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In the present research, the burned area of each province was investigated according to land use in order to manage and prioritize provinces with extent and high-risk burns. The analyses of burned areas in Iran has been fulfilled in Iran for the first time. Following the expansion of agricultural lands around forests, the number of intentional wildfires aiming at reducing forest area has increased. Only 5% of wildfires in the Mediterranean region have a natural origin. In Iran, the financial damage caused by wildfires (apart from spiritual and intangible damages such as woods, bushes, and herbal plants) between 1968 and 2007 has been estimated at about 1.93 million dollars. Therefore, prioritizing and managing fire in natural ecosystems seems to be essential. In the present study, the burned areas of Iran from 2002-03-21 to 2018-03-21 based on land use were obtained from the data available in the Global Wildfire Information System and were categorized by provincial borders. In order to prepare Iran's land use map, related data (MCD12Q1) were obtained from https://www.earthdata.nasa.gov in HDF5 format. The results showed that Khuzestan, Fars, Kermanshah, and Golestan Provinces, respectively had the largest burned areas in the country. Considering that most burned areas of the country have been recorded in agricultural lands, human factors are very influential in increasing the rate of burned areas, and maintaining distance between agricultural lands and native vegetation (fuel) is mandatory.

Emerging evidence of the impact of plant compounds on growth inhibition of micropathogens and cancer cells has opened new areas to evaluate plants’ treatment properties. Here, we aimed to investigate in vitro antioxidant, antibacterial and anticancer effects of the secondary metabolites isolated from different extracts produced by Elaeagnus angustifolia and Scrophularia striata. Antibacterial activity was evaluated against human and plant pathogenic bacteria by 3 methods of tubular dilution, well and disc diffusion. The anticancer effect of E. angustifolia extract was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Biochemical experiments showed the presence of compounds such as phenol, flavonoids, resins, quinones, steroids, terpenoids and alkaloids in extracts, with the highest antioxidant activity of the methanolic extracts. For both plants’ strains, the disc method was more effective than the well diffusion method. The highest yields were obtained from Methanolic, ethanolic and hydroalcoholic extracts for E. angustifolia and aqueous extract for Scrophularia striata. The most sensitive bacteria for E. angustifolia were Bacillus subtilis and Xanthomonas campestris against pit extracts and Clavibacter michiganensis against pulp extracts. The most sensitive bacteria for S. striata were Bacillus subtilis and Staphylococcus aureus. Methanolic and aqueous solvents showed the maximum bacterial inhibitory and bactericidal activities in the Minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) tests respectively. Additionally, E. angustifolia showed anticancer effects toward MCF7 breast cancer cells. These findings provided a better understanding of the widespread application of these plants as potential antioxidants, antibacterial and anticancer sources and safe natural medicines in health maintenance and disease treatment.

Unlike plant cell suspension culture, the proliferation of callus in bioreactors has received inadequate attention. The magnificent potential of plant callus becomes more appreciated as the research unfolds and promises interesting applications including the production of valuable metabolites, therapeutic antibodies, bioactive extracts with regenerating effects, and the generation of genetically improved plants. Issues such as the lack of 3D-access of the cells to the nutrients, using an interfering gelling substance as the support matrix, and the changes in the medium formulation during the growth phase were discouraging factors for extending research on this topic. Considering the existing drawbacks, a novel open-flow spray bioreactor (OFSB) was configured to circumvent the associated problems with the solid cell culture and promote the applicability of plant callus culture via improving the feeding strategy.Applying similar subculture conditions, the proliferation of Arnebia pulchra and Hyoscyamus niger calli as the examples of two important plant families (Boraginaceae and Solanaceae) was studied in the OFSB in comparison with similar calli that grew in Petri dishes and jars.A. pulchra and H. niger calli obtained the weight gains of (%87.3 and %106.7) in the Petri dishes, (%208.7 and %226) in the jars, and (%288.6 and %320.0) in OFSB, respectively, while no significant changes were observed in the productivity indices of the examined calli.The simple design of OFSB bypasses most of the notorious problems associated with solid plant callus culture. OFSB technical features allow the bioreactor to be used for growth optimization of various types of plant calli in a cost-effective manner.

