Enhancement of surface sterilization protocol for in vitro propagation of Impatiens sirindhorniae

Abstract

A study on sterilization of leaf, nodal, and shoot explants of Impatiens sirindhorniae, an endemic species found only in Krabi and Suratthani provinces has been conducted. Plants were collected from the reservoir of Rajjaprabha Dam, Khao Sok National Park. The explants were cleaned by using detergent solution for 3-5 min and rinsed with tap water for 10-15 min. Then, the explants were immersed in different Clorox concentrations (10, 15, and 20%) or immersed in 0.1% mercuric chloride or immersed in 0.1% mercuric chloride in combination with 10, 15, and 20% Clorox. The explants were cultured on MS agar medium for 4 weeks at 25±2°C, under 16-h photoperiod with 60 photons µmol m-2 s-1 light intensity. The results showed that nodal explants that were sterilized using mercuric chloride at concentration of 0.1% for 20 min is the most effective technique. This treatment gave the survival rate of 83.3% and all the explants developed into shoots and roots. In contrary, survival leaf and shoot explants did not develop into shoots and roots.