Evaluation of an important flavonoid silymarin in callus cultures of Sylibum marianum (L.) Gaertn

Paper Details

Research Paper 01/06/2017
Views (251) Download (14)

Evaluation of an important flavonoid silymarin in callus cultures of Sylibum marianum (L.) Gaertn

Sania, Safdar Ali Mirza, Iram Liaqat, Nazish Mazhar
Int. J. Biosci.10( 6), 20-28, June 2017.
Certificate: IJB 2017 [Generate Certificate]


Silybum marianum L. Gaertn is an important medicinal herb due to its active secondary metabolite, silymarin pronounced to be effective in disturbed liver function and ailments. The plant is under harvest pressure for sylimarin production and is being endangered. With the objective to produce silymarin in vitro, seed germination were optimized maintaining aseptic conditions. Seedlings raised from sterilized seeds in vitro on MS medium containing gibberellic acid, were used as explants for callus induction. Among different explants used, seed and hypocotyls were found substantially suitable explants for callus induction. Significant callus induction from hypocotyl explant was documented using MS medium added with 2, 4-D whereas seed explant produced callus under the influence of BAP along with 2, 4-D. Callus cultures from different explants were put to HPLC analysis to determine silymarin content. Since silymarin is a mixture of different components, all the callus cultures were same with respect to composition but different in amount of components, seed callus was observed to contain highest Silybinin content (89%) as compared to the callus cultures of other explants. The in vitro production of silymarin may prove significant source for the said valuable medicinal compound.


Ackerson A. 2005.Milk thistle better nutrition. http://findarticles/8582801.

Ali S, Tariq A, Ajaib M, Khan KM. 2015. Appraisal of β-Phellandrene in Callus Cultures of Momordica charantia L. Cultivars, Jaunpuri and Jhalri. Journal of Chemical Society of Pakistan 37 (4), 817-823.

Ali S, Khan MS, Iqbal J. 2010. Genotype independent in vitro regeneration system in elite varieties of sugarcane. Pakistan Journal of Botany 42(6), 3783-3790.

Balian S, Ahmad S, Zafar R. 2006. Anti-inflammatory activity of leaf and leaf callus of Silybum marianum (L.) Gaertn. in albino rats. Research Letters 38(3), 213-214. http://dx.doi.org/10.4103/0253-7613.25815

Ball KR, Kowdley KV. 2005. A review of Silybum marianum (milk thistle) as a treatment for alcoholic liver disease. Journal of Clinical Gastroenterology 39(6), 520-528. http://dx.doi.org/10.1097/01.mcg.0000165668.79530.a0

Cimino C, Vairo CS, Spina F, Natalucci C, Priolo N. 2006. Callus culture for biomass production of Milk thistle as a potential source of milk clotting peptidases. Electronic Journal of Biotechnology 9(3), Special Issue, 2006. http://dx.doi.org/10.2225/vol9-issue3-fulltext-14

Comelli F, Giagnoni G, Bettoni I, Colleoni M, Costa B. 2007. The inhibition of monoa-cylglycerol lipase by URB602 showed an anti-inflammatory and anti-nociceptive effect in a murine model of acute inflammation. British Journal of Pharmacology 152, 787–94. http://dx.doi.org/10.1038/sj.bjp.0707425

Helmja K, Vaher M, Puessa T, Kamsol K, Orav A, Kaijurand M. 2007. Bioactive components of the hop strobilus and Comparison of different extraction methods by capillaryelectrophoretic and chromatographic methods. Journal of Chromatography 2, 222-229. http://dx.doi.org/10.1016/j.chroma.2006.12.067

Linden JC. 2000. Secondary products from plant tissue culture. Biotechnology Vol. VI. EOLSS Pulication, Paris France. http://www.eolss.net/Eolss-sampleAllChapter.aspx

Malewicz B, Wang Z, Jiang C, Guo J, Cleary MP, Grande JP, Lu  J. 2006.  Enhancement of mammary carcinogenesis in two rodents models by silymarin dietary supplements. Journal of Carcinogenesis 27(9), 1739-1747. http://dx.doi.org/10.1093/carcin/bgl032

Mantell SH, Smith H. 1984. Culture factors that influence secondary metabolite accumulation in plant cell and tissue cultures. In: Mantell SH, Smith H. (Eds.) Plant Biotechnology.  Cambridge University Press, Cambridge, 75–108 P. http://dx.doi.org/10.1080/02648725.1984.10647794

Murashige T, Skoog F. 1962. A revised medium for rapid growth and bioassays with tabacco tissue cultures. Physiologia Plantarum 115, 493-497.

