels, greater insulin release during an oral glucose tolerance test, and higher levels of liver triglycerides than controls. Furthermore, hemp seed substitution mitigated diet-induced obesity-associated increases in intestinal permeability and circulating PAI-1 levels, while having no effects on markers of inflammation in epididymal adipose tissue, which were, however, increased in mice fed linseeds. Both hemp seeds and linseeds were able to modify the expression of several endocannabinoidome genes and markedly increased the levels of several omega-3 fatty acid-derived endocannabinoidome bioactive lipids with previously suggested anti-inflammatory actions in a tissue specific manner, despite the relatively low level of seed substitution. While neither diet markedly modified the gut microbiome, mice on the hemp seed diet had higher abundance of Clostridiaceae 1 and Rikenellaceae than mice fed linseed or control diet, respectively. Thus, hemp seed-containing foods might represent a source of healthy fats that are not likely to exacerbate the metabolic consequences of obesogenic diets while producing intestinal permeability protective effects and some anti-inflammatory actions., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Ben Necib, Manca, Lacroix, Martin, Flamand, Di Marzo and Silvestri.)

Flaxseeds are typically consumed either as whole flaxseed, ground flaxseed, flaxseed oil, partially defatted flaxseed meal, or as a milk alternative. They are considered a rich source of vitamins, minerals, proteins and peptides, lipids, carbohydrates, lignans, and dietary fiber, which have shown hypolipidemic, antiatherogenic, anticholesterolemic, and anti-inflammatory property activity. Here, an in vitro batch culture model was used to investigate the influence of whole milled flaxseed and partially defatted milled flaxseed press cake on the gut microbiota and the liberation of flaxseed bioactives. Microbial communities were profiled using 16S rRNA gene-based high-throughput sequencing with targeted mass spectrometry measuring lignan, cyclolinopeptide, and bile acid content and HPLC for short-chain fatty acid profiles. Flaxseed supplementation decreased gut microbiota richness with Firmicutes, Proteobacteria, and Bacteroidetes becoming the predominant phyla. Secoisolariciresinol, enterodiol, and enterolactone were rapidly produced with acetic acid, butyric acid, and propionic acid being the predominant acids after 24 h of fermentation. The flaxseed press cake and whole flaxseed were equivalent in microbiota changes and functionality. However, press cake may be superior as a functional additive in a variety of foods in terms of consumer acceptance as it would be more resistant to oxidative changes.

Flaxseed (FS) and sesame seed (SS) are traditional and functional foods in traditional Indian medicine for treating various disorders. The present study investigated the hepatoprotective effects of bioactive-fatty acids (FAs) from FS and SS against carbon tetrachloride (CCl 4 )-induced hepatic damage in rats. Pre and post-treatments for 28 consecutive days significantly increased the activities of in vivo antioxidant enzymes such as catalase (CAT), superoxide dismutase (SOD), and peroxidase (POX), whereas, lipid peroxidation (LPO) activity was markedly decreased in a dose-dependent manner in liver and kidneys. A significant reduction was observed in the hematological parameters like aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), and total bilirubin in the serum of post-treated animals compared to the negative control. The results were confirmed histopathologically. The results suggested that the ω-3 and ω-6 FAs from flaxseed oil (FSO) and sesame seed oil (SSO), respectively, showed potential synergistic hepatoprotective and antioxidant effects that were mediated mainly by ω-3 and ω-6 FAs present in the respective seed oils.

Background: Plant peptides have been reported to have cholesterol-lowering activities. Previous research has found that ≤1 kDa flaxseed peptide (FP 5 ) reduces cholesterol absorption and synthesis in vitro. In this research, we investigated the cholesterol-lowering activity of FP 5 in Sprague-Dawley (SD) rats fed a high-cholesterol and high-fat diet. In addition, amino acid sequences of FP 5 were determined by high-performance liquid chromatography-electrospray ionization-Orbitrap mass spectrometry., Results: FP 5 supplement significantly decreased the serum and hepatic cholesterol levels and modulated the hepatic gene and protein expression of cholesterol metabolism-related enzymes or regulators (3-hydroxy-3-methylglutaryl coenzyme A reductase, Low-Density Lipoprotein Receptor (LDLR), Cholesterol 7 α-hydroxylase, Niemann-Pick C1-like 1, ATP-binding cassette transporters G5 and G8). Eleven peptides were identified from FP 5 . These peptides were characterized as hydrophobic amino acids such as leucine (L), proline (P), glycine (G), isoleucine (I) and continuous sequences, including LP, LL, LG and II, with low molecular weights., Conclusion: FP 5 has a certain cholesterol-lowering activity in SD rats fed a high-cholesterol and high-fat diet. The possible mechanism for ameliorating hepatic cholesterol metabolism of FP 5 includes inhibiting hepatic cholesterol de novo synthesis, promoting the synthesis and excretion of bile acids, and inhibiting the reabsorption of bile acids during enterohepatic circulation. © 2022 Society of Chemical Industry., (© 2022 Society of Chemical Industry.)

