[Elicitation effects of fungal extracts on silymarin accumulation in Silybum marianum [L.] Gaertn hairy root culture]

Silymarin, extracted from the seeds of milk thistle [Silyburn marianum [L.] Gaertn] is mostly used for liver disease treatment. Hairy root cultures derived from S. marianum are able to produce silymarin. Elicitation of hairy root cultures is an important strategy for improving the production of secondary metabolites. The elicitors could be changed metabolite biosynthesis pathway and are useful for study of cell signaling pathway. In this study after preparation of S. marianum hairy root cultures, the effects of various levels of Fusarium oxysprum and Phytophtora meloni extract [0,10 and 20 mg 50 ml[-1] culture] in 4 different exposure times [0, 24, 48 and 72 h] have been investigated on flavonolignans production. The flavonolignans were analyzed by High Performance Liquid Chromatography method. Our results showed that hairy root cultures of S. marianum were consisted of a large number of flavonolignans including silychristin, silydianin, silybin, isosilybin and taxifolin. The highest production of silymarin [0.32 mg g[-1] DW] was observed in F. oxysprum elicited root cultures [10 mg/50 ml culture] after 72h [2.28- fold that of the control]. In Ph. meloni treated root cultures [20 mg/50 ml culture], the maximum silymarin production [0.13 mg g[-1], DW] was obtained after 72 h [1.9- fold that of the control]. In this experiment it has been concluded that hairy root cultures of S. marianum are susceptible to elicitation by fungal elicitors and is useful for efficient large-scale production of silymarin by hairy root cultures of S. marianum

[Hairy root induction in Burdock [arctium lappa L.]]

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. In this study, hairy roots were induced in Arctium lappa using Agrobacterium rhizogenes, strain AR15834. 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. 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. 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

Effect of salicylic acid on antioxidant activity in milk thistle hairy root cultures

Elicitors can trigger defense responses and activate specific genes involved in secondary metabolite biosynthesis. For understanding SA-treatment signaling pathway we determined defense-related compounds and enzymes in S. marianum hairy root cultures. 28 day old hairy roots were treated by salicylic acid [6 mg/50 ml culture] for different times [24, 48, 72, 96 and 120 h]. The antioxidant activity was assayed by the 1-diphenyl-2-picrylhydrazyl stable free radical method and results were calculated based on the IC[50] H[2]O[2], peroxidase and ascorbate peroxidase activity and total tocopherol content was measured in treated and non-treated hairy root cultures. The highest accumulation of H[2]O[2] was 86.60 micro mol g[-1] DW that was obtained 120 h after elicitation [2-times of non-treated hairy roots]. IC[50] increased upon treatment and was 1.15- times higher than the control 24h after elicitation. The content of total tocopherol content under SA treatment was lower than the control experiments. Maximum activity of peroxidase and ascorbate peroxidase were 0.17 and 1.50 micro mol min[-1] mg[-1] protein, 96 and 72 h after elicitation, respectively. Oxidative stress defends system can be activated by salicylic acid in Silybum marianum hairy root cultures. This study suggested that exogenous SA can increase H[2]O[2] content of hairy roots, and induce the expression of antioxidant enzymes and increase secondary metabolite accumulation

[Comparative study on anti-Helicobacter pylori effects of licorice roots collected from different regions of Iran]

Helicobacter pylori infection is common in gastrointestinal tract and lead to gastritis, peptic ulcer and finally stomach cancer. Many synthetic drugs are used to eradicate the bacteria but there are some problems in treatment due to drug resistance and frequent relapses of infection. Herbal drugs are very important in the case. Glycyrrhiza glabra is used for treatment of H. pylori infection. This plant grows in different regions of Iran, therefore, investigation of anti H. pylori effects of the plant collected from different growing area is important. The aim of this study is determination of MIC of licorice roots collected from different regions of Iran against H. pylori in order to introduce the best growing area of the plant. Roots of the plant were collected from Kerman, Kermanshah, Sirjan, Sarhadd [Fars province], Istahban [Fars province], Ghasredasht [Fars province], Najaf Abad [Isfahan province], Ganjnameh [Hamedan province], Ekbatan [Hamedan province], Mahabad, Khorram Abad and Ardabil and extracted by using ethanol 80%. MIC of each extract was determined against three clinical isolated strains of H. pylori which obtained during endoscopy procedure. The results showed that susceptibilities of three clinical isolated bacteria to the licorice extracts are different. Among the studied extracts, licorice roots collected from Ardabil showed the lowest MIC [at least: 125 ppm] and all strains were almost resistant to the extract from Mahabad. It is concluded that drug resistance is observed in herbals just like synthetic drugs. Moreover, in order to achieve best therapeutic results, before using an herbal preparation for treatment a disorder, growing area of a medicinal herb should be considered

[Polyphenolics, flavonoids and anthocyanins content and antioxidant activity of qare-qat [vaccinium arctostaphylos L.] from different areas of Iran]

