On the basis of previous studies, an antioxidant enriched high yield potential genotype (Accession VA13) was selected for this investigation. This genotype was grown in pots of a rain shelter open field of Bangabandhu Sheikh Mujibur Rahman Agricultural University, Bangladesh (AEZ-28, 24° 23′ north latitude, 90° 08′ east longitude, 8.4 m.s.l.). The seeds were sown in plastic pots (15 cm in height and 40 cm length and 30 cm width) in a randomized complete block design (RCBD) with three replications. N: P₂O₅:K₂O were applied @92:48:60 kg/ha as a split dose. First, in pot soil, @46:48:60 kg ha 1 N: P₂O₅:K₂O and second, at 7 days after sowing (DAS) @46:0:0 kg/ha N: P₂O₅:K₂O. The genotype was grouped into three sets and subjected to four salinity stress treatments that are, 100 mM NaCl, 50 mM NaCl, 25 mM NaCl, and control or no saline water (NS). Pots were well irrigated with fresh water every day up to 10 days after sowing (DAS) of seeds for proper establishment and vigorous growth of seedlings. Imposition of salinity stress treatment was started at 11 DAS and continued up to 40 DAS (edible stage). Saline water (100 mM NaCl, 50 mM NaCl and 25 mM NaCl) and fresh water were applied to respective pots once a day. At 40 DAS the leaves of Amaranthus tricolor were harvested. All the parameters were measured in six samples.

At Moderate salinity stress (MSS) and Severe salinity stress (SSS) conditions, leaf color parameters and pigments, vitamins, phenolic acids, flavonoids and antioxidant capacity of A. tricolor leaves were very high compared to control condition. Hence, salt-stressed A. tricolor leaves had a good source of natural antioxidants compared to plant grown in normal cultivation practices.

They were grown in 14 cm plastic pots containing soil mixture of peat:sand (1:1) and maintained in a controlled greenhouse conditions in a private nursery located on Alexandria-Cairo Desert Road. The soil was fertilized with 2 g l -1 Crystalon (19% N: 19% P: 19% K, Chema Industries, Egypt). The plants were maintained at natural light conditions and the temperature ranged between 15 and 28 ℃ . Plants were watered every two days with 2000 and 4000 ppm NaCl solution. The plants subjected to saline conditions (watered with 2000 and 4000 ppm NaCl solution) were treated with 7 ml/L weekly application of Seaweed extracts (SWE) (Ascophyllum nodosum, Stella Maris, Acadian Seaplants, Canada) as foliar spray until drop-off two weeks ahead of saline conditions. A foliar spray of 5-aminolevulinic acid (Sigma-Aldrich, Germany) at (3, 5 and 10 ppm) was applied weekly until drop-off to all plants during saline conditions and untreated plants were considered as controls. Experiments continued for 6 weeks in two consecutive seasons of 2016 and 2017 and the plants were distributed in three blocks and each treatment was represented by 5 replicates with a total number of 270 plants. The NaCl was added to the tank gradually and the electrical conductivity (EC) was measured using EC meter.

There were significant increases in branch length and number of branches per plant, fresh and the dry weight in Seaweed extracts (SWE) + 5-aminolevulinic acid (ALA) treated plants under saline irrigation conditions compared to control. These morphological improvements associated with several physiological changes in treated plants including increased accumulation of specific phenols (robinin, rutin, apigein, chlorogenic acid and caffeic acid) and increased antioxidant activities of leaf extracts. There were also increased the chlorophyll composition and the accumulation of sugars and proline. Improved transpiration and photosynthetic rates as well as stomatal conductance were also detected in treated plants. The expression of several genes responsible for water management, flavonoid accumulation and antioxidants accumulation was enhanced in SWE + ALA treated plants.

Under normal and saline irrigation conditions, there were changes in genes expression following SWE and ALA applications.The transcription levels of ANN1 and ANN2 increased significantly in SWE + ALA treatments compared to control under non-saline and saline conditions . However, transcription levels of MYB44 showed no significant variations among treatments. PIP1, P5CS1 and CHS relative expressions was higher in plants treated with SWE + ALA under normal and saline irrigation conditions . The transcription levels of the redox responsive genes of APX1 and GPX3 were significantly higher in SWE + ALA treated plants compared to control treatments . Increasing ALA amount from 3 to 10 ppm had no significant effects on APX1 transcription levels under non-saline conditions. In addition, the application of SWE or ALA only had no significant effects on GPX3 transcription levels under non-saline conditions.