The use of phytoregulators is a modern way of producing crops to increase yields and product quality. The components of phytoregulators have a beneficial effect on the growth and development of plants and help to withstand unfavorable environmental factors, stress, lack of mineral nutrition, soil fatigue, and the action of a wide range of pesticides. The research is aimed to investigate the influence of three phytoregulators: Cherkaz, Floravit, EcoFus on two varieties of flax: Severny and LM 98. We conducted a 3-year, field experiment with flax. In the so-called herringbone stage of the plant growth, the flax seedlings were treated the investigated phytoregulators. Flax plants were analyzed for yields. The method of near-infrared spectroscopy was used to analyze the content of lipids and proteins in flax seeds after harvesting. The oil content in flax seeds was determined. Using chromatography, we analyzed each variant of the experiment from the point of view of fatty acid composition. The application of phytoregulators increased the seed yield by 14-19% and the short fiber yield by 9-20% relative to control. The protein content in the seeds of the experimental samples Floravit and Cherkaz increased by 2.3-2.5% and by 1.5% in the experiments with EcoFus relative to the control. The lipid content in flax seeds increased by 5-5.6% (Floravit), by 3.5-3.95% (EcoFus), and by 4.7-5.1% (Cherkaz) relative to control. The oil ratio in flax seeds of the LM 98 and Severny varieties averaged 35.5-45.5%. An increase in the oil yields was 2.8-6.9% for Floravit, 2-4.1% for EcoFus, and 4.4-4.9% for Cherkaz relative to control. The application of Floravit, Cherkaz and EkoFus in flaxseed cultivation contributed to an increase in polyunsaturated fatty acids in oil: the content of ω-3 fatty acids ranged 61.90-63.10% compared to control 54.70%. Floravit proved to be the most effective of the three tested phytoregulators. The studies carried out over three years allow us to conclude that the use of new phytoregulators contributes to an increase in the quality and flaxseed yield and the flax fiber yield. Floravit was the most effective preparation and can be used for pre-sowing seed treatment and during the growing season.

Context: Flaxseed has a characteristic fatty acids composition and unique phytonutrient profile that may have health-promoting properties., Objective: This study aimed to determine the effects of 10 weeks of supplementation with the flaxseed (28 g/day) on endothelial cells (EC) function, serum lipids and proinflammatory mediators in patients with mild and severe dyslipidaemia., Materials and Methods: Eleven lean patients with severe dyslipidaemia treated with apheresis (group 1; 10 weeks treated in four phases: (i) ordinary diet, (ii) ordinary diet + flaxseed, (iii) ordinary diet (wash out), (iv) ordinary diet + placebo) and eleven obese patients with mild dyslipidaemia-not treated with apheresis (group 2; 10 weeks treated in two phases: (i) ordinary diet, (ii) low fat diet + flaxseed). Flaxseed was given blindly. Serum was collected at the end of each phase of the study. ECs were exposed in vitro to the medium supplemented with pooled serum taken from patients from both groups to detect their morphological changes using light and electron microscopy. ECs proliferation was also measured at the end of each study phase., Results: Serum vascular endothelial growth factor was decreased after flaxseed supplementation but only in group 1. ECs proliferation was increased after flaxseed supplementation only in obese patients. ECs exposed to medium supplemented with obese patients' serum revealed the following cellular abnormalities: accumulation of lipid droplets, changes of rough endoplasmic reticulum and mitochondria, and flaxseed did not reverse observed changes. At the same time, flaxseed supplementation decreases total cholesterol in both tested groups, low-density lipoprotein cholesterol in group 1 and triglycerides in group 2., Conclusions: Our findings support the potential role of flaxseed in treating dyslipidaemia but indicate only a slight impact on endothelial cell function.

High-fiber plant foods contain lignans that are converted to bioactive enterolignans, enterolactone (ENL) and enterodiol (END) by gut bacteria. Previously, we conducted an intervention study to gain mechanistic insight into the potential chemoprotective effects of flaxseed lignan supplementation (secoisolariciresinol diglucoside; SDG) compared to a placebo in 42 men and women. Here, we expand on these analyses to further probe the impact of the microbial metabolite phenotype on host gene expression in response to lignan exposure. We defined metabolic phenotypes as high- or low-ENL excretion based on the microbial metabolism of SDG. RNA-seq was used to assess host gene expression in fecal exfoliated cells. Stratified by microbial ENL excretion, differentially expressed (DE) genes in high- and low-ENL excreter groups were compared. Linear discriminant analysis using the ENL phenotypes identified putative biomarker combinations of genes capable of discriminating the lignan treatment from the placebo. Following lignan intervention, a total of 165 DE genes in high-ENL excreters and 1450 DE genes in low-ENL excreters were detected. Functional analysis identified four common upstream regulators (master genes): CD3, IFNG, IGF1 and TNFRSF1A. Our findings suggest that the enhanced conversion of flaxseed lignan to ENL is associated with a suppressed inflammatory status.