Vaccinium arctostaphylos L. [Ericaceae family], locally named Qare-Qat is a shurb or woody bush which has been extensively used in Iranian folk medicine as antidiabetic and antihypertensitive agents for many years. Phenolics found in leaves and fruits of the plant are a specific group of secondary metabolites that plays an important role of antioxidant activity. Comparative study of phenolics, anthocyanins and flavonoids contents in V. arctostaphylos methanolic extracts and evaluation of antioxidant activities. Leaves and fruits of V. arctostaphylos have been collected from the high altitude areas of Ardabil [Hoor], Gilan [Masouleh and Asalem] and Mazandaran [Kelardasht] provinces in August 2007 and stored at-80°C until metabolites analysis. Then extraction was carried out with acidified methanol and the mixture was used for antioxidant activity and phenolics content assays by UV-Vis spectrophotometer. The data indicated that fruits of Kelardasht genotype had the highest total phenolics content as gallic acid [42.73 +/- 1.5 mg g[-1] DW], anthocyanin content as cyaniding-3-glucoside [1.0 +/- 0.07 mg g[-1]] and antioxidant activity [DPPH assay] of fruits of Kelardasht [IC[50]: 0.14 +/- 0.07 mg ml[-1]] were highest in comparison to other genotypes. Leaves of Masouleh genotype had the highest flavonoid content as quercetin [2.9 +/- 0.07 mg g[-1] DW]. It could be concluded that the leaves and fruits of V. arctostaphylos have high amount of phenolics and antioxidant activity. Therefore, this plant can be a potential source of antioxidant compounds for food and pharmaceutical industry

Hairy roots as a source for production of valuable pharmaceutical materials

The plants have important role in our daily diet life. Their food value has been studied for many years. They are usually used as medicinal plants to improve national health in relation to nutrition or sanitation. Today, these metabolites extract from wild or cultivated medicinal plants. Production of secondary metabolites by traditional methods, depend on their species, require to months or years time. Furthermore, content of production affected by different factors such as climate changes or pathogens. Tissue culture of medicinal plants has been introduced as a source for production of secondary metabolites. Recently, hairy roots culture has also been a stable source of metabolite production. The hairy roots, induced by inoculation of explants with Agrobacterium rhizogenez, often growth as fast as or faster than plant cell cultures and have attractive properties for secondary metabolite production compare to related plants

[Using yeast extract as an approach to increase flavonolignans content in cell suspension culture of Milk Thistle plant via elicitation mechanism]

Silybum marianum [L.] Gaertn is a kind of medicinal plants. Silymarin is a derivate substance from the fruits of milk thistle plant which is consists of a large number of flavonolignans. Cell cultures derived from this species could be an alternative for production of flavonolignans. Elicitors cause enhancement in metabolite production by effecting on key enzymes in secondary metabolites pathways. In this study various level of Yeast extract in 5 different exposure time have been used as an biotic elicitor in order to evaluation the effect of Yeast extract on silymarin production and cell growth and nomination the best time and best consistence of the elicitor. In this study various level of Yeast extract in 5 different exposure time have been used as an biotic elicitor in order to evaluation the effect of Yeast extract on silymarin production and cell growth and nomination the best time and best consistence of the elicitor. In this work after preparation cell suspension culture of S. marianum, the effects of various level of yeast extract [1, 2. 4, 6 and 8 mg/ 50 ml culture] in 6 different exposure time [12,24, 48, 72, 144 and 216 h] on flavonolignans production by High Performance Liquid Chromatography [HPLC] have been studied. Determination and quantification of flavonolignans showed that cell suspension cultures of S.marianum were consists of a large number of flavonolignans including silychristin, silydianin, silybin, isosilybin and taxifolin. The results showed that Yeast extract cause improvement in silymarin content in the media treated with 6 [mg/ 50 ml culture] Yeast extract at the end of 72h which was 5- fold to compare the control and the maximum cell Dry weight was 5.82 g in the media treated with 4 mg/ 50 ml culture Yeast extract at the end of 24h. In this experiment it has been observed that cell suspension culture of S. marianum are susceptible to elicitation by Yeast extract and it can be extremely useful in increasing productivities in cell suspension culture of Milk thistle plant

Determination of tocopherols and fatty acids in seeds of Silybum marianum [L.] gaerth

The dried fruits of Silybum marianum [L.] Gaertn [Compositae] contain silymarin, an isomeric mixture of flavonolignans. Silymarin acts as a strong anti -hepatotoxic. This fruits contains a high amount of oil The objective of this study was to determine total lipid content, fatty acid composition and content of different kind of tocopherols in seed oils of S. marianum Fatty acid compositon and a-, y- and ?-tocopherol [vitamin E] content from seeds of Silybum maianum were investigated by gas chromatography and high performance liquid chromatography methods. and Conclusion: Total lipid percentage was approximately 25% and nine fatty acids including; palmitic acid [8.25%], palmeotic acid [0.07%], Stearic acid [6.67%], oleic acid [31.58%], Isomer oleic acid [0.53%], linoleic acid [45.36%], linolenic acid [0.87%], arashidic acid [4.11%], eicozantoic acid [0.088%] and behenic acid [2.6%] were determined a-, y- and tocopherol content were around 563.157, 88.87 and 163.791 mg Kg -1 DW respectively that could be a natural good sources of antioxidant

