Biotechnology
F. Noormand moaied
Abstract
Background and objectives: Milk thistle (Silybum marianum L.) is a significant medicinal plant that has gained a prominent place in the pharmaceutical industry. This annual or biennial plant contains bioactive compounds, particularly flavonoids, which are primarily concentrated in its seeds. The collective ...
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Background and objectives: Milk thistle (Silybum marianum L.) is a significant medicinal plant that has gained a prominent place in the pharmaceutical industry. This annual or biennial plant contains bioactive compounds, particularly flavonoids, which are primarily concentrated in its seeds. The collective active components of this plant are referred to as silymarin, a compound recognized for its anti-cancer properties. Cell culture systems provide a promising method for large-scale cultivation of plant cells to produce secondary metabolites. This research aimed to investigate the effects of genotype, micro-sample type, phytohormone treatments, and callus type on the synthesis of bioactive compounds in S. marianum calluses using high-performance liquid chromatography (HPLC).Methodology: In this research, a combination of Tween-20 solution, 70% ethanol, hydrogen peroxide, and sterile distilled water was used to disinfect Silybum marianum seeds to produce sterile seedlings. The sterilized seeds were then transferred to sterilized water and an agar culture medium (12 g/L) and kept in dark conditions at a temperature of 25°C for 15 days. After germination and initial growth, the seedlings were moved to light conditions. An experiment was conducted using two genotypes, Hungary and Borazjan, with cotyledon and hypocotyl micro-samples. The experiment utilized Murashige and Skoog (MS) medium and hormonal treatments of 2,4-Dichlorophenoxyacetic acid (2,4-D) at concentrations of 1, 2.5, and 5 mg/L, along with Benzyl Amino Purine (BAP) at 0.25 and 0.5 mg/L. This setup was maintained in dark conditions and organized as a factorial design based on a completely randomized block structure. After 30 days, the calluses were transferred to MS culture medium with half the concentration of hormones. The effective compounds were extracted from the callus through defatting with petroleum ether, followed by extraction with methanol solvents. The components of silymarin were then separated into taxifolin, silychristine, silydianin, silybin, and isosilybin using high-performance liquid chromatography (HPLC). Statistical analysis of the data was performed using SPSS software.Results: The test results demonstrated the effects of different micro-sample treatments, ecotypes, and hormone concentrations on the levels of silymarin flavonoid compounds (taxifolin, silychristin, silydianin, silybin, and isosilybin) in Silybum marianum extracts. Analysis of variance and comparison of the mean values of flavonolignans in the callus samples revealed that the highest amount of taxifolin was obtained from the hypocotyl micro-sample. The highest silychristin content was associated with the Hungarian genotype and a treatment of 5 mg/L 2,4-D. The highest silydianin concentration was observed in the Hungarian genotype treated with 1 mg/L 2,4-D and 0.25 mg/L BAP. The maximum silybin content was found in the Hungarian cultivar using the hypocotyl micro-sample, with 1 mg/L 2,4-D and 0.25 mg/L BAP. The highest isosilybin levels were achieved in the Hungarian genotype, hypocotyl micro-sample, and 1 mg/L 2,4-D. The overall highest silymarin concentration was recorded in the Hungarian genotype treated with 1 mg/L 2,4-D and 0.5 mg/L BAP.Conclusion: The results showed that improved varieties of Silybum marianum, such as the Hungarian genotype, produced higher levels of bioactive compounds compared to native ecotypes. In the production of these compounds, the type of micro-sample played a more critical role than the size and type of callus derived from it. Specifically, the hypocotyl micro-sample, which produced small, non-embryogenic calluses, yielded a higher percentage of effective substances compared to the cotyledon, which produced larger, embryogenic calluses. Additionally, lower concentrations of auxin and cytokinin hormones were more effective than higher concentrations in promoting the production of bioactive compounds.
