Silicon nanoparticles (SiNPs) stimulated secondary metabolism and mitigated toxicity of salinity stress in basil (Ocimum basilicum) by modulating gene expression: a sustainable approach for crop protection.

The underlying mechanisms through which silicon oxide nanoparticles (SiNPs) can confer salinity resistance to plants are poorly understood. This study explored the efficacy of supplementing nutrient solution with SiNPs (20-30 nm; 10 mg kg-1 soil) to stimulate metabolism and alleviate the risks associated with salinity (0.73 g kg-1 soil) in basil seedlings. For this purpose, variations in photosynthetic indices, proline osmoprotectant, antioxidant markers, phenylpropanoid metabolism, and transcriptional behaviors of genes were investigated. SiNPs increased shoot fresh weight (38%) and mitigated the risk associated with the salinity stress by 14%. SiNPs alleviated the inhibitory effects of salinity on the total chlorophyll concentration by 15%. The highest increase (twofold) in proline content was recorded in the SiNP-treated seedlings grown under salinity. The nano-supplement enhanced the activity of enzymatic antioxidants, including peroxidase (2.5-fold) and catalase (4.7-fold). SiNPs induced the expression of gamma-cadinene synthase (CDS) and caffeic acid O-methyltransferase (COMT) genes by 6.5- and 18.3-fold, respectively. SiNPs upregulated the eugenol synthase (EGS1) and fenchol synthase (FES) genes by six- and nine-fold, respectively. Salinity transcriptionally downregulated the geraniol synthase (GES) gene, while this gene displayed an upward trend in response to SiNPs by eight-fold. The nano-supplement transcriptionally stimulated the R-linalool synthase (LIS) gene by 3.3-fold. The terpinolene synthase (TES) gene displayed a similar trend to that of the GES gene. The highest expression (25-fold) of the phenylalanine ammonia-lyase (PAL) gene was recorded in seedlings supplemented with SiNPs. The physiological and molecular assessments demonstrated that employing SiNPs is a sustainable strategy for improving plant primary/secondary metabolism and crop protection.

Selenium nanoparticles conferred drought tolerance in tomato plants by altering the transcription pattern of microRNA-172 (miR-172), bZIP, and CRTISO genes, upregulating the antioxidant system, and stimulating secondary metabolism.

Drought stress is one of the major limiting factors for the production of tomato in Iran. In this study, the efficiency of selenate and Se nanoparticle (SeNP) foliar application on tomato plants was assessed to vestigate mitigating the risk associated with water-deficit conditions. Tomato plants were treated with SeNPs at the concentrations of 0 and 4 mg L-1; after the third sprays, the plants were exposed to water-deficit conditions. The foliar spraying with SeNPs not only improved growth, yield, and developmental switch to the flowering phase but also noticeably mitigated the detrimental risk associated with the water-deficit conditions. Gene expression experiments showed a slight increase in expression of microRNA-172 (miR-172) in the SeNP-treated plants in normal irrigation, whereas miR-172 displayed a downregulation trend in response to drought stress. The bZIP transcription factor and CRTISO genes were upregulated following the SeNP and drought treatments. Drought stress significantly increased the H2O2 accumulation that is mitigated with SeNPs. The foliar spraying with Se or SeNPs shared a similar trend to alleviate the negative effect of drought stress on the membrane integrity. The applied supplements also conferred drought tolerance through noticeable improvements in the non-enzymatic (ascorbate and glutathione) and enzymatic (catalase and peroxidase) antioxidants. The SeNP-mediated improvement in drought stress tolerance correlated significantly with increases in the activity of phenylalanine ammonia-lyase, proline, non-protein thiols, and flavonoid concentrations. SeNPs also improved the fruit quality regarding K, Mg, Fe, and Se concentrations. It was concluded that foliar spraying with SeNPs could mitigate the detrimental risk associated with the water-deficit conditions.

Methyl jasmonate conferred Arsenic tolerance in Thymus kotschyanus by DNA hypomethylation, stimulating terpenoid metabolism, and upregulating two cytochrome P450 monooxygenases.

