In vitro conservation protocol of Ceropegia bulbosa: An important medicinal and threatened plant species of Western Rajasthan

  • Suman Parihar Jai Narain Vyas University


In vitro regeneration protocol has been standardized for highly medicinal and threatened succulent Ceropegia bulbosa Roxb. The paper focuses on morphogenic response of nodal explant when cultured on MS media. Murashige and Skoog medium supplemented with 6-benzyladenine (BA) (2.0 mgl-1) was found optimum for axillary shoot bud induction with 83.4 % response. Further shoots were multiplied through repetitive (3-4 times) transfer of the original explant and by subculture of the in vitro generated shoots. Maximum number of shoots 5.7±0.78 with shoot length of 3.6±0.82 cm was achieved on MS medium augmented with combination of 0.25 mgl-1 BA + 0.25 mgl-1 KN + 0.1 mgl-1 IAA and additives (50.0 mgl-1 ascorbic acid, 25 mgl-1 each of citric acid, arginine and adenine sulphate). For ex vitro rooting, pulse treatment of IBA 250 mgl-1 for 3 min was found optimum. The rooted shoots were successfully hardened in the green house condition (RH 75-80% at 26-28˚C) and about 80 % shoots were transferred to the garden.

Author Biography

Suman Parihar, Jai Narain Vyas University
Assistant Professor


Adibatti, N. A., P. Thirugnanasambantham, C. Kulothungan, S. Viswanathan, L. Kameswaran, K. Balakrishna, and E. Sukumar. 1991. A Pyridine alkaloid from Ceropegia juncea. Phytochemistry 30: 2449–24501. doi: 10.1016/0031-9422(91)83683-C

Compton, M.E, and C.W Mize. 1999. Statistical considerations for in vitro research. I.Birth of an idea to collecting data. In Vitro Cell. Dev. Biol. Plant. 35, 115–121. doi: 10.1007/s11627-999-0020-2

Dhir, R., and G.vS. Shekhawat, 2014. Ecorehabilitation and biochemical studies of Ceropegia bulbosa Roxb.: a threatened medicinal succulent. Acta Physiol Plant. 36 (6): 1335-1343. doi: 10.1007/s11738-014-1512-1 

Dhir, R., Shekhawat, G. S. and A. Alam. 2014. Improved Protocol for Somatic Embryogenesis and Calcium Alginate Encapsulation in Anethum graveolens L.: A Medicinal Herb. Applied Biochemistry and Biotechnology 39: 540-549 doi: 10.1007/s12010-014-1032-x

Dhir, R., and G. S. Shekhawat. 2013. Production, storability and morphogenic response of alginate encapsulated axillary meristems and genetic fidelity evaluation of in vitro regenerated Ceropegia bulbosa: a pharmaceutically important threatened plant species. Ind Crops Prod. 47: 139–144. doi: 10.1016/j.indcrop.2013.02.005

Dhir, R., and G. S. Shekhawat. 2012. Critical review on Tecomella undulata: a medicinally potent endangered plant species of Indian Thar Deseart. Int J Curr Res. 4(6) :36–44

Goyal, D., and S. Bhadauria. 2006. In vitro propagation of Ceropegia bulbosa using nodal segments. Indian Journal of Biotechnology.243(5): 565–567.

Jana, S. and G. S. Shekhawat. 2012. RAPD analysis, antioxidant study during cell differentiation and standardization of in vitro regeneration protocol of Anethum graveolens: A medicinal herb and spice. Biologia plantarum 56(1): 9-14. doi: 10.1007/s10535-012-0009-2

Jana, S. and G. S. Shekhawat. 2010. Phytochemical analysis and antibacterial screening of in vivo and in vitro extracts of Indian medicinal herb: Anethum graveolens. Research journal of medicinal Plant 4(4): 206-212. doi: 10.3923/rjmp.2010.206.212

Jana, S. and G. S. Shekhawat. 2010. Plant growth regulators, adenine sulfate and carbohydrates regulate organogenesis and in vitro flowering of Anethum graveolens. Acta Physiologiae Plantarum 33:305–311. doi: 10.1007/s11738-010-0548-0

Khan, M. A., and D. Pradhan. 2012. Antiurolithc activity of Ceropegia bulbosa extract in rats. Der Pharmacia Sinica 3(1): 148–152.

Mabberley, D. J. 1978. The plant book. Cambridge University Press, Cambridge.