The decoction of the stem bark of Salix alba (EASA) is traditionally used in Iran against headache, low back pain and to treat piles. The present investigation was carried out to evaluate the safety of decoction of Salix alba stem bark extract after acute and sub-chronic oral administration in male rodents. For the acute study, a single administration of (EASA) was given orally to male mice at doses of 2 ,4, 8, 12,16 and 20 g/kg. In the sub-chronic toxicity study, Salix alba bark extract was administered orally as a single administration to male Wistar rats at doses of 200, 400, 800 and 1600 mg/kg/day for 28 days. Animal body weight was recorded throughout the experimental period while hematological and biochemical parameters of blood and urine, as well as the weight of organs, were evaluated. The relative weight of organs was affected significantly by the treatment at the doses of 800 and 1600 mg/kg between liver and kidney that was dose-related. Hematological changes were observed at the doses 400, 800 and 1600 mg/kg at the parameters of RBC, Hgb, HCT, PLT, and WBC. Serum AST, ALT, ALP, and total bilirubin increased significantly while total protein and triglycerides significantly decreased. Renal function indices assay in blood showed a significant modification in the 800 and 1600 mg/kg groups at the parameters of creatinine, uric acid and potassium compared to control while, in urine, protein, and calcium markedly increased at the doses of 1600 mg/kg. These results showed that there is a wide margin of safety for the therapeutic use of EASA and that the aqueous stem bark extract of Salix alba had no toxicity in oral acute high dose administration and very low toxicity in oral sub-chronic administration.

Medicinal plants used in the treatment of diseases earlier times are potential sources of new drugs. The present study was undertaken to study the chemical composition, antioxidant and antibacterial activity of certain medicinal plants of Iran by gas chromatography and mass spectrometry (GC/MS), DPPH and disk diffusion method. According to the results of GC/MS, there are 46 kinds of chemical compounds including mucilage, fatty acids, flavonoid and diterpenes in flower of Echium khuzistanicum. There are aldehydes (7.9%), phenols (7.5%), fatty acids (5.8%) and furfural (5.4%) in the methanol extract of Echinops cephalotes. Furfural, steroids, vitamin B and flavonoids are the main compounds of Marrubium anisodan. Results of the antibacterial test showed that Staphylococcus aureus and Bacillus subtilis were more sensitive to methanol extract of Echium khuzistanicum root. Pseudomonas auruginosa was more sensitive to DMSO extract of Marrubium anisodan at 600 mg/ml concentration. Maximum flavonoid and phenol contents were belonging to Echinops cephalotes. Marrubium anisodan showed the best DPPH free radical scavenging activity., Medicinal plants used inthe treatment of diseases earlier times are potential sources of new drugs. Thepresent study was undertaken to study the chemical composition, antioxidantand antibacterial activity of certain medicinal plants of Iran by gas chromatography and mass spectrometry (GC/MS), DPPH and disk diffusion method. According to theresults of GC/MS, there are 46 kinds of chemical compounds including mucilage,fatty acids, flavonoid and diterpenes in flower of Echium khuzistanicum.There are aldehydes (7.9%), phenols (7.5%), fatty acids (5.8%) and furfural(5.4%) in the methanol extract of Echinops cephalotes. Furfural,steroids, vitamin B and flavonoids are the main compounds of Marrubiumanisodan. Results of the antibacterial test showed that Staphylococcusaureus and Bacillus subtilis were more sensitive to methanol extractof Echium khuzistanicum root. Pseudomonas auruginosa was moresensitive to DMSO extract of Marrubium anisodan at 600 mg/mlconcentration. Maximum flavonoid and phenol contents were belonging to Echinopscephalotes. Marrubium anisodan showed the best DPPH free radicalscavenging activity.