The experiments were conducted under controlled conditions using plastic pots having lower diameter of 7.8 cm, upper diameter of 13.5 cm and 12 cm in height. The soil was collected from the top layer (0-20 cm) from the Botanical Garden of the university. Soil was air dried crushed and sieved through 2 mm filter autoclaved at 121 ℃ for 2 h. The soil was autoclaved to exclude soil pathogens and other microorganisms if any. The autoclaved soil was poured in pots and kept in the growth chamber. The pots were filled with 500 g uncontaminated soil and partially decayed compost (cow manure) (2:1) and was used as growing medium. The cow dung was added into the soil for better performance of earthworms. A subsample of the study soil before mixing with compost was analyzed for its physicochemical characteristics. The soil used for the experiment was sandy loam soil having pH 7.8 , EC (Electrical conductivity) (µS/cm) =184.25 , TDS (Total Dissolved Solids) (mg/kg) = 130 , N (Nitrogen) (mg/kg) = 103 , P (Phosphorus) (mg/kg) = 10.6 , K (Potassium) (mg/kg) = 0.343 , %OC = 0.894, Cd (mg/kg) = ND (not detected by AAS).The Cd treatment was given by using anhydrous CdCl₂ (Minimum assay: 95.0%) procured from Hi-Media laboratories. The CdCl₂ anhydrous was added to the soil to make different concentrations of Cd 0.50 mM, 0.75 mM, 1.00 mM, and 1.25 mM (i.e. 56 mg/Kg , 84 mg/Kg , 112 mg/Kg and 140 mg/Kg respectively). The various treatments given are as shown below:(1)C0 (Control): (Cadmium absence);(2)C1: (0.5 mM Cd);(3)C2: (0.75 mM Cd);(4)C3: (1.00 mM Cd);(5)C4: (1.25 mM Cd).Each Cd treatment was given in soils without as well as with earthworms (WTE = without, WE = with earthworms). Earthworms (3 earthworms per pot) were inoculated after seven days of Cd treatment and incubated for 7 d in soil with earthworms. The seeds after surface sterilization were sown in soil containing different concentration of Cd and earthworms in plastic pots. These pots were kept in seed germinator under controlled conditions i.e. 25 ℃ temperature and 16:8 h dark: light photoperiod (1700 lx) for 15 d. Seedlings were harvested after 15 d followed by washing with distilled water. The growth and biochemical analysis was done on these seedlings.

Increased Cd uptake in plants in presence of earthworms enhances the total antioxidative capacity, metal chelating compounds and content of other antioxidants in plants grown under metal polluted soils. Earthworms can improve plant growth by improving nutrient availability to plants through their vermicasting activity. Their role in modifying soil pH and increasing metal phytoavailability made their use ideal in phytoremediation of polluted soils. Increased uptake and accumulation of Cd in plants activates the antioxidative system of plants takes place by addition of earthworms to soil.

The gene expression for the key enzymes involved in organic acid metabolism was studied to understand the role of earthworms in organic acid metabolism in plants under Cd metal stress. It was observed that in comparison to control (C0) seedlings the expression of CS, SUCLG1, SDH and FH was enhanced 1.72, 1.58, 1.65 and 1.88 folds in seedlings given C4 treatment with 1.25 mM dose of Cd respectively . However, after supplementation of earthworms to Cd treated soils given C4 treatment resulted in further enhancement in expression of CS (2.53 fold), SUCLG1 (2.35 fold), SDH (2.13 fold) and FH (3.06 fold) .

Seeds of B. juncea (cv. RLC-1) were given pre-sowing treatment with 24-epibrassinolide (EBR) solutions (0 and 100 nM EBR/L) for 8 h. Petri-plates were lined with Whatman1 filter paper and were supplemented with different imidacloprid (IMI) concentrations (0, 150, 200, and 250 mg IMI/L). The EBR treated seeds were rinsed with distilled water and grown in Petri-plates supplemented with IMI solutions (three petri-plates for each treatment). The Petri-plates were kept in seed germinator (temperature = 25 ℃ , photoperiod = 16 h, light intensity = 175 µmol m ^-2 s^-1) and the seedlings were harvested 10 days after sowing for further analysis.

Seed soaking with 24-epibrassinolide recovers the impaired growth of B. juncea seedlings under imidacloprid stress by modulating the expression of genes encoding key enzymes including chlorophyllase, citrate synthase, succinyl Co-A ligase, succinate dehydrogenase, fumarate hydratase, malate synthase, phytoene synthase, chalcone synthase, and phenylalanine ammonialyase.

