Eco-friendly soil amendments improve growth, antioxidant activities, and root colonization in lingrain (Linum Usitatissimum L.) under drought conditions.

This study's primary purpose was to investigate the possible amelioration of limited irrigation conditions by mycorrhiza (AMF), vermicompost, and green manure for lingrain plants. This experiment was accomplished as a factorial based on the completely randomized design with three replications. The first factor was green manure (without green manure and with Trifolium pratense as green manure); the second factor consisted of Rhizophagus irregularis mycorrhiza, vermicompost, a combination of mycorrhiza and vermicompost and none of them, and also the third factor was irrigation regime (full irrigation and late-season water limitation). Green manure, vermicompost, and mycorrhiza single-use enhanced the plant's underwater limitation conditions compared to the control. However, vermicompost and green manure or mycorrhiza developed a positive synergistic effect on most traits. Combining green manure with the dual fertilizer (mycorrhiza + vermicompost) resulted in the vermicompost and mycorrhiza synergistic effects, especially under limited irrigation. Consequently, the combination of green manure, mycorrhiza, and vermicompost experienced the highest amount of leaf relative water content, root colonization, leaf nitrogen, chlorophyll a, chlorophyll b, carotenoids, antioxidant enzymes activity, grain yield, and oil yield, which would lead to more resistance of plants to limited irrigation conditions.

Salicylic acid affects mycorrhizal features, antioxidant enzyme activities and seed yield of linseed under water-deficit stress in open-field conditions.

The research aims were to study salicylic acid (SA) effects on mycorrhiza [hyphal width (HW), vesicle diameter (VD) and mycorrhizal colonization (MC)] and interaction between them on greenness index (GI), drought tolerance index (DTI), antioxidant enzymes activities, and seed yield of linseed under drought. A factorial experiment was conducted in an open-field place with mycorrhiza [non-inoculation, Funneliformis mosseae (FM), and Rhizoglomus intraradices (RI)], SA (250 µM and non-SA), and irrigation levels [100%, 70%, and 40% field capacity (FC)] as treatments. Severe drought increased VD, MC, superoxide dismutase (SOD), ascorbate peroxidase (APX), and peroxidase activities while decreased GI, DTI, and yield. The RI-linseed had higher MC, GI, SOD, and glutathione reductase (GR) activities, but FM-linseed had greater VD and yield under drought. Inoculated linseed with both mycorrhiza showed a reduction in DTI and yield under SA than non-SA. In RI-linseed, SA increased GI, MC, HW, VD, catalase and GR, but decreased in FM-plants. Mycorrhiza (particularly RI) alleviated drought (40% FC)-caused negative effects on linseed via the improvement of SOD, APX, and GI. Regardless of other treatments, SA had negative effects on HW and VD, but SA effects varied depending on mycorrhizal species so that SA increased HW, VD, and MC in RI. Due to the positive correlation between MC and HW, SA reduces FM colonization by reducing the HW of FM. Totally, SA along with RI species can mitigate the harmful effects of drought and improve tolerance via increasing MC, HW, VD, catalase, peroxidase, and GR activities.

Identification of two compounds able to improve flax resistance towards Fusarium oxysporum infection.

Plants are surrounded by a diverse range of microorganisms that causes serious crop losses and requires the use of pesticides. Flax is a major crop in Normandy used for its fibres and is regularly challenged by the pathogenic fungus Fusarium oxysporum (Fo) f. sp. lini. To protect themselves, plants use "innate immunity" as a first line of defense level against pathogens. Activation of plant defense with elicitors could be an alternative for crop plant protection. A previous work was conducted by screening a chemical library and led to the identification of compounds able to activate defense responses in Arabidopsis thaliana. Four compounds were tested for their abilities to improve resistance of two flax varieties against Fo. Two of them, one natural (holaphyllamine or HPA) and one synthetic (M4), neither affected flax nor Fo growth. HPA and M4 induced oxidative burst and callose deposition. Furthermore, HPA and M4 caused changes in the expression patterns of defense-related genes coding a glucanase and a chitinase-like. Finally, plants pre-treated with HPA or M4 exhibited a significant decrease in the disease symptoms. Together, these findings demonstrate that HPA and M4 are able to activate defense responses in flax and improve its resistance against Fo infection.