Arsenic (As) is a highly cytotoxic element impairing normal cellular functions, and its bioremediation has become one of the environmental concerns. This study explored the molecular and physiological responses of thyme (Thymus kotschyanus) seedlings to incorporating As (0 and 10 mgl-1) and methyl jasmonate (MJ; 0 and 10 µM) into the culture medium. The MJ treatment reinforced root system and mitigated the As cytotoxicity risk. MJ contributed to hypomethylation, a potential adaptation mechanism for conferring the As tolerance. Two cytochrome P450 monooxygenases, including CYP71D178 and CYP71D180 genes, were upregulated in response to As and MJ. The MJ treatment contributed to up-regulation in the γ-terpinene synthase (TPS) gene, a marker gene in the terpenoid metabolism. The As presence reduced photosynthetic pigments (chlorophylls and carotenoids), while the MJ utilization alleviated the As toxicity. The MJ supplementation increased proline accumulation and soluble phenols. The application of MJ declined the toxicity sign of As on the concentration of proteins. The activities of peroxidase, catalase, and phenylalanine ammonia-lyase (PAL) enzymes displayed an upward trend in response to As and MJ treatments. Taken collective, MJ can confer the As tolerance by triggering DNA hypomethylation, regulating CYPs, and stimulating primary and secondary metabolism, especially terpenoid.

Alteration in the callogenesis, tropane alkaloid formation, and gene expression in Hyoscyamus niger under clinorotation.

This study aimed to investigate the effects of clinorotation induced by 2-D clinostat on the growth, tropane alkaloid production, gene expression, antioxidant capacity, and cellular defense responses in the callus tissue of Hyoscyamus niger. Callus induction was conducted by putting hypocotyl explants in the MS culture medium supplemented with 1 mgL-1 2,4-D and 1 mgL-1 BAP growth regulators. The sub-cultured calli were placed on a clinostat for 0, 3, 7, and 10 days (2.24 × 10-5 g on the edge of the callus ring). Clinorotation significantly increased callus fresh weight, dry weight, protein, carbohydrate, and proline contents compared to the control, and their maximum contents were obtained after 7 and 10 days. H2O2 level enhanced under clinorotation with a 76.3% rise after 10 days compared to control and positively affected the atropine (77.1%) and scopolamine (69.2%) productions. Hyoscyamine 6-beta hydroxylase and putrescine N-methyltransferase gene expression involved in the tropane alkaloid biosynthesis were upregulated markedly with 14.2 and 17.1-folds increase after 10 days of clinorotation, respectively. The expressions of jasmonic acid, mitogen-activated protein kinase, and ethylene-responsive element-binding transcription factor were upregulated, and the activity of peroxidase and catalase showed a 72.7 and 80% rise after 10 days. These findings suggest that microgravity can enhance callogenesis by stimulating the ROS level, which can impact the antioxidant enzymes, tropane alkaloid formation, and gene expression.

Zinc oxide nanoparticles (ZnONPs) influenced seed development, grain quality, and remobilization by affecting the transcription of microRNA 171 (miR171), miR156, NAM, and SUT genes in wheat (Triticum aestivum): a biological advantage and risk assessment study.

Limited studies have been conducted on the role of microRNAs (miRs) and transcription factors in regulating plant cell responses to nanoparticles. This study attempted to address whether the foliar application of zinc oxide nanoparticles (ZnONPs; 0, 10, 25, and 50 mgL-1) can affect miRs, gene expression, and wheat grain quality. The seedlings were sprayed with ZnONPs (0, 10, 25, and 50 mgL-1) or bulk counterpart (BZnO) five times at 72 h intervals. The application of ZnONPs at 10 mgL-1 increased the number of spikelets and seed weight, while the nano-supplement at 50 mgL-1 was accompanied by severe restriction on developing spikes and grains. ZnONPs, in a dose-dependent manner, transcriptionally influenced miR156 and miR171. The expression of miR171 showed a similar trend to that of miR156. The ZnONPs at optimum concentration upregulated the NAM transcription factor and sucrose transporter (SUT) at transcriptional levels. However, the transcription of both NAM and SUT genes displayed a downward trend in response to the toxic dose of ZnONPs (50 mgL-1). Utilization of ZnONPs increased proline and total soluble phenolic content. Monitoring the accumulation of carbohydrates, including fructan, glucose, fructose, and sucrose, revealed that ZnONPs at 10 mgL-1 modified the source/sink communication and nutrient remobilization. The molecular and physiological data revealed that the expression of miR156 and miR171 is tightly linked to seed grain development, remobilization of carbohydrates, and genes involved in nutrient transportation. This study establishes a novel strategy for obtaining higher yields in crops. This biological risk assessment investigation also displays the potential hazard of applying ZnONPs at the flowering developmental phase.