In the present study, as compared to control seedlings, the expression of gene CHLASE (encoding chlorophyllase) was observed to increase by 2.66-fold under IMI toxicity, but seed soaking with EBR significantly reduced the expression of CHLASE to 1.07-fold in the seedlings under IMI toxicity . Data analysis using two-way ANOVA and Tukey's HSD showed significant difference for CHLASE expression in B. juncea seedlings (FIMI p < 0.01, FEBR p < 0.01, FIMI * EBR p < 0.001). MLR analysis of the fold change in CHLASE expression also revealed the increased expression of gene with IMI toxicity and EBR application (positive betaIMI-value), whereas interaction between IMI and EBR was observed to be negative .Further, in comparison to control seedlings, the expression of PSY (encoding phytoene synthase) and CHS (encoding chalcone synthase) was significantly enhanced by 5.22 and 4.54-folds respectively in the seedlings raised from EBR treated as well as untreated seeds grown under IMI stress . Significant differences in expression PSY (FIMI p < 0.001, FEBR P<0.05) and CHS (FIMI * EBR p < 0.001) were observed after analyzing the data using two-way ANOVA and Tukey's HSD. MLR analysis of fold change in gene expression also revealed the role of EBR in modulation of gene expression of PSY and CHS. Concentrations of IMI as well as EBR were regressed positively on the fold change in gene expression of PSY and CHS, thus revealing enhanced expressions of these genes under both the treatments. Moreover, interaction between IMI and EBR was positive for PSY expression, whereas negative interaction was observed for the expression of CHS .In the present study, the expression of PAL was also observed to enhance significantly by 6.68-fold in the seedlings raised from EBR treated seeds and grown under IMI stress . After analyzing the data using two-way ANOVA and Tukey's HSD, significant difference in the expression of PAL was observed (FIMI p < 0.01, FEBR p < 0.01, FIMI * EBR P<0.05). MLR analysis of the fold change in gene expression also confirmed the role of EBR in increasing the PAL gene expression under IMI pesticide stress. Positive beta-regression coefficients were observed for IMI, EBR, and IMI * EBR .The expression of genes encoding the key enzymes involved in organic acid metabolism was also studied to understand the role of EBR in organic acid metabolism under IMI pesticide stress. It was observed that as compared to control seedlings, the expression of CS (encoding citrate synthase, 2.35-fold), SUCLG1 (encoding succinyl-Co-A ligase, 1.57-fold), SDH (encoding succinate dehydrogenase, 2.01-fold), FH (encoding fumarate hydratase, 1.57-fold), and MS (encoding malate synthase, 1.91-fold) were increased in B. juncea seedlings raised from untreated seeds and grown under IMI pesticide toxicity . However, seed soaking with 100 nM EBR and germinating them under IMI toxicity resulted in further enhancement in expression of CS (2.61-fold), SUCLGD1 (4.18-fold), SDH (2.55-fold), FH (3.73-fold), and MS (4.03-fold). Data analysis using two-way ANOVA and Tukey's HSD showed significant differences in the expression of CS (FEBR p < 0.01, FIMI * EBR p < 0.01), SUCLG1 (FEBR p < 0.001, FIMI * EBR P<0.05), SDH (FEBR p < 0.01), FH (FEBR p < 0.001), and MS (FEBR p < 0.001). MLR analysis showed that gene expression in seedlings under IMI stress as well as after the EBR seed treatment was increased as indicated by positive beta-regression coefficients. Whereas, negative interactions were noticed between IMI and EBR treatments for the expression of all genes studied related to organic acid metabolism.

AMF inocula [Funneliformis mosseae (T.H. Nicolson & Gerd.) C. Walker & A. Schubler] consisting of spores, soil, hyphae and infected clove (Trifolium repens L.) root fragment from a stock culture of F. mosseae.The inoculated dosage was 10 g of inocula per pot containing approximately 720 spores calculated by microscopy before experiment. There was 2100 infective propagules/g in the inoculum as determined by MPN assay . Seeds were surface sterilized by immersion in 70% ethanol for 5 min, rinsed four times with distilled water, and placed on wet filter paper in Petri dishes at 28 ℃ for germination. After 3 days, the germinate seeds were transplanted into 13 cm × 13 cm plastic pots containing 0.88 kg organized soil substrate (organic manure, soil and decomposed straw = 1:2:1). Half of the pots (AM plants) were inoculated with 10 g of F. mosseae per pot. Non-AM plants received the same weight of autoclaved inocula. The inocula were placed adjacent to each seeding root. The organic substrate was collected from the greenhouse of Institute of Vegetables and Flowers, CAAS and sterilized for 4 h at 160 ℃ , with the chemical properties as follows: pH 7.26, 11.1% organic matter, 150 mg/kg available phosphorus, 451 mg/kg available nitrogen and 518 mg/kg available potassium. The experimental pots were placed in solar greenhouse at an average temperature of 28 ℃ /20 ℃ (day/night) with photon flux density of 600 µmol m^-2 s^-1 and 85% relative humidity.The seedlings uniformed in size were transferred to a growth chamber subjected to different temperature conditions at 20 days after inoculation. The experimental design consisted of four treatments crossing two mycorrhizal inoculations levels (non-AMF and F. mosseae) with two temperature levels with photon flux density of 100 µmol m^-2 s^-1 (25 ℃ /15 ℃ , 15 ℃ /10 ℃ , day/night). (1) Normal temperature (NT): 10 g of sterilized inoculua, 25 ℃ /15 ℃ (day/night); (2) AMF-inoculation (AMF): 10 g of inocula, 25 ℃ /15 ℃ (day/night); (3) low temperature (LT): 10 g of sterilized inocula, 15 ℃ /10 ℃ (day/night); (4) AMF-inoculation under low temperature (AMF + LT): 10 g of inocula, 15 ℃ /10 ℃ (day/night).The experimental design was a completely randomized block design and thirty plants were arranged in each replication. On 45 days after inoculation, phenolic compounds contents, enzymes and gene transcription were determined.

