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Abstract Flax occupies an important place in the world food supply as an oil and fiber crop of general and multi-purpose use. It should be noted that modern agriculture uses varieties of flax with high productivity and resistance to stress conditions. Modern varieties of flax respond well to soil fertility, are drought-resistant, technologically advanced and reliable, and are adapted in many countries of the world and are grown on all continents. According to FAO, flax crops annually in the world amount to more than 8 million hectares of flax. The main types of products obtained from growing flax are seeds and fiber. For many centuries, fiber flax was grown to obtain fiber, and oil flax to obtain seeds and oil. Flax growing is a specific branch of plant growing, in which the production process can be divided into two production cycles: the first is growing and harvesting flax, the second is processing of raw materials. Analysis of the results by stages of production is carried out taking into account the quality of flax products. Quality control of the resulting products is an important criterion for deciding on their further use in production for the manufacture of various goods for the population. High-quality flax fiber should contain 75-80% cellulose, no more than 4-5% lignin, low content of pectin substances. Flaxseed is of great importance in the global food industry as a functional food product. The value of flax seeds is determined by their high content of fats, polyunsaturated fatty acids and proteins. Modern technologies for growing agricultural crops involve the use of new complex fertilizers that can provide plants with essential nutrients - macro- and microelements - during the growing season. The aim of the research was to study the effect of two new complex fertilizers: Complex-Zn and Complex-Extra on the yield and quality of products of two varieties of fiber flax. Research methodology. The studies were conducted over 3 years in field conditions. The varieties of fiber flax Dobrynya and Pamyati Krepkova were studied in the experiments. Flax plants were treated with the studied complex fertilizers during the onset of the formation phase of the first pair of true leaves until the formation of 5-6 pairs of true leaves. The effect of the studied fertilizers on changes in morphological characteristics and plant growth indicators, as well as on yield, was studied. After harvesting, the content of lipids and protein in flax seeds was determined by near infrared spectroscopy. The content of cellulose, lignin, pectin substances and ash in flax fiber was determined. Main research results. In our studies, the use of complex fertilizers containing macro- and microelements contributed to the improvement of plant growth and development, increased yield and quality of flax products by the end of the growing season. Resumo O linho ocupa um lugar importante no suprimento mundial de alimentos como uma cultura de óleo e fibra de uso geral e multifuncional. Deve-se notar que a agricultura moderna utiliza variedades de linho com alta produtividade e resistência a condições de estresse. As variedades modernas de linho respondem bem à fertilidade do solo, são resistentes à seca, tecnologicamente avançadas e confiáveis, adaptando-se a muitos países e cultivadas em todos os continentes. De acordo com a FAO, as plantações de linho anualmente no mundo somam mais de 8 milhões de hectares de linho. Os principais produtos obtidos do cultivo do linho são sementes e fibras. Por muitos séculos, o linho de fibra foi cultivado para obtenção de fibras, e o linho oleaginoso era utilizado para produção de sementes e óleo. O cultivo do linho é um ramo específico da produção vegetal, cujo processo de produção pode ser dividido em dois ciclos: o primeiro é o cultivo e a colheita do linho, e o segundo é o processamento de matérias-primas. A análise dos resultados produtivos é realizada levando em consideração a qualidade dos produtos de linho. O controle de qualidade dos produtos resultantes é um critério importante para decidir sobre seu uso posterior na fabricação de vários bens para a população. A fibra de linho de alta qualidade deve conter 75-80% de celulose, não mais do que 4-5% de lignina, baixo teor de substâncias pécticas. A semente de linhaça é de grande importância na indústria alimentícia global como um produto alimentar funcional. O valor das sementes de linhaça é determinado por seu alto teor de gorduras, ácidos graxos poli-insaturados e proteínas. As tecnologias modernas para o cultivo agrícola envolvem o uso de novos fertilizantes complexos que podem fornecer às plantas nutrientes essenciais – macro e microelementos – durante a estação de crescimento. O objetivo da pesquisa foi estudar o efeito de dois novos fertilizantes complexos: Complex-Zn e Complex-Extra no rendimento e na qualidade dos produtos de duas variedades de fibra de linho. Metodologia de pesquisa. Os estudos foram conduzidos ao longo de três anos em condições de campo. As variedades de linho fibroso Dobrynya e Pamyati Krepkova foram estudadas nos experimentos. As plantas de linho foram tratadas com os fertilizantes complexos estudados durante o início da fase de formação do primeiro par de folhas verdadeiras, até a formação de 5-6 pares de folhas verdadeiras. Foi estudado o efeito dos fertilizantes estudados nas mudanças nas características morfológicas e indicadores de crescimento da planta, bem como no rendimento. Após a colheita, o conteúdo de lipídios e proteínas nas sementes de linho foi determinado por espectroscopia no infravermelho próximo. O conteúdo de celulose, lignina, substâncias de pectina e cinzas na fibra de linho foi analisado. Principais resultados da pesquisa. Em nossos estudos, o uso de fertilizantes complexos contendo macro e microelementos contribuiu para a melhoria do crescimento e desenvolvimento das plantas, aumentou o rendimento e a qualidade dos produtos de linho até o final da estação de crescimento.