Agriculture and horticulture
S. Alipour
Abstract
To investigate the effects of vermicompost and phosphate biofertilizer Barvar-2 on quantitative and qualitative yield of Silybum marianum L., a factorial experiment was conducted in a randomized complete block design with three replications in a field near Ardebil city in 2018-2019 crop year. Vermicompost ...
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To investigate the effects of vermicompost and phosphate biofertilizer Barvar-2 on quantitative and qualitative yield of Silybum marianum L., a factorial experiment was conducted in a randomized complete block design with three replications in a field near Ardebil city in 2018-2019 crop year. Vermicompost was used at five levels (0, 10, 20, 30, and 40 ton.ha-1) and biofertilizer at two levels (inoculation and non-inoculation). The results showed that seed yield, 1000-seed weight, number of seeds per capitule, number of capitules per plant, biological yield, oil yield, silymarin yield, and flower fresh weight were significantly affected by different levels of vermicompost. Also, the application of phosphate biofertilizer Barvar-2 had a significant effect on all traits except inflorescence diameter. The highest amount of traits in the vermicompost treatment was obtained in the application of 40 tons.ha-1. However, there was no statistically significant difference for the traits including number of seeds per capitule, oil, silymarin, and silybin percentage, oil yield, and number of inflorescences in the application of 40 and 30 tons vermicompost per hectare. In biofertilizer treatment, the highest amount of traits was observed in inoculation with this fertilizer. The highest silybin percentage (16.81) and silymarin yield (37.61 kg.ha-1) were obtained in the application of 40 tons vermicompost per hectare and the lowest one (16.12% and 30.56 kg.ha-1, respectively) at the control level of vermicompost.
Biotechnology
F. Noormand Moaied; N. Valizadeh; T. Samandari Giklo
Abstract
Silybum marianum L., from fam. asteraceae, is a very important medicinal plant in the pharmaceutical industry due to its active ingredient silymarin. Silymarin is very effective in treating the liver diseases, hepatitis, and cardiovascular disorders. Since the potential and production speed of this medicinal ...
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Silybum marianum L., from fam. asteraceae, is a very important medicinal plant in the pharmaceutical industry due to its active ingredient silymarin. Silymarin is very effective in treating the liver diseases, hepatitis, and cardiovascular disorders. Since the potential and production speed of this medicinal compound is low in habitats under natural conditions, the use of genetic engineering and production of transgenic plants using the various techniques such as tissue culture has been considered in this plant. The first step in this direction is to obtain an in vitro regeneration protocol. In this study, the leaf explants obtained from the Hungarian cultivar of S. marianum were evaluated for the direct regeneration affected by different combinations of hormones. Six weeks after the culture, the highest shooting (57%) was observed in the ½ MS medium containing 0.4 mg l-1 zeatin, 0.2 mg l-1 BAP, and 0.2 mg l-1 NAA. The shoots were then transferred to the root culture medium with the different hormonal treatments. The results showed that the best treatment to obtain the long and high roots was the ½ MS medium containing 0.25 mg l-1 NAA and 2 g l-1 activated charcoal. The seedlings obtained from the direct regeneration grew well after transplanting into the soil.
Kh. Azizi; J. Nazari Alam; Mohammad Feizian; R. Heydari
Abstract
This study was aimed to evaluate the quantitative and qualitative performance of different populations of Milk Thistle (Silybum marianum L.( under the influence of autumn and spring planting season. Therefore, a split plot experiment was conducted including autumn and spring planting as main plots and ...