Effects of Biogenic Zinc Oxide Nanoparticles on Growth and Oxidative Stress Response in Flax Seedlings vs. In Vitro Cultures: A Comparative Analysis.

Linum usitatissimum biosynthesizes lignans and neolignans that are diet and medicinally valuable metabolites. In recent years, zinc oxide nanoparticles (ZnONPs) have emerged as potential elicitors for the enhanced biosynthesis of commercial secondary metabolites. Herein, we investigated the influence of biogenic ZnONPs on both seedlings and stem-derived callus of L. usitatissimum. Seedlings of L. usitatissimum grown on Murashige and Skoog (MS) medium supplemented with ZnONPs (1-1000 mg/L) presented the highest antioxidant activity, total phenolic content, total flavonoid content, peroxidase and superoxide dismutase activities at 500 mg/L, while the maximum plantlet length was achieved with 10 mg/L. Likewise, the high-performance liquid chromatography (HPLC) analysis revealed the enhanced production of secoisolariciresinol diglucoside, lariciresinol diglucoside, dehydrodiconiferyl alcohol glucoside and guaiacylglycerol-ß-coniferyl alcohol ether glucoside in the plantlets grown on the 500 mg/L ZnONPs. On the other hand, the stem explants were cultured on MS media comprising 1-naphthaleneacetic acid (1 mg/L) and ZnONPs (1-50 mg/L). The highest antioxidant and other activities with an enhanced rooting effect were noted in 25 mg/L ZnONP-treated callus. Similarly, the maximum metabolites were also accumulated in 25 mg/L ZnONP-treated callus. In both systems, the dose-dependent production of reactive oxygen species (ROS) was recorded, resulting in oxidative damage with a more pronounced toxic effect on in vitro cultures. Altogether, the results from this study constitute a first comprehensive view of the impact of ZnONPs on the oxidative stress and antioxidant responses in seedlings vs. in vitro cultures.

3-Hydroxybutyrate Is Active Compound in Flax That Upregulates Genes Involved in DNA Methylation.

In mammalian cells, 3-hydroxybutyrate (3-HB) is not only an intermediate metabolite during the oxidation of fatty acids, but also an important signaling molecule. On the other hand, the information about the metabolism or function of this compound in plants is scarce. In our study, we show for the first time that this compound naturally occurs in flax. The expression of bacterial ß-ketothiolase in flax affects expression of endogenous genes of the 3-HB biosynthesis pathway and the compound content. The increase in 3-HB content in transgenic plants or after control plants treatment with 3-HB resulted in upregulation of genes involved in chromatin remodeling. The observation that 3-HB is an endogenous activator of methyltransferase 3 (CMT3), decreased DNA methylation I (DDM1), DEMETER DNA glycosylase (DME), and an inhibitor of sirtuin 1 (SRT1) provides an example of integration of different genes in chromatin remodeling. The changes in chromatin remodeling gene expression concomitant with those involved in phenolics and the lignin biosynthesis pathway suggest potential integration of secondary metabolic status with epigenetic changes.

Photoperiod sensitivity of Canadian flax cultivars and 5-azacytidine treated early flowering derivative lines.

