Assessment of genetic stability of micropropagated Curculigo latifolia from Indonesia by RAPD and ISSR

Micropropagation of Curculigo latifolia is very important because this plant has high economic value including containing Curculin and Neoculin as sweetener and taste-modifying proteins. Micropropagated plants may experience genetic changes like somaclonal variation so it is necessary to assess their genetic stability. The purpose of this study was to obtain a micropropagation method of C. latifolia that is more efficient than the previous research and to assess the genetic stability of micropropagated C. latifolia. The plant material used was sterile leaves obtained from sterile seedlings raised from C. latifolia seed germination. Leaves were grown on a callus induction medium containing 5 mg l⁻¹ indole-3-butyric acid and 3 mg l⁻¹ BA in a dark room. Callus formed were subcultured to growth selection medium and shoots formed were subcultured to Murashige and Skoog-free growth regulator medium with the addition of 6 g l⁻¹ agar and 3% sucrose. The cultures were incubated at 23ºC ± 2ºC under 1,500 lux white LED (Philip™ TL) on a 16 hour/8 hour light/dark photoperiod. The cultures were incubated for 14 weeks. Genetic stability of in vitro propagated C. latifolia from Indonesia was assessed by molecular markers, RAPD and ISSR. The micropropagation method produces an efficient protocol because each growth stage occurs in a free growth regulator medium except for callus induction. Based on the results of the assessment using molecular markers, no polymorphism in DNA amplification using RAPD and a high percentage of genetic stability (66.67%) from ISSR analysis were obtained. In sum, the in vitro micropropagation protocol created can be used commercially to produce C. latifolia on a large scale that is genetically stable.