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This study was aimed to evaluate the quantitative and qualitative performance of different populations of Milk Thistle (Silybum marianum L.( under the influence of autumn and spring planting season. Therefore, a split plot experiment was conducted including autumn and spring planting as main plots and four local populations (Shush, Poldoktar, Khoram Abad and Aleshtar) and a crop cultivar as sub plots. The study was carried out in the Faculty of Agriculture, Lorestan University, for two years 2014-2015. In addition, the native and crop populations of this species in Aleshtar were compared in a randomized complete block design. Based on the results, there was a significant difference between spring and autumn planting seasons. The highest plant height (107 cm), capitol diameter (44 mm), number of grains in capitol (78) and grain yield (1000 kg ha-1) were obtained in autumn planting. The interaction effect of planting season and population type showed that the highest content of silymarin (22 mg/g of dry weight) and silybin (3.9 mg /g of dry weight) was obtained from the crop cultivar in autumn planting. The comparison between native and crop populations in Aleshtar region showed that the highest content of silybin (4.2 mg /g of dry weight) and silymarin (more than 21 mg / g of dry weight) was recorded for native Aleshtar population compared with other populations. In general, it was determined that the highest and lowest quantitative and qualitative performance of Milk Thistle in the temperate region of Khorramabad was obtained in autumn planting, and the highest content of silymarin and silybin was recorded for the native Aleshtar population compared with other populations.
E. Zangani; S. Zehtab Salmasi; B. Andalibi; A.A. Zamani
Abstract
To evaluate the effects of sodium nitroprusside (SNP) on tolerance enhancement towards drought in Milk thistle (Silybium marianum (L.) Gaertn.), an experiment was conducted in a split split plot restriction in a randomized complete block design with three replications. The study was conducted in ...
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To evaluate the effects of sodium nitroprusside (SNP) on tolerance enhancement towards drought in Milk thistle (Silybium marianum (L.) Gaertn.), an experiment was conducted in a split split plot restriction in a randomized complete block design with three replications. The study was conducted in the research field of the University of Zanjan in 2014. Sodium nitroprusside at three levels of 0, 100 and 200 µmol l-1 was considered as the main factor, while drought stress at three levels of control and withholding irrigation at stem elongation and anthesis stages were regarded as sub factors. Finally, two genotypes of milk thistle (Hungarian and sari) comprised sub-sub factors. With increasing drought severity, leaf photosynthesis rate decreased significantly up to 45 percent compared with the control in both genotypes. Application of 100 µM SNP increased the leaf photosynthesis during stem elongation by roughly 80 and 100 percent in the Hungarian cultivar and Sari ecotype, respectively compared to non-application of SNP. Drought stress significantly decreased the pigment content in Sari ecotype, while this decrease was compensated by applying SNP during stem elongation. Drought stress in both stages of irrigation withholding reduced the grain yield in both genotypes. However, the exogenous application of 100 µM SNP compensated the reduction of grain yield, particularly in Sari ecotype. In addition, withholding of irrigation reduced the active ingredient yield; however, this reduction was compensated by increasing the content of silymarin and grain yield especially in the stage of anthesis stress with application of 100 µM SNP. Research findings showed that concentration of 100 µM SNP was more efficient to protect the milk thistle with increasing drought severity and cultivation of this plant in the water deficit systems.
H. Yadegari; I. Khammari; M. Salari; B.A. Fakheri; M. Rahimi; F. Bidarnamani
Abstract
Milk thistle (Silybum marianum L.), belonging to the Asteraceae family, is an annual plant and prefers light and fertile soils. Silymarin is the most important compound extracted from the seeds of this plant and it is used in the treatment of liver diseases, hepatitis, cirrhosis and liver cancer prevention. ...
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Milk thistle (Silybum marianum L.), belonging to the Asteraceae family, is an annual plant and prefers light and fertile soils. Silymarin is the most important compound extracted from the seeds of this plant and it is used in the treatment of liver diseases, hepatitis, cirrhosis and liver cancer prevention. In usual agricultural systems, the purpose of using fertilizers and their combination is to obtain the highest seed yield and quality of the active substances. In order to investigation of the effect of different fertilizers and their combination on some quantitative and qualitative characteristics of milk thistle, an experiment was arranged in split plot in a randomized complete block design with three replications in the Research Farm of Agricultural Institute, University of Zabol, during 2013-2014. In this study, the main factors were Hungary genotype (A1) and Mashhadi population (A2) and the sub factors were application of different fertilizers such as control treatment, manure, chemical fertilizer (urea + ammonium phosphate + potassium), mycorrhizal fungi (Glomus mossaea), combination of nitroxin, bio-sulfur and bio-superphosphate, combination of chemical fertilizer and manure, combination of mycorrhizal fungi and manure, and nano-iron chelate. The results showed that there was not any significant difference between Hungary genotype and Mashhadi population. Totally, the mycorrhizal fungi (G. mossaea) was the best treatment than other fertilizer treatments in the milk thistle plant and it caused enhancing the growth and better activity of photosynthetic pigments as well as improved seed yield and increased silymarin substance in the milk thistle plant.