Seed coat color, a trait exhibiting significant phenotypic variation, has been reported in previous studies to influence key germination parameters, including germination percentage, germination rate, and mean germination time. Looking into these relationships can help us identify plant varieties that better withstand both living threats and environmental challenges, which would make breeding programs more effective and focused. The objectives of this study were to evaluate the drought tolerance of 20 flax varieties at the germination stage, investigate the effects of gamma-aminobutyric acid (GABA) on seed germination performance under drought stress conditions, and explore potential associations between seed coat color and germination parameters under both stressed and non-stressed conditions using digital image processing. The final germination percentage, germination speed, and span of germination were assessed for seeds germinated under various germination conditions, including 23% PEG (− 0.169 MPa), 10 mM GABA, 23% PEG + 10 mM GABA, and a control treatment at 25 ± 0.5 °C. The results demonstrated substantial genetic variation across all germination parameters measured in the seeds of 20 distinct flax varieties. Genotype-specific responses to drought stress were observed, with 10 mM GABA alleviating the effects of drought. Among the varieties tested, Hermes exhibited the highest drought tolerance, while Lisette and Bonny-Doon were identified as drought-sensitive. Digital image processing analysis revealed significant differences ( p  < 0.05) in drought tolerance levels among flax varieties based on variations in RGB values of their seed coats. Although no statistically significant correlations were found in direct pairwise correlations between the color parameters converted from RGB (Red, Green, and Blue) to L*a*b* and germination parameters, multivariate PCA-biplot analysis indicated that L* and b* values had a positive influence on germination performance. In contrast, seeds with higher a* values exhibited reduced germination performance. Furthermore, the biplot analysis suggested that varieties with lighter seed coats tended to show better germination compared to those with darker seed coats. These findings highlight the potential of incorporating seed coat color parameters into flax breeding programs, suggesting their role in enhancing seed germination and overall seed quality under various stress conditions.

Background Linseed is an important oilseed crop with diverse applications in the food, nutraceutical, oil, and paint industries. Flowering time is a critical trait in linseed, as it greatly influences seed yield potential and quality across various agro-ecological zones. Trehalose-6-phosphate synthase (TPS) genes have been implicated in the regulation of flowering time in plants. Therefore, a comprehensive analysis of the TPS gene family in linseed, using comparative genomics and bioinformatics approaches, is essential for elucidating the genetic mechanisms underlying flowering time regulation in this crop. Results A total of 18 LuTPS genes, including several paralogs, were identified in the linseed genome and clustered into two distinct groups. Gene expression analysis in developing floral buds, flower, and vegetative tissues revealed that most TPS genes exhibited basal expression levels. However, LuTPS6.1, LuTPS6.2, LuTPS10.1, LuTPS1.1, LuTPS1.2, LuTPS7.2, LuTPS7.3, LuTPS7.4 , and LuTPS8. 2 showed significantly higher expression and strong correlation with key flowering-related genes such as FT, FUL, and SOC1 . Allelic variation analysis using early- and late-flowering linseed accessions revealed trait-specific SNPs in LuTPS6.1 and LuTPS10.2 . A comprehensive analysis of cis-regulatory elements (CREs ) in the promoter regions of LuTPS genes, compared to the entire linseed genome, identified several CREs that were significantly enriched in TPS gene promoters, as well as those that were consistently present across all LuTPS gene promoters. Furthermore, the genome-wide syntenic network analysis involving linseed and nine other plant genomes provided valuable insights into TPS-associated syntelogs and the evolutionary dynamics and expansion of the TPS gene family. The physical proximity of TPS genes to known flowering time QTLs/QTNs is also discussed. Conclusion This study, with TPS gene family characterization, gene expression and allelic variation highlights the potential role of the TPS genes in the regulation of flowering time in linseed. The identified enriched CREs in the promoters of TPS genes would be crucial to understand the regulation of TPS genes in growth, development and stress response. TPS-associated syntelogs provided valuable insights into the evolutionary history and expansion of the TPS gene family. Collectively, these findings represent a significant step toward understanding the complex genetic regulation of flowering time in linseed.

