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Introduction: With increasing drought stress due to climate change and water scarcity, the agricultural sector has sought innovative strategies to mitigate the detrimental effects on crop productivity. One approach that has received significant attention is the use of fertilizers and biostimulants as potential means of alleviating drought stress., Methods: In this study, five different irrigation levels including 100% (control), 80% (slight stress), 60% (mild stress), 40% (moderate stress), and 20% (severe stress) of field capacity (FC) and seaweed extract (SWE) at three concentrations (0, 5, and 10 g/L) were applied to the pots containing one-year-old licorice ( Glycyrrhiza glabra L. ) plants in a factorial completely randomized design experiment with three replications for eight weeks., Results and Discussion: The glycyrrhizic acid content increased with water stress intensity without the application of SWE until severe (20% FC) water stress treatment. The application of 10 g/L SWE under 100% FC led to a significant increase in the glycyrrhizic acid value (32.5±0.889 mg/g DW) compared with non-SWE application (30.0±1.040 mg/g DW). The maximum glabridin content (0.270±0.010 mg/g DW) was obtained under irrigation of 20% field capacity with 10 g/L SWE application. In addition, the activity of the all studied enzymes such as APX (ascorbate peroxidase), CAT (catalase), POD (peroxidase), and SOD (superoxide dismutase) were boosted by increasing the water stress levels. The use of SWE further enhanced the increase of some of these metabolites and enzymes, which, in turn, helped the plant to tolerate stress conditions through the scavenging of more ROS (Reactive oxygen species), wherein for this purpose, the SWE 10 g/L was more effective than other concentration. The plants efficiently eliminated ROS driven from drought stress by both non-enzymatic and enzymatic systems., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Fozi, Esmaeili, Alizadeh, Eghlima and Mirjalili.)

Unfavorable environmental conditions during planting can reduce seed germination and hinder seedling growth. To address this issue, manufacturers are exploring innovative and cost-effective methods, such as cold plasma discharge. This simple, low-cost, and efficient physical technique induces significant biological responses in seeds and plants without the use of traditional, environmentally hazardous chemicals. This study investigated the impact of dielectric barrier discharge (DBD) plasma and plasma-activated water (PAW), produced by gliding arc plasma, on the germination and seedling growth of My344 cotton seeds. The seeds were pre-treated with 80 W of DBD plasma for 0, 1, 2, and 3 min, and subsequently soaked for 90 min in PAW with varying pH levels of 5.82, 3.88, 3.63, and 3.38. The results showed that plasma treatment positively influenced seed germination and seedling growth. The highest germination percentage (98.89 %) was observed with 1 min of DBD treatment, followed by PAW priming at pH levels of 3.63 and 3.38. Additionally, a 3-min DBD treatment followed by soaking in PAW with a pH of 3.63 led to significant increases in stem length (76.76 %), root length (48.77 %), and wet weight (76.44 %). Furthermore, it was observed that the electrical conductivity of the seeds in all groups decreased significantly with increased PAW acidity. The physical and chemical effects of cold DBD plasma on the seed surface, as well as changes in hydrophilicity, were further examined using scanning electron microscopy (SEM), Fourier-transform infrared spectroscopy (FTIR), and water contact angle imaging., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 Published by Elsevier Ltd.)

Fenugreeks (Trigonella L. spp.), are well-known herbs belonging to the family Fabaceae, whose fresh and dried leaves have nutritional and medicinal value. In the present study, the content of phytochemical traits (essential oil, diosgenin, trigonelline, total phenol, total flavonoid, total saponins, and total tannins), bitterness value, pigments (chlorophyll, carotenoid, β-carotene, and anthocyanin), vitamins (group B vitamins and ascorbic acid), minerals, and antioxidant activity of thirty cultivated populations belonging to ten Trigonella species were evaluated. The species and populations were significantly different in all studied parameters. A significant positive and negative correlation (p < 0.05) was also observed between the studied parameters. In total, T. teheranica, T. elliptica, and T. foenum-graecum were distinguished as superior species. The results showed that fenugreeks leaves can be considered as a valuable source of food and phytochemical compounds. The obtained data can be help to expand the inventory of wild and cultivated Trigonella species for further exploitation of rich chemotypes in the new foods and specific applications., (© 2024. The Author(s).)

Aims: To identify promising fungal endophytes that are able to produce glycyrrhizin and enhance it in licorice and the mechanisms involved., Methods and Results: Fifteen fungal endophytes were isolated from Glycyrrhiza glabra L. rhizomes among which SGGF14 and SGGF21 isolates were found to produce glycyrrhizin by 4.29 and 2.58 µg g-1 dry weight in the first generation of their culture. These isolates were identified as Fusarium solani and Alternaria tenuissima, respectively, based on morphological characteristics and sequence analysis of internal transcribed spacer, TEF1, ATPase, and CAL regions. Subsequently, G. glabra plants were inoculated with these fungal isolates to examine their effect on glycyrrhizin production, plant growth parameters and the expression of key genes involved in glycyrrhizin pathway: SQS1, SQS2, bAS, CAS, LUS, CYP88D6, and CYP72A154. Endophytes were able to enhance glycyrrhizin content by 133%-171% in the plants. Natural control (NC) plants, harboring all natural endophytes, had better growth compared to SGGF14- and SGGF21-inoculated and endophyte-free (EF) plants. Expression of SQS1, SQS2, CYP88D6, and CYP72A154 was upregulated by inoculation with endophytes. LUS and CAS were downregulated after endophyte inoculation. Expression of bAS was higher in SGGF21-inoculated plants when compared with NC, EF, and SGGF14-inoculated plants., Conclusions: Two selected fungal endophytes of G. glabra can produce glycyrrhizin and enhance glycyrrhizin content in planta by modulating the expression of key genes in glycyrrhizin biosynthetic pathway., (© The Author(s) 2024. Published by Oxford University Press on behalf of Applied Microbiology International.)

Background: Fenugreeks (Trigonella L. spp.), belonging to the legume family (Fabaceae), are well-known multipurpose crops that their materials are currently received much attention in the pharmaceutical and food industries for the production of healthy and functional foods all over the world. Iran is one of the main diversity origins of this valuable plant. Therefore, the aim of the present study was to explore vitamins, minerals, and fatty acids profile, proximate composition, content of diosgenin, trigonelline, phenolic acids, total carotenoids, saponins, phenols, flavonoids, and tannins, mucilage and bitterness value, and antioxidant activity of the seed of thirty populations belonging to the ten different Iranian Trigonella species., Results: We accordingly identified notable differences in the nutrient and bioactive compounds of each population. The highest content (mg/100 g DW) of ascorbic acid (18.67 ± 0.85‒22.48 ± 0.60) and α-tocopherol (31.61 ± 0.15‒38.78 ± 0.67) were found in the populations of T. filipes and T. coerulescens, respectively. Maximum content of catechin was found in the populations of T. teheranica (52.67 ± 0.05‒63.50 ± 0.72 mg/l). Linoleic acid (> 39.11% ± 0.61%) and linolenic acid (> 48.78 ± 0.39%) were the main polyunsaturated fatty acids, with the majority in the populations of T. stellata (54.81 ± 1.39‒63.46 ± 1.21%). The populations of T. stellata were also rich in trigonelline (4.95 ± 0.03‒7.66 ± 0.16 mg/g DW) and diosgenin (9.06 ± 0.06‒11.03 ± 0.17 mg/g DW)., Conclusions: The obtained data provides baseline information to expand the inventory of wild and cultivated Iranian Trigonella species for further exploitation of rich chemotypes in the new foods and specific applications., (© 2024. The Author(s).)

