Foliar application of Phenylalanine functionalized multi-walled carbon nanotube improved the content of volatile compounds of basil grown in greenhouse.

Carbon nanotubes are among the elicitors that have different effects on plants. Basil as a useful and valuable plant has significant medicinal properties; The aim of this research is to study the effect of different concentrations of functionalized multi-walled carbon nanotubes with phenylalanine and non-functionalized in concentrations of (0, 50, 100, 150 and 200 mg.l-1) and activated carbon on total phenol and flavonoid content, antioxidant capacity, the content of H2O2, reactive oxygen species detection, antioxidant enzyme activity, and the concentration of volatile compounds of basil in the greenhouse culture, in an experiment in the form of a completely randomized design with three replications, and in the faculty of sciences of Urmia university's laboratory. The highest content of total phenol, flavonoid, anthocyanin, antioxidant capacity and hydrogen peroxide content were observed in the 200 mg.l-1 functionalized carbon nanotube. The highest percentage of alpha-Copaene, trans-alpha-Bergamotene, alpha-Guaiene, Bicyclogermacrene, 1,10-di-epi-Cubenol and alpha-Eudesmol compounds at 150 mg.l-1 of functionalized carbon nanotube and the highest percentage of compounds 1,8-cineole and eugenol was observed at 100 mg.l-1 of functionalized carbon nanotube. The compounds of linalool, camphor and anethole also showed their highest amount in treatments of 200, 150 and 50 mg.l-1 of carbon nanotube, respectively. In general, the observations of this research indicated that the use of functionalized carbon nanotubes as a stimulant has increased the antioxidant capacity of basil and on the other hand, it has led to an improving in the content of secondary metabolites.

Response of Saponaria officinalis L. hairy roots to the application of TiO2 nanoparticles in terms of production of valuable polyphenolic compounds and SO6 protein.

Saponaria officinalis L. is a perennial plant from the Caryophyllaceae family whose various parts are used in traditional medicine as the treatment agent of skin diseases, blood purifier, diuretic, sudorific, and bile purifier. The cultivation system of hairy roots is a proper alternative for improving the valuable pharmaceutical compounds production compared to other in-vitro methods. The extensive nanotechnology applications in hairy roots cultivation is a sustainable production foundation to produce such active elements. In this study, the effect of various concentrations of titanium dioxide nanoparticles (TiO2 NPs) (0, 10, 20, 30, 50 mg L-1) with two treatments (24 and 48 h) was examined on the growth level, antioxidant capacity, total phenol and flavonoid contents, antioxidant enzyme activities, certain polyphenol compounds and SO6 protein in hairy roots of S. officinalis. According to the results, the maximum (3.09 g) and minimum (0.96 g) fresh weight (FW) of hairy roots were observed in treated culture media with 10 and 20 mg L-1 of TiO2 NPs after 24 and 48 h of exposure times, respectively. The highest rate of total phenol (9.79 mg GLA g-1 FW) and total flavonoid contents (1.06 mg QE g-1 FW) were obtained in the treated hairy roots with 50 and 30 mg L-1 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-1 concentration showed the highest production level of SO6 protein.

Inductive effect of titanium dioxide nanoparticles on the anticancer compounds production and expression of rosmarinic acid biosynthesis genes in Dracocephalum kotschyi transformed roots.

Methoxylated flavonoids, mainly xanthomicrol and cirsimaritin that can be extracted from Dracocephalum kotschyi Boiss, have anticancer, antispasmodic and antiplatelet effects. The production of these valuable pharmaceutical compounds is one of the major goals of biotechnology studies. In this work, induced transformed roots were influenced by various concentrations of titanium dioxide nanoparticles (TiO2 NPs) at 24 or 48 h exposure time. The effects of TiO2 NPs were assessed on growth rate, activity of antioxidant enzymes, total phenol and flavonoid content (TPC and TFC) and rosmarinic acid (RA) and some flavonoids accumulation. The gene expression level of phenylalanine ammonia-lyase (pal) and rosmarinic acid synthase (ras) genes were assessed by real time PCR analysis. The transformed roots biomass was substantially increased in elicited roots in comparison with the control. The TPC, TFC and antioxidant enzymes activitywere affected by TiO2 NPs concentration and exposure time. Valuable flavonoids with anticancer characteristics along with xanthomicrol, cirsimaritin and isokaempferide exhibited an increase (70, 34.28 and 7.81-fold, respectively) versus the control. The maximum content of RA (530.5 µg g-1 FW), which was 4.30 times as great as that of control was detected in samples treated with TiO2 NPs (50 mg L-1) 24 h after elicitation. Real-time PCR analysis revealed a considerable increase in pal and ras expression rate engaged by TiO2 NPs levels and exposure time. Overall D. kotschyi transformed roots elicitation by TiO2 NPs led to a massive increment in the production of valuable anticancer flavonoids such as xanthomicrol, cirsimaritin and RA as polyphenol.

