Abstract
An efficient in vitro plant regeneration protocol for Swertia chirata Buch.-Ham. ex Wall (Gentianaceae), a critically endangered Himalayan medicinal herb, was developed using shoot tip explants derived from in vitro grown seedlings. Media with 2% sucrose and various types of hormones markedly influenced in vitro propagation of S. chirata. An in vitro shootlet production system using Murashige and Skoog (MS) medium with various hormones such as BAP, KN and TDZ was established. BAP at 1.0 mg/l and KN, 0.1 mg/l induced highest number of multiple shoots (42.16 ± 1.05) per explant. Micro-proliferated shoots were transferred to elongation medium amended with GA3 (0.1 mg/l) and hormone free basal medium, after which they were transferred to rooting medium. The highest frequency of rooting (22.48 ± 1.08) was obtained in half-strength MS medium supplemented with NAA, 0.1 mg/l after testing with different auxins at various concentrations within 4 weeks of transfer to the rooting medium. Hardening was successfully attained under controlled conditions inside the plant tissue culture room. This method could effectively be applied for the conservation and clonal propagation to meet the pharmaceutical demands.
Similar content being viewed by others
Abbreviations
- BAP:
-
6-Benzylaminopurine
- 2,4-D:
-
2,4-Dichlorophenoxyacetic acid
- IBA:
-
Indole-3-butyric acid
- IAA:
-
Indole-3-acetic acid
- KN:
-
Kinetin
- NAA:
-
α-Naphthalene acetic acid
- TDZ:
-
Thidiazuron
References
Agastian P, Lincy W, Ignacimuthu S (2006) In vitro propagation of Justicia gendarussa Burm f.—a medicinal plant. Indian J Biotechnol 5:246–248
Anis M, Faisal M (2005) In vitro regeneration and mass multiplication of Psoralea corylifolia—an endangered medicinal plant. Int J Biotechnol 4:261–264
Anonymous (1952) Wealth of India. A dictionary of Indian raw materials and industrial products, vol 10. CSIR, New Delhi
Arora R, Bojwani SS (1989) In vitro propagation and low temperature storage of Saussurea lappa C B Clarke, an endangered medicinal plant. Plant Cell Rep 8:44–47. doi:10.1007/BF00735776
Asthana PK, Sharma NK, Kulshreshtha DK, Chatterjee SKA (1991) Xanthone from Swertia chirayita. Phytochemistry 30:1037–1039. doi:10.1016/0031-9422(91)85308-M
Baskaran P, Jayabalan N (2007) Rapid micropropagation of Psoralea corylifolia L. using nodal explants cultured in organic additive-supplemented medium. J Hortic Sci Biotechnol 82:908–913
Bhat SR, Chandel KPS, Malik SR (1995) Plant regeneration from various explants of cultivated Piper species. Plant Cell Rep 14:395–402. doi:10.1007/BF00238605
Bijelovic A, Rosic N, Miljus-djukic J, Ninkovic S, Grubisic D (2004) In vitro regeneration and transformation of Blackstonia perfoliata. Biol Plant 48(3):333–338. doi:10.1023/B:BIOP.0000041083.41294.31
Chakravarty AK, Mukhopadhayay S, Das B (1991) Swertane terpenoids from Swertia chirata. Phytochemistry 30:4052–4087. doi:10.1016/0031-9422(91)83473-X
Chakravarty AK, Mukhopadhyay S, Masuda K, Ageta H (1992) Chiratenol, a novel rearranged hopane tritepenoid from Swertia chirata. Tetrahedron Lett 31:7649–7652. doi:10.1016/S0040-4039(00)97322-2
Chakravarty AK, Mukhopadhyay S, Moitra SK, Das B (1994) Syringareinol, a hepatoprotective agent and other constituents from Swertia chirata. Indian J Chem B 33:405–408