Currently, the stable, uniform, and highly efficient production of raw materials for pharmaceutical companies has received special attention. To meet these criteria and reduce harvesting pressure on the natural habitats of licorice (Glycyrrhiza glabra L.), cultivation of this valuable plant is inevitable. In the present study, to introduce the glycyrrhizic acid (GA)- and glabridin-rich genotypes from cultivated Iranian licorice, forty genotypes from eight high-potential wild populations were cultivated and evaluated under the same environmental conditions. The GA content varied from 5.00 ± 0.04 mg/g DW (TF2 genotype) to 23.13 ± 0.02 mg/g DW (I5 genotype). The highest and lowest glabridin content were found in the K2 (0.72 ± 0.021 mg/g DW) and M5 (0.02 ± 0.002 mg/g DW) genotypes, respectively. The rutin content in the leaves of the studied genotypes varied from 1.27 ± 0.02 mg/g DW in E4 to 3.24 ± 0.02 mg/g DW in BO5 genotypes. The genotypes from the Ilam population were characterized by higher vegetative growth and yield traits in the aerial parts and roots. The average root dry yield was 2.44 tons per hectare (t/ha) among the studied genotypes and a genotype from Ilam (I5) yielded the maximum value (3.08 ± 0.034 t/ha). The highest coefficient of variation among the genotypes was observed for leaf width (CV = 34.9%). The GA and glabridin-rich genotypes introduced in this study can be used in the future breeding programs to release new bred licorice cultivars., (© 2024. The Author(s).)

Drought is known to be the most important constraint to the growth and yield of agricultural products in the world, and plant symbiosis with arbuscular mycorrhizal fungi (AMF) can be a way to reduce drought stress negative impacts. A two-year experiment to investigate the factorial combination of mycorrhizal fungi (Glomus mosseae, Glomus intraradices, Control) and phosphorus fertilizer (application and non-application of phosphorus) on fruit yield and phenolic acids changes bitter gourd under different irrigation regimes as a split factorial based on a randomized complete block design. Three irrigation regimes, including irrigation after 20%, 50%, and 80% available soil water content depletion (ASWD), were considered in the main plots. The results showed that under water deficit stress, fruit yield and physiological (photosynthesis rate (Pn), transpiration rate (Tr), stomatal conductance (Gs), RWC, total chlorophyll, and root colonization) parameters decreased compared to 20% ASWD, and biochemical (proline, soluble sugar, MDA, CAT, SOD, phenol) parameters and fruit phenolic acids (caffeic acid, coumaric acid, ferulic acid) increased. However, the inoculation of AMF and phosphorus fertilizer in three irrigation regimes decreased MDA content, but physiological and biochemical parameters and fruit phenolic acids were increased. In this study, the factorial combination of AMF and sufficient phosphorus improved the resistance of bitter gourd to water deficit, and this not only improved fruit yield but also increased fruit phenolic acids under 80% ASWD, which can be an innovation in the management of water resources and the production industry of medicinal plants with high antioxidant properties in water deficit areas., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Masson SAS.)

Neurodegenerative disorders are characterized by mitochondrial dysfunction and subsequently oxidative stress, inflammation, and apoptosis that contribute to neuronal cytotoxicity and degeneration. Huntington's (HD), Alzheimer's (AD), and Parkinson's (PD) diseases are three of the major neurodegenerative diseases. To date, researchers have found various natural phytochemicals that could potentially be used to treat neurodegenerative diseases. Particularly, the application of natural phenolic compounds has gained significant traction in recent years, driven by their various biological activities and therapeutic efficacy in human health. Polyphenols, by modulating different cellular functions, play an important role in neuroprotection and can neutralize the effects of oxidative stress, inflammation, and apoptosis in animal models. This review focuses on the current state of knowledge on phenolic compounds, including phenolic acids, flavonoids, stilbenes, and coumarins, as well as their beneficial effects on human health. We further provide an overview of the therapeutic potential and mechanisms of action of natural dietary phenolics in curing neurodegenerative diseases in animal models., (© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Fenugreek (Trigonella foenum-graecum L.) is a multi-use annual forage legume crop that is widely used in food products such as syrup, bitter run, curries, stew, and flavoring. In the present study, morphological traits, proximate composition (moisture, crude fibre, protein, fat, carbohydrate, and energy value), total phenol and total flavonoid contents, and antioxidant properties of 31 Iranian agro-ecotypic populations of the plant was investigated. Among the leaf and seed samples studied, the seeds exhibited the high ash (3.94 ± 0.12%), fat (7.94 ± 0.78%), crude fibre (10.3 ± 0.25%), protein (35.41 ± 1.86%), and carbohydrate (50.5 ± 1.90%) content. In general, more energy value (kcal/100 g) was also obtained from the seed (318.88 ± 1.78-350.44 ± 1.27) than leaf samples (45.50 ± 1.32-89.28 ± 0.85). Antioxidant activity and power of leaf samples were ranged from 67.95 ± 0.05‒157.52 ± 0.20 μg/ml and from 45.17 ± 0.01‒361.92 ± 0.78 μmol Fe +2 per g dry weigh, respectively. Positive linear correlations between antioxidant activity and total phenolic compounds were observed. A significant correlation between proximate composition (dependent variable) and some morphological features (independent variable) was observed. Considerable variability in the studied traits among the plant samples can be interestingly used in further food and production systems., (© 2024. The Author(s).)

The ability of a data fusion system composed of a computer vision system (CVS) and an electronic nose (e-nose) was evaluated to predict key physiochemical attributes and distinguish red-fleshed kiwifruit produced in three distinct regions in northern Iran. Color and morphological features from whole and middle-cut kiwifruits, along with the maximum responses of the 13 metal oxide semiconductor (MOS) sensors of an e-nose system, were used as inputs to the data fusion system. Principal component analysis (PCA) revealed that the first two principal components (PCs) extracted from the e-nose features could effectively differentiate kiwifruit samples from different regions. The PCA-SVM algorithm achieved a 93.33% classification rate for kiwifruits from three regions based on data from individual e-nose and CVS. Data fusion increased the classification rate of the SVM model to 100% and improved the performance of Support Vector Regression (SVR) for predicting physiochemical indices of kiwifruits compared to individual systems. The data fusion-based PCA-SVR models achieved validation R 2 values ranging from 90.17% for the Brix-Acid Ratio (BAR) to 98.57% for pH prediction. These results demonstrate the high potential of fusing artificial visual and olfactory systems for quality monitoring and identifying the geographical growing regions of kiwifruits., (© 2023. The Author(s).)