Effect of Polyploidy Induction on Natural Metabolite Production in Medicinal Plants.

Polyploidy plays an important role in plant diversification and speciation. The ploidy level of plants is associated with morphological and biochemical characteristics, and its modification has been used as a strategy to alter the quantitative and qualitative patterns of secondary metabolite production in different medicinal plants. Polyploidization can be induced by many anti-mitotic agents, among which colchicine, oryzalin, and trifluralin are the most common. Other variables involved in the induction process include the culture media, explant types, and exposure times. Due to the effects of polyploidization on plant growth and development, chromosome doubling has been applied in plant breeding to increase the levels of target compounds and improve morphological characteristics. Prompted by the importance of herbal medicines and the increasing demand for drugs based on plant secondary metabolites, this review presents an overview of how polyploidy can be used to enhance metabolite production in medicinal plants.

Improved biotechnological production of paclitaxel in Taxus media cell cultures by the combined action of coronatine and calix[8]arenes.

Paclitaxel (PTX), a widely used anticancer agent, is found in the inner bark of several Taxus species, although at such low levels that its extraction is ecologically unsustainable. Biotechnological platforms based on Taxus sp. cell cultures offer an eco-friendlier approach to PTX production, with yields that can be improved by elicitation. However, the also limited excretion of target compounds from the producer cells to the medium hampers their extraction and purification. In this context, we studied the effect of treating T. media cell cultures with the elicitor coronatine (COR) and calix[8]arenes (CAL), nanoparticles that can host lipophilic compounds within their macrocyclic scaffold. The highest taxane production (103.5 mg.L-1), achieved after treatment with COR (1 µM) and CAL (10 mg.L-1), was 15-fold greater than in the control, and PTX represented 82% of the total taxanes analyzed. Expression levels of the flux-limiting PTX biosynthetic genes, BAPT and DBTNBT, increased after the addition of COR, confirming its elicitor action, but not CAL. The CAL treatment significantly enhanced taxane excretion, especially when production levels were increased by COR; 98% of the total taxanes were found in the culture medium after COR + CAL treatment. By forming complexes with PTX, the nanoparticles facilitated its excretion to the medium, and by protecting cells from PTX toxicity, its intra-and extra-cellular degradation may have been avoided. The addition of COR and CAL to T. media cell cultures is therefore a bio-sustainable and economically viable system to improve the yield of this important anticancer compound.

Improved tropane alkaloid production and changes in gene expression in hairy root cultures of two Hyoscyamus species elicited by silicon dioxide nanoparticles.

Species of Hyoscyamus are rich sources of medicinally important tropane alkaloids, which have anticholinergic, antispasmodic and sedative effects and are competitive inhibitors of acetylcholine. The application of nanotechnology and nanomaterials for elicitation is rapidly expanding and recent research indicates that silicon dioxide nanoparticles (SiO2 NPs) can be used as an efficient elicitor to increase the production of hyoscyamine and scopolamine in Hyoscyamus species. Thus, in this work, the effect of SiO2 NPs (0, 25, 50, 100 and 200 mg L-1) with two treatment times (24 and 48 h) on the growth rate, total phenol and flavonoid content (TPC, TFC), antioxidant enzyme activity, tropane alkaloid yield and pmt (putrescine N-methyltransferase) and h6h (hyoscyamine 6-hydroxylase) gene expression levels in hairy roots of two Hyoscyamus species (H. reticulatus and H. pusillus) was investigated. The highest TPC and TFC accumulation was obtained in H. reticulatus elicited by SiO2 NPs (100 and 200 mg L-1), respectively, at 24 h of treatment. High-performance liquid chromatography (HPLC) revealed the highest amount of hyoscyamine (140.15 µg g-1 FW) and scopolamine (67.71 µg g-1 FW) accumulated in H. reticulatus transformed roots treated with 100 mg L-1 SiO2 NPs at 24 h, with a respective increase of 1212% and 272% compared to non-treated roots. In H. pusillus, the highest hyoscyamine (7.42 µg g-1 FW) and scopolamine (15.56 µg g-1 FW) production (about 82% and 241% higher, respectively, compared to the lowest amounts) was achieved with 25 and 100 mg L-1 SiO2 NPs, respectively, at 48 h of treatment. Semi-quantitative RT-PCR analysis determined the highest expression level of pmt and h6h genes in H. reticulatus transformed roots supplemented with 100 mg L-1 SiO2 NPs.

