Abstract
An efficient and rapid plant regeneration system via direct organogenesis was established for felty germander (Teucrium polium L.), an endangered and valuable medicinal plant. Induction of shoot organogenesis was obtained using nodal explants and preconditioned intact seedlings on the MS medium supplemented with various concentrations and combinations of plant growth regulators. The highest number of shoots (7.14/explants) was obtained from nodal explants cultured on MS containing 2.0 mg·L-1 BAP and 0.5 mg·L-1 NAA. For preconditioning, Best response in terms of number of adventitious shoots per explant (6.85) and greatest shoot length (5.43 cm) was obtained with seedlings perpetrated in 0.5 mg·L-1 TDZ. The rooted plantlets were acclimatized successfully in the greenhouse with a survival rate of 100%. Significant differences (p < 0.05) were found between the greenhouse-grown tissue culture raised plants in total flavonoid contents. Methanolic extracts of plants raised on TDZ (0.5 mg·L-1) containing medium showed the highest amount of flavonoid contents (1.67 mg Ru·g-1 DW) and a higher antioxidant activity (52.31%) in comparison with the field-grown mother plant. Randomly amplified polymorphic DNA (RAPD) analysis of plants raised preconditioning procedure revealed there were no somaclonal variations among them. Our results indicated that the cytokinin-preconditioning mediated micropropagation protocol could be used for the production of a large number of true-to-type plants of T. polium, which may contain higher quantities of valuable phytochemicals than those of wild plants.
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Abdollahi, M., H. Karimpour, and H.R. Monsef-Esfehani. 2003. Antinociceptive effects of Teucrium polium L total extract and essential oil in mouse writhing test. Pharmacol. Res. 48:31–35.
Afifi, F.U., B. Al-khalidi, and E. Khalil. 2005. Studies on the in vivo hypoglycemic activities of two medicinal plants used in the treatment of diabetes in Jordanian traditional medicine following intranasal administration. J. Ethnopharmacol. 100:314–318.
Ahuja, M.R. 1993. Micropropagation of woody plants. Kluwer Academic Publishers, Dordrecht, Netherlands p. 311–336.
Al-Qudah, T.S., R.A. Shibli, and F.Q. Alali. 2011. In vitro propagation and secondary metabolites production in wild germander (Teucrium polium L.). In Vitro Cell Dev. Biol.-Plant 47:496–505.
Baluchnejadmojarad, T., M. Roghani, and F. Roghani-Dehkordi. 2005. Antinociceptive effect of Teucrium polium leaf extract in the diabetic rat formalin test. J. Ethnopharmacol. 97:207–310.
Belide, S., R.R. Sajjalaguddam, and A. Paladugu. 2010. Cytokinin preconditioning enhances multiple shoot regeneration in Pongamia pinnata (L.) Pierre — A potential, non-edible tree seed oil source for biodiesel. Electron. J. Biotechn. 13:1–9.
Benavente-García, O., J. Ccastillo, J. Lorente, A. Ortuno, and J.A. Del Rio. 2000. Antioxidant activity of phenolics extracted from Olea europaea L. leaves. Food Chem. 68:457–62.
Bouhouche, N. and T. Ksiksi. 2007. An efficient in vitro plant regeneration system for the medicinal plant Teucrium stocksianum Boiss. Plant Biotechnol. Rep. 1:179–184.
Chung, K.T., T.Y. Wong, Y.W. Huang, and Y. Lin. 1998. Tannins and human health: A review. Crit. Rev. Food Sci. 38:421–464.
Cline, M.G. 1996. Exogenous auxin effects on lateral bud outgrowth in decapitated shoots. Ann. Bot. 78:255–266
Corredoira, E., A. Ballester, and A.M. Vieitez. 2008. Thidiazuron-induced high-frequency plant regeneration from leaf explants of Paulownia tomentosa mature trees. Plant Cell Tiss. Org. 95:197–208.
Danova, K., E. Čellárová, A. Macková, Z. Daxnerová, and V. Kapchina-Toteva. 2010. In vitro culture of Hypericum rumeliacum Boiss. and production of phenolics and flavonoids. In Vitro Cell Dev. Biol.-Plant 46:422–429.
Djeridane, A., M. Yousfi, B. Nadjemi, D. Boutassouna, P. Stocher, and N. Vidal. 2006. Antioxidant activity of some Algerian medicinal plants extracts containing phenolic compounds. Food Chem. 97: 654–660.
Doyle, J.J. and J.L. Doyle. 1990. Isolation of plant DNA from fresh tissue. Focus 12:13–15.
El-Oualidi, J., O. Verneau, S. Puech, and J.Y. Dubission. 1999. Utility of rDNA ITS sequences in the systematic of Teucrium section polium (Lamiaceae). Plant Syst. Evol. 215:49–70.
Frabetti, M., P. Gutierrez-Pesce, E. Mendoza-de Gyves, and E. Rugini. 2009. Micropropagation of Teucrium fruticans L., an ornamental and medicinal plant. In Vitro Cell Dev. Biol.-Plant 45:129–134.
