Biotechnology
Marieh Dorrazehi; Maryam Allahdou; Barat Ali Fakheri; Leila Mehravaran
Abstract
Background and objectives: Medicinal plants produce bioactive compounds with high antioxidant value. Due to the importance of Withania coagulans in the pharmaceutical industry and also due to the cutting of this shrub species and its use in the pharmaceutical industry using traditional methods, the risk ...
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Background and objectives: Medicinal plants produce bioactive compounds with high antioxidant value. Due to the importance of Withania coagulans in the pharmaceutical industry and also due to the cutting of this shrub species and its use in the pharmaceutical industry using traditional methods, the risk of extinction of this valuable species will increase, so we should look for the method - to be alternatives to increase the production of secondary metabolites of this plant. Using elicitors as a technique in vitro increases plant production of effective compounds (secondary metabolites). It has been reported that fungal stimulants are also used to induce the production or increase of secondary metabolites in medicinal plants. Therefore, in this study, the effect of cellulase elicitor on the growth index and biochemical and physiological traits of Withania coagulans (Stocks) Dunal) was investigated in cell suspension culture.Methodology: Seeds of Withania coagulans were collected from the Mehrestan region of Saravan city, and after disinfection, they were cultivated in ½ MS medium to produce explant. Production explants were cultured in an MS culture medium containing auxin and cytokinin hormones (2mg/l 2, 4, D, and 0.5 mg/l Kinetin) to produce callus. Three sub-cultures of production called into MS culture medium containing transfer previous regulating hormones were done. Investigated treatments include six elicitor treatments: 1: Control 2: Elicitor with a concentration of 200 μg/ml and 24 hours of exposure to the Elicitor, 3: Elicitor with a concentration of 200 μg/ml and 48 hours of exposure to the Elicitor, 4: Elicitor with a concentration of 200 μg/ml and 72 hours of exposure Elicitor, 5: Elicitor with a concentration of 7.5 μg/ml and 14 days of exposure to the Elicitor and 6: Elicitor with a concentration of 10 μg/ml and 14 days of exposure to the cellulase elicitor in a completely randomized design with three replications in the central laboratory of the university of Zabol was applied. Investigated traits were: callus growth index, antioxidant activity, proline content, soluble carbohydrates, malondialdehyde, total alkaloid, and activity of antioxidant enzymes (catalase, ascorbate peroxidase, guaiacol peroxidase, superoxide dismutase, and polyphenol oxidase).Results: The effect of different cellulase treatments on all traits was significant except polyphenol oxidase enzyme activity. In general, different elicitor treatments improved all the measured traits except the content of total carbohydrates. However, the increase in the trait depended on the concentration and period of exposure to the Elicitor. So, in most of the measured traits, the elicitor treatment with a concentration of 10 micrograms/ml and a period of 14 days caused an increase in all treatments. All cellulase elicitor treatments increased the antioxidant activity of Withania coagulans cells in cell suspension culture. The highest antioxidant activity was observed in the sixth and fifth treatments and the lowest in the second and first treatments (control). The addition of Elicitor in the early stage of cell growth in cell suspension culture (eighth day) despite the lower concentrations of Elicitor (7.5 and 10 μg/ml) leads to a further increase in callus growth, antioxidant activity, Proline, and total alkaloid increased then the addition of Elicitor in exponential phase, despite the high concentration of Elicitor (200 μg/ml). A higher concentration of elicitors causes a hypersensitive response, which leads to cell death; hence, an optimal Elicitor level is required to induce growth.Conclusion: Cellulase elicitor improved the growth index and antioxidant potential of Withania coagulans through physiological and biochemical traits. According to the obtained results, the use of cellulase elicitor can be considered an important strategy to increase the growth, effective compounds, and antioxidant properties of Withania coagulans for commercial production.
Improvement and breeding
B. Asghari; S. Mafakheri; M. M. Zarrabi
Abstract
Background and Objectives: Hibiscus sabdariffa L., commonly known as Roselle, is a plant cultivated in many regions of the world regions. Roselle has been used as a medicinal herb in traditional medicine for controlling and treating different diseases and physical ailments. These therapeutic effects ...
