In-vitro propagation, callus culture and bioactive lignan production in Phyllanthus tenellus Roxb: a new source of phyllanthin, hypophyllanthin and phyltetralin

This is the first report on identification and quantification of important hepatoprotective and anticancer polyphenolic lignans such as phyllanthin (PH), hypophyllanthin (HPH), niranthin (NH) and phyltetralin (PT) in natural plant and in vitro cultures of Phyllanthus tenellus Roxb. The identification of lignans was carried out by Liquid Chromatography-High Resolution Mass Spectrometry (LC-HRMS) and quantified using High-Performance Liquid Chromatography (HPLC). In addition, an efficient protocol has been developed for multiple shoot induction in nodal explants of in vitro derived shoots of P. tenellus. Maximum number of shoot regeneration (7.83 +/- 0.15) was achieved on medium incorporated with 1.0 mg/l 6-Benzylaminopurine (BAP). The medium containing Indole-3-acetic acid (IAA) 2 mg/l was superior for induction of rooting in in vitro raised shoots. The plantlets were acclimatized to the field condition with 100% survival. The quantitative HPLC analysis showed that the lignan content was variable with the auxins and cytokinins incorporated in the medium. The lignan content was higher in callus grown on Murashige and Skoog (MS) medium+2.0 mg/l Naphthaleneacetic acid (NAA). The reported protocol can be used for mass propagation and application of biotechnological approaches for improvement of P. tenellus. The results indicate intriguing possibilities for the utilization of P. tenellus plant parts as an alternative source and of callus culture to scale up bioactive lignan production for pharmaceutical applications.