Mycoremediation of oil contaminant by Pleurotus florida (P.Kumm) in liquid culture.

This study investigated the potential functions of Pleurotus florida (an edible mushroom) in the biodegradation of gas oil at concentrations of 0 (control), 2.5, 5, and 10% (V: V) for 30 days. The gas oil increased dry weight and protein concentration in all treatments (by an average of 19.5 and 108%, respectively). Moreover, the pH, surface tension (ST), and interfacial tension (IFT) were reduced by the mushroom supplementation. The lowest surface tension (31.9 mN m-1) and the highest biosurfactant production belonged to the 10% gas oil treatment (0.845 ± 0.03 mg mL-1). The results demonstrated that the adsorption isotherm agreed well with the Langmuir isotherm. The maximum Langmuir adsorption capacity was calculated at 0.743 mg g-1 wet biomass of P. florida. The fungal supplementation efficiently remedied the total petroleum hydrocarbons (TPHs) by an average of 55% after 30 days. Gas chromatography (GC) analysis revealed that P. florida effectively detoxified C13-C28 hydrocarbons, Pristane, and Phytane, implying its high mycoremediation function. The toxicity test showed that mycoremediation increased the germination by an average of 35.82% ± 8.89 after 30 days. Laccase activity increased significantly with increasing gas oil concentration in the treatments. The maximum laccase activity was obtained in the 10% gas oil treatment (142.25 ± 0.72 U L-1). The presence of pollutants was also associated with induction in the tyrosinase activity when compared to the control. These results underline the high mycoremediation capacity of P. florida through the involvement of biosurfactants, laccase, and tyrosinase.

Synthesis and characterization of chitosan encapsulated zinc oxide (ZnO) nanocomposite and its biological assessment in pepper (Capsicum annuum) as an elicitor for in vitro tissue culture applications.

Nanotechnology paves the way for introducing nanoscale fertilizers, pesticides, and elicitors. This study intends to address the synthesis of chitosan/zinc oxide nanocomposite (CS-ZnONP) and its biological assessment in in-vitro conditions. The zinc oxide nanoparticles (ZnONPs) were successfully coated with the chitosan (CS) polymer through a cost-effective approach. Transmission electron microscopy and Fourier transform infrared spectroscopy assessments proved the surface capping of chitosan polymer on ZnONP. The nanocomposite was more capable of improving growth and biomass than the bare ZnONPs. The application of the nanocomposite increased the concentration of chlorophylls (51%), carotenoids (70%), proline (2-fold), and proteins (about 2-fold). The supplementation of culture medium with the nanomaterials upregulated enzymatic antioxidant biomarkers (catalase and peroxidase). The activity of the phenylalanine ammonia-lyase enzyme also displayed a similar significant upward trend in response to the nano-supplements. The CS-ZnONP treatment considerably enhanced the accumulation of alkaloids (60.5%) and soluble phenols (40%), implying stimulation in secondary metabolism. The micropropagation test revealed that the CS-ZnONP treatment improved the organogenesis performance. Overall, the nanocomposite can be considered a highly potent biocompatible elicitor.

Nitric oxide and selenium nanoparticles confer changes in growth, metabolism, antioxidant machinery, gene expression, and flowering in chicory (Cichorium intybus L.): potential benefits and risk assessment.

This experiment was conducted to provide a better insight into the plant responses to nitric oxide (NO) and selenium nanoparticle (nSe). Chicory seedlings were sprayed with nSe (0, 4, and 40 mg l-1), and/or NO (0 and 25 µM). NO and/or nSe4 improved shoot and root biomass by an average of 32%. The nSe40 adversely influenced shoot and root biomass (mean = 26%), exhibiting moderate toxicity partly relieved by NO. The nSe and NO treatments transcriptionally stimulated the dehydration response element B1A (DREB1A) gene (mean = 29.6-fold). At the transcriptional level, nSe4 or NO moderately upregulated phenylalanine ammonia-lyase (PAL) and hydroxycinnamoyl-CoA quinate transferase (HCT1) genes (mean = sevenfold). The nSe4 + NO, nSe40, and nSe40 + NO groups drastically induced the expression of PAL and HCT1 genes (mean = 30-fold). With a similar trend, hydroxycinnamoyl-CoA Quinate/shikimate hydroxycinnamoyl transferase (HQT1) gene was also upregulated in response to nSe and/or NO (mean = 25-fold). The activities of nitrate reductase and catalase enzymes were also induced in the nSe- and/or NO-treated seedlings. Likewise, the application of these supplements associated with an increase in ascorbate concentration (mean = 31.5%) reduced glutathione (mean = 35%). NO and/or nSe enhanced the PAL activity (mean = 36.4%) and soluble phenols (mean = 40%). The flowering was also influenced by the supplements in dose and compound dependent manner exhibiting the long-time responses. It appears that the nSe-triggered signaling can associate with a plethora of developmental, physiological, and molecular responses at least in part via the fundamental regulatory roles of transcription factors, like DREB1A as one the most significant genes for conferring tolerance in crops.