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Background and Objectives: Hibiscus sabdariffa L., commonly known as Roselle, is a plant cultivated in many regions of the world regions. Roselle has been used as a medicinal herb in traditional medicine for controlling and treating different diseases and physical ailments. These therapeutic effects are attributed to active compounds such as carotenoids, fatty acids, and phenolic compounds, including phenolic acids and flavonoids. One common method to stimulate plants for increased production of active compounds is subjecting them to various types of stresses, such as drought or salinity, along with plant growth regulators. This study investigated salicylic acid's effect on the phytochemical content and various biological properties of Roselle plants under water scarcity conditions.Methodology: The experiment was conducted as a factorial experiment in a completely randomized design, including irrigation intervals at three levels: three days (D1), five days (D2), and seven days (D3), and foliar application of salicylic acid at three levels: 0 (SA0), 0.75 mM (SA1), and 1.25 mM (SA2), with three replicates in a research greenhouse at the International Imam Khomeini University. The required seeds for cultivation were obtained from Pakanbazr Company in Isfahan. The imposition of drought stress through irrigation intervals of 3, 5, and 7 days started at the 4-leaf stage of the plants, and the plant spraying with salicylic acid was performed at the 6-leaf stage, three days before the start of irrigation treatments. The measurement of electrolyte leakage and malondialdehyde content was carried out to determine membrane stability and lipid peroxidation level. Additionally, the measurement of proline content and the activity of three antioxidant enzymes, including peroxidase, catalase, and ascorbate peroxidase, was performed in different samples. Furthermore, the total phenolic, flavonoid, and anthocyanin contents, as the main secondary metabolites of Roselle plants, were measured. The biological and medicinal properties, including the free radical scavenging activity (DPPH) and α-glucosidase inhibitory activity, were also evaluated. The obtained data were analyzed using SAS statistical software version 9.4. Mean comparisons were conducted using Duncan's multiple range test at a significance level of 5%.Results: The results showed both electrolyte leakage and malondialdehyde content increased significantly with increasing levels of applied drought stress on Roselle plants. However, overall, applying salicylic acid treatments led to a decrease in both indicators. The measurement of proline content in Roselle samples showed that the highest amount of proline was observed in plants subjected to the highest drought stress and treated with the 1.5 mM salicylic acid solution. The activity of all three antioxidant enzymes increased with the intensity of the applied drought stress and the concentration of salicylic acid solutions. Roselle's highest levels of phenolic and anthocyanin content were observed in samples sprayed with 1.5 mM salicylic acid solution. The change in anthocyanin content of Roselle plants also demonstrated that increasing the severity of water scarcity resulted in higher levels of this compound. The highest DPPH free radical scavenging activity was observed in Roselle samples grown under the highest drought severity and sprayed with the highest concentration of salicylic acid (61.93%). The highest antioxidant activity was also observed in samples treated with the highest concentration of salicylic acid solution in all irrigation regimes. The highest α-glucosidase inhibitory activity (69.2% and 67.7%) was associated with Roselle samples grown under the highest applied drought severity, sprayed with 1.5 mM and 0.75 mM salicylic acid solutions, respectively.Conclusion: It can be concluded that the use of salicylic acid as a growth regulator can contribute to the enhancement of phytochemical content, especially secondary metabolites such as phenolic and flavonoid compounds, anthocyanins, and the elevation of defensive parameters such as the activation of antioxidant enzymes, as well as an increase in osmolyte content like proline in Roselle plants. These effects assist the plants in exhibiting greater resistance to drought stress and mitigating the negative impacts of this type of stress on the plant.
Improvement and breeding
A. Negari; M. Jami Al-Ahmadi; G. Zamani
Abstract
Medicinal plants are a rich and valuable source of secondary metabolites that are strongly influenced by the environmental factors, especially drought stress. In this regard, a greenhouse experiment was conducted on Thymus vulgaris L. as split plots in a randomized complete block design with three replications. ...