Iron oxide nanoparticles: a novel elicitor to enhance anticancer flavonoid production and gene expression in Dracocephalum kotschyi hairy-root cultures.

BACKGROUND: Dracocephalum kotschyi Boiss. is a valuable source of rosmarinic acid (RA) and methoxylated hydroxyflavones (such as xanthomicrol and cirsimaritin) with antioxidative and antiplatelet effects and with antiproliferative potential against various cancer cells. The extensive application of nanotechnology in hairy root cultures is a new sustainable production platform for producing these active constituents. In the present study, hairy roots derived from 4-week-old leaves and Agrobacterium rhizogenes strain ATCC15834 were used to investigate the impact of various concentrations of iron oxide nanoparticles (Fe NPs) in two elicitation time exposures (24 and 48 h) on growth, antioxidant enzyme activity, total phenolic and flavonoid content (TPC and TFC), and some polyphenols. Gene expression levels of phenylalanine ammonia-lyase (pal) and rosmarinic acid synthase (ras) were also analyzed. RESULTS: Iron nanoparticles enhanced biomass accumulation in hairy roots. The treatment time and Fe NP dosage largely improved the activity of antioxidant enzymes, TPC and TFC. The highest RA (1194 µg g-1 FW) content (9.7-fold), compared to controls, was detected with 24 h of exposure to 75 mg L-1 Fe NP, which was consistent with the expression of pal and ras genes under the influence of elicitation. The xanthomicrol, cirsimaritin, and isokaempferide content was increased 11.87, 3.85, and 2.27-fold, respectively. CONCLUSION: Stimulation of D. kotschyi hairy roots by Fe NPs led to a significant increase in the induction and production of important pharmaceutical compounds such as rosmarinic acid and xanthomicrol. © 2019 Society of Chemical Industry.

Fruit phytochemical composition and color parameters of 21 accessions of five Rosa species grown in North West Iran.

BACKGROUND: The genus Rosa comprises economically important horticultural plants belonging to the family Rosaceae. Recently, the use of different Rosa species has increased owing to their multipurpose properties (ornamental, food and medicinal uses). In this study, 21 accessions of Rosa genotypes were compared for fruit phytochemical composition and color parameters. RESULTS: The highest antioxidant activity (37.86 mg AAE g-1 FW) and total phenolic (8.17 mg GAE g-1 FW), total flavonoid (2.53 mg QUE g-1 FW), total carotenoid (20.21 mg g-1 FW) and ascorbic acid (84.27 mg g-1 FW) contents were observed in G20 (R. canina), G8 (R. canina), G9 (R. canina), G5 (R. damascena) and G10 (R. moschata) respectively. Chlorogenic acid and gallic acid were found as the main phenolic constituents of Rosa fruits. High amounts of apigenin, rutin, quercetin, p-coumaric acid, cinnamic acid, chlorogenic acid, caffeic acid and gallic acid were obtained in fruit extracts of G6, G14, G6, G8, G19, G9, G19 and G12 respectively. Moreover, the level of color parameters also varied among genotypes. The highest values of a*, b*, L* and chroma were obtained in G4 (R. canina). Based on hierarchical clustering analysis with heat-map, five groups of accessions were identified. CONCLUSION: Different Rosa genotypes are rich in certain phytochemical compounds, with significant variations in their levels being observed. Hence evaluation of Rosa genetic resources can supply valuable data for screening accessions containing high levels of individual phenolics, antioxidants and other bioactive compounds for use in breeding programs and food and pharma industries. © 2019 Society of Chemical Industry.

Classification of barberry genotypes by multivariate analysis of biochemical constituents and HPLC profiles.

