Podophyllotoxin production via cell and adventitious root cultures of Podophyllum peltatum

Abstract

Podophyllum peltatum is an important medicinal plant that produces podophyllotoxin (PTOX) with anticancer properties. We established the embryogenic cell and adventitious root culture systems in P. peltatum and analyzed PTOX production. For the growth of embryogenic cell clumps in shake flask culture, the most efficient concentration of 2,4-dichloroacetic acid (2,4-D) was 6.78 μM, and the growth of embryogenic cell clumps was 15.9-fold increased in Murashige and Skoog MS liquid medium with 6.78 μM 2,4-D after 3 wk of culture. To induce adventitious roots, half-strength MS medium showed the best results for adventitious root induction compared to full strength MS medium and MS medium lacking NH 4 NO 3 . Optimal indole-3-butyric acid concentration for adventitious root formation was 14.78 μM. In liquid medium, the frequency of adventitious root formation from root segments was 87.7% and the number of laterally formed adventitious roots was 22.3 per segment. PTOX production in embryogenic cells and adventitious roots was confirmed by liquid chromatography and electrospray ionization-tandem mass spectrometry analysis. Highperformance liquid chromatography analysis revealed that adventitious roots contained higher PTOX than embryogenic cell clumps. Elicitor treatment (20 μM methyl jasmonate) strongly enhanced the production of PTOX in both embryogenic cell clumps and adventitious roots. This observation suggests that both embryogenic cell and adventitious root culture can be adopted to produce PTOX.

An efficient regeneration system via direct and indirect organogenesis for the medicinal plant Dysosma versipellis (Hance) M. Cheng and its potential as a podophyllotoxin source

“…After equal culture times (8 weeks), the podophyllotoxin content was significantly higher in the cultured rhizome (3.20 mg g -1 ) than in the calli (1.17 mg g -1 ). Similar results were achieved for P. peltatum (Anbazhagan et al 2008). They found that podophyllotoxin content in embryogenic cells (coming from embryogenic callus) was 0.0865 mg g -1 , and 0.362 mg g -1 in adventitious roots.…”

“…In addition, they found that phenylalanine (0.2 g l -1 ) enhanced the production of podophyllotoxin in calli to 5 mg g -1 . Anbazhagan et al (2008) showed that elicitor treatment (20 lM methyl jasmonate) strongly enhanced the production of podophyllotoxin in both embryogenic cell clumps and adventitious roots. In our study, the podophyllotoxin content in the calli growing for 8 weeks reached one-fifth of that in wild plants that had grown for 2 years.…”

Effects of some elicitors on tanshinone production in adventitious root cultures of Perovskia abrotanoides Karel

“…Jasmonates are signal molecules and plant regulators that mediate various developmental processes and induce plant defense responses to pathogens and insects (Chen et al, 2007;Yu et al, 2002;Rhee et al, 2010). In particular, exogenous application of MJ as an elicitor can be effective to stimulate the production of various secondary metabolites belonging to different structural groups in plant cell and tissue cultures (Yoo et al, 2011;Zhao et al, 2010;Rhee et al, 2010), including the formation of decursinol angelate in A. gigas root culture (Rhee et al, 2010), ginsenosides in Panax ginseng hairy root and adventitious root culture (Yu et al, 2002;Hong et al, 2012), podophyllotoxin in Podophyllum peltatum adventitious root and cell cultures (Anbazhagan et al, 2008), saikosaponins in Bupleurum kaoi adventitious root culture (Chen et al, 2007) and hyoscyamine and scopolamine in A. belladonna hairy root culture (Eskandari et al, 2012).…”

Investigation and Expression of the Secoisolariciresinol Dehydrogenase Gene Involved in Podophyllotoxin Biosynthesis

“…One of these sources is the rhizomes of the plant Podophyllum peltatum [6]. Due to a combination of the slow growth-rate of the plant and increasing demand for PPT [7][8][9], various other approaches for the chemical synthesis and in vitro production of PPT have been explored [1,[10][11][12][13][14]. These methods have so far proven to be inadequate for the commercial production of PPT due to low yields and high costs.…”