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Medicinal plants are a rich and valuable source of secondary metabolites that are strongly influenced by the environmental factors, especially drought stress. In this regard, a greenhouse experiment was conducted on Thymus vulgaris L. as split plots in a randomized complete block design with three replications. The main plot included three levels of moisture supply (40, 65, and 90% of potting soil field capacity (FC)) and the sub-plot was the elicitors levels including (1) control: without elicitor, (2) 150 µM cyclodextrin (CYC), (3) 75 nM coronatine (COR) + 150 µM CYC, (4) 150 nM COR + 150 µM CYC, (5) 150 µM methyl jasmonate (MJ) + 150 µM CYC, and (6) 300 µM MJ + 150 µM CYC. Except for the essential oil percentage, the other traits including the leaves proline content, electrolyte leakage, relative water content, plant height, canopy diameter, dry and fresh weight of shoots, dry weight of leaves and stems, and weight of essential oil had a very significant correlation with each other. The highest amount of leaves proline was obtained form the interaction of elicitors (levels 4 and 6) and the 40% FC level of moisture supply (3.88 and 3.94 μmol g-1 fresh leaf weight, respectively). The highest amount of plant height (28.3 cm), canopy diameter (17 cm), and relative water content (79%) was observed at the 90% FC level of moisture supply. The highest percentage of essential oil was obtained in the treatment of non-application of elicitor (2.67%) and the highest essential oil weight per plant (0.092 g plant-1) was observed in the treatment of non-application of elicitor at the 65% FC level of moisture supply. The mild stress increased the essential oil yield and the elicitor treatments decreased it.
Improvement and breeding
E. Chamani; F. Narimaniyan; Y. Pourbairami Hir; H.R. Heydari
Abstract
To investigate the effects of different nitrogen sources including ammonium nitrate, asparagine, and glutamine in five concentrations of 0, 50, 100,150, and 200 mg l-1 on some physiological and biochemical characteristics of rosemary (Rosmarinus officinalis L.), an experiment was conducted in a completely ...
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To investigate the effects of different nitrogen sources including ammonium nitrate, asparagine, and glutamine in five concentrations of 0, 50, 100,150, and 200 mg l-1 on some physiological and biochemical characteristics of rosemary (Rosmarinus officinalis L.), an experiment was conducted in a completely randomized design with five repliacations. The indices including plant height, shooting rate, number of leaves, necrotic leaf size, chlorophylls a, b, and total chlorophyll content, and the content of carotenoids, phenols, and flavonoids were measured. The results showed that the nitrogen sources significantly (P≤0.01) affected the all measured indices except the necrotic leaf size. The results of means comparison showed that the highest shooting rate and number of leaves were obtained at 100 mg l-1 of glutamine and asparagine and the highest plant height was obtained at 200 mg l-1 of ammonium nitrate. Also, the highest amount of photosynthetic pigments was obtained at 50 mg l-1 of asparagine and the highest amount of phenols and flavonoids was related to 200 mg l-1 of glutamine. In general, the results showed that the various nitrogen sources studied could positively and significantly improve the morphological and physiological characteristics of rosemary under in vitro conditions.
Improvement and breeding
S.G. Sajjadi; A. Tavassoli; M. Dadmehr
Abstract
To investigate the effects of hydrogel on growth, yield, and some secondary metabolites of roselle (Hibiscus sabdariffa L.) under drought stress conditions, an experiment was conducted in a farm located in the south of Dalgan city, Sistan and Baluchestan province in the 2017-2018 crop year. This research ...
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To investigate the effects of hydrogel on growth, yield, and some secondary metabolites of roselle (Hibiscus sabdariffa L.) under drought stress conditions, an experiment was conducted in a farm located in the south of Dalgan city, Sistan and Baluchestan province in the 2017-2018 crop year. This research was carried out as a split plot experiment in a randomized complete block design with three replications. The experimental treatments included drought stress as main plot at four levels (irrigation at FC (field capacity), 85% of FC, 70% of FC, and 55% of FC), and hydrogel consumption as sub plot at three levels (A200 hydrogel consumption, silicate hydrogel consumption, and no hydrogel consumption). The quantitative traits including plant height, number of sub-branches per plant, number of flower per plant, plant fresh and dry weight, sepal fresh and dry yield, and harvest index and qualitative traits including content of anthocyanin and sepal phenolic compounds were measured. The results showed that the highest amount of quantitative traits was obtained from full irrigation treatment with the consumption of A200 hydrogel. But there was no significant difference between this treatment and the treatments of full irrigation with silicate hydrogel consumption and full irrigation without hydrogel. Also, no significant difference was observed between the above three treatments and the treatments of both types of hydrogels application in 85% and 70% of FC. The qualitative traits increased with increasing the stress intensity. The hydrogel consumption in comparison with not consuming it led to a decrease in the amount of anthocyanin and sepal phenolic compounds due to the modification of stress conditions by hydrogel.
