An efficient protocol for in vitro clonal propagation of natural sweetener plant (Stevia rebaudiana Bertoni)

Ashish Ojha¹*, V. N. Sharma¹ and Vinay Sharma²

¹Department of Biotechnology, Plant Tissue Culture and Research Development Laboratory, MIET group of Institutions, Meerut- 250005, U. P. India.

²Department of Biotechnology and Bioscience, Banasthali University, Banasthali, Rajasthan, India.

*Corresponding author. E-mail: ashish_ojha1087@rediffmail.com.

Accepted 18 May, 2010.

Abstract

Leaf segments of Stevia were cultured on MS medium supplemented with varying concentrations (0.5, 1.0, 1.5, 2.0, 2.5 mg/L^-1) of growth regulators (BAP and 2, 4-D). Ninety one percent aseptic cultures were obtained when sterilized with 0.1% HgCl₂ for 10 min. The highest amount of callus was obtained in MS medium supplemented with1.0 mg/L^-1 BAP+0.5 mg/L^-1 2, 4-D, respectively. On the other hand, 2.5 mg/L^-1 BAP + 0.5 2,4-D showed lowest performance. Highest shoot was obtained in 2.0 mg/L^-1 BAP. These shoots was transfer into different concentration of IBA, NAA and IAA (0.5. 1.0, 1.5, and 2.0) used. Highest rooting percentage was recorded on MS medium with 0.5 mg/L^-1 IAA. The rooted plantlets were transfer into mist chamber with relative humidity for 2 - 4 week after these plants were hardened and successfully established in soil.

Key words: Stevia rebaudiana, sweetener, callus, leaf.