An efficient in vitro protocol was introduced for the conservation of Nepeta asterotricha , a vulnerable and endangered medicinal species found in the central of Iran for the first time. Growth, phytochemical, and biological traits of in vitro regenerated plant (RP) and acclimated plant (AP) were compared to the mother plant (MP). In addition, the genetic stability of AP was assessed by using inter-simple sequence repeats (ISSR) markers. The highest number of lateral branches (4.25) was obtained from the medium with 3 mg/mL kinetin (KIN), while the highest length of lateral branches (13.25 cm) was achieved on the medium culture fortified with 3 mg/mL thidiazuron (TDZ) and 6-benzylaminopurine (BAP). The highest number of leaves (20.25) and main branch length (12.25 cm) were obtained from the medium containing 3 mg/mL TDZ. The highest number of roots (46.25) and root length (2.25 cm) was measured from the medium fortified with 1 mg/mL indole-3-butyric acid (IBA) and 0.6 mg/mL indole-3-acetic acid (IAA), respectively. RP was successfully acclimated (85%) in vivo. Molecular analysis showed that the AP was true to the type of the MP. cis -Sabinene hydrate (26.8-57.7), 1,8-cineole (6.2-24.1), 4aα,7β,7aα-nepetalactone (4.1-12.3), and terpinene-4-ol (3.2-15.0) were the major essential oils compounds. The studied samples contained rosmarinic acid (2.55-5.97 mg/g DW), cichoric acid (1.68-12.7 mg/g DW), chlorogenic acid (1.91-64.21 mg/g DW), rutin (0.59-1.09 mg/g DW), apigenin (0.52-0.72 mg/g DW), betulinic acid (0.17-2.20 mg g DW), oleanolic acid (0.84-5.37 mg/g DW) and ursolic acid (3.46-15.70 mg/g DW). Acclimated plant exhibited the highest antioxidant activity (IC 50  = 196.4 μg/mL), while the methanolic extract of MP displayed the highest antibacterial activity (MIC = 8 mg/mL) against Staphylococcus aureus ., Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-024-01416-x., Competing Interests: Conflict of interestThe authors declare that they have no conflict of interest., (© Prof. H.S. Srivastava Foundation for Science and Society 2024. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.)

Rosa canina L. (Rosaceae), commonly known as the rose hip, is originated from Europe, Africa, and Asia with a long history in medicinal applications. This study aimed to analyze the morphological traits, fatty acids profile, and content of phenolic compounds, anthocyanins, vitamin C, total carotenoid, total phenol, total flavonoid, and antioxidant activity of the fruits of eleven Iranian R. canina ecotypes (RCEs). The highest coefficient of variation was obtained in 1000 seed weight (46.57%). The seed oil varied from 8.08 ± 0.17% to 16.91 ± 0.35%. Linoleic (35.41 ± 0.78% to 49.59 ± 0.96%) and eicosanoic (17.67 ± 0.06% to 25.36 ± 0.54%) acids were the predominant fatty acids in the studied samples. The anthocyanin content in the fruits was ranged from 0.98 ± 0.03 to 4.41 ± 0.04 mg cyanidin 3-glucoside/100 g of dry weight (mg C3G/100 g DW). The high content of vitamin C (103.51 ± 1.24-419.70 ± 3.12 mg/100 g DW), total carotenoid (111.22 ± 0.78-206.98 ± 1.25 mg β-carotene equivalents per g of dry weight (mg β-CARE/g DW)), total phenol (52.87 ± 0.82-104.52 ± 0.23 mg GAE/g DW), and total flavonoid (14.20 ± 0.12-25.18 ± 0.47 mg RE/g DW) were observed in the studied samples. Catechin (20.42 ± 0.47-19.22 ± 0.13 µg/g DW) was the major phenolic compound. The high antioxidant activity in the fruits of the plant was recorded in the studied RCEs (IC 50  = 12.54 ± 0.18-26.33 ± 0.13 μg/ml). A significant correlation between some phytochemical compounds (dependent variable) and morphological features (independent variable) was found. Based on our findings, the fruit of the studied ecotypes can be used for future breeding programs and drug development., (© 2023. The Author(s).)

Nisin, derived from Lactococcus lactis, is a well-known natural food preservative. In the present study, the gene of nisin was transformed to carrot by Agrobacterium tumefaciens strain LBA4404 harboring the recombinant binary vector pBI121 containing neomycin phosphotransferase II (nptII) gene, peptide signal KDEL, and Kozak sequence. The integration of nisin and nptII transgenes into the plant genome was confirmed by polymerase chain reaction (PCR) and dot blot analysis. The gene expression was also performed by RT-PCR and Enzyme-Linked Immunosorbent Assay. The level of nisin expressed in one gram of transgenic plant ranged from 0.05 to 0.08 μg/ml. The stability of nisin varied in orange and peach juices depending on the temperature on the 70th day. The leaf protein extracted from the transgenic plant showed a significant preservative effect of nisin in peach and orange juice. A complete inhibition activity against Staphylococcus aureus and Escherichia coli in orange juice was observed within 24 h. After 24 h, log 1 and log 2 were obtained in a peach juice containing Staphylococcus aureus and Escherichia coli, respectively. Results of HPLC indicated that Chlorogenic and Chicoric acid compounds were increased in transgenic plants, but this increase was not significant. The study of determining the genetic stability of transgenic plants in comparison with non-transgenic plants showed high genetic stability between non-transgenic plants and transgenic plants. This study confirmed the significant inhibitory effect of nisin protein on gram-positive and gram-negative bacteria., (© 2023. The Author(s).)

Perilla frutescens (L.) Britt is a renowned medicinal plant with pharmaceutically valuable phenolic acids and flavonoids. The present study was aimed to study the eliciting effect of silver and copper nanoparticles (AgNPs and CuNPs, 50 and 100 mg/L), and methyl jasmonate (MeJa, 50 and 100 μM) on the biochemical traits, the accumulation of phenolic compounds and antioxidative capacity of P. frutescens cell suspension culture. Suspension cells were obtained from friable calli derived from nodal explants in Murashige and Skoog (MS) liquid medium containing 1 mg/L 2,4-D and 1 mg/L BAP. The 21 days old cell suspension culture established from nodal explant derived callus supplemented with 100 mg/L MeJa resulted in the highest activity of catalase and guaiacol peroxidase enzymes, and CuNPs 100 mg/L treated cells indicated the maximum content of total phenol, total anthocyanin, superoxide dismutase, malondialdehyde, and H 2 O 2 . Also, the highest content of ferulic acid (1.41 ± 0.03, mg/g DW), rosmarinic acid (19.29 ± 0.12, mg/g DW), and phenylalanine ammonia-lyase (16.81 ± 0.18, U/mg protein) were observed with 100 mg/L CuNPs, exhibiting a total increase of 1.58-fold, 2.12-fold, and 1.51-fold, respectively, higher than untreated cells. On the other hand, AgNPs 100 mg/L treated cells indicated the most amounts of caffeic acid (0.57 ± 0.03, mg/g DW) and rutin (1.13 ± 0.07, mg/g DW), as well as the highest scavenging potential of free radicals. Overall, the results of the present study can be applied for the large-scale production of valuable phenolic acids and flavonoids from P. frutescens through CuNPs and AgNPs 100 mg/L elicited cell suspension cultures., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Masson SAS. All rights reserved.)

Glabridin is a well-known active isoflavone found in the root of licorice ( Glycyrrhiza glabra L.) that possess a wide range of biological activity. Plant cells, hairy roots, and fungal endophytes cultures are the most important alternative methods for plant resources conservation and sustainable production of natural compounds, which has received much attention in recent decades. In the present study, an efficient culture condition was optimized for the biomass accumulation and glabridin production from fungal endophyte Aspergillus eucalypticola SBU-11AE isolated from licorice root. Type of culture medium, range of pH, and licorice root extract (as an elicitor) were tested. The results showed that the highest and lowest biomass production was observed on PCB medium (6.43 ± 0.32 g/l) and peptone malt (5.85 + 0.11 g/l), respectively. The medium culture PCB was produced the highest level of glabridin (7.26 ± 0.44 mg/l), while the lowest level (4.47 ± 0.02 mg/l) was obtained from the medium peptone malt. The highest biomass (8.51 ± 0.43 g/l) and glabridin (8.30 ± 0.51 mg/l) production were observed from the PCB medium adjusted with pH = 6, while the lowest value of both traits was obtained from the same medium with pH = 7. The highest production of total glabridin (10.85 ± 0.84 mg/l) was also obtained from the culture medium treated with 100 mg/l of the plant root extract. This information can be interestingly used for the commercialization of glabridin production for further industrial applications., Competing Interests: No potential conflict of interest was reported by the author(s)., (© 2023 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group on behalf of the Korean Society of Mycology.)