Effect of ß-carotene on the differentiation potential of ciliary epithelium-derived MSCs isolated from mouse eyes on alginate-based scaffolds.

Retinal degenerative diseases are considered a major challenge all over the world, and stem cell therapy is a promising approach to restore degenerative cells due to RD. MSCs are multipotent stem cells found in a variety of tissues. They are capable of differentiating into various retinal cell types, so it can be a good candidate for various degenerative disorders like retinal degenerations. ß-carotene is an antioxidant that could accelerate the stem cell differentiation while using the proper scaffold. In this study, we evaluated the effect of ß-carotene on the differentiation potential of ciliary epithelium-derived MSCs isolated from mouse eyes on alginate-based scaffolds. MSCs were isolated from mouse ciliary epithelium, cultured in DMEM medium supplemented with 10% FBS, and identified by detecting their surface antigens. Three 3D culture systems, alginate, alginate/gelatin, and gelatin hydrogels were prepared, and their structures were checked via SEM. MSCs were cultured on 3D and 2D culture system scaffolds following treated with differentiation medium containing 50 µM ß-mercaptoethanol, 1 × minimum essential medium-nonessential amino acids and 20% of knockout serum replacement and ß-carotene. MSCs viability and differentiation ability were examined by MTT and ICC, respectively. The expression changes of several retinal specific genes (Nestin, RPE65, and Rhodopsin) were also evaluated by qPCR. Over 80% of cells isolated from mouse ciliary epithelium were positive for MSC-specific markers. The viability rates of MSCs grown on all alginate-based scaffolds were above 70%. MSCs cultured on alginate-based scaffold in the differentiation medium containing ß-carotene expressed higher levels of rhodopsin protein compared to a 2D culture. Also, the expressions of Nestin, Rhodopsin, and RPE65 genes were upregulated in ß-carotene-treated MSCs grown on alginate-based scaffolds. Our results indicate that the addition of ß-carotene to the differentiation medium, along with applying alginate-based scaffolds, could induce higher differentiation in mouse ciliary epithelium-derived MSCs into specialized retinal cells.

Comparative efficacy of selenate and selenium nanoparticles for improving growth, productivity, fruit quality, and postharvest longevity through modifying nutrition, metabolism, and gene expression in tomato; potential benefits and risk assessment.

This study attempted to address molecular, developmental, and physiological responses of tomato plants to foliar applications of selenium nanoparticles (nSe) at 0, 3, and 10 mgl-1 or corresponding doses of sodium selenate (BSe). The BSe/nSe treatment at 3 mgl-1 increased shoot and root biomass, while at 10 mgl-1 moderately reduced biomass accumulation. Foliar application of BSe/nSe, especially the latter, at the lower dose enhanced fruit production, and postharvest longevity, while at the higher dose induced moderate toxicity and restricted fruit production. In leaves, the BSe/nSe treatments transcriptionally upregulated miR172 (mean = 3.5-folds). The Se treatments stimulated the expression of the bZIP transcription factor (mean = 9.7-folds). Carotene isomerase (CRTISO) gene was transcriptionally induced in both leaves and fruits of the nSe-treated seedlings by an average of 5.5 folds. Both BSe or nSe at the higher concentration increased proline concentrations, H2O2 accumulation, and lipid peroxidation levels, suggesting oxidative stress and impaired membrane integrity. Both BSe or nSe treatments also led to the induction of enzymatic antioxidants (catalase and peroxidase), an increase in concentrations of ascorbate, non-protein thiols, and soluble phenols, as well as a rise in the activity of phenylalanine ammonia-lyase enzyme. Supplementation at 3 mgl-1 improved the concentration of mineral nutrients (Mg, Fe, and Zn) in fruits. The bioaccumulated Se contents in the nSe-treated plants were much higher than the corresponding concentration of selenate, implying a higher efficacy of the nanoform towards biofortification programs. Se at 10 mgl-1, especially in selenate form, reduced both size and density of pollen grains, indicating its potential toxicity at the higher doses. This study provides novel molecular and physiological insights into the nSe efficacy for improving plant productivity, fruit quality, and fruit post-harvest longevity.