Tissue-type plasminogen activator (tPA) is a serine protease that plays a crucial role in the fibrinolytic system. We increased the activity of tPA by splicing the active site of dodder-cuscutain gene to human tPA. The chimeric cDNA of tPA was constructed by Splicing by Overlap Extension Polymerase Chain Reaction (SOEing-PCR) method and transferred to the hairy roots of tobacco using different strains of Agrobacterium rhizogenes. Chimeric-tPA was purified by lysine-sepharose chromatography and specific aptamers were designed using SELEX method. Multi wall carbon nanotubes were functionalized with selected aptamers, packed in a column, and used for purification. The results demonstrated that selected aptamer having KD values of 0.320 nM and IC50 of 28.9 nM possessed good affinity to tPA, and the chimeric-tPA was properly purified by aptamer-chromatography. Hairy roots expressing chimeric-tPA and normal-tPA produced 900 and 450 ngmg−1 of total protein, respectively. The activities of chimeric–tPA and normal–tPA were 90 and 60 IUml−1, respectively. Compared to the normal-tPA, chimeric-tPA showed more activity.

Sensitive detection of biomarkers will mean accurate and early diagnosis of diseases. A tissue plasminogen activator (tPA) has a crucial role in many cardiovascular diseases and it is related to many processes such as angiogenesis in cancer cells. Therefore, sensitive determination of tPA is important in diagnosis and clinical research. tPA monoclonal antibody was covalently attached onto single-wall carbon nanotubes (SWCNTs) using diimide-activated imidation coupling. Functionalized SWCNTs were immobilized onto a glassy carbon electrode and the modification process was investigated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), SEM, cyclic voltammetry (CV), and electrochemical impedance spectroscopy (EIS). Cyclic voltammograms (CVs) in a scan rate of 100 mVs−1 was studied and comparisons were made between the modified glassy carbon electrodes (immobilized with antibodies) as a working electrode before and after the formation of tPA-antibody complex. Results of the SDS-PAGE demonstrated that the antibody was covalently and site directly attached to the SWCNTs. The fabricated biosensor provided a good linear response range from 0.1 to 1.0 ng mL−1 with a low detection limit of 0.026 ng mL−1. The immunosensor showed selectivity, reproducibility, good sensitivity, and acceptable stability. Satisfactory results were observed for early and sensitive determination of tPA in human serum samples. For the first time, such specific biosensor is currently being fabricated for tPA in our laboratories and successfully could determine tPA in myocardial infraction and breast cancer patients.

A label-free electrochemical sensor is reported in the present work for the analysis of activase. Firstly, Au electrode was modified by electrochemical deposition of gold nanoparticles to increase the surface area of the Au electrode, which resulted in capturing large amounts of other modifiers. Next, the modified electrode with gold nanoparticles was coated with 3-mercaptopropionic acid (MPA) and l -lysine ( l -Lys) amino acid via self-assembled monolayers (SAMs). The morphologies of the Au nanoparticles and the fabrication process of the sensor were characterized by scanning electron microscopy (SEM), cyclic voltammetry (CV), and electrochemical impedance spectroscopy (EIS), respectively. Under the optimal conditions, this sensor displayed an excellent correlation between the charge transfer resistance changes and the amount of the absorbed activase protein, and also demonstrated a good linear range from 0.2 to 1.0 ng mL − 1 of activase, with a detection limit of 0.01 ng mL − 1 . The present sensor has the advantages of simplicity (preparing a self-assembled monolayer), cheapness (not using antibodies), fast detection (with an incubation time of 15 min), and good sensitivity.

A simple, sensitive, selective, and rapid optical nanobiosensor based on FRET was designed to detect tropane alkaloids as anti-cholinergic agents in natural and transgenic hairy roots extracts of Atropa belladonna. To achieve that, conjugation of tioglycolyic acid capped cadmium telluride quantum Dots, M2 muscarinic receptor (Cd/Te QDs-M2R) and conjugation of scopolamine-rhodamine123 (Sc-Rho123) were performed. More specifically, proportional amounts of M2 muscarinic receptor and quantum dots (QDs) were conjugated while scopolamine (as a tropane alkaloid) and rhodamine123 were also combined and these moieties functioned as donor and acceptor pairs, respectively. The system response was linear over the range of 0.01-4µmolL-1 of scopolamine hydrochloride concentration with a detection limit of 0.001µmolL-1. The developed nanobiosensor was successfully used for in vitro recognition of scopolamine as an anti-cholinergic agent in the investigated plant extracts. In addition, Agrobacterium rhizogenesis mediated gene transfer technique was employed to generate hairy roots and to enhance the production of tropane alkaloids in the studied medicinal plant.