Linum album is a well-known rich source of anticancer compounds, i.e., podophyllotoxin (PTOX) and other lignans. These compounds play an important role in the plant's defensive system. The RNA-Seq data of flax (L. usitatissimum) were analyzed under various biotic and abiotic stresses to comprehend better the importance of lignans in plant defense responses. Then, the association between the lignan contents and some related gene expressions was experimented with HPLC and qRT-PCR, respectively. Transcriptomic profiling showed a specific expression pattern in different organs, and just the commonly regulated gene EP3 was detected with a significant increase under all stresses. The in silico analysis of the PTOX biosynthesis pathway identified a list of genes, including laccase (LAC11), lactoperoxidase (POD), 4-coumarate-CoA ligase (4CL), and secoisolariciresinol dehydrogenase (SDH). These genes increased significantly under individual stresses. The HPLC analysis showed that the measured lignan contents generally increased under stress. In contrast, a quantitative expression of the genes involved in this pathway using qRT-PCR showed a different pattern that seems to contribute to regulating PTOX content in response to stress. Identified modifications of critical genes related to PTOX biosynthesis in response to multiple stresses can provide a baseline for improving PTOX content in L. album., (© 2023. The Author(s).)

Green synthesized nanoparticles (GSNPs) display fascinating properties compared to physical and chemical synthesized ones. GSNPs are currently used in numerous applications such as food packaging, surface coating agents, environmental remediation, antimicrobial, and medicine. In the present study, the aqueous leaf extract of Perilla frutescens L. having suitable capping, reducing, and stabilizing compounds was used for green synthesis of silver nanoparticles (Pf-AgNPs). The bioreductant capacity of aqueous leaf extract of P. frutescens for Pf-AgNPs was determined by different confirmatory techniques including UV-Visible spectroscopy, XRD, FESEM, EDX, zeta potential, DLS, SERS, and FTIR analysis. The results exhibited that Pf-AgNPs had optimal size (< 61 nm), shape (spherical), and stability (- 18.1 mV). The antioxidant activity of Pf-AgNPs with both DPPH and FRAP assays was significantly higher compared to P. frutescens extract. Furthermore, Pf-AgNPs had high antimicrobial activity against Escherichia coli and Staphylococcus aureus (MIC = 0.78 mg/mL), and Candida albicans (MIC = 8 mg/mL) while the plant extract showed low antimicrobial activity against both bacterial strains and the fungus tested. Pf-AgNPs and P. frutescens extract also exhibited moderate toxicity on MCF-7 cancer cells with IC 50 values of 346.2 and 467.4 µg/mL, respectively. The results provide insights into using the biosynthesized Pf-AgNPs as an eco-friendly material for a wide range of biomedical applications., (© 2023. The Author(s).)

The seeds of Trachyspermum ammi were gathered at the ripening stage from different regions of Iran and grouped into 14 populations (P1-P14) accordingly. The essential oil (EO) extraction yielded in the 3.16-5% range. EOs were analyzed by gas chromatography-flame ionization detection (GC-FID) and gas chromatography-mass spectrometry (GC-MS) and 11 constituents were identified. Thymol (59.92-96.4%), p-cymene (0.55-21.15%), γ-terpinene (0.23-17.78%), and carvacrol (0.41-2.77%) were the major constituents. The highest contents of thymol and carvacrol were found in the Ghayen population (P2). Also, P2 and P8 (Estahban) had the highest value of total phenol (TPC) 43.2 mg gallic acid equivalent (GAE)/g DW, and total flavonoids (TFC) 8.03 mg quercetin equivalent (QE)/g DW, respectively. P1 (Kalat) had the highest total coumarin (TCC) value (0.26 mg coumarin equivalent CE/g DW). Based on EO constituents, principal component analysis (PCA) and cluster analysis classified populations into two chemotypes of thymol/p-cymene/γ-terpinene and thymol/carvacrol. The highest positive correlation coefficient was between α-terpinene and limonene (0.96), while the highest negative correlation was between thymol and p-cymene (-0.984). The antioxidant activities of extracts and EOs were evaluated by phosphomolybdenum (total antioxidant capacity; TAC), diphenylpicrylhydrazyl (DPPH IC 50 ), and ferric ion reducing antioxidant power (FRAP) assays. Also, the antimicrobial activity of EOs was studied against Escherichia coli and Staphylococcus aureus. P8 with high thymol, EO content (%v/w), TFC, and antibacterial and antioxidant activities is recommended but further studies are needed to confirm the chemotype introduction., (© 2022. The Author(s).)

Taxus baccata L. cell culture is a promising commercial method for the production of taxanes with anti-cancer activities. In the present study, a T. baccata cell suspension culture was exposed to white light and 2-aminoindan-2-phosphonic acid (AIP), a phenylalanine ammonia lyase (PAL) inhibitor, and the effects of this treatment on cell growth, PAL activity, total phenol content (TPC), total flavonoid content (TFC), taxane production and the expression of some key taxane biosynthetic genes (DXS, GGPPS, T13OH, BAPT, DBTNBT) as well as the PAL were studied. Light reduced cell growth, whereas AIP slightly improved it. Light increased PAL activity up to 2.7-fold relative to darkness. The highest TPC (24.89 mg GAE/g DW) and TFC (66.94 mg RUE/g DW) were observed in cultures treated with light and AIP. Light treatment also resulted in the maximum content of total taxanes (154.78 μg/g DW), increasing extracellular paclitaxel and cephalomannin (3.3-fold) and intracellular 10-deacetyl paclitaxel (2.5-fold). Light significantly increased the expression level of PAL, DBTNBT, BAPT, and T13αOH genes, whereas it had no effect on the expression of DXS, a gene active at the beginning of the taxane biosynthetic pathway. AIP had no significant effect on the expression of the target genes. In conclusion, the light-induced activation of PAL transcription and altered expression of relevant biosynthetic genes reduced cell growth and increased the content of total phenolic compounds and taxanes. These findings can be applied to improve taxane production in controlled cultures and bioreactors., Competing Interests: Declaration of Competing Interest The authors declare no conflict of interest., (Copyright © 2022. Published by Elsevier B.V.)

The genus Salvia L. belongs to the Lamiaceae family including several known species rich in natural compounds that are extensively used in pharmaceutical, food, and cosmetic industries. Salvia multicaulis populations contain a broad diversity of flavonoids and phenolic acids. The present study aimed to explore biological and pharmacological effects including antimicrobial and antioxidant activities of nineteen S. multicaulis populations ( SMPs ) grown in Iran for the first time. High content of rosmarinic acid (RA) in SMP12 (Gazan) (5.65 ± 0.33 mg/g DW) caused high antimicrobial activity against two bacteria ( Staphylococcus aureus , Escherichia coli ) and the fungus Candida albicans , while methanolic extract of SMP1 (Taleghan) showed high antioxidant activity due to high content of salvianolic acid A (SAA) and quercetin (0.53 ± 0.04 and 0.49 ± 0.12 mg/g DW, respectively). Altogether these results can be considered for further commercial exploitations to meet the demands of the food and pharmaceutical industries.[Formula: see text].