Chaudhuri PK, Daniewshi WM (1996) Unambiguous assignments of the 1H and 13C chemical shifts of the major bitter principles of Swertia chirata by 2D NMR study and characterization of other constituents. Pol J Chem 69:1514–1519
Chaudhuri RK, Amita P, Timir BJ (2007) Production of genetically uniform plants from nodal explants of Swertia chirata Buch-Ham ex Wall—an endangered medicinal herb. In vitro Cell Dev Biol Plant 43:467–472. doi:10.1007/s11627-007-9095-9
Dalal SR, Shah RC (1956) Swerchirin, a novel xanthone from Swertia chirata. Chem Ind 7:664
Dubey NK, Kumar R, Tripathi P (2004) Global promotion of herbal medicine: India’s opportunity. Curr Sci 1(86):10
Dutta Gupta S, Ibaraki Y (eds) (2006) Plant tissue culture engineering. Agritech Publications, New York
Earle ED, Langhans RW (1974) Propagation of Chrysanthemum in vitro I. Multiple plantlets from shoot tips and the establishment of tissue cultures. J Am Soc Hortic Sci 99:128–132
Elghag HM (1991) In vitro propagation of Catha edulis. Hortic Sci 26:212
Figueiredo SFL, Albarello N, Viana VRC (2001) Micropropagation of Rollina mucosa (Jacq) Baill. In vitro Cell Dev Biol Plant 37:471–475. doi:10.1007/s11627-001-0083-1
George EF (1993) Plant propagation by tissue culture. Exegetics Ltd, Edington
Hatjimanoli M, Favre-Bonvin J, Koauadji M, Mariotte AM (1988) Monohydroxy-and 2, 5 dihydroxy terephthalic acids, 2 unusual phenolics isolated from Centaurium erythraea and identified in Gentianaceae members. J Nat Prod 51:997. doi:10.1021/np50059a030
Hisato K, Toshiki N, Kinya M, Masanobu T, Masahiro M (1995) Plant regeneration from mesophyll protoplasts of lisianthus (Eustoma grandiflorum) by adding activated charcoal into protoplast culture medium. Plant Cell Tissue Organ Cult 43:59–65. doi:10.1007/BF00042672
Hossain M, Biswas BK, Karim MR, Rahman S, Islam R, Joarder O (1994) In vitro organogenesis of elephant apple (Feronia limonia). Plant Cell Tissue Organ Cult 39:265–268. doi:10.1007/BF00035980
Joshi P, Dhawan V (2007a) Assessment of genetic fidelity of micropropagated Swertia chirayita plantlets by ISSR marker assay. Biol Plant 51(1):22–26. doi:10.1007/s10535-007-0005-0
Joshi P, Dhawan V (2007b) Axillary multiplication of Swertia chirayita (Roxb. Ex Fleming) H. Karst., a critically endangered medicinal herb of temperate Himalayas. In vitro Cell Dev Biol Plant 43:631–638. doi:10.1007/s11627-007-9065-2
Kirtikar KR, Basu BD (eds) (1984) Indian medicinal plants, vol III. LM Basu Publishers, Allahabad, pp 1664–1666
Kukreja AK, Mathur AK, Zaim M (1990) Mass production of virus free patchouli plants [Pogostemon cablin (Blanco)] by in vitro culture. Trop Agric 67:101–104
Loc NH, Duc DT, Kwon TH, Yang MS (2005) Micropropagation of zedoary (Curcuma zeodoria Roscoe)—a valuable medicinal plant. Plant Cell Tissue Organ Cult 81:119–122. doi:10.1007/s11240-004-3308-2
Mederos-Molina S (2002a) Micropropagation of Hypericum canariense L. for the production of hypericin. In: Nagata Ebizuka (ed) Biotechnology in agriculture and forestry: Medicinal and aromatic plants, tome XII, vol 51. Springer, Berlin, pp 95–117
Mederos-Molina S (2002b) In vitro propagation of Maytenus canariensis (Loes.) Kand & Sund from apical meristem culture. Plant Tissue Cult 12:99–108
Miura H (1991) Swertia spp. In vitro culture, regeneration and the production of secondary metabolites. In: Bajaj YPS (ed) Biotechnology in agriculture and forestry. Medicinal and aromatic plants, vol 15. Heidelberg, Berlin, pp 451–463