Species of Ulva are distributed worldwide in various aquatic habitats and their morphological characters change considerably in response to environmental conditions, which makes identification in this genus problematic. All Ulva species so far described from the Persian Gulf, Iran, have been identified by classical taxonomic methods. In this study, Ulva spp. collected from the Persian Gulf were identified by combined morphological and molecular methods. We sequenced the internal transcribed spacer ITS2 and the rbcL gene in 15 specimens, representing typical morphotypes of over 50 samples. Maximum likelihood, maximum parsimony and Bayesian inference (BI) phylogenetic reconstructions using ITS and rbcL revealed clades, which we identified as Ulva chaugulii, U. paschima and U. ohnoi. These three species have not been recorded from the Persian Gulf, Iran before. The morphology of U. chaugulii differed considerably from its species description, showing further evidence for extreme intraspecific variation in morphology in this cosmopolitan genus.

Glycyrrhiza glabra (G. glabra) has been used as a flavoring and sweetener agent, in addition to its therapeutic properties. It is rich in phytoestrogen and may prevent osteoporosis caused by estrogen deficiency; however, there is no evidence for its effects on proliferation and osteogenesis in mesenchymal stem cells. So, we were encouraged to investigate whether the ethyl acetate extract of licorice root as a source of phytoestrogen can act similar to estrogen in cell culture. Furthermore, the analysis of the licorice extract (LE) based on HPLC-DAD-ESI-MS indicated that LE comprises phytoestrogen compounds, such as glabridin and glabrene. In this study, the effects of LE on proliferation of human bone-marrow mesenchymal stem cells (hBM-MSCs) were investigated using MTT assay. In addition, its effects on the osteogenesis were evaluated using alkaline phosphatase activity (ALP), calcium deposition, and bone specific gene expression such as ALP, osteocalcin, Runx2, and BMP-2. The quantitative gene expression was studied by real-time RT-PCR. Our results showed a significant increase in proliferation in presence of LE in concentration 10-50 µg/mL. The differentiation of hBM-MSCs increased in doses of LE (10-25 µg/mL) compared to the control group. The effects of LE were similar to those of 17β-estradiol (E2) (10-8 M) and were abolished by ICI 182,780 an antagonist of estrogen receptor (ER) (10-7), indicating that the stimulatory effects of LE occur through estrogen receptor-mediated mechanism . Taking these into account, LE may be a potential candidate for prevention of osteoporosis in menopausal women.

Phenolic compounds that present in the several industries are harmful and dangerous for human health. In this study we have studied the potential of Atropa belladonna hairy roots in phenol removal of wastewater. The optimal conditions for the removal process were evaluated using different phenol (10-500 mg.1(-1)) and H2O2 (1-15 Mm) concentrations. In the presence of H2O2, Roots were able to remove phenol concentrations up to 500 mg.1(-1). in the wide range of pH (4-9), reaching high removal efficiency. When roots were re-used for five consecutive cycles, phenol removal efficiency decreased from 98-62%, in the last cycle. After the removal process, the solutions were obtained from the experiment were estimated for their toxicity using a test with Lactaca sativa L. seeds. Results showed that the treated solution was less toxic than the parent solution.