BACKGROUND: Early flowering and maturing flax (Linum usitatissimum L.) cultivars are better adapted than lines with a longer reproductive phase for the short growing season of the northern Canadian Prairies. We examined the role of long days (LD) and short days (SD) on the time taken to flower in five established flax cultivars and three mutant-derived F10 lines. The photoperiod sensitivity of these eight different genotypes was determined using a reciprocal transfer experiment involving weekly transfers between LD and SD environments. RESULTS: The genotypes tested had varying degrees of photoperiod sensitivity and demonstrated reduced time to flowering if exposed to LD environments prior to a critical time point. The duration of each of the three phases of vegetative growth differed among the genotypes studied. Transfers from SD to LD shortened the vegetative stage, reduced time to flowering, and extended the reproductive phase in the genotypes studied. Mutant-derived lines RE1/2/3 flowered significantly earlier compared to CDC Sorrel, CDC Bethune, Flanders, Prairie Thunder, and Royal. Modelling of the flowering times indicated that transferring the cultivars from SD to LD increased the photoperiod sensitive time; however, different reproductive phases for mutant lines were not defined as parsimonious models were not identified. Expression of the putative flax homologs for CONSTANS (CO), FLOWERING LOCUS T (FT), and GIGANTEA (GI) was examined in the leaves of Royal and RE1/2/3 plants at 10, 15, 19 and 29 days after planting. Expression of putative FT homologs was detected in all three early-flowering lines but expression was negligible, or not detected, in Royal. CONCLUSIONS: Models defining the three phases of reproductive development were established for the five cultivars studied; however, it was not possible to identify these phases for the three early flowering and photoperiod insensitive epimutant-derived lines. A putative flax homolog of FT, a key regulator of flowering time, is more highly expressed in RE plants, which may condition the day-length insensitivity in the early flowering 'epimutant' lines.

Biogenic zinc oxide nanoparticles-enhanced biosynthesis of lignans and neolignans in cell suspension cultures of Linum usitatissimum L.

Zinc oxide nanoparticles (NPs) have emerged as a novel elicitor for enhanced biosynthesis of secondary metabolites in in vitro plant cell cultures. The current study was aimed to explore elicitation abilities of ZnO-NPs for enhanced accumulation of lignans and neolignans in cell cultures of Linum usitatissimum. We optimized concentration of zinc oxide NPs before carrying out a full-fledged experiment. Subsequently, an optimum dose of 100 mg/l was introduced into the culture medium on day 0, days 0 and 15, and finally days 0 and 25. We observed that repeated elicitation stimulated various parameters and physiological responses in Linum usitatissimum cell cultures than one-time elicitation. Repeated elicitation of cell cultures on day 0 and 15 resulted in highest fresh weight (412.16 g/l) and lignans production (secoisolariciresinol diglucoside 284.12 mg/l: lariciresinol diglucoside 86.97 mg/l). Contrarily, repeated elicitation on day 0 and 25 resulted in highest DW (13.53 g/l), total phenolic production (537.44 mg/l), total flavonoid production (123.83 mg/l) and neolignans production (dehydrodiconiferyl alcohol glucoside 493.28 mg/l: guaiacylglycerol-ß-coniferyl alcohol ether glucoside 307.69 mg/l). Enhancement in plant growth and secondary metabolites accumulation was several fold higher than controls. Furthermore, a linear relationship existed between total phenolic and flavonoid contents which in turn was correlated with higher antioxidant activities.

24-Epibrassinolide alters DNA cytosine methylation of Linum usitatissimum L. under salinity stress.

Salinity is a common environmental challenge limiting worldwide agricultural crop yield. Plants employ epigenetic regulatory strategies, such as DNA methylation which relatively allows rapid adaptation to new conditions in response to environmental stresses. Brassinosteroids (BRs) are a novel group of phytohormones recognized as transcription and translation regulators which are able to mitigate the impact of environmental stresses on the plants. In the current investigation, the influence of salinity and 24-epibrassinolide (24-epiBL) was investigated on the extent and pattern of cytosine DNA methylation using methylation-sensitive amplified polymorphisms (MSAP) technique in flax. Upon NaCl (150 mM) exposure, total methylation of CCGG sequences was decreased in comparison to control plants, while 24-epiBL (10-8 M) induced total methylation under salinity stress. Sequencing and analysis of six randomly selected MSAP fragments detected genes involved in various biological and molecular processes such as vitamine B1 biosynthesis, protein targeting and localization, post-translational modification and gene regulation. In conclusion, 24-epiBL seed priming could play critical role in regulation of cellular and biological processes in response to salt stress by epigenetic modification and induction of methylation.

Serratia sp. CP-13 augments the growth of cadmium (Cd)-stressed Linum usitatissimum L. by limited Cd uptake, enhanced nutrient acquisition and antioxidative potential.

