Volume 3, Issue 5, October 2015, Page: 279-284
Rapid Method for In vitro Multiplication of Hypericin Rich Shoots of Hypericum perforatum
Hemant Sood, Department of Biotechnology and Bioinformatics, Jaypee University of Information Technology, Waknaghat, Solan (HP), India
Kirti Shitiz, Department of Biotechnology and Bioinformatics, Jaypee University of Information Technology, Waknaghat, Solan (HP), India
Neha Sharma, Department of Biotechnology and Bioinformatics, Jaypee University of Information Technology, Waknaghat, Solan (HP), India
Received: Sep. 28, 2015;       Accepted: Oct. 11, 2015;       Published: Oct. 24, 2015
DOI: 10.11648/j.jps.20150305.16      View  3431      Downloads  78
Hypericum perforatum is a high value medicinal herb possessing a broad range of pharmacological activities. These medicinal properties are attributed to the presence of major compound hypericin. In current study, the effect of different growth hormones and media combinations along with various growth parameters including light and temperature were analyzed for hypericin production in in vitro grown shoot cultures of H. perforatum. Rapid method for micropropagation with an average of 36.3 shoots per explant in 5-6 days was developed. Highest hypericin content of 0.119 µg/mg was detected in shoots grown on MS medium containing sucrose (3%), IBA (3mg/l), KN (1mg/l) and agar (0.9%) at 25±2 °C under light conditions after two months of incubation. In addition, sustention of in vitro grown shoots after hardening under greenhouse conditions was also observed showing comparable hypericin content in hardened and field grown plants after 2 years. Therefore, current study reports the rapid method for in vitro multiplication for hypericin rich shoots of H. perforatum under optimized light and temperature conditions. To the best of our knowledge, this is the first report on comparative analysis of hypericin production in hardened and field grown shoots of H. perforatum. Hence, the outcome is an endeavor to meet the increasing industrial demands by providing quality rich raw material using tissue culture techniques.
Hypericin, Hypericum perforatum, Micropropagation, HPLC
To cite this article
Hemant Sood, Kirti Shitiz, Neha Sharma, Rapid Method for In vitro Multiplication of Hypericin Rich Shoots of Hypericum perforatum, Journal of Plant Sciences. Vol. 3, No. 5, 2015, pp. 279-284. doi: 10.11648/j.jps.20150305.16
Copyright © 2015 Authors retain the copyright of this article.
This article is an open access article distributed under the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Agostinis, P., Vantieghem, A., Merlevede, W., de Witte, P. A., 2002. Hypericin in cancer treatment: more light on the way. The International Journal of Biochemistry & Cell Biology, 34: 221-41.
Bais, H. P., Vepachedu, R., Lawrence, C. B., Stermitz, F. R., Vivanco, J. M., 2003. Molecular and biochemical characterization of an enzyme responsible for the formation of hypericin in St. John’s Wort (Hypericum perforatum L.). The Journal of Biological Chemistry, 278: 32413–32422.
Baskaran, P., Ncube, B., Van Staden, J., 2012. In Vitro Propagation and Secondary Product Production by Merwilla plúmbea (Lindl.) Speta. Plant Growth Regulation, 67: 235-245.
Coste, A., Vlase, L., Halmagyi, A., Deliu, C., Coldea, G., 2011. Effects of plant growth regulators and elicitors on production of secondary metabolites in shoot cultures of Hypericum hirsutum and Hypericum maculatum. Plant Cell Tissue and Organ Culture, 106: 279–288.
Cui, X. H., Chakrabarty, D., Lee, E. J., Paek, K. Y., 2010. Production of adventitious roots and secondary metabolite by Hypericum perforatum L. in a bioreactor. Bioresource Technology, 101: 4708–4716.
Franklin, G., Dias, A. C. P., 2006. Organogenesis and embryogenesis in several Hypericum perforatum genotypes. In Vitro Cellular and Developmental Biology, 42: 324–330.
Gioti, E. M., Skalkos, D. C., Fiamegos, Y. C., Stalikas, C. D., 2005. Single-drop liquid-phase microextraction for the determination of hypericin, pseudohypericin and hyperforin in biological fluids by high performance liquid chromatography. Journal of Chromatography A, 1093: 1-10.
Hostanska, K., Reichling, J., Bommer, S., Weber, M., Saller, R., 2003. Hyperforin a constituent of St John's wort (Hypericum perforatum L.) extract induces apoptosis by triggering activation of caspases and with hypericin synergistically exerts cytotoxicity towards human malignant cell lines. European Journal of Pharmaceutics and Biopharmaceutics, 56: 121-32.