INTRODUCTION: Recently, there has been a growing interest in the use of edible barberry and their extracts as a source of natural antioxidants in food and pharmaceutical industries. OBJECTIVE: The aim of this study was to evaluate the biochemical constituents of 18 samples of barberry fruits and classification of barberry genotypes by multivariate analysis. METHODS: Total phenolic, total flavonoid, total anthocyanin, total tannin, total carbohydrate contents and antioxidant activity were determined using Folin-Ciocalteu, aluminum chloride, colorimetric, vanillin, anthron and DPPH (2,2'-diphenyl-1-picrylhydrazyl) assays, respectively. High-performance liquid chromatography (HPLC) system is used for quantitative determination of phytochemical constituents. The multivariate data analysis (principal component analysis and hierarchical cluster analysis) and heat map data visualisation techniques were performed to classify barberry genotypes using Minitab and GraphPad Prism software, respectively. RESULTS: The highest amounts of total phenolics and flavonoids were obtained in fruit extracts of G3 (Berberis vulgaris). The highest total anthocyanin content and antioxidant activity were observed in G8 (B. vulgaris) and G16 (B. vulgaris), respectively. HPLC analysis of phytochemicals (gallic acid, caffeic acid, chlorogenic acid, p-coumaric acid, cinnamic acid, rutin, apigenin, and quercetin) revealed that gallic acid and p-coumaric acid were found as the most abundant phytochemical compounds. Based on multivariate analysis and heat map visualisation techniques, Berberis genotypes were classified into three main clusters. CONCLUSIONS: These results showed that barberry species (especially B. vulgaris and B. carataegina) are promising sources of natural antioxidants and biochemical compounds beneficial to be used in the food industry and that the multivariate analysis was a suitable approach to classify the barberry samples.

Influence of nano-zinc oxide on tropane alkaloid production, h6h gene transcription and antioxidant enzyme activity in Hyoscyamus reticulatus L. hairy roots.

The use of nanotechnology and biotechnology to improve the production of plant bioactive compounds is growing. Hyoscyamus reticulatus L. is a major source of tropane alkaloids with a wide therapeutic use, including treatment of Parkinson's disease and to calm schizoid patients. In the present study, hairy roots were obtained from two-week-old cotyledon explants of H. reticulatus L. using the A7 strain of Agrobacterium rhizogenes. The effects of different concentrations of the signaling molecule nano-zinc oxide (ZnO) (0, 50, 100 and 200 mg/L), with three exposure times (24, 48 and 72 h), on the growth rate, antioxidant enzyme activity, total phenol contents (TPC), tropane alkaloid contents and hyoscyamine-6-beta-hydroxylase (h6h) gene expression levels were investigated. Growth curve analysis revealed a decrease in fresh and dry weight of ZnO-treated hairy roots compared to the control. ANOVA results showed that the antioxidant activity of the enzymes catalase, guaiacol peroxidase and ascorbate peroxidase was significantly higher in the ZnO-treated hairy roots than in the control, as was the TPC. The highest levels of hyoscyamine (37%) and scopolamine (37.63%) were obtained in hairy roots treated with 100 mg/L of ZnO after 48 and 72 h, respectively. Semi-quantitative RT-PCR analysis revealed the highest h6h gene expression was in hairy roots treated with 100 mg/L of ZnO after 24 h. It can be concluded that ZnO is as an effective elicitor of tropane alkaloids such as hyoscyamine and scopolamine due to its enhancing effect on expression levels of the biosynthetic h6h gene.

Sodium nitroprusside stimulated production of tropane alkaloids and antioxidant enzymes activity in hairy root culture of Hyoscyamus reticulatus L.

Hyoscyamus reticulatus L. is a herbaceous biennial belonging to the solanaceae family. Hyoscyamine and scopolamine as main tropane alkaloids accumulated in henbane are widely used in medicine to treat diseases such as parkinson's or to calm schizoid patients. Hairy roots media manipulation which uses elicitors to activate defense mechanisms is one of the main strategies for inducing secondary metabolism as well as increasing the production of valuable metabolites. Cotyledon-derived hairy root cultures were transformed by Agrobacterium rhizogenes. Sodium nitroprusside (SNP), a nitric oxide donor), was used in various concentrations (0, 50, 100, 200 and 300 µM) and exposure times (24 and 48 h). Treatment with SNP led to a significant reduction in fresh and dry weight of hairy roots, compared to control cultures. ANOVA results showed that elicitation of hairy root cultures with SNP at different concentrations and exposure times significantly affected the activity of as antioxidant enzymes such as catalase (CAT), peroxidase (POD) and ascorbate peroxidase (APX). The highest hyoscyamine and scopolamine production (about 1.2-fold and 1.5-fold increases over the control) was observed at 50 and 100 µM SNP at 48 and 24 hours of exposure time, respectively. This is the first report of SNP elicitation effects on the production of tropane alkaloids in hairy root cultures.