Selenium nanoparticles induced variations in growth, morphology, anatomy, biochemistry, gene expression, and epigenetic DNA methylation in Capsicum annuum; an in vitro study.

This study aimed to explore whether supplementation of the culture medium with selenium nanoparticles (nSe) can influence growth, biochemistry, expression of transcription factors, and epigenetic DNA methylation in Capsicum annuum. The seeds were grown in hormone-free MS culture medium supplemented with nSe (0, 0.5, 1, 10, and 30 mgL-1) or corresponding doses of bulk type selenate (BSe). Incorporation of nSe into the medium caused variations in morphology and growth in a manner dependent on the dose and Se type. The low doses of nSe displayed growth-promoting effects, whereas nSe at 10 and 30 mgL-1 were associated with severe toxicity and abnormality in leaf and root development. MSAP analysis confirmed the substantial variation in cytosine DNA methylation in response to the toxic dose of nSe exhibiting epigenetic modification. The nSe toxicity was associated with DNA hyper-methylations. The nSe treatments transcriptionally upregulated the bZIP1 transcription factor by an average of 3.5 folds. With a similar trend, the upregulation (mean = 9.8 folds) in the expression of the WRKY1 transcription factor resulted from the nSe application. The nSe0.5 or nSe1 treatments resulted in a significant induction (mean = 48%) in nitrate reductase activity. A high dose of nSe led to an increase in proline concentration. The nSe treatments were also associated with modifications in activities of peroxidase and catalase enzymes. Besides, the nSe utilization increased the activity of phenylalanine ammonia-lyase enzyme (mean = 76%) and concentrations of soluble phenols (mean = 51%). The toxic dose of nSe also caused abnormalities in the structure of the stem apical meristem. The nSe toxicity was also associated with inhibition in the differentiation of xylem tissues. These findings provide novel insights into the nSe-associated molecular variations in conferring the modified growth, anatomy, and metabolism.

Red elemental selenium nanoparticles mediated substantial variations in growth, tissue differentiation, metabolism, gene transcription, epigenetic cytosine DNA methylation, and callogenesis in bittermelon (Momordica charantia); an in vitro experiment.

To gain a better insight into the selenium nanoparticle (nSe) benefits/toxicity, this experiment was carried out to address the behavior of bitter melon seedlings to nSe (0, 1, 4, 10, 30, and 50 mgL-1) or bulk form (selenate). Low doses of nSe increased biomass accumulation, while concentrations of 10 mgL-1 and above were associated with stem bending, impaired root meristem, and severe toxicity. Responses to nSe were distinct from that of bulk in that the nano-type exhibited a higher efficiency to stimulate growth and organogenesis than the bulk. The bulk form displayed higher phytotoxicity than the nano-type counterpart. According to the MSAP-based analysis, nSe mediated substantial variation in DNA cytosine methylation, reflecting the epigenetic modification. By increasing the concentration of nSe, the expression of the WRKY1 transcription factor linearly up-regulated (mean = 7.9-fold). Transcriptions of phenylalanine ammonia-lyase (PAL) and 4-Coumarate: CoA-ligase (4CL) genes were also induced. The nSe treatments at low concentrations enhanced the activity of leaf nitrate reductase (mean = 52%) in contrast with the treatment at toxic concentrations. The toxic concentration of nSe increased leaf proline concentration by 80%. The nSe supplement also stimulated the activities of peroxidase (mean = 35%) and catalase (mean = 10%) enzymes. The nSe-treated seedlings exhibited higher PAL activity (mean = 39%) and soluble phenols (mean = 50%). The nSe toxicity was associated with a disrupted differentiation of xylem conducting tissue. The callus formation and performance of the explants originated from the nSe-treated seedlings had a different trend than that of the control. This experiment provides new insights into the nSe-associated advantage/ cytotoxicity and further highlights the necessity of designing convincing studies to introduce novel methods for plant cell/tissue cultures and agriculture.