Jain, S. C., Pancholi, B., Jainb, R., 2012. In-vitro callus propagation and secondary metabolite quantification in Sericostoma pauciflorum. Iranian Journal of Pharmaceutical Research, 11: 1103–1110.
Li, W., Fitzloff, J. F., 2001. High performance liquid chromatographic analysis of St. John’s Wort with photodiode array detection. Journal of Chromatography B, 765: 99–105.
Linde, K., Ramirez, G., Mulrow, C. D., Pauls, A., Weidenhammer, W., Melchart, D., 1996. St John's wort for depression--an overview and meta-analysis of randomised clinical trials. British Medical Journal, 313: 253-8.
Liu, X. N., Zhang, X. Q., Sun, J. S., 2007. Effects of cytokinins and elicitors on the production of hypericins and hyperforin metabolites in Hypericum sampsonii and Hypericum perforatum. Plant Growth Regulation, 53: 207–214.
Mulinacci, N., Giaccherini, C., Santamaria, A. R., Cantato, R., Ferrari, F., Valleta, A., Vincieri, F. F., Pasqua, G., 2008. Anthocyanins and Xanthones in the calli and regenerated shoots of Hypericum perforatum var. angustifolium (sin, Frohlich) Borkh. Plant Physiology and Biochemistry, 46: 414–420.
Murashige, T., Skoog, F., 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum, 15: 473–497.
Nahrstedt, A., Butterweck, V., 1997. Biologically active and other chemical constituents of the herb of Hypericum perforatum L. Pharmacopsychiatry, 30: 129-34.
Narasimhan, S., Nair, G. M., 2004. Effect of auxins on berberine synthesis in cell suspension culture of Coscinium fenestratum (Gaertn.) Colebr - A critically endangered medicinal liana of Western Ghats. Indian Journal of Experimental Biology, 42: 616-619.
Palmer, C. D., Keller, W. A., 2010. Plant regeneration from petal explants of Hypericum perforatum L. Plant Cell Tissue and Organ Culture, 105: 129–134.
Patial, V., Devi, K., Sharma, M., Bhattacharya, A., Ahuja, P. S., 2012. Propagation of Picrorhiza kurroa Royle ex Benth: an important medicinal plant of Western Himalaya. Journal of Medicinal Plant Research, 6: 4848–4860.
Pretto, F. R., Santarem, E. R., 2000. Callus formation and plant regeneration from Hypericum perforatum leaves. Plant Cell, Tissue and Organ Culture, 62: 107–113.
Santoro, M. V., Nievas, F., Zygadlo, J., Giordano, W., Banchio, E.,2013. Effects of growth regulators on biomass and the production of secondary metabolites in Peppermint (Mentha piperita) micropropagated in vitro. American Journal of Plant Sciences, 4: 49-55.
Siatka, T., Kasparová, M., 2008. Effects of auxins on growth and scopoletin accumulation in cell suspension cultures of Angelica archangelica L. Ceská a Slovenská Farmacie, 57: 17-20.
Soelberg, J., Jørgensen, L. B., Ja¨ger, A. K., 2007. Hyperforin accumulates in the translucent glands of Hypericum perforatum. Annals of Botany, 99: 1097–1100.
Yadollah-Damavandi, S., Chavoshi-Nejad, M., Jangholi, E., Nekouyian, N., Hosseini, S., Seifaee, A., Rafiee, S., Karimi, H., Ashkani-Esfahani, S., Parsa, Y., Mohsenikia, M., 2015. Topical Hypericum perforatum improves tissue regeneration in full-thickness excisional wounds in diabetic rat model. Evidence-based Complementary and Alternative Medicine , doi: 10.1155/2015/245328.
You, M., Kim, D. W., Jeong, K. S., Bang, M. A., Kim, H. S., Rhuy, J., Kim, H. A., 2015. St. John’s Wort (Hypericum perforatum) stimulates human osteoblastic MG-63 cell proliferation and attenuates trabecular bone loss induced by ovariectomy. Nutrition Research and Practice, 9(5): 459-465, 2015.
Zobayed, S. M. A., Afreen, F., Kozai, T., 2005. Temperature stress can alter the photosynthetic efficiency and secondary metabolite concentrations in St. John's wort. Plant Physiology and Biochemistry, 43: 977-84.
Zobayed, S. M. A., Murch, S. J., Rupasinghe, H. P. V., Saxena, P. K., 2004. In vitro production and chemical characterization of St. John’s wort (Hypericum perforatum L. cv ‘New stem’). Plant Science, 166: 333–340.
Zobayed, S. M. A., Saxena, P. K., 2003. In vitro grown roots: a superior explant for prolific shoot regeneration of St. John’s wort (Hypericum perforatum L. cv ‘New Stem’) in a temporary immersion bioreactor. Plant Science, 165: 463–470.
Browse journals by subject