Mathur, S., and G. S. Shekhawat. 2013. Establishment and characterization of Stevia rebaudiana (Bertoni) cell suspension culture: an in vitro approach for production of Steviosides. Acta Physiologiae Plantarum 35(3): 931-939 doi: 10.1007/s11738-012-1136-2

Murashige, T., and F. Skoog. 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Acta Physiol Plant 15: 473–497. doi: 10.1111/j.1399-3054.1962.tb08052.x

Nadkarni, K. M. 1976. Indian Materia Medica. Popular Prakashan, Bombay, India, 303–304

Patel, A. K., M. Phulwaria, M. K. Rai, A. K. Gupta, S. Shekhawat, and N. S. Shekhawat. 2014. In vitro propagation and ex vitro rooting of Caralluma edulis (Edgew.) Benth. & Hook. f.: an endemic and endangered edible plant species of the Thar Desert. Sci Hortic. 165: 175–180. doi: 10.1016/j.scienta.2013.10.039

Phulwaria, M., N. S. Shekhawat, J. S. Rathore, and R. P. Singh. 2013. An efficient in vitro regeneration and ex vitro rooting of Ceropegia bulbosa Roxb.—A threatened and pharmaceutical important plant of Indian Thar Desert. Ind Crops Prod. 42: 25–29. doi: 10.1016/j.indcrop.2012.05.013

Shekhawat, G. S., A. Batra, and S. Mathur. 2002. A reliable in vitro protocol for rapid mass propagation of Azadirachta indica Juss. Journal of Plant Biology 29(1): 109-112

Shekhawat, G. S., S. Mathur, and A. Batra. 2009. Role of phytohormones and various nitrogen inorganic and organic nutrients in induction of somatic embryogenesis in cell culture derived from leaflets of Azadirachta indica A. Juss. Biologia Plantarum 53 (4): 707-710. doi 10.1007/s10535-009-0127-7

Shekhawat, M. S., and N. S. Shekhawat. 2011. Micropropagation of Arnebia hispidissima (Lehm). DC. and production of alkannin from callus and cell suspension culture. Acta Physiol Plant. 33: 1445–1450. doi: 10.1007/s11738-010-0680-x

Singh, M., M. S. Rathore, D. Panwar, J. S. Rathore, H. R. Dagla, and N. S. Shekhawat. 2009 Micropropagation of selected genotype of Aloe vera L.-ancient plant for modern industry. J. Sustain. Forest. 28: 935-950. doi: 10.1080/10549810903344660

Singh, S. K., M. K. Rai, and L. Sahoo. 2012. An improved and efficient micropropagation of Eclipta alba through transverse thin cell layer culture and assessment of clonal fidelity using RAPD analysis. Industrial Crops and Products 37: 328–333. doi: 10.1016/j.indcrop.2011.12.005

Sukumar, E., G. R. Hamsaveni, and R. R. Bhima. 1996. Pharmacological actions of cerpegin, a novel pyridine alkaloid from Ceropegia juncea. Fitoterapia 66: 403–406

Swarnkar, S. and S. S., Katewa. 2008. Ethnobotanical observation on tuberous plants from tribal area of Rajasthan (India). Ethnobotanical Leaflets 12, 647-666.

Thiyagarajan, M. and P. Venkatachalam. 2012. Large scale in vitro propagation of Stevia rebaudiana (bert) for commercial application: pharmaceutically important and antidiabetic medicinal herb. Industrial Crops and Products 37: 111–117. doi: 10.1016/j.indcrop.2011.10.037

Wang, J., W. Gao, L. Zhang, and L. Huang. 2013. Establishment and quality assessment of tissue cultures in Glycyrrhiza uralensis Fisch. Appl Biochem Biotechnol. 169: 588–594. doi: 10.1007/s12010-012-0012-2

Walter, S. K. and J. Gillet. 1998. 1997 IUCN Red list of threatened plants. World Conservation Monitring Centre, Cambridge, UK

Yan, H. C., Liang, L. and Li. Y., Yang. 2010. In vitro and ex vitro rooting of Siratia grosvenorii, a traditional medicinal plant. Acta Physiol Plant. 32:115–120. doi: 10.1007/s11738-009-0386-0.
How to Cite
PARIHAR, Suman. In vitro conservation protocol of Ceropegia bulbosa: An important medicinal and threatened plant species of Western Rajasthan. Plant Science Today, [S.l.], v. 4, n. 1, p. 21-27, jan. 2017. ISSN 2348-1900. Available at: <>. Date accessed: 23 may 2017. doi:
Research Articles


Ceropegia bulbosa; khedula; herbal medicine; ex vitro root

Most read articles by the same author(s)

Obs.: This plugin requires at least one statistics/report plugin to be enabled. If your statistics plugins provide more than one metric then please also select a main metric on the admin's site settings page and/or on the journal manager's settings pages.