Micropropagation of Phyllanthus fraternus Webster (Euphorbiaceae) from field-derived shoot tip explant and assessment of its genetic fidelity

Abstract

Phyllanthus fraternus Webster is an important medicinal plant of family Euphorbiaceae. According to a report from the National Medicinal Plants Board, New Delhi, this plant is considered under high-volume trade or consumption category (>100 million tones/year) and is having high demand in domestic and international market for making herbal formulations. Pharmaceutical industries need huge volume of raw materials of this valuable species. Therefore, the development of alternative plant regeneration tools for this medicinal plant is urgently needed. In vitro micropropagation techniques offer a viable tool for rapid mass multiplication and germplasm conservation of important medicinal plants for meeting the pharmaceutical needs. In the current paper, an efficient protocol for in vitro propagation of P.fraternus from field-derived mature shoot tip explants has been presented. The shoot tip explants induced multiple shoots on cytokinin-containing medium. The best response was achieved on MS medium supplemented with 1 mg/L 6-benzylaminopurine and 0.5 mg/L kinetin (Kn), in which 99 % of cultures produced 21.07 ± 0.58 shoots per explants. Regenerated shoots were rooted on MS medium supplemented with 0.5 mg/L IBA. Plantlets were successfully acclimatized and established in soil. Regenerated plants were grown normally in the field without showing any morphological variations. Genetic fidelity of acclimatized plants was assessed by using inter-simple sequence repeat analysis. The molecular analysis complemented and compared well and showed genetic stability in the plants regenerated through mature shoot tips. This protocol can be used for the huge production of raw material of the species which could be utilized by the industries for the discovery of potential new drug candidate.