The induction of polyploidy is an efficient technique for creating a diversity of genetic, phenotypic, and phytochemical novelties in plant taxa. Sage (Salvia officinalis L.) is a well-known medicinal plant rich of valuable bioactive molecules such as triterpenic and phenolic acids. In the present study, the effect of in vitro and in vivo polyploidization on morphological characteristics, anatomical structures, phytochemical traits, and expression level of the genes involved in the biosynthesis of major triterpenic acids (ursolic, betulinic, and oleanolic acids) of the plant was studied. The sterile seeds treated with different concentrations (0, 0.05, 0.1, and 0.2%) of colchicine for 24 and 48 h were considered for polyploidy induction. Flow cytometry and chromosome counting were used to confirm the ploidy level of diploid (2n = 2x = 14, 2C DNA = 1.10 pg) and tetraploid (2n = 4x = 28, 2C DNA = 2.12 pg) plants after seven months. The highest polyploidy induction was obtained by applying 0.1% (w/v) colchicine for 48 h with an efficiency of 19.05% in vitro tetraploidy. Polyploids showed differences in leaf shape and color, leaf and stem thickness, trichrome density, root length, plant height, and number of leaves compared to diploid plants. There was also a significant decrease in rosmarinic acid content in polyploid (plants) as compared to diploid plants. Although a significant decrease in ursolic acid content was observed in polyploids, betulinic acid content associated with the expression levels of genes encoding enzymes being active in triterpene biosynthesis such as squalene epoxidase (SQE) and lupeol synthase (LUS). The expression of SQE and LUS was significantly increased in in vitro tertaploids (2.9-fold) and in vivo mixoploids (2.4-fold). The results confirm the idea that induced polyploidy can randomly alter breeding traits of plants as well as the content of bioactive compounds., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Modern lifestyle factors, such as physical inactivity, obesity, smoking, and exposure to environmental pollution, induce excessive generation of free radicals and reactive oxygen species (ROS) in the body. These by-products of oxygen metabolism play a key role in the development of various human diseases such as cancer, diabetes, heart failure, brain damage, muscle problems, premature aging, eye injuries, and a weakened immune system. Synthetic and natural antioxidants, which act as free radical scavengers, are widely used in the food and beverage industries. The toxicity and carcinogenic effects of some synthetic antioxidants have generated interest in natural alternatives, especially plant-derived polyphenols (e.g., phenolic acids, flavonoids, stilbenes, tannins, coumarins, lignins, lignans, quinines, curcuminoids, chalcones, and essential oil terpenoids). This review focuses on the well-known phenolic antioxidant rosmarinic acid (RA), an ester of caffeic acid and ( R )-(+)-3-(3,4-dihydroxyphenyl) lactic acid, describing its wide distribution in thirty-nine plant families and the potential productivity of plant sources. A botanical and phytochemical description is provided of a new rich source of RA, Satureja khuzistanica Jamzad (Lamiaceae). Recently reported approaches to the biotechnological production of RA are summarized, highlighting the establishment of cell suspension cultures of S. khuzistanica as an RA chemical biofactory.

The analysis of plant cell structure provides valuable information about its morphological, physiological, and biochemical characteristics. Nowadays, scanning electron microscope (SEM) is widely used to provide high-resolution images at the surface of biological samples. However, biological specimens require preparation, including dehydration and coating with conductive materials for imaging by SEM. There are several techniques for providing images with maximum maintenance of cell structure and minimum cellular damage, but each requires the use of expensive and hazardous materials, which can be damaging to the cell in many cases. Therefore, the provision of new and effective preparation methods based on maintaining cell structure for imaging can be very practical. In the present study, a fast and cost-effective protocol was first performed for chemical fixation and preparation of the plant cells for imaging by SEM. Taxus baccata and Zhumeria majdae cells were chemically fixed using glutaraldehyde and then successfully dried with different percentages of ethanol including 70, 80, 90, and 100%. In addition, SEM was performed for imaging the cell surface in different micro-scales. This protocol can be used by plant cell biologists and biotechnologists who are interested in studying structural and biochemical responses of treated or stressed plant cells by SEM., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Narcissus tazetta L., a bulbous plant belongs to the Amaryllidaceae family, contains alkaloid galantamine (GAL) with acetylcholinesterase inhibitory activity which has been recently considered to treat Alzheimer's disease (AD). In the current work, the effect of photoperiod (16/8 h light/dark and 24 h dark) and various concentrations of NAA, BAP, and GA 3 (0, 0.5, 1 and 2 mg l ‒1 ) on the in vitro mass bulblet regeneration of N. tazetta was studied. The GAL production ability of the regenerated bulblets was assessed by HPLC-UV-MS. Light treatments significantly affected the number of bulblet and leaf, the ratio of bulblet/leaf, and leaf length. The maximum number of bulblet (31.0 ± 1.58) and leaf (13.3 ± 1.33) was recorded from the cultures fortified with NAA and BAP (2 mg l ‒1 ) kept in 16/8 h light/dark, while the maximum leaf length (2.1 ± 0.92 cm) was measured on the MS medium containing 0.5 mg l ‒1 NAA and 2 mg l ‒1 BAP incubated in the same photoperiod. The average ratio of bulblet proliferation per explant was significantly different between studied photoperiod (1.1 ± 0.86) and 24 h dark (0.62 ± 0.31). The regenerated bulblets contained 40 and 20 µg g ‒1 DW GAL underexposed photoperiod and 24 h dark, respectively. This information could be useful in the commercial production of GAL as a valuable anti-AD compound through in vitro mass bulblet proliferation of N. tazetta ., Competing Interests: Conflict of interestThe authors declare that they have no conflict of interest., (© Springer Nature B.V. 2020.)

Triterpenic acids (TAs), a large group of natural compounds with diverse biological activity, are produced by several plant taxa. Betulinic, oleanolic, and ursolic acids are the most medicinally important TAs and are mainly found in plants of the mint family. Metabolic engineering is strongly dependent on identifying the key genes in biosynthetic pathways toward the products of interest. In this study, gene expression tracking was performed by transcriptome mining, co-expression network analysis, and tissue-specific metabolite-expression analysis in order to identify possible key genes involved in TAs biosynthetic pathways. To this end, taxa-specific degenerate primers of six important genes were designed using an effective method based on the MEME algorithm in a phylogenetically related group of sequences and successfully applied in three members of the Lamiaceae (Rosmarinus officinalis, Salvia officinalis, and Thymus persicus). Based on the results of in-depth data analysis, genes encoding squalene epoxidase and oxido squalene cyclases are proposed as targets for boosting triterpene production. The results emphasize the importance of identifying key genes in triterpene biosynthesis, which may facilitate genetic manipulation or overexpression of target genes.