AIMS: The current study was aimed to evaluate the beneficial effects and bioremediation potential of a Cd-tolerant bacterial strain, Serratia sp. CP-13, on the physiological and biochemical functions of Linum usitatissimum L., under Cd stress. METHODS AND RESULTS: The bacterial strain was isolated from the wastewater collection point of Chakera, Faisalabad, Pakistan, as this place contains industrial wastewater of the Faisalabad region. The Serratia sp. CP-13, identified through 16S rRNA gene sequence analysis, exhibited a significant phyto-beneficial potential in terms of in vitro inorganic phosphate solubilization, indole-3-acetic acid production and 1-aminocyclopropane-1-carboxylic acid deaminase activity. Effects of Serratia sp. CP-13 inoculation on L. usitatissimum were evaluated by growing the plants in CdCl2 (0, 5 or 10 mg kg-1 dry soil)-spiked soil. Without inoculation of Serratia sp. CP-13, Cd stress significantly reduced the plant biomass as well as the quantity of proteins and photosynthetic pigments due to enhanced H2 O2 , malondialdehyde (MDA) contents and impaired nutrient homeostasis. Subsequently, Serratia sp. CP-13 increased the plant fresh and dry biomass, plant antioxidation capacity, whereas it decreased the lipid peroxidation under Cd stress. In parallel, Serratia sp. inoculation assisted the Cd-stressed plants to maintain an optimum level of nutrients (K, Ca, P, Mg, Fe and Mn). CONCLUSIONS: The isolated bacterial strain (Serratia sp. CP-13) when applied to Cd-stressed L. usitatissimum inhibited the Cd uptake, reduced Cd-induced lipid peroxidation, maintained the optimum level of nutrients and thereby, enhanced L. usitatissimum growth. The analysis of bio-concentration and translocation factor revealed that L. usitatissimum with Serratia sp. CP-13 inoculation sequestered Cd in plant rhizospheric zone. SIGNIFICANCE AND IMPACT OF THE STUDY: Serratia sp. CP-13 inoculation is a potential candidate for the development of low Cd-accumulating linseed and could be used for phytostabilization of Cd-contaminated rhizosphere/soil colloids.

Hydrogen sulfide directs metabolic flux towards the lignan biosynthesis in Linum album hairy roots.

Hydrogen sulfide (H2S) has been recently found as an important signaling molecule especially in root system architecture of plants. The regulation of root formation through H2S has been reported in previous works; while the profiling of metabolites in response to H2S is not clearly discussed. To this end, different concentrations of sodium hydrosulfide (an H2S donor) were applied to the culture of Linum album hairy roots. Subsequently, the amino acid profiles, soluble carbohydrates, and central intermediates of phenylpropanoid pathway with two branches of lignans and flavonoids were assessed by spectroscopy and high performance liquid chromatography techniques. An analysis of the signaling molecules (nitric oxide, hydrogen peroxide, and salicylic acid) was also conducted as they proposed to act in conjunction with H2S. The H2S activated antioxidant systems and caused a shift from flavonoid to lignan production (podophyllotoxin and 6-methoxypodophyllotoxin); although, some of the flavonoids increased in a dose-dependent manner. The H2S decreased the contents of phenylalanine and tyrosine as substrates of the phenylpropanoid pathway, but increased proline and histidine as an osmolyte and antioxidant, respectively. These findings propose that H2S modulates other signaling molecules, regulates free amino acids, and mediates biosynthesis of lignans and flavonoids in the phenylpropanoids biosynthesis pathway.

Early plant growth and bacterial community in rhizoplane of wheat and flax exposed to silver and titanium dioxide nanoparticles.