H. Kordi; M. Aghdasi; M. Khalafi
Abstract
Silybum marianum L. is an annual or biannual herbaceous species from Asteraceae which is important in medicinal industry. The main components of this species are various flavonolignans consisting of silibinin, isosilibinin, silychristin, silydianin and taxifolin that are known as silymarin. This species ...
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Silybum marianum L. is an annual or biannual herbaceous species from Asteraceae which is important in medicinal industry. The main components of this species are various flavonolignans consisting of silibinin, isosilibinin, silychristin, silydianin and taxifolin that are known as silymarin. This species is used in treatment of heart diseases, diabetes, blood cholesterol, liver diseases (jaundice, cirrhosis and hepatitis), and gallbladder disease. In this research, different organs of Silybum marianum (young and old leaves, stem, root, inflorescence and seeds) were separately collected during the different months in the Gorgan region and then total flavonoid, silymarin content, and the amount of silymarin components were measured by HPLC method. Our results showed that the highest amounts of total flavonoid were observed in the seed and inflorescence samples and then in the stem samples. Meanwhile the effect of time on flavonoid content and also the effect of sampling month and organ factors on flavonoid content were significant. Data from the HPLC analyses revealed that the silymarin compositions, observed in this study, were similar and the main difference among samples was the amount of silymarin composition. The highest amount of silydianin was observed in the stem sample; however the amount of other components were higher in the seed sample. According to the results, Gorgan is a suitable region for the cultivation of Silybium marinum and since the stem of this species contain significant amounts of silymarin, it could be considered as an optimal source of the production of pharmaceutical substances.
S. Abdolah zareh; E. Fateh; A. Aynehband
Abstract
To study the effects of different sowing dates and different fertilizing methods (chemical &organic) on yield and amount of active substance of milk thistle (Silybum marianum (L.) Gaerate), a field experiment was done at Agricultural College of Shahid Chamran University during 2009-2010. Experimental ...
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To study the effects of different sowing dates and different fertilizing methods (chemical &organic) on yield and amount of active substance of milk thistle (Silybum marianum (L.) Gaerate), a field experiment was done at Agricultural College of Shahid Chamran University during 2009-2010. Experimental design was split plot on RCBD with three replications. Main plot was sowing dates including (14Nov, 1 Dec and 15 Dec) and sub plot was different fertilization levels including: F1: control, F2:100% chemical fertilizer (100-120-150 kg/ha NPK, respectively), F3: %25 organic fertilizer + %75 chemical fertilizer (75-90-112.5 kg/ha NPK + 7.5 ton/ha animal manure), F4: %50 organic fertilizer + %50 chemical Fertilizer (50-60-75 kg/ha NPK + 15 ton/ha animal manure), F5: %75 organic Fertilizer + %25 chemical fertilizer (25-30-37.5 kg/ha NPK + 22.5 ton/ha animal manure), F6: %100 organic fertilizer (30 ton/ha animal manure). Results showed that sowing date had significant effect on the percentage of phenol and flavenoid and the amount of silymarin. Late sowing date (15 dec) decreased grain silymarin content, while the highest percentage of phenol and flavenoid was obtained at 15 Dec. Also, the highest percentage of flavenoid, flavenoid yield and silymarin yield were obtained at 100% chemical fertilizer and integrated fertilizer. In this regard, F2 treatment had no advantage compared to integrated fertilization levels. Also, the highest percentage and yield of phenol was obtained by integrated fertilizer methods. The highest silymarin content was related to F6, and the highest grain antioxidant activity was related to F4 and F6, respectively.