The combined use of elicitors can be an effective way to increase the production of secondary metabolites (SMs) in plant cell, tissue and organ cultures. This study investigated the effects of a salicylic acid (SA) pretreatment and different glucose levels on the growth, biochemical traits and taxane production in a Taxus baccata callus culture. For this purpose, after a pretreatment with SA (5 μM), three-month-old calli were cultured on B5 medium fortified with different concentrations of glucose (0, 0.5, 1, 2 and 3%), and they were compared with calli cultured on a B5 medium supplemented only with glucose. When the calli were harvested at 21 days, their fresh weight (g), dry weight (g) and cell viability (%) had decreased significantly (p < 0.05) with the higher glucose concentrations. The glucose treatment increased the hydrogen peroxide (H 2 O 2 ) and malondialdehyde (MDA) content, and caused oxidative stress in treated tissues. The lower H 2 O 2 content and oxidative stress was associated with an increased antioxidant enzyme activity in the SA-pretreated samples, which resulted in less membrane damage and improved growth and cell viability under the glucose treatment compared to the control. By reducing the activity of phenylalanine ammonia-lyase (PAL) and polyphenol oxidase (PPO), the SA pretreatment reduced the production and oxidation of phenolic compounds under the glucose treatment; this decrease was associated with less browning of tissues and higher viability. Increases in taxol (5.1-fold) and total taxanes (3.5-fold) in the SA-pretreated calli cultured on the medium containing 2% glucose, compared to the control, indicated that the two treatments had a significant effect on taxane production in the T. baccata callus culture., (Copyright © 2018 Elsevier Masson SAS. All rights reserved.)

Genetic and the essential oil composition variability among twelve Perovskia abrotanoides populations (PAbPs) growing wild in Iran were assessed by ISSR markers, GC-FID and GC/MS, respectively. Nine selected ISSR primers produced 119 discernible bands, of them 96 (80.7%) being polymorphic. Genetic similarity values among populations ranged between 0.07 and 0.79 which indicated a high level of genetic variation. Polymorphic information content, resolving power and marker index generated by ISSR primers were, 0.31, 6.14, and 3.32, respectively. UPGMA grouped PAbPs into four main clusters. Altogether, 38 chemical compounds were identified in the oils, and a relatively high variation in their contents was found. Camphor (11.9 - 27.5%), 1,8-cineole (11.3 - 21.3%), α-bisabolol (0.0 - 13.1%), α-pinene (5.9 - 10.8%), and δ-3-carene (0.1 - 10.5%) were the major compounds. Oxygenated monoterpenes (32.1 - 35.8%) and monoterpene hydrocarbons (25.7 - 30.4%) were the main groups of compounds in the oils studied. Cluster analysis and principal-component analysis were used to characterize the samples according to oil components. Four main chemotypes were found to be Chemotype I (camphor/1,8-cineol), Chemotype II (1,8-cineole/camphor), Chemotype III (camphor/1,8-cineol/α-bisabolol), and Chemotype IV (camphor/δ-3-carene/α-bisabolol). The information, provided here on P. abrotanoides populations, will be useful to introduce this plant into agricultural systems., (© 2018 Wiley-VHCA AG, Zurich, Switzerland.)

Plant secondary metabolism evolved in the context of highly organized and differentiated cells and tissues, featuring massive chemical complexity operating under tight environmental, developmental and genetic control. Biotechnological demand for natural products has been continuously increasing because of their significant value and new applications, mainly as pharmaceuticals. Aseptic production systems of plant secondary metabolites have improved considerably, constituting an attractive tool for increased, stable and large-scale supply of valuable molecules. Surprisingly, to date, only a few examples including taxol, shikonin, berberine and artemisinin have emerged as success cases of commercial production using this strategy. The present review focuses on the main characteristics of plant specialized metabolism and their implications for current strategies used to produce secondary compounds in axenic cultivation systems. The search for consonance between plant secondary metabolism unique features and various in vitro culture systems, including cell, tissue, organ, and engineered cultures, as well as heterologous expression in microbial platforms, is discussed. Data to date strongly suggest that attaining full potential of these biotechnology production strategies requires being able to take advantage of plant specialized metabolism singularities for improved target molecule yields and for bypassing inherent difficulties in its rational manipulation.

Methyl jasmonate (MJ), a plant-derived stress hormone, has been shown to be a promising anti-cancer agent with high selectivity toward cancerous cells. The aim of the present study was to design a MJ loaded nanoemulsion (NE) to overcome the low MJ water solubility and also improve its anti-cancer efficiency. Box-Behnken design (BBD) was employed to optimize the composition effect of three independent manufacturing variables on two responses including average droplet size and poly dispersity index (PDI). ANOVA analysis indicated that both of the studied responses were well fitted by resultant quadratic models with the coefficient of determinations (R2) 0.994 and 0.975, respectively. The actual average droplet size 75.06 nm and PDI 0.017 obtained for the optimum MJNE was in good agreement with those values predicted with numerical optimization. Physicochemical characterization indicated that the optimum MJNE was transparent, isotropic, spherical and sterically stabilized. MTT assay indicated that MJNE was more efficacious in killing cancer cells than MJ solution. Cell cycle analysis revealed that MJNE induced a stronger sub-G1 arrest than MJ solution. A considerable absence of toxicity was achieved for MJNE and blank NE in HUVEC normal cells. These results may provide strong support to develop a NE delivery system as a promising carrier for improving the safety and anti-cancer efficacy of MJ.

An in vitro approach to the production of rosmarinic acid (RA), a medicinally important caffeic acid ester, in a cell suspension culture (CSC) of Satureja khuzistanica Jamzad (Lamiaceae) has been investigated for the first time. The CSC was established from friable calli derived from shoot tip explants in Gamborg's B5 liquid medium supplemented with 30 g/L sucrose, 20 mg/L L-glutamine, 200 mg/L casein hydrolysate, 5 mg/L benzyladenine (BA) and 1 mg/L indole-3-butyric acid (IBA). The effect of nitrogen source (KNO3 and (NH4)2SO4) and their different concentrations on the fresh and dry weight (g/L), as well as RA content (mg/g dry weight) were measured. CSC growth measurements indicated a maximum specific cell growth rate of 1.5/day, a doubling time of 7.6 days and a high percentage of cell viability (96.4 %) throughout the growth cycle. Maximum cell fresh weight (353.5 g/L), dry weight (19.7 g/L) and RA production (180.0 mg/g) were attained at day 21 of culture. Cell growth and RA content were affected by nitrogen deficiency. Media containing 8.3 mM of total nitrogen (¼ of B5 standard medium) led to a minimum cell fresh weight (243.0 g/L), dry weight (17.4 g/L) and RA content (38.0 mg/g) after 21 days. The established CSC provided useful material for further optimization experiments aimed at a large-scale production of RA.

Rosmarinic acid (RA), an ester of caffeic acid and 3,4-dihydroxyphenyl lactic acid, is widely distributed in the plant kingdom. Interest in it is growing due to its promising biological activities, including cognitive-enhancing effects and slowing the development of Alzheimer's disease, cancer chemoprotection or anti-inflammatory activity, among others. In order to meet the increasing demand for this compound, several biotechnological approaches to its production based on plant cell and hairy root cultures have been developed. Empirical strategies are currently being combined with metabolic engineering tools to increase RA production in plant cell platforms in a more rational way. Discussed here are the latest advances in the field, together with recent trends in plant biotechnology, such as the application of single use technology and the use of biosensors in downstream processes.

