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Background LEA proteins are widely distributed in the plant and animal kingdoms, as well as in micro-organisms. LEA genes make up a large family and function in plant protection against a variety of adverse conditions. Results Bioinformatics approaches were adopted to identify LEA genes in the flax genome. In total, we found 50 LEA genes in the genome. We also conducted analyses of the physicochemical parameters and subcellular location of the genes and generated a phylogenetic tree. LuLEA genes were unevenly mapped among 15 flax chromosomes and 90% of the genes had less than two introns. Expression profiles of LuLEA showed that most LuLEA genes were expressed at a late stage of seed development. Functionally, the LuLEA1 gene reduced seed size and fatty acid contents in LuLEA1 -overexpressed transgenic Arabidopsis lines. Conclusion Our study adds valuable knowledge about LEA genes in flax which can be used to improve related genes of seed development.

Seed development plays an important role during the life cycle of plants. Linseed flax is an oil crop and the seed is a key organ for fatty acids synthesis and storage. So it is important to understand the molecular mechanism of fatty acid biosynthesis during seed development. In this study, four small RNA libraries from early seeds at 5, 10, 20 and 30 days after flowering (DAF) were constructed and used for high-throughput sequencing to identify microRNAs (miRNAs). A total of 235 miRNAs including 114 known conserved miRNAs and 121 novel miRNAs were identified. The expression patterns of these miRNAs in the four libraries were investigated by bioinformatics and quantitative real-time polymerase chain reaction (qPCR) analysis. It was found that several miRNAs, including Lus-miRNA156a was significantly correlated with seed development process. In order to confirm the actual biological function of Lus-miRNA156a, over-expression vector was constructed and transformed to Arabidopsis. The phenotypes of homozygous transgenic lines showed decreasing of oil content and most of the fatty acid content in seeds as well as late flowering time. The results provided a clue that miRNA156a participating the fatty acid biosynthesis pathway and the detailed molecular mechanism of how it regulates the pathway needs to be further investigated.

Efficient regeneration of explants devoid of intrinsic somaclonal variations is a cardinal step in plant tissue culture, thus, a vital component of transgenic technology. However, recalcitrance of economically important crops to tissue culture-based organogenesis ensues a setback in the use of transgenesis in the genetic engineering of crop plants. The present study developed an optimized, genotype-independent, nonconventional tissue culture-independent in planta strategy for the genetic transformation of flax/linseed. This apical meristem-targeted in planta transformation protocol will accelerate value addition in the dual purpose industrially important but recalcitrant fiber crop flax/linseed. The study delineated optimization of Agrobacterium tumefaciens- mediated transformation and stable T-DNA (pCambia2301: GUS : nptII ) integration in flax. It established successful use of a stringent soilrite-based screening in the presence of 30 mg/L kanamycin for the identification of putative transformants. The amenability, authenticity, and reproducibility of soilrite-based kanamycin screening were further verified at the molecular level by GUS histochemical analysis of T 0 seedlings, GUS and nptII gene-specific PCR, genomic Southern hybridization for stable integration of T-DNA, and expression analysis of transgenes by sqRT-PCR. This method resulted in a screening efficiency of 6.05% in the presence of kanamycin, indicating amenability of in planta flax transformation. The strategy can be a promising tool for the successful development of transgenics in flax.

Flax (Linum usitatissimum L.) is a self-pollinating, annual, diploid crop grown for multi-utility purposes for its quality oil, shining bast fiber, and industrial solvent. Being a cool (Rabi) season crop, it is affected by unprecedented climatic changes such as high temperature, drought, and associated oxidative stress that, globally, impede its growth, production, and productivity. To precisely assess the imperative changes that are inflicted by drought and associated oxidative stress, gene expression profiling of predominant drought-responsive genes (AREB, DREB/CBF, and ARR) was carried out by qRT-PCR. Nevertheless, for normalization/quantification of data obtained from qRT-PCR results, a stable reference gene is mandatory. Here, we evaluated a panel of four reference genes (Actin, EF1a, ETIF5A, and UBQ) and assessed their suitability as stable reference genes for the normalization of gene expression data obtained during drought-induced oxidative stress in flax. Taking together, from the canonical expression of the proposed reference genes in three different genotypes, we report that EF1a as a stand-alone and EF1a and ETIF5A in tandem are suitable reference genes to be used for the real-time visualization of cellular impact of drought and oxidative stress on flax.

