PJB-2017-86
In vitro propagation of Silybum marianum (L.) Gaertn. and genetic fidelity assessment of micro-propagated plants
Yu-Wei Lv, Rui-Jie Wamg, Ya-Wei Lv, Ze-Shang Yang and Ying-Juan Wang
Abstract
Silybum marianum (milk thistle), an annual and biennial herbaceous plant, has been used to treat liver disease for 2000 years. An efficient system for micropropagation from leaf explants of wild-grown S. marianum was successfully established and the genetic fidelity of micropropagated plants was assessed using PCR-based random amplified polymorphic DNA (RAPD). The highest frequency (98.9%) of callus induction was obtained from explants cultivated for 4 weeks on Murashige and Skoog (MS) medium supplemented with 1.5 mg l-1 N6-Benzylaminopurine (BA) and 2.0 mg l-1 a-Naphthaleneacetic acid (NAA). The optimal medium for shoots organogenesis was MS supplemented with 3.0 mg l-1 Gibberellic acid (GA3) and 1.0 mg l-1 NAA, while shoot proliferation was accomplished in MS supplemented with 1.0 mg l-1 BA and 0.3 mg l-1 NAA. Rooting (94.5%) of these shoots was achieved after 4 weeks on 1/2 MS medium supplemented with 1.0 mg l-1 IBA. A total of 92% of the plants were surviving 6 weeks after transplantation of plantlets to soil. RAPD analysis revealed that the genetic fidelity between the parent and the in vitro-raised plantlets exhibited 100% similarity. This confirmed the true-to-type nature of the in vitro-raised clones
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