Indirect somatic embryogenesis and evaluation of genetic fidelity of in vitro propagated plants of Bunium persicum (Boiss.) Fedtsch by using DNA markers

A reproducible and efficient protocol was developed for inducing indirect somatic embryogenesis using callus originated from different explants of Bunium persicum viz. both immature and mature zygotic embryos, fruit pedicel and cotyledons excised from in vitro germinated seedlings. Calli derived from different explants was inoculated on Murashige and Skoog (MS) basal medium of half and full strength supplemented with various concentrations of thiadiazuron, indole butyric acid and with activated charcoal. Callus derived from fruit pedicel gave best response for somatic embryogenesis on MS basal full strength + 0.1% activated charcoal. Half strength MS basal + 1.00 mg/l TDZ + 0.05 mg/l IBA giving as much as 84.33 per cent of somatic embryogenesis and 25.78 was the average number of somatic embryos per callus clump of 200 mg. Maximum of 70.61% somatic embryos showed germination when transferred to half strength MS basal medium. Histological observations revealed that somatic embryos displayed a standard development pattern, from globular to finally cotyledonary stage embryos intervened by heart shaped and torpedo shaped embryos. Molecular characterization of somatic embryo-derived plants was carried out by employing Random Amplified Polymorphic DNA (RAPD) and Inter Simple sequence Repeat (ISSR) primers. DNA extracted from shoots of 15 Bunium persicum plantlets regenerated from somatic embryos and mother plant gave monomorphic banding pattern upon amplification, which indicates that somatic embryogenesis is a suitable technique of micropropagation.