[Investigating on the quality of wild licorice roots collected from different regions of Iran]

Glycyrrhiza glabra L. has been used in respiratory diseases, gastric and duodenal ulcers and liver insufficiencies for many years. The plant grows in different regions of Iran. Since our country is considered one of the greatest exporting countries of the plant roots, investigation of licorice quality from different parts of Iran and determination of the best area for its growing seems to be important. According to validated references, glycyrrhizic acid percentage and water-soluble extractives are determinative of quality; therefore, in this investigation, these two factors have been measured in the plants collected from different regions of Iran. Roots of the plant were collected from Kerman, Kermanshah, Sirjan, Sarhadd [Fars province], Istahban [Fars province], Ghasredasht [Fars province], Najaf Abad [Isfahan province], Ganjnameh [Hamedan province], Ekbatan [Hamedan province], Mahabad, Khorram Abad and Ardabil and water-soluble extractives using maceration method and glycyrrhizic acid percentage with HPLC technique were determined in each sample. The results showed that the amount of water-soluble extractives and glycyrrhizic acid are high in samples from Kermanshah, Sarhadd and Kerman while these two factors are the lowest in the plant roots collected from Ekbatan and Ganjnameh. The plant roots collected from Kermanshah, Sarhadd and Kerman have the highest quality and are the best for medicinal purposes, sweetening and flavoring agent and for exporting as well

Review on Jojoba [Simmondsia chinensis [Link] Sch.] Oil

Jojoba is a woody evergreen shrub or small multi-stemmed tree native to the semiarid regions of southern Arizona, southern California and northwestern Mexico. The fruit is a green capsule which encloses up to three seeds. Jojoba wax [called oil] makes up 50% of the seed's dry weight. Jojoba is being cultivated to provide a renewable source of unique high-quality oil. Most jojoba oil produced in the U.S. today is sold at a high price for use in cosmetics and hair care products. As many as 300 products containing jojoba have appeared in the U.S. market in recent years. As the supply of oil increases and price decreases, more uses will become economically feasible. For example, the viscosity index of jojoba oil is much higher than that of petroleum oil; therefore, it may be used as a high temperature, high pressure lubricant. The stability of jojoba oil makes it attractive to the electronic and computer industries and since jojoba oil contains no cholesterol or triglycerides and is not broken down by normal metabolic pathways, it may become an important low-calorie oil for human consumption. The oil can be used as an antifoam in antibiotics production and as a treatment for skin disorders. Other proposed uses include candles, plasticizers, detergents, fire retardants, transformer oil, and for the leather industry. The meal contains up to 30% protein, but toxic compounds [simmondsins] make it currently hazardous as an animal feed. Much of the interest in joj'oba worldwide is the result of the plant's ability to survive in a harsh desert environment

Evaluation of phenotypic coefficient and flavonolignan content in dried fruits of cultivated and endemic Silybum marianum [L.] gaertn

Silybum marianum [L.] Gaertn. has been used for centuries as an herbal medicine for treatment of liver disease. Fruits of S. marianum contain silymarin, which is composed of the flavonolignans, silybin, isosilybin, silydianin and silychristin. The aim of this research was to investigate the effects of environmental conditions on silymarin content and morphological characteristic of S. marianum. The dried fruits of S. marianum were collected from 13 area of Iran and cultivated at greenhouse conditions. Morphological characteristic were studied and silymarin content analyzed by HPLC. The results showed that silybin is the main component of silymarin and was affected by environmental conditions. The amount of silymarin was very different in cultivated and endemic plants in comparison with together. The silymarin content had a significant positive correlation with seed yield/plant, stem diameter and number of capsules/plant

Determination of flavonalignans of dried fruits of silybum marianum [L.] gaertn collected from different areas of Iran by spectrophotometry, TLC and HPLC

Silymarin complex consisted of five flavonolignans [silybum A and B Isosylibin A and B, silychristin, silydianin and taxifolin] isolated from dried fruits of Silybum marianum [L]Gaertn silybin is the main component of silymarin complex. Hepatoprotective activity of silymarin may be related to its antioxidant property. This study was conducted to understand the environmental condition on silymarin methabolism. Silymarin extraction and flavolignans assay by spectrophotometery, TLC and HPLC was performed on milk thistle seeds collected from different areas of Iran, Hungarian seeds cultivated in greenhouse and field in Karaj. Valasht and Borazjan had the highest levels of silymain content by spectrophotometery method. In TLC method all 5 component were identified. We analyzed the flavolignans contents by HPLC and results showed that borazjan had the highest level of silymarin. To produce commercially silymarin we need to search for the best genotype