B. Mosallanejad; R. Avizeh; H. Najafzadeh Varzi; M. Pourmehdi
Abstract
In the present study, effect of silymarin was evaluated as a protective drug of liver against acute hepatotoxicity due to administration of mebendazole in German shepherd dogs (mixed breeds). Twenty five healthy dogs were randomly allotted to five equal groups. Dogs in group A were given mebendazole ...
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In the present study, effect of silymarin was evaluated as a protective drug of liver against acute hepatotoxicity due to administration of mebendazole in German shepherd dogs (mixed breeds). Twenty five healthy dogs were randomly allotted to five equal groups. Dogs in group A were given mebendazole with single dose 150 mg⁄kg, p.o.; group B consisted of dogs that received silymarin with single dose 30 mg⁄kg, p.o. concurrent with mebendazole administration; groups C, D and E were treated like group B, but silymarin was administered 2, 12 and 24 h after administration of mebendazole respectively. The serum concentrations of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), lactate dehydrogenase (LDH) and total and direct bilirubin were measured before administration of mebendazole and 2, 12, 24 and 72 h later as indices of liver injury. A single oral administration of mebendazole significantly elevated serum concentrations of ALT, AST, ALP, LDH in all cases of group A (P<0.05), after 24 h. In both groups of B and C, levels of serumenzyme activities remained within the normal values. The difference was significant between groups B and C with group A (P<0.05). Levels of serumenzyme activities were higher than normal values in three cases of the group D and in all dogs of the group E. This study showed that silymarin could protect liver tissue against oxidative stress in dogs with mebendazole intoxication particularly in the first 2 hours after exposure.
H. Najafzadeh Varzi; M. Sabzevarizadeh; R. Hajhosseini; Gh. Khajeh
Abstract
Rhabdomyolysis is an injury of skeletal muscle and it causes leakage of cellular contents including myoglobine. Myoglobinuria may produce acute renal failure due to the oxidative reactions. In the present study, the effect of silymarin and vitamin C (as natural antioxidant) was compared on myoglobinuric ...
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Rhabdomyolysis is an injury of skeletal muscle and it causes leakage of cellular contents including myoglobine. Myoglobinuria may produce acute renal failure due to the oxidative reactions. In the present study, the effect of silymarin and vitamin C (as natural antioxidant) was compared on myoglobinuric renal toxicity in rats after glycerol injection. This study was carried in 7groups of rats: Group 1: control, group 2: glycerol, group 3: glycerol + silymarin by injection, group 4: glycerol + silymarin and vitamin C by injection, group5: glycerol + vitamin C by injection, group 6: glycerol + silymarin by orally and group7: glycerol + vitamin C by orally. Serum of rats was isolated 24 hours after glycerol injection. Serum concentrations of BUN, creatinine, sodium and potassium were determined as renal function indexes. Also myoglobine, gama glutamyl transferase and alkaline phosphatase levels were measured by kit. According to the results, the glycerol caused muscular injury and significantly elevated myoglobine. This elevation was controlled by injection of vitamin C. The changes of other serum factors were better controlled by injection of vitamin C than that of silymarin. Myoglobinuric renal failure was modulated by oxidative stress as vitamin C showed a protective effect. However, Silymarin did not have any effects on muscular injury and renal toxicity while it was effective when co-administrated with vitamin C.
H. Najafzadeh; S. Esmailzadeh; H. Morovvati; R. Avizeh; M. Ezati
Abstract
Drug-induced nephrotoxicity is an important factor for renal failure in dogs. Aminoglycosides including gentamicin could produce nephrotoxicity in dogs. The oxidative stress might be the case. Silymarin (extract seed of silybum marianum) is a potent antioxidant. In present study, the effect of silymarin ...