Multi-walled carbon nanotubes improved growth, anatomy, physiology, secondary metabolism, and callus performance in Catharanthus roseus: an in vitro study.

This study was conducted to monitor the physiological and molecular responses of Catharanthus roseus (rose periwinkle) to multi-walled carbon nanotube (MWCNT) incorporation into the culture medium. The seeds were grown on hormone-free MS medium supplemented with 0, 50, 100, and 150 mgL-1of MWCNT. The supplementations of culture medium with MWCNTs led to significant increases in plant growth indexes such as leaf width, leaf area, leaf fresh weight, root length, and total plant biomass). Slight increases were also observed in chlorophyll a (Chla), Chlb, and carotenoid contents (mean = 18.6%) in MWCNT-treated seedlings. Protein concentrations increased by an average of 34% relative to the control. The application of MWCNT resulted in twofold increases in the catalase and peroxidase activities. A similar trend was also observed in the phenylalanine ammonia lyase activities (by an average of 36.5%), soluble phenols (by 23%), and alkaloids (by 1.7-fold). Moreover, upregulations (mean = 37-fold) in the transcriptions of the DAT gene resulted from the MWCNT supplementations. Exposure to MWCNT improved cell sizes and xylem conducting tissue in treated seedlings. The applications of MWCNTs also stimulated the callus initiation and performance, implying their effects on proliferation and possible differentiation. This study has provided evidence of role MWCNT play in improving plant performance and production of pharmaceutical secondary metabolites.

Differential physiology and expression of phenylalanine ammonia lyase (PAL) and universal stress protein (USP) in the endangered species Astragalus fridae following seed priming with cold plasma and manipulation of culture medium with silica nanoparticles.

KEY MESSAGE: Seed priming with cold plasma in combination with manipulation of culture medium with silica nanoparticle provokes anatomical, physiological and molecular changes, thereby reinforcing the plant growth and protection. ABSTRACT: This study addressed responses of Astragalus fridae to seed priming with cold plasma (0.84 W/cm2; 0, 30, 60, and 90 s) and applications of SiO2 nanoparticle (nSi; 0, 5, 40, and 80 mgl-1) in culture medium (an in vitro study). FE-SEM confirmed nSi uptake and translocation. Bulk Si at high concentrations reduced biomass accumulation (mean = 45%), while nSi did not make significant differences. The growth-enhancing effects of plasma by 41.5% were promoted by the nSi supplementation and reached 71%. Plasma did not make significant changes in Chla, while led to the slightly higher (mean = 14%) Chlb. The presence of nSi at high doses caused slight reductions in Chlb (mean = 25%) which were mitigated by plasma. The plasma and/or nSi treatments modified activities of phenylalanine ammonia lyase (PAL) in both roots (mean = 32%) and leaves (mean = 44%). With a similar trend, both individual and combined treatments of plasma and nSi provoked inductions in peroxidase activities in roots and leaves. The simultaneous treatments of plasma and nSi had the highest expression rates of PAL gene. The individual treatments of plasma did not make a significant difference in the expression of universal stress protein (USP) gene, whereas the nSi-treated seedlings exhibited the higher expression rates of USP. Leaf thicknesses and development of the vascular system (xylem and phloem) were reinforced in response to plasma and nSi. The findings provide evidence on potential benefits and phytotoxicity of nSi and plasma which may be employed as a theoretical basis for possible exploitation.

Differential growth, nutrition, physiology, and gene expression in Melissa officinalis mediated by zinc oxide and elemental selenium nanoparticles.

Regarding the rapid progress in the production and consumption of nanobased products, this research considered the behavior of Melissa officinalis toward zinc oxide nanoparticles (nZnO), nanoelemental selenium (nSe), and bulk counterparts. Seedlings were irrigated with nutrient solution containing different doses of nZnO (0, 100, and 300 mg l-1) and/or nSe (0, 10, and 50 mg l-1). The supplements made changes in growth and morphological indexes in both shoot and roots. The mixed treatments of nSe10 and nZnO led to a drastic increase in biomass, activation of lateral buds, and stimulations in the development of lateral roots. However, the nSe50 reduced plants' growth (45.5%) and caused severe toxicity which was basically lower than the bulk. Furthermore, the nSe and nZnO improved K, Fe, and Zn concentrations in leaves and roots, except for seedlings exposed to nSe50 or BSe50. Moreover, the nSe and nZnO supplementations in a dose-dependent manner caused changes in leaf non-protein thiols (mean = 77%), leaf ascorbate content (mean = 65%), and soluble phenols in roots (mean = 28%) and leaves (mean = 61%). In addition, exposure to nZnO and/or nSe drastically induced the expression of rosmarinic acid synthase (RAS) and Hydroxy phenyl pyruvate reductase (HPPR) genes. Besides, the nSe, nZnO, or bulk counterparts influenced the activities of nitrate reductase in leaves and peroxidase in roots, depending on dose factor and compound form. The comparative physiological and molecular evidence on phytotoxicity and potential advantages of nSe, nZnO, and their bulk counterparts were served as a theoretical basis to be exploited in food, agricultural, and pharmaceutical industries.