Silver and titanium dioxide nanoparticles (AgNPs and TiO2NPs) are highly useful, but they are also a significant reason for concern as they exert toxicity. The goal of research was to assess the role of three kinds of NPs in concentrations of 100 mg L-1 on early growth plants (wheat, flax) and bacterial community in rhizoplane. Titanium (IV) oxide anatase (TiO2NPs1) and titanium (IV) oxide nanopowder (TiO2NPs2) are commercial products. A suspension of AgNPs was prepared via a procedure of reduction with tannic acid. The response of Monocot and Dicot growth form plants to the tested NPs was different. Germination and seedling growth of wheat treated with TiO2NPs1 was better. The response of flax to NPs was noted as an increase of chlorophyll content. The bacterial community in wheat rhizoplane was not significantly modified, but there was a declining trend. In turn, a difference in the surface charge of NPs had an influence on the total bacterial community in Dicot rhizoplane. Positively charged TiO2NPs2 significantly decreased the quantity of total bacteria in contrast to negatively charged AgNPs and TiO2NPs1 which increased it. A qualitative analysis did not confirm the influence of the surface charge of NPs on an increase/decrease in the quantity of Pseudomonas and Bacillus bacteria, but did show that there was no toxicity of the tested NPs to the plant growth-promoting bacteria community. The rhizoplane microbiome was dependent on the species of plant, and the bacteria found in the communities are sensitive to NPs to a varying degree.

[CAX3 Gene is Involved in Flax Response to High Soil Acidity and Aluminum Exposure].

Understanding the molecular mechanisms of plant response to unfavorable conditions is necessary for the effective selection of tolerant genotypes. Earlier, using high-throughput transcriptome sequencing of flax plants after exposure to aluminum ions (Al^(3+)) and high soil acidity, we detected stress-induced alteration in the expression of several genes, including CAX3, which encodes Ca^(2+)/H^(+)-exchanger involved in calcium ion transport. Here we describe CAX3 mRNA levels in flax cultivars either tolerant (Hermes and TMP1919) or sensitive (Lira and Orshanskiy) to Al^(3+). Stress-induced increased expression of CAX3 was detected only in aluminum-tolerant flax cultivars. The product of CAX3 gene may participate in flax response to high soil acidity and high Al^(3+) concentration through Ca^(2+)-mediated intracellular regulation.

Effect of 1-aminocyclopropane-1-carboxylic acid (ACC)-induced ethylene on cellulose synthase A (CesA) genes in flax (Linum usitatissimum L. 'Nike') seedlings.

INTRODUCTION: Cellulose microfibril is a major cell wall polymer that plays an important role in the growth and development of plants. The gene cellulose synthase A (CesA), encoding cellulose synthases, is involved in the synthesis of cellulose microfibrils. However, the regulatory mechanism of CesA gene expression is not well understood, especially during the early developmental stages. OBJECTIVE: To identify factor(s) that regulate the expression of CesA genes and ultimately control seedling growth and development. METHODS: The presence of cis-elements in the promoter region of the eight CesA genes identified in flax (Linum usitatissimum L. 'Nike') seedlings was verified, and three kinds of ethylene-responsive cis-elements were identified in the promoters. Therefore, the effect of ethylene on the expression of four selected CesA genes classified into Clades 1 and 6 after treatment with 10-4 and 10-3 M 1-aminocyclopropane-1-carboxylic acid (ACC) was examined in the hypocotyl of 4-6-day-old flax seedlings. RESULTS: ACC-induced ethylene either up- or down-regulated the expression of the CesA genes depending on the clade to which these genes belonged, age of seedlings, part of the hypocotyl, and concentration of ACC. CONCLUSION: Ethylene might be one of the factors regulating the expression of CesA genes in flax seedlings.

Investigation of Linum flavum (L.) Hairy Root Cultures for the Production of Anticancer Aryltetralin Lignans.

Linum flavum hairy root lines were established from hypocotyl pieces using Agrobacterium rhizogenes strains LBA 9402 and ATCC 15834. Both strains were effective for transformation but induction of hairy root phenotype was more stable with strain ATCC 15834. Whereas similar accumulation patterns were observed in podophyllotoxin-related compounds (6-methoxy-podophyllotoxin, podophyllotoxin and deoxypodophyllotoxin), significant quantitative variations were noted between root lines. The influence of culture medium and various treatments (hormone, elicitation and precursor feeding) were evaluated. The highest accumulation was obtained in Gamborg B5 medium. Treatment with methyl jasmonate, and feeding using ferulic acid increased the accumulation of aryltetralin lignans. These results point to the use of hairy root culture lines of Linum flavum as potential sources for these valuable metabolites as an alternative, or as a complement to Podophyllum collected from wild stands.

Genome-wide identification, characterization, and expression profile of aquaporin gene family in flax (Linum usitatissimum).