M. Dorafshan; M. Soltani Howyzeh; V. Shariati
Abstract
Identification of the genes biosynthesizing medicinal plants-specific metabolites is now performed with great speed and accuracy using new transcriptome study technologies such as RNA sequencing. The present study was carried out to find the specific genes in the biosynthetic pathway of triterpenes and ...
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Identification of the genes biosynthesizing medicinal plants-specific metabolites is now performed with great speed and accuracy using new transcriptome study technologies such as RNA sequencing. The present study was carried out to find the specific genes in the biosynthetic pathway of triterpenes and sesquiterpenes in the fruit tissue of colocynth (Citrullus colocynthis L.). After RNA extraction from the tissue of colocynth fruits harvested from Andimeshk region in Khuzestan province in 2017, RNA sequencing technique was performed using the Illumina HiSeq2500 platform. The bioinformatics steps including de novo assembly, using the Evidential-gene software, and functional annotation, using the KAAS database, were performed. In the KAAS database, 17359 unigenes were annotated in 134 plant pathways. Among the different and important secondary metabolites pathways in the fruit tissue of colocynth, 39 unigenes and 8 orthologous genes were assigned to the triterpenes and sesquiterpenes gene pathways. Transcriptome analysis of this medicinal plant with the aim of identifying the genes of secondary metabolites biosynthetic pathways underlies various research and practical aspects such as biosynthetic pathway engineering of herbal medicines.
A. Khakpoor; M. Zolfaghari; K. Sorkheh
Abstract
Medicinal plant licorice (Glycyrrhiza glabra L.) has been used by the pharmaceutical industries for its important secondary metabolites. This study was carried out to determine the best harvest time for obtaining the highest content of extract, glycyrrhizin, total phenols, total flavonoids, and phenolic ...
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Medicinal plant licorice (Glycyrrhiza glabra L.) has been used by the pharmaceutical industries for its important secondary metabolites. This study was carried out to determine the best harvest time for obtaining the highest content of extract, glycyrrhizin, total phenols, total flavonoids, and phenolic compounds of licorice rhizome of the Behbahan area in Khuzestan province in two seasons of autumn and spring. The results of this study showed that the extract content of rhizomes and also glycyrrhizin, total phenols, and flavonoids content in autumn were higher than those in spring. The results of the independent T-test for glycyrrhizin, as an important and valuable secondary metabolite of licorice, showed a significant difference between autumn and spring seasons. Among the phenolic compounds studied, the highest amount was given to ferulic and coumaric acids in autumn, and the lowest one to caffeic acid in spring. Considering the importance of these secondary metabolites in the pharmaceutical, health and food industries, as well as the widespread use of phenolics, glycyrrhizin, and flavonoids of licorice in herbal drugs, it is necessary to determine the best time for the highest production and accumulation of these metabolites in the licorice rhizomes to take an effective step to enrich the industries mentioned.
S. Syadat Jamian; M. Aghaalikhani; A. Mokhtassi-Bidgoli
Abstract
To investigate the effect of planting density on quantitative and qualitative characteristics of globe artichoke (Cynara cardunculus L. var. Scolymus (L.) Fiori) under weedy and weed-free conditions, a field experiment was conducted at Tarbiat Modares University, Tehran in 2016 growing season. A factorial ...
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To investigate the effect of planting density on quantitative and qualitative characteristics of globe artichoke (Cynara cardunculus L. var. Scolymus (L.) Fiori) under weedy and weed-free conditions, a field experiment was conducted at Tarbiat Modares University, Tehran in 2016 growing season. A factorial arrangement of the experimental treatments was studied in a randomized complete block design with three replications. The factors included four crop planting densities (2, 4, 6 and 8 plants m-2) under two conditions of presence and absence of weeds. Different traits including leaf number per plant, leaf area index, total dry weight, chlorophylls a, b and total chlorophyll, caffeic acid, chlorogenic acid and total polyphenol contents were measured. The results showed that leaf number and dry weight were significantly affected by plant density and weed competition in artichoke, so that the weed competition reduced the artichoke dry weight by 22.4%. The highest leaf area index (2.29) and dry weight of artichoke (253.44 g m-2) were observed at 6 plants m-2, while the highest number of leaves per plant (16.33) belonged to the density of 2 plants m-2. Among qualitative traits chlorophylls a, b and total chlorophyll were only affected by weed presence while caffeic acid, chlorogenic acid and total polyphenol contents were only affected by plant density. The effect of crop plant density on weed leaf area and dry weight was also significant. Overall, the highest performance of the main secondary metabolites in artichoke including caffeic acid, chlorogenic acid and total polyphenol (5.99, 9.55 and 5.89 g m-2, respectively) were obtained at 6 plants m-2, completely affected by crop dry matter yield.