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.

Flaxseeds are a functional food representing, by far, the richest natural grain source of lignans, and accumulate substantial amounts of other health beneficial phenolic compounds (i.e., flavonols, hydroxycinnamic acids). This specific accumulation pattern is related to their numerous beneficial effects on human health. However, to date, little data is available concerning the relative impact of genetic and geographic parameters on the phytochemical yield and composition. Here, the major influence of the cultivar over geographic parameters on the flaxseed phytochemical accumulation yield and composition is evidenced. The importance of genetic parameters on the lignan accumulation was further confirmed by gene expression analysis monitored by RT-qPCR. The corresponding antioxidant activity of these flaxseed extracts was evaluated, both in vitro, using ferric reducing antioxidant power (FRAP), oxygen radical absorbance capacity (ORAC), and iron chelating assays, as well as in vivo, by monitoring the impact of UV-induced oxidative stress on the lipid membrane peroxidation of yeast cells. Our results, both the in vitro and in vivo studies, confirm that flaxseed extracts are an effective protector against oxidative stress. The results point out that secoisolariciresinol diglucoside, caffeic acid glucoside, and p-coumaric acid glucoside are the main contributors to the antioxidant capacity. Considering the health benefits of these compounds, the present study demonstrates that the flaxseed cultivar type could greatly influence the phytochemical intakes and, therefore, the associated biological activities. We recommend that this crucial parameter be considered in epidemiological studies dealing with flaxseeds.

Background Cinnamoyl-CoA reductase (CCR) is the first important and committed enzyme in the monolignol synthesis branch of the lignin biosynthesis (LB) pathway, catalyzing the conversion of cinnamoyl-CoAs to cinnamaldehydes and is crucial for the growth of Linum usitatissimum (flax), an important fiber crop. However, little information is available about CCR in flax ( Linum usitatissimum L.). Results In this study, we conducted a genome-wide analysis of the CCR gene family and identified a total of 22 CCR genes. The 22 CCR genes were distributed across 9 chromosomes, designated LuCCR1 - LuCCR22 . Multiple sequence alignment and conserved motif analyses revealed that LuCCR7/13/15/20 harbor completely conserved NADP-specific, NAD(P)-binding, and CCR signature motifs. Furthermore, each of these LuCCRs is encoded by 5 exons separated by 4 introns, a characteristic feature of functional CCRs . Phylogenetic analysis grouped LuCCRs into two clades, with LuCCR7/13/15/20 clustering with functional CCRs involved in LB in dicotyledonous plants. RNA-seq analysis indicated that LuCCR13/20 genes are highly expressed throughout all flax developmental stages, particularly in lignified tissues such as roots and stems, with increased expression during stem maturation. These findings suggest that LuCCR13/20 play crucial roles in the biosynthesis process of flax lignin. Additionally, LuCCR2/5/10/18 were upregulated under various types of abiotic stress, highlighting their potential roles in flax defense-related processes. Conclusions This study systematically analyzes the CCR gene family (CCRGF) of flax ( Linum usitatissimum L.) at the genomic level for the first time, so as to select the whole members of the CCRGF of flax and to ascertain their potential roles in lignin synthesis. Therefore, in future work, we can target genetic modification of LuCCR13/20 to optimize the content of flax lignin. As such, this research establishes a theoretical foundation for studying LuCCR gene functions and offers a new perspective for cultivating low-lignin flax varieties.

A two-level factorial design in Design-Expert ® software was applied to statistically model and optimize the effects of key endogenous enzymes on the release of total phenolic content (TPC) and total flavonoid content (TFC) during flaxseed germination. The model demonstrated high predictive accuracy (R² > 0.99) and identified germination day (GD), β-glucosidase (β-GL), peroxidase (POX), and catalase (CAT) as critical variables with significant synergistic interactions influencing TPC and TFC biosynthesis. Under these optimal factors, five-day germinated sprouts showed the highest levels of bioactive compounds, with TPC and TFC increasing by 7.6-fold and 38.27-fold, respectively. Compared to dry seeds, High-performance liquid chromatography (HPLC) analysis confirmed marked increases in sinapic acid (6.4-fold), gallic acid (6.1-fold), and p-coumaric acid (5.5-fold). Antioxidant activity also improved, as evidenced by reduced IC 50 values for DPPH (2.26-fold) and ABTS (2.6-fold) assays. Enzyme activity analysis revealed early-stage activation of polyphenol oxidase (PPO) and phenylalanine ammonia-lyase (PAL), supporting the enzymatic role in phenolic compound biosynthesis. Additionally, 5-day sprout extracts exhibited notable antibacterial and antidiabetic activities. These findings provide a robust, model-guided framework for enhancing flaxseed’s nutritional and functional value through controlled germination, with direct applications in the development of health-promoting functional foods.