Morphological and molecular characterization of germplasm is important for the sustainable exploitation of crops. Linseed or flax (Linum usitatissimum) is a multipurpose crop grown in many environments for food, feed, fibre and industry. In Ethiopia, a centre of diversity for linseed, it is valued for food and export. Here, we aimed to develop and use a set of morphological descriptors to determine levels and patterns of diversity in Ethiopian germplasm from the tropical highlands (3–15°N, > 2,000 m a.s.l.) in 198 Ethiopian traditional varieties. The Ethiopian traditional varieties included plants with both fibre and oil-seed stem-branching morphotypes, although most were relatively small-seeded. Traditional variety oil quality was assessed; oil content was as low as 30 % compared to 47 % reported elsewhere. Days-to-flowering and days-to-maturity varied widely and were highly heritable. Ethiopian linseed had dominant and recessive yellow seed genotypes; some had a recessive conjoined or conjoined-seed character. The descriptors developed here will be useful for genetic mapping and selection of breeding lines. The results show the range of characters which can be exploited in breeding lines appropriate for smallholder and commercial farmers in Ethiopia, producing a sustainable, secure, high-value crop meeting agricultural, economic and cultural needs.

Linseed flax is a multipurpose crop. It is cultivated for its seeds and particularly for its oil. The main contributors for this crop are Canada, France and Belgium. In general, straws of linseed flax are buried in the fields or burnt. However, these solutions are not good practices for the environment and from an economical point of view. In this study, straws of linseed flax (six batches in total) with different dew retting durations and harvesting techniques were studied to possibly use them for producing innovative geotextiles. Two different fibre extraction processes were investigated. A first process (A) involved horizontal breaker rollers and then a breaking card. A second one (B) consisted in using vertical breaker rollers, and an “all fibre” extraction device (fibre opener) followed by sieving. The chemical composition of fibres in parietal constituents appeared to be globally equivalent to the one of textile flax with a pectic content decreasing as a function of the dew retting duration. This contributed to an increase in the cellulose content. The fibre content was situated in a range from 29% to 33%, which corresponds to a good yield for linseed flax fibre. The level of purity can reach values of up to 90% for method A (without extra-sieving) and 96% for method B (with extra-sieving), and the length of the fibres (larger for method A than for method B) and their tensile properties make them suitable for structural geotextile yarn manufacturing.

Soluble flaxseed gum (SFG) was extracted from flax (Linum usitatissimum) hulls using hot water, and its functional groups and antioxidant properties were investigated using infrared spectroscopy and different antioxidant assays (2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid (ABTS), reducing power capacity, and β-carotene bleaching inhibition assay), respectively. The antioxidant capacity of SFG showed interesting DPPH radical-scavenging capacity (IC50 SFG = 2.5 mg·mL−1), strong ABTS radical scavenging activity (% inhibition ABTS = 75.6% ± 2.6% at 40 mg·mL−1), high reducing power capacity (RPSFG = 5 mg·mL−1), and potent β-carotene bleaching inhibition activity (IC50 SFG = 10 mg·mL−1). All of the obtained results demonstrate the promising potential use of SFG in numerous industrial applications, and a way to valorize flaxseed hulls.

•Inflammatory bowel disease (IBD) refers to diseases that cause inflammation of the intestinal wall.•Flaxseed is a rich source of omega-3 fatty acids (α-linolenic acid; ALA), phytoestrogens and soluble fiber.•flaxseed and flaxseed oil, attenuate inflammatory markers, disease severity, blood pressure, and WC. The present study aimed to evaluate the possible effect of grounded flaxseed and flaxseed oil on serum levels of inflammatory markers, metabolic parameters, and the severity of disease in patients with UC. In this open-labeled randomized controlled trial, 90 UC patients were randomly assigned to one of the 3 groups for 12 weeks: grounded flaxseed (GF; 30 g/day), flaxseed oil (FO; 10 g/day) and control group. The weight, waist circumference, systolic and diastolic blood pressure, serum inflammatory markers (interleukin-6 (IL-6), interferon gamma (INF-γ), transforming growth factor beta (TGF-β), and Erythrocyte Sedimentation Rate (ESR)), and fecal calprotectin were measured at the baseline and end of the study. Totally, 75 patients (43 men and 32 women) with a mean age of 31.54 ± 9.84 years participated in the present study. Comparing the change of the variables indicated a significant decrease in fecal calprotectin (P < 0.001), Mayo score (P < 0.001), ESR (P < 0.001), INF-γ (P < 0.001), IL-6 (P < 0.001), waist circumference (P = 0.02), Diastolic Blood Pressure (DBP) (P < 0.001), and Systolic Blood Pressure (SBP) (P < 0.001) and a significant increase in TGF-β (P < 0.001) and Inflammatory Bowel Disease Questionnaire-Short form (IBDQ-9) score (P < 0.001) in the GF and FO groups compared to the control. No difference was obvious between the FO and GF groups except for TGF-β. The present study showed that both flaxseed and flaxseed oil, attenuate inflammatory markers, disease severity, blood pressure, and WC. However, the effect of flaxseed on weight and BMI was not evident.