Mukherji B (ed) (1953) Indian pharmaceutical codex, Indigenous drugs, vol I. CSIR, New Delhi, pp 64–65
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue. Physiol Plant 15:473–497. doi:10.1111/j.1399-3054.1962.tb08052.x
Pattnaik SK, Chand PK (1996) In vitro propagation of the medicinal herbs Ocimum americanum L. Syn, O. canum Sims (hoary basil) and Ocimum sanctum L. (holy basil). Plant Cell Rep 15:846–850. doi:10.1007/BF00233154
Piatczaka E, Wielanekb M, Wysokinskaa H (2005) Liquid culture system for shoot multiplication and secoiridoid production in micropropagated plants of Centaurium erythraea Rafn. Plant Sci 168:431–437. doi:10.1016/j.plantsci.2004.08.013
Purohit AN (1997) Medicinal plants—upgrading technology for trading the traditions. In: Proceedings in national seminar on harvesting herbs 2000. HAPPRC, Srinagar
Purohit SD, Dave A, Kukda G (1994) Micropropagation of safed musli (Chlorophytum borivilianum), a rare medicinal herb. Plant Cell Tissue Organ Cult 39:93–96. doi:10.1007/BF00037596
Rai LK, Prasad Pankaj, Sharma E (2000) Conservation threats to some important medicinal plants of the Sikkim Himalaya. Biol Conserv 93:27–33. doi:10.1016/S0006-3207(99)00116-0
Ray S, Majumder HK, Chakravarty AK, Mukhopadhyay S, Gil RR, Cordell GA (1996) Amarogentin, a naturally occurring secoiridoid glycoside and a newly recognized inhibitor of topoisomerase I from Leishmania donovani. J Nat Prod 59:27–29. doi:10.1021/np960018g
Sapna S, Ravi TK (2007) Approaches towards development and promotion of herbal drugs. Pharmacognosy reviews vol 1(1), Jan–May
Sen J, Sharma AK (1991) Micropropagation of Withania somnifera from germinating seeds and shoot tips. Plant Cell Tissue Organ Cult 26:71–73. doi:10.1007/BF00036108
Sharma AK, Prasad RN, Chaturvedi HC (1981) Clonal propagation of Bougainvillea glabra “Magnifica” through shoot apex culture. Plant Cell Tissue Organ Cult 1:33–38. doi:10.1007/BF02318901
Skrzypczak L, Wesolowska M, Skrzypczak E (1993) Gentiana species: in vitro culture, regeneration and production of secoiridoid glucosides. In: Bajaj YPS (ed) Biotechnology in agriculture and forestry. Medicinal and aromatic plants IV, vol 21. Springer, pp 172–186
Sudha GC, Seeni S (1994) In vitro multiplication and field establishment of Adhatoda beddomei CB Clarke, a rare medicinal plant. Plant Cell Rep 13:203–207. doi:10.1007/BF00239893
Vishwanath MP, Jayanthi M (1997) Micropropagation of two species of Rauvolfia (Apocynaceae). Curr Sci 72:961–965
Wawrosch C, Maskay N, Kopp B (1999) Micropropagation of the threatened Nepalese medicinal plant Swertia chirayita Buch-Ham. ex. Wall. Plant Cell Rep 18:997–1001. doi:10.1007/s002990050697
Acknowledgment
The authors are grateful to the University Grants Commission, New Delhi, India, for providing financial assistance (Sanction No. F.31-238/2005(SR) dated 05.04.2006).
Author information
Authors and Affiliations
Corresponding author
Additional information
Communicated by E. Lojkowska.
Rights and permissions
About this article
Cite this article
Balaraju, K., Agastian, P. & Ignacimuthu, S. Micropropagation of Swertia chirata Buch.-Hams. ex Wall.: a critically endangered medicinal herb. Acta Physiol Plant 31, 487–494 (2009). https://doi.org/10.1007/s11738-008-0257-0
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11738-008-0257-0