African Journal of

  • Abbreviation: Afr. J. Biotechnol.
  • Language: English
  • ISSN: 1684-5315
  • DOI: 10.5897/AJB
  • Start Year: 2002
  • Published Articles: 12269

Full Length Research Paper

Efficient low cost seesap (CSUP) technique for micropropagation of newly introduced sweetener plant, Stevia rebaudiana Bertoni

Pubudini Lakshani Thilakarathne
  • Pubudini Lakshani Thilakarathne
  • Faculty of Science and Technology, Horizon Campus, 482/B, Suhada Mawatha, Millenium Drive, Chandrika Kumaranathunga Mawatha, Malabe, Sri Lanka.
  • Google Scholar
Sriyani Edussuriya Peiris
  • Sriyani Edussuriya Peiris
  • Sri Lanka Institute of Information Technology, New Kandy Road, Malabe, Sri Lanka.
  • Google Scholar
Pathirana Chathurangi Lankika
  • Pathirana Chathurangi Lankika
  • Sri Lanka Institute of Information Technology, New Kandy Road, Malabe, Sri Lanka.
  • Google Scholar

  •  Received: 28 April 2019
  •  Accepted: 12 June 2019
  •  Published: 19 June 2019


Stevia rebaudiana is a non-caloric sweetening herb, which has a high potential for commercialization. During this study low cost media sterilization, seesap (CSUP) technique which applies sodium hypochlorite was used as an alternative to autoclaving. In vitro cultures were established using nodal segments collected from stevia mother plants. These in vitro shoots were cultured on Murashige & Skoog (MS) medium supplemented with 16 combinations of 6- benzyl amino purine (BAP) and kinetin (KN) with 3% sugar at 5.8 pH, to investigate the in vitro shoot multiplication. The developed shoots were transferred to half MS medium supplemented with 0, 1 and 2 mg/L Indole butyric acid (IBA) with 3, 4 and 5% sugar in nine combinations at 5.8 pH, to investigate the rooting. The well rooted stevia plantlets were transferred to Jiffy™ bags for hardening. The results showed that the combination of 1.0 mg/L BAP and 1.5 mg/L KN resulted in the highest number of shoots (11.8 shoots/shoot).  The combination of 2.0 mg/L IBA with 5% sugar in half MS medium resulted in the highest root induction (7.2 roots / plant) and root elongation (1.02 cm). The plants grown in culture medium containing 5% sucrose gave 100% survival rate at acclimatization.


Key words: Stevia, micropropagation, shoot induction, root induction, acclimatization, low cost seesap (CSUP).