Graphical abstract Drought stress increased the amount of glycyrrhizin in the stolons of Glycyrrhiza glabra. The expression of sequalene synthase (SQS) and β-amyrin synthase (bAS) genes were increased under drought stress, whereas expression of cycloartenol synthase (CAS) was not changed., Highlights • We applied various drought stresses in liquorice. • Drought stress increased the expression of SQS and bAS, whereas expression of CAS was not changed. • Drought stress increased the amount of glycyrrhizin in the stolons of G. glabra. • Applying special irrigation regimes can principally improve secondary metabolite production., Glycyrrhiza glabra is an important medicinal plant throughout the world. Glycyrrhizin is a triterpenoid that is among the most important secondary metabolites produced by liquorice. Drought stress is proposed to enhance the levels of secondary metabolites. In this study, the effect of drought stress on the expression of important genes involved in the glycyrrhizin biosynthetic pathway was examined. Drought stress at the seedling stage was applied to 8-day-old plants using polyethylene glycol. Subsequently, the samples were collected 0, 4, 8 or 24 h post-treatment. At the adult plant stage, 10-month-old plants were subjected to drought stress by discontinuing irrigation. Subsequently, samples were collected at 2, 16 and 28 days after drought imposition (S2d, S16d and S28d, respectively). We performed semi-quantitative RT-PCR assays to evaluate the gene expression levels of sequalene synthase (SQS), β-amyrin synthase (bAS), lupeol synthase (LUS) and cycloartenol synthase (CAS) during stress. Finally, the glycyrrhizin content of stolons was determined via HPLC. The results revealed that due to osmotic stress, the gene expression levels of SQS and bAS were increased, whereas those of CAS were relatively unchanged at the seedling stage. At the adult plant stage, the expression levels of SQS and bAS were increased under drought stress conditions, whereas the gene expression level of CAS remained relatively constant. The glycyrrhizin content in stolons was increased only under severe drought stress conditions (S28d). Our results indicate that application of controlled drought stress up-regulates the expression of key genes involved in the biosynthesis of triterpenoid saponins and directly enhances the production of secondary metabolites, including glycyrrhizin, in liquorice plants.

Different parts of three plants ( Primula auriculata , Fumaria vaillantii and Falcaria vulgaris ) were extracted with three different solvents to yield 72 crude extracts. The phytochemical analysis (chemical screening, GC–MS) of three plants was investigated for their antioxidant and antibacterial activity using nine Gram-positive and Gram-negative bacteria. The principal antioxidant and antimicrobial components were determined using HPLC with UV detection. All extracts possessed antibacterial activity especially methanolic extracts from flowers of P. auriculata . The DPPH-radical scavenging assay exhibited high antioxidant activities in three plants (more than 80% at 50 μg). The F. vulgaris showed high content of carvacrol (29.8%) as main component. The contents of carvacrol and fumaric acid in the methanolic–water extracts were 1119 and 1966 mg/l respectively. Our results indicate that these plants would be able to promise sources of natural products with potential antibacterial and antioxidant activity.

Background: Arctium lappa is a medicinal plant, which natively grows in Iran. Arctiin and arctigenin are two of the most important secondary metabolites produced in this plant and used as anticancer reagents. Application of biotechnological methods for increasing the production of these metabolites is very essential. Recently, induction of hairy roots in plants to improve the production of desirable secondary metabolites has been considered. Objective: In this study, hairy roots were induced in Arctium lappa using Agrobacterium rhizogenes, strain AR15834. Methods: The leaf explants and seedlings of Burdock were used for the hairy roots induction. The leaf explants were turned brown and died when cultured in vitro. To improve the viability of these explants, different antioxidants including ascorbic acid (ASC), citric acid (CIT), polyvinylpyrrolidone (PVP) and L-cysteine were added to the MS medium, in different concentrations either alone or in combination with each other. Polymerase Chain Reaction (PCR) for rolB gene confirmed the morphological identification of the transformed hairy roots. Results: Although the best antioxidant reagent was PVP 0.5% (w/v), it did not stop tissues browning of the leaf explants when they were co-cultured with A. rhizogenes for the hairy roots induction. When the two or three weeks old seedling were used for the hairy roots induction, the roots were observed 2 weeks after the bacterial infection with 5% frequency. Conclusion: In this study, A. rhizogenes induced hairy roots in Burdock. To the best of our knowledge, this is a first report on the hairy roots induction in Arctium lappa.