Phytochemical and Morphological Evidences for Shikonin Production by Plant Cell Cultures of Onosma sericeum Willd

ABSTRACT Shoot regeneration, callus growth, and biosynthesis of shikonin in callus cultures of Onosma sericeum were examined. Plant tissue culture was used as an alternative method for increasing the production of shikonin, a secondary metabolite. The isolated cultures were subjected to abiotic factors such as light, plant growth regulators, and nutritional factors. Identification was carried out by High- Performance Liquid Chromatography (HPLC) after 10th subculture. Nodal explants were incubated in Murashige and Skoog (MS) medium along with different combination of growth hormones. Shoot regeneration from calli were achieved on MS basal medium supplemented with 3 mg/l 6-benzylaminopurine (BAP) and 0.5 mg/l Naphthalene acetic acid (NAA) under light cycle. Shikonin was formed in dark culture. Calli grown on MS (ammonium ion-free) medium supplemented with 3 mg/l BAP and 0.5 mg/l NAA contained the maximum shikonin level (15.26 µg/mg DW). Minimum shikonin content (9.85 µg/mg DW) was observed in calli cultured on MS (ammonium ion-free) medium supplemented with 3 mg/l BAP and 0.5 mg/l indole-3-acetic acid (IAA). In establishing cell culture, the ammonium ion, and light cycle inhibited shikonin formation. This is the first report on the establishment of isolated cultures of O. sericeum for shikonin production and callus growth.

The ontogenetic trends of microtuber formation in potato (Solanum tuberosum L.).

The main aim of this investigation is the understanding of how microtubers are formed out of auxiliary buds in induced conditions. In the induced buds, meristematical cells with high cytoplasmic and nuclear stain ability expand deeper into the inner sections of the buds, comparing with non-induced buds, which the area is restricted to the apical regions. The first sign of microtuberization is the increase in size of cortical parenchyma cells in lower section and also increase in mitosis divisions in inner sections of the meristems. Most of the growth rates occur in the induced medium along the length and the width of the cells. It is also considered that the diametric growth of the tubers and the base of the leaves on the tubers begin their radial growth. The cortical parenchyma cells begin forming amidon grains during their vacuolar extension at extending the internodes much earlier than pith parenchyma cells. The extension of cells in sub-apical region plays an important role in the longitudinal growth of tubers. In the first stages, the growth of tubers results from the change in the dimensions of cortical and pith parenchyma cells, due to the reproduction of apical meristem and later, mainly from the growth of the productive tissue in the pith parenchyma. Longitudinal growth is initially grater than the growth in diameter; however with shift in the position of vacuoles and their arrangement across rather than along the tubers, the growth pattern begins to change and lateral growth catches on and exceeds longitudinal growth. In fully grown tubers, pith parenchyma cells are larger than the cells in cortical parenchyma.

Shoot micropropagation and microtuberization in potato (Solanum tuberosum L.) by the semi-continuous bioreactor.

The aim of this investigation is introducing the use of continuous and semi-continuous bioreactors and their functions at shoot multiplication and microtuberization of potato. The study shows that the explants have several nodes and when they are suspended continuously under the liquid culture medium, the shoot micropropagations and microtuberization was inhibited. The surfaces of the explants were formed callus and subsequently, they were died. However, in the semicontinuous bioreactor, with the periodical pumping of the nutritional medium the explants aren't continuously suspended under the nutritional solution. The shoot micropropagation, the leaf growth and, the root formation are suitable. In the microtuber inductive medium, the numerous of the tubers are induced. They were sessile tubers. The dormancy of the tubers are long-term, spourting after 3-4 months in the room conditions.