AMF-inoculated cucumber seedlings had significant higher fresh weight and dry weight than non-AMF inoculated control plants under both normal (25/15 ℃ ) and low temperature (15/10 ℃ ) treatment. Under chilling stress, AMF inoculation significantly improved the content of related secondary metabolites including phenols, flavonoids, lignin, DPPH activity and phenolic compounds compared with the non-AMF control. Furthermore, large increments were observed in a number of enzymatic activities related to secondary metabolism and antioxidant system in AMF-inoculated seedlings under low temperature, such as glucose-6-phosphate dehydrogenase (G6PDH), shikimate dehydrogenase (SKDH), phenylalanine ammonia-lyase (PAL), cinnamyl alcohol dehydrogenase (CAD), polyphenol oxidase (PPO), guaiacol peroxidase (G-POD), caffeic acid peroxidase (CA-POD) and chlorogenic acid peroxidase (CGA-POD). As well, the expression of stress-related marker genes was enhanced in AMF-inoculated seedlings in comparison with the non-AMF control. Furthermore, AMF symbiosis decreased hydrogen peroxide (H₂O₂) content under low temperature.

Consistent with the changes of enzyme activities, the relative transcriptional level of genes related to secondary metabolism increased significantly in response to AM fungi inoculation in cucumber roots . Moreover, under chilling stress, the expression levels of WRKY30, PR-1, C4H, CCOMT, CAD, G6PDH, PAL, LPO, and POD genes in the AMF-inoculated seedling were 2.46, 2.90, 1.84, 2.47, 1.96, 2.52, 1.89 and 1.93 folds respectively compared with those under low temperature alone.

Seeds were treated with sulfuric acid (98%, 10 min) and then surface sterilized with 70% ethanol (v/v) for 1 min and sodium hypochlorite solution (10%, at 10 min). After sterilization, seeds were germinated on MS media (Murashige and Skoog, 1962) containing 7 g/L agar (Duchefa, Netherlands). Cultures were maintained under 16/8 h light/dark. Explants were taken from 4-week-old leaves for inoculation with bacteria strain.ATCC15834 strain of A. rhizogenes was supplied by microbial unit of the National Research Center for Genetic Engineering and Biotechnology, Tehran-Iran. Bacterial cells cultivated on LB (Luria-Bertani) culture medium (Bertani, 1952) on rotary shaker (at 26 ℃ and 180 rpm for 48 h) in the darkened state.The leaves were wounded and inoculated with bacterial suspension for 5 min and transferred to MS media containing 7 g/L agar in darkness at 25 ℃ . After 48 h treated Explants were cultured on the 1/2 MS media containing cefotaxime (500 mg/L) and indole-3-butyric acid (IBA) (2 mg/L). Hairy roots emerged at wounded sites, after 4-weeks of incubation, and then each hairy root line was isolated from explants tissue and was subcultured weekly in new media (1/2 MS hormone-free media) with appropriate antibiotic. The concentration of cefotaxime was decreased gradually and eliminated from the culture medium after 8 subcultures and axenic root cultures were obtained. Then hairy root lines were transferred to the 250 mL Erlenmeyer flasks containing 30 mL hormone- free 1/2 MS liquid medium and incubated on a rotary shaker (120 rpm) at 25 ℃ and subcultured every two week. Hairy root line, which showed sufficient growth in 1/2 MS liquid medium, was selected for further investigations.The genomic DNA was extracted from transformed hairy root lines and plant intact roots with CTAB method . Gene-specific primers from rol B were used for amplification of the 780-bp segment in PCR analysis. The primers sequences were, F:5'-ATGGATCCCAAATTGCTATTCCCCCACGA-3'and R:5'-TTAGGCTTCTTTCATTCGGTTTACTGCAGC-3'. Thirty-five PCR cycles were performed with 5 min initial denaturation at 94 ℃ , annealing steps at 60 ℃ for 80 s, extension at 72 ℃ for 90 s, and final extension step of 72 ℃ for 10 min. The amplimer were analyzed by 1% agarose gel electrophoresis.To investigate the effects of SiO₂ NPs, various concentrations (0, 25, 50, 100 and 200 mg/L) of this elicitor were added to the hairy roots culture medium (1/2 MS + 3% sucrose, pH = 5.7) at the end of log phase of growth stages (21-days-old cultures). Hairy roots were incubated with elicitor for 24 and 48 h of exposure time. Hairy roots were harvested 7 days after elicitation and dried on sterile filter paper to remove excess surface moisture and were weighed before freezing by liquid nitrogen and stored at -80℃ until used to measure growth, biochemical and phytochemical analysis.

The effect of silicon dioxide nanoparticles on production of phenolic compounds and expression rate of pal and ras genes involved in rosmarinic acid biosynthesis pathway has been investigated in D. kotschyi. SiO₂ nanoparticles, used as an abiotic elicitor in our study, has appropriate optical, electrical and catalysts properties and has many applications in various industries as well as agriculture. This study clearly suggested that, in the presence of this nanoparticle, induction, production and accumulation of valuable compounds and corresponding antioxidant activity increased in hairy roots.

According to the results, expression levels of the pal and ras genes were influenced by elicitor concentration and exposure time. The elicitation by SiO₂ NP of 100 mg/L after 48 h of exposure time dramatically increased pal expression compared to the control. Briefly, with increasing SiO₂ NP concentrations after 48 h of exposure time, the expression level of pal was also significantly induced . Similarly, ras expression was significantly raised at 48 h after treatment by increasing SiO₂ NP concentration and enhanced to the greatest extent in 50 mg/L concentration. After 24 h of exposure time, the minimum level of ras expression was observed in the 200 mg/L SiO₂ . Amplification products of real-time PCR were assessed with 1.8% agarose gel which was corresponded to the predicted size.