Membrane intrinsic proteins (MIPs) form transmembrane channels and facilitate transport of myriad substrates across the cell membrane in many organisms. Majority of plant MIPs have water transporting ability and are commonly referred as aquaporins (AQPs). In the present study, we identified aquaporin coding genes in flax by genome-wide analysis, their structure, function and expression pattern by pan-genome exploration. Cross-genera phylogenetic analysis with known aquaporins from rice, arabidopsis, and poplar showed five subgroups of flax aquaporins representing 16 plasma membrane intrinsic proteins (PIPs), 17 tonoplast intrinsic proteins (TIPs), 13 NOD26-like intrinsic proteins (NIPs), 2 small basic intrinsic proteins (SIPs), and 3 uncharacterized intrinsic proteins (XIPs). Amongst aquaporins, PIPs contained hydrophilic aromatic arginine (ar/R) selective filter but TIP, NIP, SIP and XIP subfamilies mostly contained hydrophobic ar/R selective filter. Analysis of RNA-seq and microarray data revealed high expression of PIPs in multiple tissues, low expression of NIPs, and seed specific expression of TIP3 in flax. Exploration of aquaporin homologs in three closely related Linum species bienne, grandiflorum and leonii revealed presence of 49, 39 and 19 AQPs, respectively. The genome-wide identification of aquaporins, first in flax, provides insight to elucidate their physiological and developmental roles in flax.

The use of phosphomannose isomerase selection system for Agrobacterium-mediated transformation of tobacco and flax aimed for phytoremediation.

A plant selection system based on the phosphomannose isomerase gene (pmi) as a selectable marker is often used to avoid selection using antibiotic resistance. Nevertheless, pmi gene is endogenous in several plant species and therefore difficult to use in such cases. Here we evaluated and compared Agrobacterium-mediated transformation of Linum usitatissimum breeding line AGT-952 (without endogenous pmi gene) and Nicotiana tabacum var. WSC-38 (with endogenous pmi gene). Transformation was evaluated for vectors bearing transgenes that have the potential to be involved in improved phytoremediation of contaminated environment. Tobacco regenerants selection resulted in 6.8% transformation efficiency when using a medium supplemented with 30 g/L mannose with stepwise decrease of the sucrose concentration. Similar transformation efficiency (5.3%) was achieved in transformation of flax. Relatively low selection efficiency was achieved (12.5% and 34.8%, respectively). The final detection of efficient pmi selection was conducted using PCR and the non-endogenous genes; pmi transgene for flax and todC2 transgene for tobacco plants.

miR319, miR390, and miR393 Are Involved in Aluminum Response in Flax (Linum usitatissimum L.).

Acid soils limit agricultural production worldwide. Major reason of crop losses in acid soils is the toxicity of aluminum (Al). In the present work, we investigated expression alterations of microRNAs in flax (Linum usitatissimum L.) plants under Al stress. Flax seedlings of resistant (TMP1919 and G1071/4_k) and sensitive (Lira and G1071/4_o) to Al cultivars and lines were exposed to AlCl3 solution for 4 and 24 hours. Twelve small RNA libraries were constructed and sequenced using Illumina platform. In total, 97 microRNAs from 18 conserved families were identified. miR319, miR390, and miR393 revealed expression alterations associated with Al treatment of flax plants. Moreover, for miR390 and miR393, the alterations were distinct in sensitive and resistant to Al genotypes. Expression level changes of miR319 and miR390 were confirmed using qPCR analysis. In flax, potential targets of miR319 are TCPs, miR390-TAS3 and GRF5, and miR393-AFB2-coding transcripts. TCPs, TAS3, GRF5, and AFB2 participate in regulation of plant growth and development. The involvement of miR319, miR390, and miR393 in response to Al stress in flax was shown here for the first time. We speculate that these microRNAs play an important role in Al response via regulation of growth processes in flax plants.

Improvement of adventitious root formation in flax using hydrogen peroxide.