Nair AJ, Sudhakaran PR, Madhusudana JR, Ramakrishna SV. 1992. Berberine synthesis by callus and cell suspension cultures of Coscinium fenestratum, Plant Cell Tissue and Organ Culture 29, 7–10. http://dx.doi.org/10.1007/BF00036139

Neumann J, Zeindl-Eberhart E, Kirchner T, Jung A. 2009. Frequency and type of KRAS mutations in routine diagnostic analysis of metastatic colorectal cancer. Pathology Research and Practice 205(12), 858-862. http://dx.doi.org/10.1016/j.prp.2009.07.010

Pradhan SC, Girish C. 2006.  Hepatoprotective herbal drug, silymarinfrom experimental pharmacology to clinical medicine.  Indian Journal of Medical Research 124(5), 491-504. PMID:17213517

Praveen N, Naik PM, Manohar SH, Nayeem A, Murthy HN. 2009. In vitro regeneration of Brahmi shoots using semisolid and liquid cultures and quantitative analysis of bacoside A. Acta Physiology Plant 31, 723–728. http://dx.doi.org/10.1016/j.phymed.2007.03.010

Rady MR, Matter MA, Ghareeb HA, Hanafy MS, Saker MM, Eid SA, Hammoda FM, Imbaby SI, Nazief NH. 2014. In vitro cultures of Silybum marianum and silymarin accumulation, Journal of Genetic Engineering and Biotechnology 12(1), 75-79. http://dx.doi.org/10.1016/j.jgeb.2013.11.003

Ramachandra RS, Ravishankar GA. 2002. Plant cell cultures: chemical factories of secondary metabolites. Biotechnology Advances 20, 101–153.  http://dx.doi.org/10.1016/S07349750(02)00007-1

Sánchez-Sampedro A, Kim HK, Choi YH, Verpoorte R, Corchete P. 2007. Metabolomic alterations in elicitor treated Silybum marianum suspension cultures monitored by nuclear magnetic resonance spectroscopy. Journal of Biotechnology 130, 133–42. http://dx.doi.org/10.1016/j.jbiotec.2007.03.007

Sanchez-Sampedro MA, Fernandez-Tarago J, Corchete P. 2005. Yeast extract and methyl jasmonate induced silymarin production in cell culture of Silybum marianum L. Gaerth. Journal of Biotechnology. 119, 60-69. http://dx.doi.org/10.1016/j.jbiotec.2005.06.012

Sersen F, Vencel T, Annus J. 2006. Silymarin and its components scavenge phenylglyoxylicketyl radicals. Fitoterapia 77, 525–529. http://dx.doi.org/10.1016 /j.fitote. 2006.06.005

Shilpashree HP, Ravishankar R. 2009. In vitro Plant Regeneration and accumulation of flavonoids in Hypericum mysorense. International Journal of Integrated Biology 8, 43-49.

Tanveer H, Ali S, Asi MR. 2012. Appraisal of an important flavonoid, quercetin in callus cultures of Citrullus colocynthis. International Journal of Agriculture Biology 14, 528–532. http://www.fspublishers.org

Vanhala L, Eeva M, Lapinjoki S, Hiltunen R, Oksman-Caldentey KM. 1998. Effect of growth regulators on transformed root cultures of Hyoscyamus muticus. Journal of Plant Physiology 153, 475–481. http://dx.doi.org/10.1016/S0176-1617(98)801776

Verpoorte R, Contin A, Memelink J. 2002. Biotechnology for the production of plant secondary metabolites. Photochemistry Reviews 1, 13–25. http://dx.doi.org/10.1023/A:1015871916833

Weathers PJ, Bunk G, McCoy MC.  2005.  The effect of phytohormones on growth and artemisinin production in Artemisia annuahairy roots. In vitro Cellular and Developmental Biology Plant 41:47–53. http://dx.doi.org/10.1079/IVP2004604

Wink M. 1988. Plant breeding: importance of plant secondary metabolites for protection against pathogens and herbivores. Theoretical and Applied Genetics 75, 225–233. http://dx.doi.org/10.1007/BF00303957