Lettuce plants (Lactuca sativa L. cv. Romana Lentissima a Montare 4, FOUR-BLUMEN s.r.l., Piacenza, PC, Italy) were cultivated under the greenhouse of Agronomy and Crop Sciences Research and Education Center, University of Teramo, Mosciano Sant' Angelo (42° 53′ N and 13° 55′ E, 15 m; above sea level) from June to July 2013. The greenhouse is covered with a single layer of ethylene-vinyl acetate (PATILUX) provided by P.A.T.I. S.p.A. (San Zenone degli Ezzelini, TV, Italy); it has a natural ventilation system, and it is not provided of artificial lights, fans, and heaters. The % of reduction with respect to outdoors conditions in terms of total (W m ^-2) and PAR (µmol m^-2 s^-1) radiation amounted to 7.4% and 12.6%, respectively. Moreover, the plastic film, as expected, causes a reduction of the irradiance (µmol m^-2 s^-1) by 64, 32 and 24% on average in the ultraviolet, PAR and near infrared regions, respectively. Starting from transplanting air temperature was constantly monitored with sensors connected to a data logger (EM50 Data Collection System, Decagon Devices Inc., Pullman, WA, USA) .Seeds were sown on a nursery potting soil (Huminsubstrat N3, Neuhaus, Klasmann-Deilmann, Geeste, Germany), composed of 90% peat, 10% clay; pH 6; NPK 14:16:18, 1.3 kg m^-3; conductivity 35 mS m^-1. On 18 June, uniform sized seedlings of lettuce at the 3-leaf stage were transplanted into individual plastic pots (14 × 14 cm) filled with peat-based compost (peat:vermiculite:perilte 1:1:1, v/v); the composition of the peat moss is given as follows (percentage on dry matter): organic carbon 40%, organic nitrogen 0.1%, organic matter 80%. At 8 and 10-leaf stages, a treatment with fungicide Ortiva (a.i. Azoxystrobin 23.2%, Syngenta Crop Protection S.p.A., Milano, Italy) at the dose of 0.08 mL m^-2 was applied.The experiment was arranged on a complete randomized block design. Two nutrient-deficiency conditions and two abiotic stressful conditions were imposed starting from 4 days after transplanting (DAT), i.e. no phosphorus fertilization (named 0_P), no nitrogen fertilization (0_N), limitation of the photosynthetic active radiation (PAR, range from 400 to 700 nm) (LR) and water availability constraint (WR), plus one unstressed CONTROL. Each treatment was replicated three times and each replication consisted in 49 pots (7 rows, 7 pots per row) for a total of 147 pots per treatment; to avoid edge effects, plants were daily re-randomized with the accuracy in maintaining the same sun orientation. All the plots with no-phosphorus limitation (CONTROL, 0_N, LR and WR) were fertilized with simple superphosphate at the rate of 40 kg P₂O₅ ha^-1. All the plots with no-nitrogen limitation (CONTROL, 0_P, LR and WR) were fertilized with two applications, at 4 and 7 DAT, at the total rate of 90 kg N ha^-1 with calcium nitrate (Ca(NO₃)2). In order to standardize the amount of calcium to the plants, 0_N treatment was fertilized with 156 kg ha^-1 of calcium oxide (as the commercial product Brexil Ca, Valagro S.p.a., Piazzano di Atessa, CH, Italy). At transplanting plants were fertilized with potassium chloride at the dose of 100 kg K₂O ha^-1, and KSC Mix (Timac Agro Italia, Milano, Italy) at the dose of 0.02 kg ha^-1 composed as follows: 15% water-soluble magnesium oxide (MgO); 28% water-soluble sulphurous anhydride (SO₃); 0.5% water-soluble boron (B); 0.5% water-soluble copper (Cu) chelated by EDTA; 2.5% water-soluble iron (Fe) chelated by EDTA; 2% water-soluble manganese (Mn) chelated by EDTA; 0.2% water-soluble molybdenum (Mo); 1.5% water-soluble zinc (Zn) chelated by EDTA.Shade treatments (LR) were accomplished using a shade net in order to obtain a 85% of reduction in PAR wavelengths. PAR intensity was hourly measured with a PAR Photon Flux Sensor (Decagon Devices Inc., Pullman, WA, USA), connected to a data logger (EM50 Data Collection system, Decagon Devices Inc., Pullman, WA, USA). Per cent of shading was determined by comparing the average PAR values of net with the average PAR values of the un-shaded treatments. Nets were wrapped around a rigid and removable structure placed above the vegetation so that they covered the incoming light from the top and sides to 5 cm below the bottom of the pots. To allow air circulation, light was not limited from below.Water stress (WR) was imposed by maintaining soil volumetric water content at 30% of water holding capacity (WHC) which corresponded to 0.240 and 0.072 m³ m^-3 for no-water stressed and water stressed plants, respectively. Water loss, due to evapotranspiration, was constantly monitored with soil moisture sensors installed in 5 randomly selected pots, for each replicates (EC-5, Decagon Devices, Inc., Pullman, WA, USA); the sensors were connected to a data logger - EM50 Data Collection system (Decagon Devices Inc., Pullman, WA, USA). The pots were manually re-watered with tap water (pH 7.2, EC 0.23 mS cm^-1) every day at 18:00 h.