Flax (Linum usitatissimum L.) is an important crop for the production of oil and fiber. In vitro manipulations of flax are used for genetic improvement and breeding while improvements in adventitious root formation are important for biotechnological programs focused on regeneration and vegetative propagation of genetically valuable plant material. Additionally, flax hypocotyl segments possess outstanding morphogenetic capacity, thus providing a useful model for the investigation of flax developmental processes. Here, we investigated the crosstalk between hydrogen peroxide and auxin with respect to reprogramming flax hypocotyl cells for root morphogenetic development. Exogenous auxin induced the robust formation of adventitious roots from flax hypocotyl segments while the addition of hydrogen peroxide further enhanced this process. The levels of endogenous auxin (indole-3-acetic acid; IAA) were positively correlated with increased root formation in response to exogenous auxin (1-Naphthaleneacetic acid; NAA). Histochemical staining of the hypocotyl segments revealed that hydrogen peroxide and peroxidase, but not superoxide, were positively correlated with root formation. Measurements of antioxidant enzyme activities showed that endogenous levels of hydrogen peroxide were controlled by peroxidases during root formation from hypocotyl segments. In conclusion, hydrogen peroxide positively affected flax adventitious root formation by regulating the endogenous auxin levels. Consequently, this agent can be applied to increase flax regeneration capacity for biotechnological purposes such as improved plant rooting.

New strategies for the use of Linum usitatissimum cell factories for the production of bioactive compounds.

In this work, suspension-cultured cells of Linum usitatissimum L. were used to evaluate the effect of two types of cyclodextrins, ß-glucan and (Z)-3-hexenol separately or in combination on phytosterol and tocopherol production. Suspension-cultured cells of L. usitatissimum were able to produce high levels of phytosterols in the presence of 50 mM methylated-ß-cyclodextrins (1325.96 ± 107.06 µg g dry weight(-1)) separately or in combination with ß-glucan (1278.57 ± 190.10 µg g dry weight(-1)) or (Z)-3-hexenol (1507.88 ± 173.02 µg g dry weight(-1)), being cyclodextrins able to increase both the secretion and accumulation of phytosterols in the spent medium, whereas ß-glucan and (Z)-3-hexenol themselves only increased its intracellular accumulation. Moreover, the phytosterol values found in the presence of hydroxypropylated-ß-cyclodextrins were lower than those found in the presence of methylated-ß-cyclodextrins in all cases studied. However, the results showed that the presence of methylated-ß-cyclodextrins did not increase the tocopherols production and only an increase in tocopherol levels was observed when cells were elicited with 50 mM hydroxypropylated-ß-cyclodextrins in combination with ß-glucan (174 µg g dry weight(-1)) or (Z)-3-hexenol (257 µg g dry weight(-1)). Since the levels of tocopherol produced in the combined treatment were higher than the sum of the individual treatments, a synergistic effect between both elicitors was assumed. To sum up, flax cell cultures elicited with cyclodextrins alone or in combination with ß-glucan or (Z)-3-hexenol were able produce phytosterols and tocopherols, and therefore, these elicited suspension-cultured cells of L. usitatissimum can provide an alternative system, which is at the same time more sustainable, economical and ecological for their production.

DDT degradation efficiency and ecotoxicological effects of two types of nano-sized zero-valent iron (nZVI) in water and soil.

Nano-scale zero-valent iron (nZVI) has been conceived for cost-efficient degradation of chlorinated pollutants in soil as an alternative to e.g permeable reactive barriers or excavation. Little is however known about its efficiency in degradation of the ubiquitous environmental pollutant DDT and its secondary effects on organisms. Here, two types of nZVI (type B made using precipitation with borohydride, and type T produced by gas phase reduction of iron oxides under H2) were compared for efficiency in degradation of DDT in water and in a historically (>45 years) contaminated soil (24 mg kg(-1) DDT). Further, the ecotoxicity of soil and water was tested on plants (barley and flax), earthworms (Eisenia fetida), ostracods (Heterocypris incongruens), and bacteria (Escherichia coli). Both types of nZVI effectively degraded DDT in water, but showed lower degradation of aged DDT in soil. Both types of nZVI had negative impact on the tested organisms, with nZVI-T giving least adverse effects. Negative effects were mostly due to oxidation of nZVI, resulting in O2 consumption and excess Fe(II) in water and soil.