IN VITRO PLANT REGENERATION OF THEVETIA NERIIFOLIA, JUSS FROM INTERNODE EXPLANTS VIA INDIRECT ORGANOGENESIS
Keywords:
Callus, Micropropagation, Organogenesis, Regeneration, Thevetia neriifoliaAbstract
Objective: Present study was carried out to develop an efficient in vitro protocol for micro propagation of plantlets via indirect organogenesis from internode explants of Thevetia neriifolia, a medicinal plant with immense biological potentialities.
Methods: For callus induction, internodes from 4 weeks old seedlings were cultured in Murashige and Skoog (MS) medium supplemented with varying concentrations (0.5-2.0mg/L-1) of Auxins (2,4-D, IBA) and Cytokinins (KIN, BA).
Results: Creamy white callus obtained initially was gradually transformed into the light green meristematic callus during subsequent subculturing in most of the combinations. The best organogenic response was achieved with a combination of IBA+BA (0.5+1.0mg/L-1) with 2.722±0.752 shoots per explant. Shoot initiation was induced/enhanced by transferring the non meristematic IBA+KIN calluses to the above IBA+BA medium. However, better response for maximum shoot proliferation (6.091±1.231) was achieved when cytokinin BA (1.0mg/ L-1) was supplied individually, with 100% efficacy. Good roots were produced in IBA (1.0mg/L-1) medium and the rooted plantlets were transferred to paper cups and acclimatized (72%) subsequently in green houses.
Conclusion: This rapid and successful protocol for micro-propagation definitely facilitates to promote the mass cultivation of this potent medicinal plant.
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References
Joshi SG. Family X Apocynaceae In: Medicinal Plants, 1st Ed. New Delhi, Oxford & IBH Publishing Company Pvt. Ltd; 2000. p. 40-51.
Zavaleta AN. Medicinal Plants of the Borderlands: A Bilingual Resource Guide Author House, USA, Bloomington; 2012.
Kumar A, Tandon P. Investigation on in vitro Laticifer Differentiation in Thevetia peruviana L. Phytomorphol 1990;40(1-2):113-7.
Kumar A. Somatic embryogenesis and high frequency plantlet regeneration in callus cultures of thevetia peruviana. Plant Cell Tiss Org Cult 1992;31:47-50.
Zibbu G, Batra A. Effect of adeninesulphate on organogenesis via leaf culture in an ornamental plant: thevetia peruviana (pers) schum. Int J Pharm Bio Sci 2010;1(2):1-9.
Taha HS, Farag HS, Shams AK, Abdel-Azim SN, Hanna GA, Ewais EE, et al. In vivo and in vitro studies on thevetia species growing in egypt i: isolation, identification, and quantification of cardiac glycosides in in vivo and in vitro cultures of immature seeds. J Amer Scien 2010;6(11):390-5.
Murashige T, Skoog F. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 1962;15:473-9.
Sharma A, Kumar A. Somatic embryogenesis and plant regeneration from leaf-derived cell suspension of a mature tree-thevetia peruviana l. Plant Cell Rep 1994;14:171-4.
Siwach P, Grover K, Gill AR. The influence of plant growth regulators, explant nature and sucrose concentration on in vitro callus growth of thevetia peruviana schum. Asian J Biotech 2011;3:280-92.
Chen CC, Chen SJ, Sagare AP, Tsay HS. Adventitious shoot regeneration from stem internode explants of adenophora triphylla (thunb.) a. dc. (campanulaceae)-an important medicinal herb. Bot Bull Acad Sin 2001;42:1-7.
Kaewpooa M, Te-chatob S. Influence of explant types and plant growth regulators on multiple shoot formation from jatropha curcas. Sci Asia 2009;35:353–7.
Haq R, Naz S, Aslam F, Manzoor F. Comparison of in vitro response of micropropagation and callogenesis of medicinal plant, vinca rosea L. J Agric Res 2013;51(1):9-17.
Salmu U, Rahman MSM, Islam S, Haque N, Jubair TA, Haque AK, et al. The influence of different hormone concentration and combination on callus induction and regeneration of rauwolfia serpentine L. Benth Pak J Biol Sci 2008;11:1638-41.
Khanam N, Khoo C, Khan AG. Effects of cytokinin/auxin combination on organogenesis, shoot regeneration and tropane alkaloid production in duboisia myoporoides. Plant Cell Tiss Org Cult 2000;62:125-33.
Ranyaphiaa RA, Maoaa AA, Borthakurbb SK. Direct organogenesis from leaf and internode explants of in vitro raised wintergreen plant (gaultheria fragrantissima). Sci Asia 2011;37:186–94.
Taha HS, Farag SH, Shams KA, Abdel-Azim NS, El-Nasr SMM. In vivo and in vitro studies on thevetia species growing in egypt ii: establishment of in vitro tissue culture system and production of cardiac glycosides. J Am Scien 2011;7(3):1-12.