In the present study, an efficient protocol has been developed for callus induction and production of RA in callus culture of Satureja khuzistanica for the first time. In-vitro callus induction was achieved from young shoot tip explants cultured on MS and B5 media supplemented with different concentrations of IBA (0.1, 1.0, 2.0 and 5.0 mgL(-1)) solely or in combination with cytokinins BAP and KIN (1.0, 2.0 and 5.0 mgL(-1)). B5 medium supplemented with 1.0 mgL(-1) IBA plus 5.0 mgL(-1) BAP and MS medium fortified with 2.0 mgL(-1) IBA and 2.0 mgL(-1) BAP were the most favorable media for callus formation with the highest induction rate (96%). Maximum growth index (2.89 and 2.63) and maximum callus biomass (2.34 and 2.33 g fresh weight) were obtained from the callus cultured on B5 medium supplemented with 1.0 mgL(-1) IBA plus 5.0 mgL(-1) BAP and MS medium fortified with 1.0 mgL(-1) IBA plus 1.0 mgL(-1) KIN, respectively. Determination and quantification of RA in cultured calli were performed by HPLC UV/MS analysis. Calli induced from the plant and maintained on supplements of IBA and BAP in the absence of light produced RA 7.5% based on dry weight (DW). No differentiation was observed in any callus during the course of this study.

The biological activity and composition of the essential oil of Sclerorhachis leptoclada Rech. f. an endemic species from northeast of Iran was studied. The essential oil was isolated from the aerial flowering parts of the plant and analyzed by GC and GC-MS. Fifty-four compounds accounting for 95.9% of the total oil were characterized. The main constituents were (E)-nerolidol (14.5%), terpinen-4-ol (13.3%), camphor (6.1%), 1,8-cineole (4.8%) and p-cymene (4.5%). The antimicrobial activity of the essential oil of S. leptoclada was tested against eight microbial strains and a fungi. The results of the bioassays showed that the Gram-positive bacteria, Bacillus subtilis and Staphylococcus epidermidis, were the most sensitive to the oil than others with the MIC value of 1.8 mg/mL. The tested fungi, Saccharomyces cerevisiae was highly inhibited by the oil of S. leptoclada with MIC value of 10 mg/mL. In the case of cytotoxicity, IC50 values estimated to be 312, 1250, 625 and 1250 μg oil/mL respectively, for the Vero, SW480, MCF7, and JET 3 cancer cell lines.

In this study, we report the antioxidative and protective effect of essential oil of Dracocephalum multicaule on K562 cells. Our results demonstrated that monoterpenoids, including oxygenated and hydrocarbons, 71.5% and 28.3%, respectively, were the principal essential oils of D. multicaule. Perilla aldehyde (71.5%) and limonene (28.1%) were identified as the main components. Antioxidant studies based on the 2,2-diphenylpicrylhydrazyl assay indicated that the D. multicaule essential oil possesses a marked antioxidant and radical-scavenging activity with an IC₅₀ value of 438.2 μg/mL. Pretreatment with essential oil and main constituents protected K562 cells 49.5% against H₂O₂-induced oxidative damage throughout increasing the activities of antioxidant enzymes and glutathione content in K562 cells. Collectively, D. multicaule essential oil and its main compounds especially in combinatory condition at a ratio of 7:3 with high antioxidant properties may be able to protect cells against oxidative stress induced by H₂O₂ through antioxidative mechanisms.

The objective of this study was to investigate in vitro antimicrobial activity and composition of the essential oil of Cyclotrichium leucotrichum growing wild in Iran. The essential oil was obtained by hydro-distillation and analysed by GC-FID and GC/MS. Fifty-nine components representing 98.9% of the total oil were characterised. The essential oil which has 1,8-cineol (14.8%), elemol (12.6%), spathulenol (9.4%), E-caryophyllene (5.7%) and hinesol (5.7%) as its main components, exhibited moderate activity against seven bacteria and a yeast, Candida albicans, with minimum inhibitory concentration values ranging from 0.5 to 64 mg mL(-1) and minimum bactericidal concentration values ranging from 2 to >64 mg mL(-1), respectively. The best inhibitory effects were against three gram-positive bacteria and tested yeast, C. albicans.

In this study, a total of 25 endophytic fungi were successfully isolated from the inner bark of Taxus baccata grown in Iran by the aseptic technique. Genomic DNA was extracted from isolated endophytic fungi and subjected to polymerase chain reaction (PCR) analysis for the presence of the Taxus taxadiene synthase (ts) gene, which encodes the enzyme catalyzing the first committed step of taxol biosynthesis. Four of 25 isolated endophytic fungi isolates showed PCR positive for the ts gene. Subsequently, taxol and 10-deacetylbaccatin III (10-DAB III) were extracted from culture filtrates and mycelia of the PCR positive isolates and analyzed by high-performance liquid chromatography and mass spectrometry. The analysis showed that one isolate (SBU-16) produced taxol (6.9 ± 0.2 μg L(-1) ) and its intermediate compound, 10-DAB III (2.2 ± 0.1 μg L(-1) ). The isolate SBU-16 was identified as Stemphylium sedicola SBU-16, according to its morphological characteristics as well as the internal transcribed spacer nuclear rDNA gene sequence analysis. Interestingly, this is the first report of the genus Stemphylium as a taxol-producing taxon., (© 2011 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.)

Zataria multiflora Boiss. is an aromatic shrub belonging to the Lamiaceae family. Its aerial parts are used in the traditional medicine and in the pharmaceutical and food industries. The terpenoid and genetic profiles of 18 accessions of Z. multiflora, collected in different locations in Iran, have been analyzed by GC/FID and GC/MS or by AFLP (amplified fragment length polymorphism) analyses, respectively. Altogether, 56 compounds were identified in the essential oils, with the major constituents being thymol (6.0-54.9%), carvacrol (0.7-50.6%), linalool (1.2-46.8%), and p-cymene (1.6-14.8%). On the basis of the essential-oil composition, the 18 accessions were divided into four groups. The first group was characterized by a high content of carvacrol, thymol, and linalool, the second group was dominated by carvacrol, thymol, and p-cymene, the third group was characterized by a high concentration of thymol and a low content of carvacrol and p-cymene, and the forth group contained linalool and carvacrol as the main components. The AFLP results revealed that the average genetic similarity (GS) between the accessions was 0.61, ranging from 0.40 to 0.77. The UPGMA (unweighted pair-group method with arithmetic mean) cluster analysis divided all accessions into five groups at a similarity level of 0.60. The two clusters generated, the first based on the essential-oil compositions and the second on the AFLP data, showed a different pattern of relationships among the accessions. The knowledge of the Z. multiflora chemotype diversity, illustrated in this study, will allow an improvement of the homogeneity of the plant material for the production of different types of essential oils, depending on the demands of the pharmaceutical and food industries for specific uses., (Copyright © 2011 Verlag Helvetica Chimica Acta AG, Zürich.)

Introduction: Recently, there have been growing attention on the modification and optimisation of new extraction and quantification methods, caused by the lack of environmentally friendly methodologies for the extraction of phytochemicals from complex matrices. In the case of pharmaceutical compounds, not only the extraction procedure but also the analysis method should be efficient, precise, fast and easy., Objectives: The essential pharmaceutical characteristics and trace concentration of withanolides led us to modify and optimise the previously reported extraction and quantification procedure for withaferin A (WA) as a candidate for withanolides., Material and Methods: The WA from the air-dried aerial part of Withania somnifera Dunal. was extracted using a microwave-assisted extraction (MAE) technique. Four variables affecting the extraction procedure were optimised using the central composite design approach. The method of high-performance thin-layer chromatography assay was validated and applied for the quantification of each experiment., Results: The optimum values of factors were: extraction time (150 s), extraction temperature (68°C) and 17 mL of methanol : water in the ratio 25 : 75 as extracting solvent. The solvent system consisted of ethyl acetate : toluene : formic acid : 2-propanol (7.0 : 2.0 : 0.5 : 0.5, v/v/v/v), and densitometric scanning at 220 nm was applied for the analysis. The dynamic linear range, LOD, LOQ and recovery with the inter-day, and intra-day RSDs of the developed method indicated the validity of the method., Conclusion: A pressurised MAE method for extracting WA from the plant's aerial part was optimised using factorial-based design. The net effect of time, temperature, solvent volume and its ratio suggests that the yield of WA increases until each factor reaches its optimum value, and decreases with further increase in temperature or solvent ratio., (Copyright © 2010 John Wiley & Sons, Ltd.)

