Establishment of somatic embryogenesis and podophyllotoxin production in liquid shake cultures of Podophyllum hexandrum Royle

- A high frequency somatic embryogenesis was established in Podophyllum hexandrum for podophyllotoxin (PTOX) production.
- Embryogenic callus cultured on 0.75 strength MS medium with 4% sucrose and 3 g/l PVP yielded higher number of somatic embryos.
- Highest PTOX accumulation (2.8 mg/l) was recorded in somatic embryos cultured in 0.75 strength MS medium with 8% sucrose.
- The protocol established in this study will be helpful for large scale production of podophyllotoxin.

The objective of this study was to establish a simple and efficient method for high frequency of somatic embryogenesis and plant regeneration of Podophyllum hexandrum and podophyllotoxin (PTOX) production. Embryogenic callus (100 mg FW) cultured on 0.75 strength MS medium with 4% sucrose and 3 g/l PVP produced maximum number of somatic embryos (1089) after five weeks of culture under dark condition. When these somatic embryos were subcultured to the same MS medium with 1 mg/l ABA, the embryos matured after 2 weeks of culture. The MS medium supplemented with1.0 mg/l GA3 led to high frequency germination (91.1%). The accumulation of PTOX varied in somatic embryos, matured embryos and germinated embryos. Highest PTOX accumulation (2.8 mg/l) was recorded in somatic embryos cultured in 0.75 strength MS medium with 8% sucrose. Regenerated plantlets were successfully acclimatized in the growth chamber. The protocol established in this study will be helpful for large scale production of podophyllotoxin.