Goel, M.K., A.K. Kukreja, and N.S. Bisht. 2009. In vitro manipulations in St. John’s wort (Hypericum perforatum L.) for incessant and scale up micropropagation using adventitious roots in liquid medium and assessment of clonal fidelity using RAPD analysis. Plant Cell Tiss. Org. 96:1–9.
Gurel, S., M.C. Baloglu, E. Gurel, H.A. Oktem, and M. Yucel. 2011. Two-stage pretreatment of seedlings improves adventitious shoot regeneration in sugar beet (Beta vulgaris L.). Plant Cell Tiss. Org. Cult. 106:261–268.
Gurel. S., E. Topal, and E. Gurel. 2003. The effect of pretreating seedlings with TDZ on direct shoot regeneration from petiole explants of sugar beet (Beta vulgaris L.). Asia-Pac. J. Mol. Biol. 11:57–62.
Haidara, K., A. Alachkar, and A. Moustafa. 2011. Teucrium polium plant extract provokes significant cell death in human lung cancer cells. Health 3:366–369.
Hatano, T., H. Kagawa, T. Yasuhara, and T. Okuda. 1988. Two new flavonoids and other constituents in licorice root: Their relative astringency and radical scavenging effect. Chem. Pharm. Bull. 36:2090–2097.
Ivanova, M. and J. Van Staden. 2008. Effect of ammonium ions and cytokinins on hyperhydricity and multiplication rate of in vitro regenerated shoots of Aloe polyphylla. Plant Cell Tiss. Org. 92:227–231.
Jahan, A.A., M. Anis, and I.M. Aref. 2011. Preconditioning of axillary buds in thidiazuron-supplemented liquid media improves in vitro shoot multiplication in Nyctanthes arbor-tristis L. Appl. Biochem. Biotechnol. 163:851–859.
Joshi, M. and U. Dhar. 2003. In vitro propagation of Saussurea obvallata (DC.) Edgew. — An endangered ethnoreligious medicinal herb of Himalaya. Plant Cell Rep. 21:933–939.
Karalija, E. and A. Parić. 2011. The effect of BA and IBA on the secondary metabolite production by shoot culture of Thymus vulgaris L. Biol. Nyssana 2:29–35.
Krens, F.A., A. Trifonova, L.C.P. Keizer, and R.D. Hall. 1996. The effect of exogenously-applied phytohormones on gene transfer efficiency in sugar beet (Beta vulgaris L.). Plant Sci. 116:97–106.
Krieken, W.M., H. Breteler, M.H. Visser, and D. Mavridou. 1993. The role of the conversion of IBA into IAA on root regeneration in apple: Introduction of a test system. Plant Cell Rep. 12:203–206.
Leonardi, C., A. Ruggen, and S.I. Malfa. 2001. Hormone effects on in vitro proliferation and rooting of Grevillea explants. Sci. Hort. 90:335–341.
Li, J.J., Y.M. Wu, T. Wang, and J.X. Liu. 2009. In vitro direct organogenesis and regeneration of Medicago sativa. Biol. Plant. 53:325–328.
Lincy, A. and B. Sasikumar. 2010. Enhanced adventitious shoot regeneration from aerial stem explants of ginger using TDZ and its histological studies. Turk. J. Bot. 34:21–29.
Ljubuncic, P., S. Dakwar, I. Portnaya, U. Cogan, H. Azaizeh, and A. Bomzon. 2006. Aqueous extracts of Teucrium polium possess remarkable antioxidant activity in vitro. Evid. Based Complementary Altern. Med. 3:329–338.
Lu, Y.R. and L. Yeap Foo. 2001. Antioxidant activities of polyphenols from sage (Salvia officinalis). Food Chem. 75:197–202.
Maleki Band, S., M. Ghadimzadeh, M. Jafari, and I. Bernousi. 2011. Direct shoot regeneration from stem nodal explants of two wild Medicago species-Medicago scutellata and Medicago rigidula. Austr. J. Crop Sci. 5:668–673.
Mallikarjuna, K. and G. Rajendrudu. 2007. High frequency in vitro propagation of Holarrhena antidysenterica from nodal buds of mature tree. Biol. Plant 51:525–529.
Martin, K.P. 2003. Rapid in vitro multiplication and ex vitro rooting of Rotula aquatica Lour., A rare rheophytic woody medicinal plant. Plant Cell Rep. 21:415–420.
Mehta, U.J., S.M. Barreto, and S. Hazra. 2004. Effect of thidiazuron in germinating tamarind seedlings. In Vitro Cell Dev. Biol.-Plant 40:279–283.
Mendoza-de Gyves, E., J.I. Royani, and E. Rugini. 2007. Efficient method of micropropagation and in vitro rooting of teak (Tectona grandis L.) focusing on large-scale industrial plantations. Ann. Forest. Sci. 64:73–78.