Phytochemical Composition and Antioxidant Activity of Petals of Six Rosa Species from Iran.

Background: The petals of Rosa species are used in the food industry and various traditional medicinal products, but few studies exist on the phytochemical composition and antioxidant activity of petals of Rosa L. species grown in Iran. Objective: Phytochemical characteristics and antioxidant activity and some phenolic compounds of petals of six Rosa L. species were studied. Methods: Total phenolic content, total flavonoid content, and antioxidant activity were determined using Folin-Ciocalteu reagent, aluminum chloride method, ferric-reducing antioxidant power (FRAP), and 2,2-diphenyl-1-picrylhydrazyl (DPPH) assays, respectively. An HPLC system was used for quantitative analysis of phytochemical compounds. Hierarchical cluster analysis (HCA) and principal component analysis (PCA) were performed among the variables analyzed using Minitab software. Also, heat maps were used to visualize phytochemical characteristics and antioxidant activity in each species using GraphPad Prism software. Results: The amount of total phenol content, total flavonoid content, and antioxidant activity were in the range of 25.13-52.01 mg gallic acid equivalents/g dry weight (DW), 0.61-0.82 mg quercetin equivalents/g DW, 11.47-20.93 µmol Fe++/g DW (FRAP), and 31.66-74.44% (DPPH), respectively. The p-coumaric acid (647.28 µg/g DW) and chlorogenic acid (24.37-135.23 µg/g DW) were found to be the most abundant phenolic compounds in the extracts of rose petals. The HCA and PCA revealed three distinct categories of species based on phytochemical composition and antioxidant activity. Conclusions: These results showed that phytochemical characteristics of different rose species widely correlated with species type and are promising sources of natural antioxidants beneficial for use in the food or pharmaceutical industries. Highlights: Iran is one of the main centers for genetic diversity of Rosa L. The petals of Rosa species are used in the food industry and various traditional medicinal products, but few studies exist on the phytochemical composition and antioxidant activity of petals of Rosa L. species grown in Iran. Antioxidant activity and phytochemical compound of Six Rosa L. species petals grown in Iran were studied. Phenolic compounds in petals of Rosa were analyzed by HPLC. The color parameters, amount of total phenolic, total flavonoids, antioxidant activity and some individual phenolic compounds were significantly variable amongst Rosa species.

Enhanced production of hyoscyamine and scopolamine from genetically transformed root culture of Hyoscyamus reticulatus L. elicited by iron oxide nanoparticles.

The medicinal plant Hyoscyamus reticulatus L. is a rich source of hyoscyamine and scopolamine, the tropane alkaloids. The use of hairy root cultures has focused significant attention on production of important metabolites such as stable tropane alkaloid production. Elicitation is an effective approach to induce secondary metabolite biosynthetic pathways. Hairy roots were derived from cotyledon explants inoculated with Agrobacterium rhizogenes and elicited by iron oxide nanoparticles (FeNPs) at different concentrations (0, 450, 900, 1800, and 3600 mg L-1) for different exposure times (24, 48, and 72 h). The highest hairy root fresh and dry weights were found in the medium supplemented with 900 mg L-1 FeNPs. Antioxidant enzyme activity was significantly increased in induced hairy roots compared to non-transgenic roots. The highest hyoscyamine and scopolamine production (about fivefold increase over the control) was achieved with 900 and 450 mg L-1 FeNPs at 24 and 48 h of exposure time, respectively. This is the first report of the effect of FeNP elicitor on hairy root cultures of a medicinal plant. We suggest that FeNPs could be an effective elicitor in hairy root cultures in order to increase tropane alkaloid production.

High-frequency in vitro plantlet regeneration from apical bud as a novel explant of Carum copticum L.