The aim of the present study was to evaluate the morphogenetic efficiency of the new flax (Linum usitatissimum L.) cultivar Silesia. The plant material consisted of five ecotypes of Silesia seeds selected by determining the fatty acid composition in individual plants. Thus, five ecotypes with different fatty acid compositions were applied for germination, and the resulting seedlings were used for callus induction. The observed in vitro morphogenic response of explants to the used callus induction medium was 100%, followed by varying development of shoots, with the highest value being 17 shoots per callus, with an average length of 7.15 cm (for ecotype 1). The biochemical analyses showed photosynthetic pigments were significantly affected by the tested ecotypes. The main conclusion is that the selection of plant material as a source of tissue cultures is crucial for efficient regeneration and organogenesis. One factor potentially influencing these processes is fatty acid composition and the ratio of linoleic to α-linolenic acid. A ratio ranging from 1.36 to 1.68 in the source material (seeds) used for initiation of tissue cultures resulted in the highest efficiency of shoot regeneration and number of obtained shoots per callus. A strong negative correlation (−0.78) was observed between shoot regeneration efficiency and the ratio of linoleic to α-linolenic acid in seeds from which the explants were obtained. Moreover, an efficient protocol of micropropagation from callus tissue was established for the new flax cultivar Silesia. An interesting insight into the metabolism of the obtained regenerants allowed us to determine the relationship between the content of chlorophyll and rhizogenesis efficiency. For these two parameters, the calculated correlation coefficient was 0.66. A strong relationship (high correlation coefficient: 0.79) was also established between flavonoid content and length of obtained regenerants, pointing to the developmental role of flavonoids.

Lignans are the main secondary metabolites synthetized by Linum species as plant defense compounds but they are also valuable for human health, in particular, for novel therapeutics. In this work, Linum austriacum in vitro cultures, cells (Cc), adventitious roots (ARc) and hairy roots (HRc) were developed for the production of justicidin B through elicitation with methyl jasmonate (MeJA) and coronatine (COR). The performances of the cultures were evaluated for their stability, total phenols content and antioxidant ability. NMR was used to identify justicidin B and isojusticidin B and HPLC to quantify the production, highlighting ARc and HRc as the highest productive tissues. MeJA and COR treatments induced the synthesis of justicidin B more than three times and the synthesis of other compounds. RNA-sequencing and a de novo assembly of L. austriacum ARc transcriptome was generated to identify the genes activated by MeJA. Furthermore, for the first time, the intracellular localization of justicidin B in ARc was investigated through microscopic analysis. Then, HRc was chosen for small-scale production in a bioreactor. Altogether, our results improve knowledge on justicidin B pathway and cellular localization in L. austriacum for future scale-up processes.

While nanoparticle uptake by plants has primarily been studied for environmental remediation, its purposeful use to enhance plant structural and mechanical properties remains unexplored. We hypothesized that cellulose nanofibers (CNFs), introduced into the growth medium, could be absorbed by flax stems, reinforcing their cell walls formation and, as a consequence, improve the mechanical performance. Thus, in this paper it is proved that flax plants treated with a 0.2% w/v CNF solution after root excision showed increased stem diameter, reduced pith size, and significantly accelerated root regeneration (~ 7 cm in 20 days) compared to controls treated with autoclaved deionized water. Analyses of the CNF-treated stems showed an increase in the mechanical performance of the stems revealing up to 50% increase in energy to fracture, 22% increase in Young’s modulus, and approximate 33% improvement in stiffness at a reduced density compared to the non-treated stems. To explain these effects, morphology analysis of stems was conducted. Fourier-transform infrared spectroscopy (FTIR) and differential scanning calorimetry (DSC) revealed enhanced hydrogen bonding, cellulose crystallinity, and thermal stability. This study developed a novel in vivo approach by incorporating CNFs into plants during early growth to trigger structural reinforcement, which in turn improves mechanical performance.