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.
A. Saghalli; M. Farkhari; A, Salavati; Kh. Alamisaeid; A.R. Abdali Mashhadi
Abstract
This study was aimed to evaluate various ecotypes of Silybum marianum L., collected from different parts of Iran, as well as the Budakalasz variety and two ecotypes from England. The experiment was performed in a Randomized Complete Block Design (RCBD) in the agricultural farm of the Ramin Agriculture ...
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This study was aimed to evaluate various ecotypes of Silybum marianum L., collected from different parts of Iran, as well as the Budakalasz variety and two ecotypes from England. The experiment was performed in a Randomized Complete Block Design (RCBD) in the agricultural farm of the Ramin Agriculture and Natural Resources University of Khuzestan. The evaluated traits included yield, yield components, and morphological and phenological traits. Analysis of variance showed statistically significant difference among the traits except phenological traits. Molasani 2 and Shush 2 with 4.2 and 1.7 tons/hectare had the highest and lowest yield, respectively. In addition, the yield of Budakalasz variety was estimated to be 2.2 tons/hectare. The smallest canopy among the study ecotypes belonged to Shush 2. Among the phenological traits studied, only the flowering period was statistically different (α<0.05) among the ecotypes. The number of capitula per plant, number of seeds per main capitula and 1000-seed weight according to stepwise regression and path analysis were the most effective traits on yield. Days to stem elongation indirectly influenced the yield via its effects on the number of capitula per plant and the seed number of main capitula. A total of 97 percent of yield variance was explained by four variables entered in the regression model. The resulted dendrogram based on squared Euclidean distance and Ward’s algorithm divided ecotypes into three main clusters. This grouping did not perfectly match with the geographical distribution pattern. The results indicated that there was high diversity in the morphological traits of milk thistle ecotypes, so that the ecotypes could be applied in breeding programs.
A. Nobakht
Abstract
This experiment was conducted to evaluate the effects of different levels of Silybum marianum L. aerial parts powder on performance, egg quality, blood biochemical parameters, and blood cells of laying hens. A number of 240 Hy-line (W36) laying hens from 65 to 76 weeks were used in five treatments and ...
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This experiment was conducted to evaluate the effects of different levels of Silybum marianum L. aerial parts powder on performance, egg quality, blood biochemical parameters, and blood cells of laying hens. A number of 240 Hy-line (W36) laying hens from 65 to 76 weeks were used in five treatments and four replicates (12 birds per each replicate) in a completely randomized design. The powder of Silybum marianum L. was 0, 0.5, 1, 1.5 and 2% of the diet. The results showed that the diet containing 2% of Silybum marianum L., in comparison with control group, increased the amounts of egg weight, egg production percentage, egg mass and feed intake (p<0.05).Whereas, the highest amount of egg weight and egg specific gravity and eggshell percentage were observed with 1% of Silybum marianum L. (p<0.01). The use of Silybum marianum L. in diets caused the values of blood triglyceride, cholesterol, albumin and total protein to be decreased (p<0.05). The diet containing 2% Silybum marianum L. had increasing effects on hematocrit, hemoglobin and white blood cells (p<0.05). The overall results indicated that in laying hens, the use of Silybum marianum L. up to 2% of diet significantly improved the performance, egg quality traits, biochemical parameters, and blood cells.
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.
M. Shokrpour; S.A. Mohammadi; M. Moghaddam; S.A. Ziai; A. Javanshir
Volume 24, Issue 3 , November 2008, , Pages 278-292
Abstract
To investigate the relationships between morphological, phytochemical and molecular markers in milk thistle, a set of 32 ecotypes collected from Iran along with two introduced varieties, Budakalaszi and CN seeds, were evaluated. Canonical correlation analysis between 8 morphological attributes and 7 ...
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To investigate the relationships between morphological, phytochemical and molecular markers in milk thistle, a set of 32 ecotypes collected from Iran along with two introduced varieties, Budakalaszi and CN seeds, were evaluated. Canonical correlation analysis between 8 morphological attributes and 7 flavonolignan compounds forming silymarin revealed that first two canonical variables showed high canonical correlations. The loadings of the canonical correlations indicated that ecotypes having higher values for 1000 seed weight and lower values for flowering time, plant height, capsule diameter and seed yield would have higher silychristin and silybin and lower silydianin contents. In other words, larger seeds would have higher silybin and lower silydianin. Out of 415 polymorphic markers, 37 and 29 markers showed significant association with flavonolignans markers and morphological attributes, respectively. The informative markers showed 54 and 45% of the variation for taxifolin and silychristin, respectively. In the case of morphological traits, more than 40% of 1000 seed weight, flowering date and plant height variation were determined by informative AFLP markers. Results of the study clarified that some of qualitative and quantitative properties of essential oil in milk thistle can be well predicted by morphological and also molecular markers.