Efficient plant regeneration was achieved from callus derived from immature-cotyledon explants of oleaster (Elaeagnus angustifolia L.). Calli were obtained on MS media containing 3% sucrose and different concentrations of TDZ. The highest rate of green, compact and nodular callus was formed on MS medium supplemented with 1 mg/l of TDZ. Shoot organogenesis was achieved when the callus was transferred onto MS media containing 3% sucrose and BA alone (05–4 mg/l) or BA (0.5 and 1 mg/l) combined with NAA or IAA (0.5 and 1 mg/l). Maximum organogenesis was obtained with 1 mg/l BA in combination with 0.5 mg/l NAA. Rooting of the shoots was achieved on MS medium supplemented with 0.2 mg/l IBA. Regenerated plantlets were acclimatized and successfully transplanted to soil.

Medicinal plants are a source for a wide variety of natural antioxidants. In the study reported here, we have conducted a comparative study between five medicinal plants having the same geographic origin: the Hamadan region in the west of Iran and growing in the same natural conditions. The amount of total phenolics and total flavonoids for parts of these plants used in Iranian popular medicine were evaluated. Furthermore, antioxidant activities for these parts using vitamin C equivalent antioxidant capacity (VCEAC) test were also evaluated. The results show that the antioxidant activities varied greatly among the different plant parts used in this study and some plants are rich in natural antioxidants especially leaves of Lavandula officinalis and of Melissa officinalis. A positive correlation between total phenolic or flavonoid contents and VCEAC was found with a correlation coefficient of R2 = 0.961 and R2 = 0.817, respectively. These findings show that phenolics in these plants provide substantial antioxidant activity.

The aim of this study was to use the, hairy root system for increasing the scopolamine content in Atropa belladonna. Agrobacterium rhizogenes ATCC15834 was utilized to produce hairy roots. The culture was carried out in a 1.5-l bioreactor using the inoculum size of 0.5 g fr. wt of 10-day-old hairy roots and various parameters, including agitation, aeration, conductivity and the consumption of sucrose were evaluated. Results revealed that the highest amount of scopolamine production (1.59 mg/g-1 dry wt) occurred in the bioreactor with aeration and agitation 1.25 vvm (volume per volume per minute) and 70 rpm, respectively. Study of conductivity and the consumption of sucrose showed that the highest amount of sucrose consumption and the highest amount of minerals consumption also was at 1.25 vvm and 70 rpm. Transgenic hairy root lines were confirmed by polymerase chain reaction (PCR).

Glycyrrhizin and isoliquiritigenin production by hairy root culture of Licorice (Glycyrrhiza glabra) was investigated using Agrobacterium rhizogenes strain AR15834. Hairy roots were induced by inoculation of the leaf and stem explants with A. rhizogenes. Presence of therolB gene in the hairy root lines was performed by polymerase chain reaction (PCR) and using rolB gene specific primers. The amount of glycyrrhizin and isoliquiritigenin in roots was measured by high-performance liquid chromatography (HPLC). The highest transformation frequency was obtained (50%) in leaf explants. The obtained results on hairy roots of G. glabra showed two distinguishable phenotypes, typical hairy roots and callus-like roots. In contrast to the previous studies, hairy root cultures of G. glabra synthesized glycyrrhizin in this research. In all of the hairy root lines, biomass and glycyrrhizin production was more than normal roots. The highest amount of glycyrrhizin and isoliquiritigenin was produced in callus-like roots. Key words: Agrobacterium rhizogenes, Glycyrrhiza glabra, glycyrrhizin, hairy root, isoliquiritigenin.

This research focuses on the analysis of the chemical composition lemon balm essential oil. The essential oil of flowers was obtained by steam distillation with a Clevenger apparatus. The chemical components of the essential oil of lemon balm were analyzed by capillary GC and GC/MS and 12 substances were identified. The flower oil contained trans-carveol (28.89%), citronellol (25.24%), δ-3-carene (5.26%), citronellal (4.9%), geraniol (2.2%), 1-octene-3-ol (2.03%) and spathulenol (2.06%). The trans-carveol in flowers was dominant among components.