OBJECTIVE: To develop an in vitro regeneration system to increase the recovery of Carum copticum L. plantlets as a part of developing a metabolic engineering program. METHODS: The efficacy of different concentrations and combinations of 6-benzyladenine, indole-3-acetic acid and indole butyric acid on direct shoot regeneration and rooting of ajowan from apical bud explants were assessed. All explants were cultured on Murashige and Skoog (MS) medium supplemented with different combinations of 6-benzyl amino purine (BAP) (0, 2.2, 4.4, 8.8 µmol/L) and indole-3-acetic acid (IAA) (0, 0.5, 1.1, 2.2 µmol/L). RESULTS: The maximum shoot regeneration frequency (97.5%) and the highest number of shoots produced from apical buds (34 shoots per explant) were obtained on MS medium fortified with BAP (4.4 µmol/L) and IAA (0.5 µmol/L). Low shoot regeneration frequency was observed in BAP free treatments. The effects of different strengths of MS medium and various concentrations of IAA and indole-3- butyric acid on rooting rate, length and average number of roots were also investigated. Application of indole-3- butyric acid (6 µmol/L) in full-strength MS medium, was more effective than IAA and resulted in highest shoot regeneration frequency with the rooting rate of 100% and highest mean number of roots per shoot (41.8). The rooted plantlets were acclimatized successfully in greenhouse conditions with a survival rate of 90%. CONCLUSION: In this study, a simple and reliable regeneration and acclimatization protocol for Carum copticum has been presented. This protocol can be found very advantageous for a variety of purposes, including mass multiplication of Carum species, medicinal plant breeding studies and transgenic plant production.

Overproduction of valuable methoxylated flavones in induced tetraploid plants of Dracocephalum kotschyi Boiss.

BACKGROUND: Ploidy manipulation is considered an efficient method to increase production potential of medicinally important compounds. Dracocephalum kotschyi Boiss. is an endangered medicinal plant of Iran. Various concentrations of colchicine (0.05, 0.10, 0.20, and 0.50% w/v) were applied to shoot apical meristems of D. kotschyi seedlings in two and four-leaf stages to induce tetraploidy. RESULTS: According to the results, 0.5% (w/v) of colchicine can be effective for polyploidy induction in D. kotschyi. Putative tetraploids were selected by morphological and microscopic characteristics and their ploidy level was confirmed by flow cytometry analysis and chromosome counting. The chromosome number of original diploid plant was confirmed to be 2n = 2× = 20 whereas that of the tetraploid plant was 2n = 4× = 40. Tetraploid and mixoploid plants showed different morphological, physiological and microscopic characteristics from those of diploid counterparts. The total content of flavonoids was increased from 1583.28 in diploids to 1890.07 (µg/g DW) in stable tetraploids. CONCLUSION: High-Performance Liquid Chromatography with Diode-Array Detection (HPLC-DAD) confirmed over accumulation of methoxylated hydroxyflavones in solid tetraploid plants of D. kotschyi.

Transient Expression of cor Gene in Papaver somniferum.

INTRODUCTION: Papaver somniferum is the commercial source of morphine and codeine. The isolation of effective genes involved in the morphine biosynthesis of P. somniferum is very important in the production of specific metabolites achieved using metabolic engi-neering techniques. In this pathway, the key enzyme COR is involved in the conversion of codeinone to codeine and morphinone to morphine. METHODS: the gene encoding of this enzyme was isolated using primers designed on the base of gene sequence available on (NCBI) for P. somniferum. This gene correct size around (960 bp) was first subcloned into pTZ57RIT vector then cloned into expression vectors (pBI121) between BamHI and SacI sites to allow the expression of cor gene driven by the cauliflower mosaic virus 35S pro-moter. The result was confirmed through different molecular methods e.g. PCR and en-zyme digestion by BamHI and SacI. The recombinant plasmid was transformed into the E. coli strain DH5α using a freeze-thaw method. Having selected positive colones on selection medium, plasmid was extracted by miniprep method and recombinant plasmids were selected based on PCR and digestion. The construct was then mobilized in Agrobacterium tumefaciens C58/pGV3850 (KmR RifR). After gene transformation to P. somniferum plants, the agroinfiltration method was also used for transient expression of COR enzyme. RESULTS: evaluation results showed that morphine and codeine were detectable in the leaves of transgenic plants containing cor transgene and there was significant difference in the final production. After completing this experiment for three times, results showed that in 11 sets from 15 sets of leaves experiment tested, main alkaloids (codeine, morphine, papaverin, noscapine and thebaine) were detectable. CONCLUSION: Whereas no signal was detected in non-infiltrated control leaves or in leaves infiltrated with non-recombinant bacteria for morphine and codeine, others such as thebaine and papaverine were detectable.