M. Amini; S. Yousefzadeh; K. Sadat-Asilan
Abstract
In order to study the effects of iron, zinc and manganese sulfate on essential oil percentage, yield and composition in hyssop (Hyssopus officinalis L.), an experiment was conducted as randomized complete block design with three replications during 2016.Experimental treatments consisted of control, foliar ...
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In order to study the effects of iron, zinc and manganese sulfate on essential oil percentage, yield and composition in hyssop (Hyssopus officinalis L.), an experiment was conducted as randomized complete block design with three replications during 2016.Experimental treatments consisted of control, foliar application of iron, zinc, manganese, iron+zinc, iron+manganese, zinc+manganese and iron+zinc+ manganese. Results showed that foliar application of microelements had significant effects on all traits studied except cis-pinocamphone. Foliar application of iron + zinc + manganese treatment produced the maximum essential oil percentage (0.6%) and essential oil yield (30.44 Kgha-1). Foliar application of iron + zinc + manganese treatment increased essential oil percentage up to 50% compared to the control treatment. β-pinene, trance-pinocamphone, pinocarvone and cis-pinocamphone formed 75 to 90% of the oil components. Foliar application of iron + zinc + manganese and control treatments produced the highest (89.4%) and the lowest (77.1%) values of total β-pinene, trance-pinocamphone, pinocarvone and cis-pinocamphone, respectively. Therefore, results showed that the foliar application of Zn+ Fe+ Mn could be used as a suitable management strategy to reach the optimum essential oil yield and composition in hyssop.
E. Vatankhah; B. Kalantari; B. Andalibi
Abstract
Jasmonates are key signaling compounds in plant responses to biotic and abiotic stresses as well as in development. We studied the role of methyl jasmonate (MeJA) in alleviating NaCl-induced salt stress in Mentha piperita L. through evaluating its growth, mineral and phenolics contents, essential oil ...
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Jasmonates are key signaling compounds in plant responses to biotic and abiotic stresses as well as in development. We studied the role of methyl jasmonate (MeJA) in alleviating NaCl-induced salt stress in Mentha piperita L. through evaluating its growth, mineral and phenolics contents, essential oil (EO) quantity and quality. Accordingly, plants were exposed to the NaCl concentrations of 1.86 (control), 5, 75 and 10 dS/m for two weeks, 24 h after foliar application of 0, 60 and 120 µM MeJA. The results showed that salt stress decreased fresh and dry weights of aerial parts and concentrations of potassium (K+), calcium (Ca2+), magnesium (Mg2+), K+/ Na+ ratio and phenolic compounds, while sodium (Na+) content increased significantly under salt stress. Also, EO yield increased with the increase of salinity concentration then decreased at the salinity concentration of 10 dS/m. However, exogenous application of MeJA enhanced the contents of K+, Ca2+, Mg2+, K+/ Na+ ratio and phenolics, while Na+ content in salt stressed plants declined. In addition, pretreatment with 60 µM MeJA improved the growth and EO yield. The highest values of menthone and menthol were obtained at the salinity concentration of 7.5 dS/m and concentrations of 60 and 120 µM MeJA. In summary, our results indicated that application of MeJA could reduce the adverse of salinity stress in M. piperita.
M. Noori; Sh. Gharanjik; A. Safipour Afshar; F. Saeid Nematpur
Abstract
The hairy roots induced by Agrobacterium rhizogenes due to the stability and mass production in hormone free culture provide an appropriate tissue for secondary metabolite production, and the content of metabolites produced could be affected by bacterial strain. In the current study, the effect of different ...
