In vitro regeneration of periwinkle (Catharanthus roseus L.) and fidelity analysis of regenerated plants with ISSR Markers

Catharanthus roseus is an important multipurpose medicinal plant. In this study, in vitro proliferation and root induction of periwinkle were optimized and regenerated plants were subsequently surveyed for genetic homogeneity using the inter simple sequence repeat (ISSR) markers. Shoot tips and nodal segments were cultured in Murashige and Skoog (MS) medium supplemented with different concentrations of benzylamino-purine (BAP), gibberellic acid (GA3), and indol-3-butyric acid (IBA) hormones. ISSR profiling of regenerated plants as well as the mother plant were surveyed with five primers. The highest establishment rate (80.67%) was obtained in the MS medium containing 1.0 mg L-1 GA3 and 1.0 mg L-1 BAP. Highest proliferation rate (5.20 shoots/explant) and average shoot length (6.30 cm) were observed in 1.5 mg L-1 BAP + 0.5 mg L-1 IBA. Moreover, the best rooting response (85.30%) was observed on half strength MS containing 1.0 mg L-1 IBA. Genetic fidelity analysis using ISSR markers showed the monomorphic banding pattern of the micro-propagated plants and the mother plant, which highlighted their genetic uniformity. This implies that periwinkle micropropagation through shoot tip is the most reliable method for true-to-type production of C. roseus in a large scale.