With the exception of light reduction, the other kind of limitation negatively influenced lettuce fresh yield; nevertheless, the reduction of PAR availability induced a decrease in the content of the main investigated phenolic compounds resulting in a strong reduction of total phenolic content as well as antiradical activity. Conversely, the scarcity of N nutrition allowed to obtain the highest total polyphenols content (TPC) and TEAC (Trolox Equivalent Antioxidant Capacity), although no differences were found in terms of the main phenolic compounds. Drought seems to improve the accumulation of caffeic, caftaric and chicoric acids in the bound forms as well as TPC and antiradical activity of the same fractions, while the reduction in P fertilization did not significantly influence lettuce leaves composition in terms of phytochemicals.

Seeds were sterilized in 1% (v/v) sodium hypochloride (Sigma-Aldrich, USA) for 10 min, then drained and washed with distilled water until they reached neutral pH. They were placed in distilled water and soaked for 6 h at 25 ℃ . Seeds were dark germinated for 8 days in a growth chamber (SANYO MLR-350H) on Petri dishes (125 mm) lined with absorbent paper. Seedlings were watered with 5 ml of Milli-Q water daily. Sprout (8-day-old) samples were gently collected, weighed (fresh mass), rapidly frozen and kept in polyethylene bags at -20 ℃ . For each treatment, three replicates were performed.Elicitation conditions were selected in previous screening studies. For the experiments, temperature (4 ℃ and 40 ℃ - TC and TH, respectively), H₂O2 (20 mM and 200 mM - Ox1 and Ox2, respectively), mannitol (200 mM and 600 mM - Os1 and Os2, respectively) and NaCl (100 mM and 300 mM - S-Os1 and S-Os2, respectively) were selected as abiotic elicitors. All solutions were freshly prepared before each application. Mannitol (Os1, Os2), NaCl (S-O1, S-O2) and H₂O2 (Ox1) treatments were applied by watering daily (not soaking) 2-day-old sprouts with 5 ml of test solution. For Ox2 (200 mM H₂O2) treatment 2-day-old seedlings were only once watered with 5 ml of 200 mM H₂O2 and then cultivated under standard conditions. For temperature conditioning treatment, 2-day-old sprouts were incubated at 4 ℃ and 40 ℃ (TC and TH, respectively) for 1 h and then cultivated under standard conditions. Sprout (8-day-old) samples were gently collected, weighed (fresh mass), rapidly frozen and kept in polyethylene bags at -20 ℃ .

Application of abiotic elicitors (environmental shocks) was an effective method for improvement of sprout pro-health potential via an increase of phenolic contents and subsequent elevation of antioxidant potential. Innovative application of elicitors on 2-day-old sprouts (not seed) allowed the elimination of the unfavorable influence of the factors employed on germination yield and biomass production. Assuming that the optimal germination conditions are those which most effectively increase the antioxidant potential without any negative influence on biomass accumulation and nutritional quality the elicitation with 20 mM H₂O₂ for the future applications is recommended.

The experiment was carried out in 2013. Leaves of five raspberry cultivars ('Polana', 'Polka', 'Tulameen', 'Laszka' and 'Glen Ample') were collected at the time of cultivation. Three organic and neighborhood conventional farms were used for experimental purposes. From one cultivar (one field plot), 3-4 plants were chosen, which were analyzed separately. One sample consisted of 10 leaves. The farm was treated as a replication. [organic farm no. 1 Localization: akroczym(52° 26″ N 20° 36″ E), Type of Soil: sandy middle soil IVa and IVb category (15% floatable particles) pH 5.5, Kind of Fertilizer: cow manure, Dose of Fertilizers and Time of Given: 35 t/ha one year before raspberry planting, Plant Protection System: Grevit 200 SL; organic farm no. 2 Localization: Zaluski (52° 37″ N 20° 22″ E), Type of Soil: sandy middle soil, sandy-clay IV category (20% floatable particles), pH 5.5, Kind of Fertilizer:cow manure, Dose of Fertilizers and Time of Given: 30 t/ha one year before raspberry planting, Plant Protection System: no protection; organic farm no. 3 Localization: Radzanow(51° 33″ N 20° 51″ E), Type of Soil: sandy middle soil IVa and III category (10% floatable particles), pH 6.0, Kind of Fertilizer:sheep manure, green manure, Dose of Fertilizers and Time of Given: 10 t/ha and 15 t/ha one year before raspberry planting, Plant Protection System: Bioczos 33 SL, Grevit 200 SL; conventional farm no. 1 Localization: Czerwinsk nad Wisla (52° 23″ N 20° 20″ E), Type of Soil: sandy-loamy middle soil IV and III category (20% floatable particles), pH 5.5, Kind of Fertilizer: Hydrocomplex 12-11-18; Superba 8-11-36, Dose of Fertilizers and Time of Given: (200 kg/ha, 150 kg/ha) in autumn a year before raspberry planting; 3 doses in time of cultivation, Plant Protection System: Signum 33 WG, Miros 20 SP; conventional farm no. 2 Localization: Czerwinsk nad Wisla (52° 23″ N 20° 20″ E), Type of Soil: sandy-loamy middle soil IV and III category (25% floatable particles), pH 5.5, Kind of Fertilizer: amonium nitrate, polyphosphate, magnesium sulphate, Dose of Fertilizers and Time of Given: in autumn a year before raspberry planting; 3 doses in time of cultivation, Plant Protection System: Calypso 480 SC, Miros 20 SP, Zato 50 WG; conventional farm no. 3 Localization: Czerwinsk nad Wisla(52° 25″ N 20° 23″ E), Type of Soil: sandy-clay middle soil II and III category (20% floatable particles) pH 6.0, Kind of Fertilizer:Rosafert 5-12-24-3, Dose of Fertilizers and Time of Given: 250 kg/ha in autumn a year before raspberry planting; 4 doses in time of cultivation, Plant Protection System: Calypso 480 SC, Miros 20 SP, Zato 50 WG].