Mishra, J., M. Singh, L.M.S. Palni, and S.K. Nandi. 2011. Assessment of genetic fidelity of encapsulated microshoots of Picrorhiza kurrooa. Plant Cell Tiss. Org. 104:181–186.
Mok, M.C., R.C. Martin, and M.O.K. Dws. 2000. Cytokinins: Biosynthesis, metabolism and perception. In Vitro Cell Dev. Biol.-Plant 36:102–107.
Moncaleán, P., A. Rodríguez, and B. Fernández. 2001. In vitro response of Actinidia deliciosa explants to different BA incubation periods. Plant Cell Tiss. Org. 67:257–266.
Murashige, T. and F. Skoog. 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15: 473–497.
Nadjafi, F., M. Bannayana, L. Tabrizia, and M. Rastgoo. 2006. Seed germination and dormancy breaking techniques for Ferula gummosa and Teucrium polium. J. Arid Environ. 64:542–547.
Nagao, Y., N. Ito, T. Kohno, H. Kuroda, and E. Fujita. 1982. Antitumor activity of Rabdosia and Teucrium diterpenoids against P 388 lymphocytic leukemia in mice. Chem. Pharm. Bull. 30:727–729.
Nematollahi-Mahani, S.N., M. Rezazadeh-Kermani, M. Mehrabani, and N. Nakhaee. 2007. Cytotoxic effects of Teucrium polium on some established cell lines. Pharm. Biol. 45:295–298.
Nikolic, R., N. Mitic, S. Zivkovic, D. Grubisic, M. Neskovic. 2007. Cytokinins and urea derivatives stimulate seed germination in Lotus corniculatus L. Arch. Biol. Sci. 59:125-128
Orhan, I. and M. Aslan. 2009. Appraisal of scopolamine-induced antiamnesic effect in mice and in vitro antiacetylcholinesterase and antioxidant activities of some traditionally used Lamiaceae plants. J. Ethnopharmacol. 122:327–32.
Palacioa, L., J.J. Canterob, R.M. Cusidoc, and M.E. Goleniowskia. 2012. Phenolic compound production in relation to differentiation in cell and tissue cultures of Larrea divaricata (Cav.). Plant Sci. 193–194:1–7.
Patnaik, J., S. Sahoo, and B.K. Debata. 1999. Somaclonal variation in cell suspension culture-derived regenerants of Cymbopogon martinii (Roxb.) Wats var. motia. Plant Breed. 118:351–354.
Punyarani, K. and G.J. Sharma. 2010. Micropropagation of Costus speciosus (Koen.) using nodal segment culture. Not. Sci. Biol. 2:58–62.
Rani, V. and S.N. Raina. 2000. Genetic fidelity of micropropagated plants: A critical reappraisal. In Vitro Dev. Biol.-Plant 14:459–462.
Rechinger, K.H. 1982. Teucrium L., p. 25–27. In: K.H. Rechinger (ed.). Flora Iranica. Akademische Druck und Verlagsanstalt, Wienna, Austria.
Safdari, Y. and S.K. Kazemitabar. 2010. Direct shoot regeneration, callus induction and plant regeneration from callus tissue in Mose Rose (Portulaca grandiflora L.). Plant Omics 3:47–51.
Shan, Z., K. Raemakers, E.N. Tzitzikas, Z. Ma, and R.G. Visser. 2005. Development of a highly efficient, repetitive system of organogenesis in soybean. Plant Cell Rep. 24:507–512.
Sharififar, F., G. Dehghn-Nudeh, and M. Mirtajaldini. 2009. Major flavonoids with antioxidant activity from Teucrium polium L. Food Chem. 112:885–888.
Sujana, P. and C.V. Naidu. 2011. High frequency rapid plant regeneration from shoot tip and nodal explants of Mentha piperita (L.)-An important multipurpose medicinal plant. J. Phytol. 3:09–13.
Sumlu, S., H.H. Atar, and K.M. Khawar. 2010. Breaking seed dormancy of water lily (Nymphaea alba L.) under in vitro conditions. Biotechnol. Biotechnol. Equip. 24:1582–1586.
Uranbey, S. 2005. Thidiazuron induced adventitious shoot regeneration in henbane (Hyoscyamus niger L.) Biol. Plant. 49:427–430.
Van Staden, J., C.W. Fennell, and N.J. Taylor. 2006. Plant stress in vitro: The role of phytohormones. Acta Hort. 725: 55–61.
Zayova, E., I. Stancheva, M. Geneva, M. Petrova, and I. Dimitrova. 2013. Antioxidant activity of in vitro propagated Stevia rebaudiana Bertoni plants of different origins. Turk. J. Biol. 37:106–113.
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Rad, F.A., Jafari, M., Khezrinejad, N. et al. An efficient plant regeneration system via direct organogenesis with in vitro flavonoid accumulation and analysis of genetic fidelity among regenerants of Teucrium polium L.. Hortic. Environ. Biotechnol. 55, 568–577 (2014). https://doi.org/10.1007/s13580-014-0611-7
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DOI: https://doi.org/10.1007/s13580-014-0611-7