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Drug-induced nephrotoxicity is an important factor for renal failure in dogs. Aminoglycosides including gentamicin could produce nephrotoxicity in dogs. The oxidative stress might be the case. Silymarin (extract seed of silybum marianum) is a potent antioxidant. In present study, the effect of silymarin and vitamin E compared to gentamaicin-induced nephrotoxicity. The study has been conducted on dogs in 5 groups: group 1 has been kept as control and received saline. Gentamicin was injected in other groups once daily for 9 days. Vitamin E and silymarin were given in group 3 and 4 respectively. In group 5, vitamin E and silymarin were co-administrated. Renal tissue was microscopically examined by routine method and Hematoxillin-Eosine color. In histopathological examination of renal tissue, all groups (except group 1) had several changes especially glomerulonephritis; but this abnormality was not seen in group 4. The results showed, the silymarin and vitamin E have similar effect on gentamicin–induced nephrotoxicity in dogs; but in histopathological examination, the silymarin had better effect than vitamin E or their combination.
H. Morovvati; H. Najafzadeh; R. Avizeh; N. Khadivi Kashani
Volume 24, Issue 3 , November 2008, , Pages 363-373
Abstract
Drug-induced ototoxicity is one of the important factors for less auditory in dogs. Aminoglycosides including gentamicin can produce ototoxicity in dogs. The oxidative stress can be related to it. Silymarin, in the seed extract of Silybum marianum, is a potent antioxidant. In present study, the effect ...
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Drug-induced ototoxicity is one of the important factors for less auditory in dogs. Aminoglycosides including gentamicin can produce ototoxicity in dogs. The oxidative stress can be related to it. Silymarin, in the seed extract of Silybum marianum, is a potent antioxidant. In present study, the effect of silymarin and vitamin E was compared on gentamaicin-induced ototoxicity. The study was done on dogs in 5 groups: group1 was kept as control and received saline. Gentamicin was prescribed in other groups once for 9 days. Vitamin E and silymarin were given in group 3 and 4, respectively. In group 5, vitamin E and silymarin were co-administrated. After 9 days, the dogs were euthanized and ear samples were collected. One ear of dogs was colored by silver nitrate and another was decalcified by decalcificative solution. The decalcified samples were routinely sectioned and colored by Hematoxillin- Eosine (H&E) and studied by light microscope. Hair cells were not determined by silver nitrate. Organ of coarti body, tectorial membrane, limbus spiral, spiral ganglion, interdental cells, supporting cells and type I and II of hair cells were identified in samples and colored by H&E. These histological compartments were determined in control and silymarin groups. They were not observed in other groups, because gentamicin strongly destroyed cells. Thus, silymarin decreases gentamicin-induced changes in inner ear probably by its antioxidative property.
S. Zare Kia; R. Omidbaigi
Abstract
Milk thistle (Silybum marianum) is an annual range medicinal plant that wildly grows in some places of Iran. This research is the first ecological survey on milk thistle. The purpose is was to study ecological characteristics and phonologic stages of milk thistle. The climate of the area ...
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Milk thistle (Silybum marianum) is an annual range medicinal plant that wildly grows in some places of Iran. This research is the first ecological survey on milk thistle. The purpose is was to study ecological characteristics and phonologic stages of milk thistle. The climate of the area is wet. Annual average precipitation is 879.63 millimeters and annual average temperature is 16.29 centigrade. Results showed that the plant density and canopy cover in area was 30,000 plants per hectare and 33.2 % respectively. The mean length of root was 23 cm. The phonological studies showed that growth of roset leaves of milk thistle started from the end of September and continued till mid March. Stems appeared at the end of March, flowering took place from the end of April and the fruits ripped in mid May. On the basis of soil analysis, milk thistle prefers clay loam soils. Amounts of P, K, Ca, N, Organic matters, pH and EC were 4.25 ppm, 1584 ppm, 1.54 meq/lit, 0.16%, 4.26% and 0.1692 mMoh/cm respectively. Silymarin in the seed was 1.67%. Silybin in silymarin was 6.17%. Two different kinds of insect named: Oxythyrea cincletta and Epicormetis hirta poda observed on this plant.