Compared with conventional raspberry leaves, organic raspberry leaves were characterized by a significantly higher content of dry matter, total polyphenols, total phenolic acids, chlorogenic acid, caffeic acid, salicylic acid and quercetin-3-O-rutinoside; moreover, the organic leaves were characterized by higher antioxidant activity. Among examined cultivars, 'Polka' c. was characterized by the highest antioxidant status. However, raspberry leaves from conventional farms contained more total carotenoids, violaxanthin, alpha-carotene, beta-carotene, total chlorophyll and individual forms of chlorophylls: a and b.

Different concentrations of TiO₂ NPs (0, 10, 20, 30, and 50) were prepared for hairy root treatments. 0.5 g of .S. officinalis hairy roots were transferred to 250 mL Erlenmeyer flasks containing 15 mL of liquid MS culture medium with three replicates. Then, they were placed in an incubator shaker at 110 rpm and 25 &#8451 in dark conditions. On the 22nd day, the liquid MS culture media containing different concentrations of nano titanium dioxide was added to Erlenmeyer flasks. 24 and 48 h after treatment, the hairy roots were taken out and transferred to the MS culture medium lacking elicitor.

The highest rate of total phenol (9.79 mg GLA/g FW) and total flavonoid contents (1.06 mg QE/g FW) were obtained in the treated hairy roots with 50 and 30 mg/L of nano elicitor in 24 and 48 h of treatments, respectively. The maximum level of most polyphenols, such as rosmarinic acid, cinnamic acid, and rutin, was produced in 24 h of treatment. The use of TiO₂ NP for 48 h with 50 mg/L concentration showed the highest production level of SO6 protein.

The seeds were germinated in seedling plastic tray filled up with a mixture of peat moss and coco peat (1:1). The trays were placed in a glass house at the University of Massachusetts, Amherst, USA, in September 2015. After two weeks, seedlings were transplanted into plastic pots (30 cm diameter and 30 cm height, three seedlings per pot). The pots growth media consisted of 2:1:1 ratio of sieved field soil, sand, and leaf mold, respectively. The medium in each pot was supplemented with 5.8 mg P, 3.3 mg N, and 13.8 mg K to warrant the plant growth; the electrical conductivity was 0.3 mS/cm (Beckman EC meter instrument. cedar grove, New jersey, USA); the pH was 8.04. Plants were maintained at the temperature of 18-25 ℃ ; glass house conditions were set as follows: photoperiod, 16 h; relative humidity, 60-70%; light intensity, 180 µmol m 2 s^-1. The source of light was a high pressure sodium lamp in addition to the day light in order to attain 16 h of light per day. Fourty days after planting (establishment period), the uniform sized plants were treated with four different salinity levels including: 0, 30, 60, and 90 mM NaCl. The irrigation with saline solution (250 mL in each pots) was performed every two days . Total amount of NaCl per pot during experiment was 8.75 L. To avoid osmotic shock the salinity treatment started with 15 mM NaCl, and was progressively increased (every two days) to reach the maximum salinity level in each treatment. To prevent water and nutrient leaching, a plastic dish was inserted under each pot and the leached water was given back to pots.

Salinity has negative impact on plant production and stimulates several physiological and biochemical modifications in thyme species. Here, a major decline in dry matter, relative water content (RWC), photosynthetic pigment contents, K⁺, Ca⁺ as well as increase in Na⁺ and EL was observed. Our work showed that T. daenensis was similar to T.vulgaris in terms of tolerance to high levels of salinity stress and both species were moderately tolerant to severe salt stress. It should be noted that T. daenensis, as an Iranian endemic species, competes with commercial species like T. vulgaris as a rich source of phenolic compounds. Furthermore, the relative salt tolerance of both species could be related to the exclusion of Na⁺ from the vascular system in order to protect tissues from salt toxicity and also to the increase of phenolic content and radical scavenging activity. Thus, the plant behavior under salinity stress may be used to boost the production of bioactive compounds to be used on an industrial level for the manufacture of nutraceuticals, functional foods and cosmetics.