Withania somnifera, commonly known as Ashwagandha, is an important medicinal plant that has been used in Ayurvedic and indigenous medicine for over 3,000 years. In view of its varied therapeutic potential, it has also been the subject of considerable modern scientific attention. The major chemical constituents of the Withania genus, the withanolides, are a group of naturally occurring C28-steroidal lactone triterpenoids built on an intact or rearranged ergostane framework, in which C-22 and C-26 are appropriately oxidized to form a six-membered lactone ring. In recent years, numerous pharmacological investigations have been carried out into the components of W. somnifera extracts. We present here an overview of the chemical structures of triterpenoid components and their biological activity, focusing on two novel activities, tumor inhibition and antiangiogenic properties of withaferin A and the effects of withanolide A on Alzheimer's disease. The most recent attempts in biotechnological production of withanolides are also discussed.

For successful conservation and breeding of a medicinal species, it is important to evaluate its genetic diversity as well as its content of phytochemical compounds. The aim of the present study was to investigate the genetic variation of Iranian natural populations of W. somnifera and W. coagulans, using the RAPD (random amplified polymorphic DNA) markers, and their withaferin A content. Using 16 RAPD primers, a total of 282 RAPD bands were achieved. The highest and lowest percentages of polymorphism were observed with primers OPAD-15 (100.0%) and OPC-06 (75.0%), respectively. Cluster analysis of the genotypes was performed based on data from polymorphic RAPD bands, using Dice's similarity coefficient and the UPGMA clustering method. Variations in the RAPD results were found to reflect geographical distribution and genetic factors of the plant populations. The HPTLC analysis of the studied samples revealed the presence of withaferin A in W. coagulans and W. somnifera. Moreover, the concentration of withaferin A had a range from 2.2 to 32.5 microg/g DW and was higher in the aerial part than in the root in all used samples. The results of the present study show that there is a high level of variation in the Iranian natural population of Withania, which is significant for conservation and breeding programs to improve production of withaferin A.

Taxol is one of the most effective anti-cancer drugs ever developed. The natural source of taxol is the inner bark of several Taxus species, but it accumulates at a very low concentration and with a prohibitively high cost of extraction. Another problem is that the use of inner bark for taxol production implies the destruction of yew trees. For all these reasons, the growing demand for taxol greatly exceeds the supply that can be sustained by isolation from its natural source and alternative sources of the drug are being sought. Although taxol has been prepared by total synthesis, the process is not commercially viable. Taxol can also be semisynthetically produced via the conversion of baccatin III or 10-deacethylbaccatinIII found in Taxus needles but the cost and difficulty of the extraction process of the semisynthetic precursors are also very high. The most promising approach for the sustainable production of taxol and related taxoids is provided by plant cell cultures at an industrial level. Taxol is currently being clinically used against different tumour processes but due to the difficulty of its extraction and formulation, as well as the growing demand for the compound, new taxol analogues with improved properties are being studied. In this revision we discuss current research in the design of new taxol-related compounds, the chemical structure/anti-cancer activity relationship and new formulations of the drug. We also consider the optimizing strategies to improve taxol and related taxoid production in cell cultures, as well as the current knowledge of taxol metabolism, all of which are illustrated with examples, some of them from our own research.

Thymus species are well known as medicinal plants because of their biological and pharmacological properties. Thymus caramanicus is an endemic species grown in Iran. Variation in the quantity and quality of the essential oil of wild population of T. caramanicus at different phenological stages including vegetative, floral budding, flowering and seed set are reported. The oils of air-dried samples were obtained by hydrodistillation. The yields of oils (w/w%) at different stages were in the order of: flowering (2.5%), floral budding (2.1%), seed set (2.0%) and vegetative (1.9%). The oils were analyzed by GC and GC-MS. In total 37, 37, 29 and 35 components were identified and quantified in vegetative, floral budding, full flowering and seed set, representing 99.3, 98.6, 99.2 and 97.8% of the oil, respectively. Carvacrol was the major compound in all samples. The ranges of major constituents were as follow: carvacrol (58.9-68.9%), p-cymene (3.0-8.9%), γ-terpinene (4.3-8.0%), thymol (2.4-6.0%) and borneol (2.3-4.0%). Antibacterial activity of the oils and their main compounds were tested against seven Gram-positive and Gram-negative bacteria by disc diffusion method and determining their minimum inhibitory concentration (MIC) values. The inhibition zones (IZ) and MIC values for bacterial strains, which were sensitive to the essential oil of T. caramanicus, were in the range of 15-36mm and 0.5-15.0mg/ml, respectively. The oils of various phenological stages showed high activity against all tested bacteria, of which Bacillus subtilis and Pseudomonas aeruginosa were the most sensitive and resistant strains, respectively. Thus, they represent an inexpensive source of natural antibacterial substances that exhibited potential for use in pathogenic systems., (Copyright © 2008. Published by Elsevier Ltd.)

Scopolamine is an alkaloid widely used in medicine for its anticholinergic activity. The aim of this review is to show that metabolic engineering techniques constitute a suitable tool to improve the production of tropane alkaloids, focusing in particular on scopolamine. We present an overview of results obtained by various research groups, including our own, who have studied the overexpression of genes involved in the biosynthesis of scopolamine in different plant species that produce tropane alkaloids. Experiments carried out to improve production in hairy root cultures will also be described, as well as those attempting to biotransform hyoscyamine into scopolamine in roots and transgenic tobacco cells.

The chemical composition of the essential oil obtained from the aerial flowering parts of Ziziphora clinopodioides subsp. bungeana (Juz.) Rech. f. was analyzed by GC and GC-MS. Thirty-two components representing 97.1% of the total oil were identified. Oxygenated monoterpenes (94.3%) were the predominant fraction of the oil with pulegone (65.2%), isomenthone (11.9%), 1,8-cineole (7.8%) and piperitenone (6.5%) as the main constituents. Antibacterial activity of the oil and also its two main components (pulegone and 1,8-cineole) were tested against seven bacteria. It was found that the oil exhibited interesting antibacterial activity against Staphylococcus epidermidis, S. aureus, Escherichia coli and Bacillus subtilis with MIC values of 3.75 mg/ml.

Trichlorfon is an organophosphorus insecticide, which is extensively being used for protection of fruit crops. Trichlorfon is a thermal labile compound, which cannot be easily determined by gas chromatography (GC) and has no suitable group for sensitive detection by high performance liquid chromatography (HPLC). In this study, a 31P nuclear magnetic resonance (31P NMR) has been described for monitoring of trichlorfon without any separation step. The quantitative works of 31P NMR spectroscopy has been performed in the presence of an internal standard (hexamethylphosphoramide). Limit of detection (LOD) for this method has been found to be 55 mg L(-1), without any sample preparation, and the linear working range was 150-5500 mg L(-1). Relative standard deviation (R.S.D.%) of the method for three replicates within and between days was obtained < or =9%. The average recovery efficiency was approximately 99-112%. This method was applied for monitoring trichlorfon in a commercial insecticide sample and tomato sample.