This experiment studied the effect of 3 forms of presentation of linseed fatty acids (FA) on methane output using the sulfur hexafluoride tracer technique, total tract digestibility, and performance of dairy cows. Eight multiparous lactating Holstein cows (initial milk yield 23.4 +/- 2.2 kg/d) were assigned to 4 dietary treatments in a replicated 4 x 4 Latin square design: a control diet (C) consisting of corn silage (59%), grass hay (6%), and concentrate (35%) and the same diet with crude linseed (CLS), extruded linseed (ELS), or linseed oil (LSO) at the same FA level (5.7% of dietary DM). Each experimental period lasted 4 wk. All the forms of linseed FA significantly decreased daily CH4 emissions (P < 0.001) but to different extents (-12% with CLS, -38% with ELS, -64% with LSO) compared with C. The same ranking among diets was observed for CH4 output expressed as a percentage of energy intake (P < 0.001) or in grams per kilogram of OM intake (P < 0.001). Methane production per unit of digested NDF was similar for C, CLS, and ELS but was less for LSO (138 vs. 68 g/kg of digested NDF, respectively; P < 0.001). Measured as grams per kilogram of milk or fat-corrected milk yield, methane emission was similar for C and CLS and was less for ELS and LSO (P < 0.001), LSO being less than ELS (P < 0.01). Total tract NDF digestibility was significantly less (P < 0.001) for the 3 supplemented diets than for C (-6.8% on average; P < 0.001). Starch digestibility was similar for all diets (mean 93.5%). Compared with C, DMI was not modified with CLS (P > 0.05) but was decreased with ELS and LSO (-3.1 and -5.1 kg/d, respectively; P < 0.001). Milk yield and milk fat content were similar for LSO and ELS but less than for C and CLS (19.9 vs. 22.3 kg/d and 33.8 vs. 43.2 g/kg, on average, respectively; P < 0.01 and P < 0.001). Linseed FA offer a promising dietary means to depress ruminal methanogenesis. The form of presentation of linseed FA greatly influences methane output from dairy cows. The negative effects of linseed on milk production will need to be overcome if it is to be considered as a methane mitigation agent. Optimal conditions for the utilization of linseed FA in ruminant diets need to be determined before recommending its use for the dairy industry.

Flax ( Linum usitatissimum L.), one of the oldest cultivated crops, continues to be widely grown for oil, fiber and food. Furthermore, the plants show a metal tolerance dependent on species so is ideal for research. Present study was conducted to find out the influence of copper (Cu) toxicity on plant biomass, growth, chlorophyll content, malondialdehyde (MDA) contents, proline production, antioxidative enzymes and metal up taken by L. usitatissimum from the soil grown under mixing of Cu-contaminated soil with natural soil by 0:1 (control), 1:0, 1:1, 1:2 and 1:4. Results revealed that, high concentration of Cu in the soil affected plant growth and development by reducing plant height, plant diameter and plant fresh and dry biomass and chlorophyll contents in the leaves compared with the control. Furthermore, Cu in excess causes generation of reactive oxygen species (ROS) such as superoxide radical (O – ) and hydroxyl radicals (OH), which is manifested by high malondialdehyde (MDA) and proline contents also. The increasing activities of superoxidase dismutase (SOD) and peroxidase (POD) in the roots and leaves of L. usitatissimum are involved in the scavenging of ROS. Results also showed that L. usitatissimum also has capability to revoke large amount of Cu from the contaminated soil. As Cu concentration in the soil increases, the final uptake of Cu concentration by L. usitatissimum increases. Furthermore, the soil chemical parameters (pH, electrical conductivity and cation exchange capacity) were increasing to highest levels as the ratio of Cu concentration to the natural soil increases. Thus, Cu-contaminated soil is amended with the addition of natural soil significantly reduced plant growth and biomass, while L. usitatissimum is able to revoke large amount of Cu from the soil and could be grown as flaxseed and a potential candidate for phytoremediation of Cu.