The seeds were germinated in seedling plastic tray filled up with a mixture of peat moss and coco peat (1:1). The trays were placed in a glass house at the University of Massachusetts, Amherst, USA, in September 2015. After two weeks, seedlings were transplanted into plastic pots (30 cm diameter and 30 cm height, three seedlings per pot). The pots growth media consisted of 2:1:1 ratio of sieved field soil, sand, and leaf mold, respectively. The medium in each pot was supplemented with 5.8 mg P, 3.3 mg N, and 13.8 mg K to warrant the plant growth; the electrical conductivity was 0.3 mS/cm (Beckman EC meter instrument. cedar grove, New jersey, USA); the pH was 8.04. Plants were maintained at the temperature of 18-25 ℃ ; glass house conditions were set as follows: photoperiod, 16 h; relative humidity, 60-70%; light intensity, 180 µmol m 2 s^-1. The source of light was a high pressure sodium lamp in addition to the day light in order to attain 16 h of light per day. Fourty days after planting (establishment period), the uniform sized plants were treated with four different salinity levels including: 0, 30, 60, and 90 mM NaCl. The irrigation with saline solution (250 mL in each pots) was performed every two days . Total amount of NaCl per pot during experiment was 8.75 L. To avoid osmotic shock the salinity treatment started with 15 mM NaCl, and was progressively increased (every two days) to reach the maximum salinity level in each treatment. To prevent water and nutrient leaching, a plastic dish was inserted under each pot and the leached water was given back to pots.

Salinity has negative impact on plant production and stimulates several physiological and biochemical modifications in thyme species. Here, a major decline in dry matter, relative water content (RWC), photosynthetic pigment contents, K⁺, Ca⁺ as well as increase in Na⁺ and EL was observed. Our work showed that T. daenensis was similar to T.vulgaris in terms of tolerance to high levels of salinity stress and both species were moderately tolerant to severe salt stress. It should be noted that T. daenensis, as an Iranian endemic species, competes with commercial species like T. vulgaris as a rich source of phenolic compounds. Furthermore, the relative salt tolerance of both species could be related to the exclusion of Na⁺ from the vascular system in order to protect tissues from salt toxicity and also to the increase of phenolic content and radical scavenging activity. Thus, the plant behavior under salinity stress may be used to boost the production of bioactive compounds to be used on an industrial level for the manufacture of nutraceuticals, functional foods and cosmetics.

3-year old single shoot V. vinifera plants (cultivar Pinot noir 18 Gm grafted on Kober 5BB, 51 plants) potted in 3L pots in a sandy loam soil were used. All plants were well watered (200 mL per day) at the beginning of the experiment (04.06.2010; DAY 0; 5 plants) and water was supplied to all control plants once every day (250 mL per day), whereas water supply of stressed plants was stopped. Physiological measurements and sampling of leaves took place on 07.06.2010 (DAY 3; 5 control, 5 stressed plants), 10.06.2010 (DAY 6; 5 control, 5 stressed plants) and 12.06.2010 (DAY 8; 5 control, 10 stressed plants). Due to very hot weather conditions in June 2010 the experiment was stopped after 8 days and 12 available control plants were used to restart the drought treatment with 6 control and 6 stressed plants on 11.06.2010 and all plants were measured on 15.06.2010 (DAY 5). The mean leaf temperatures at midday were: 25 ℃ (04.06.2010; DAY 0), 31.9 ℃ (07.06.2010; DAY 3), 30.8 ℃ (15.06.2010; DAY 5), 35.8 ℃ (10.06.2010; DAY 6) and 35.7 ℃ (12.06.2010; DAY 8). The mean PAR radiation per day (measured from 6:00 am till 7:00 pm) was 144.1 µmol m^-2 s^-1. Each plant was used only once for physiological measurements and sampling of leaves.On every day of the experiment (day 0, 3, 5, 6, 8) the pot weight and the volumetric soil moisture content (ThetaProbe ML2x and handheld data logger Moisture Meter HH2, Delta-T Devices, Cambridge, United Kingdom) was recorded. The water potential (PWSC Model 3000, Soilmoisture Equipment Corporation, Santa Barbara, USA) was determined for the 6th leaf (representing the insertion level of the shoot from the basis) of every plant and measurement day. Chlorophyll fluorescence and gas exchange parameters of light adapted leaves were determined with the 4th and 5th leaf, whereas dark adaptation was performed only with the 5th leaf. Immediately after these non-invasive measurements, the 5th leaf was harvested, frozen in liquid nitrogen and further used for the measurement of polyphenols, selected primary metabolites and volatiles (VOCs).

The content of different groups of primary and secondary metabolites is significantly influenced by severe drought stress in grapevine leaves. The content of the majority of the metabolites (around 60% of primary metabolites, around 85% of polyphenols and about 40% of the detected and identified VOCs) increased upon drought stress treatment. Among these especially the primary metabolites citric acid and glyceric acid were strongly influenced by the short as well as the prolonged drought stress treatment, whereas all polyphenols were only induced upon the prolonged drought stress treatment.