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The hairy roots induced by Agrobacterium rhizogenes due to the stability and mass production in hormone free culture provide an appropriate tissue for secondary metabolite production, and the content of metabolites produced could be affected by bacterial strain. In the current study, the effect of different strains of A. rhizogenes including A4, A13 and 15834 on the percentage of hairy root induction of Echinacea purpurea (L.) moench. in the leaf and hypocotyl explants were studied. In another experiment, total phenolics and polysaccharides content of hairy roots were measured compared to the control (untransformed) roots. The results showed that the percentage of hairy root induction was significantly affected by bacterial strains, so that in E. purpurea leaves incubated by strains A13, 15834 and A4, the hairy root induction was 60%, 40% and 0, and in hypocotyl explants was 85%, 45% and 70%, respectively. Maximum total phenolic compounds were observed in A4 strain. As well, in comparison to control, the polysaccharide content in hairy root induced by strains 15834 and A13 increased and by A4 strain decreased significantly. Differences observed in this study about hairy root induction and secondary metabolites production could be due to different ability of A. rhizogenes strains in T-DNA transferring to plant cells.
F. Tabe bordbar; S. Moharramipour
Abstract
The Diamond back moth Plutella xylostella L. is one of the major pests of plants in Brassicaceae that has become resistant to many insecticides. In recent years, plant secondary metabolites play an important role in controlling pests because of their insecticidal and antifeedant properties. A laboratory ...
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The Diamond back moth Plutella xylostella L. is one of the major pests of plants in Brassicaceae that has become resistant to many insecticides. In recent years, plant secondary metabolites play an important role in controlling pests because of their insecticidal and antifeedant properties. A laboratory experiment was conducted to investigate the effect of methanolic extracts of Matteuccia struthiopteris (L.) on diamondback moth third instar larvae. Findings showed significant increase in mortality as concentrations increased, so that, concentrations to cause 25% (LC25) and 50% (LC50) mortality in population were 12.01% and 28.58%, respectively. In this research, the effect of sublethal concentration of methanolic extract was studied on demography parameters such as life table, reproductive and population of P. xylostella. The obtained results showed that different concentration had a significant effect on the duration of different life stage, fecundity and adult longevity. Data analysis demonstrated that increased concentration decreased the net reproductive rate (R0), intrinsic rate of natural increase (rm), finite rate of increase (λ) but increased mean generation time (T) and doubling time (DT) significantly. These findings indicate that M. struthiopteris has the potential to be used as a reliable method in integrated management of this pest.
F. Nasibi; Kh. Manuchehri Kalantari
Abstract
Ultraviolet spectrum of sunlight has high energy. It was divided into three bands A, B, and C, due to the wavelength. Plants protected themselves from this kind of radiation, by two mechanisms, enzymatic and non-enzymatic. These mechanisms can be used for medicinal plants. Many of the secondary metabolites ...
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Ultraviolet spectrum of sunlight has high energy. It was divided into three bands A, B, and C, due to the wavelength. Plants protected themselves from this kind of radiation, by two mechanisms, enzymatic and non-enzymatic. These mechanisms can be used for medicinal plants. Many of the secondary metabolites including flavonoids, anthocyanin and alkaloids are medicinally important and they as well have a vital role in scavenging of the free radical created by UV radiation. Therefore, UV radiation seems to be able to increase these compounds in plants. In this research the effect of UV-B and UV-C on the amounts of these compounds (flavonoids, anthocyanin and alkaloids ) in hyoscyamus niger and hyoscyamus muticus was studied. The seeds were collected from their habitat, sown in pots in growth cabinet at 21±1ºC and light period of 16/8 light /dark. After 3 weeks, these plants were treated with UV-B and UV-C for 30 minutes every day, during 2 weeks. After 2 weeks, plant materials were harvested and freezed using liquid nitrogen. The samples were used for determination these parameters. Flavonoids were measured by HPLC method. The amount of flavonoids increased in comparison to the control. Anthocyanins concentration were measured using spectrophotometer. The extinction coefficient was used to calculate the concentration. This compound was shown to increase 35% and 50% in UV-B and UV-C, respectively in comparison to the control. The calculation of reducing sugar showed that these sugars decreased in both species when treated either with UV-B or UV-C.
