Abstract
Purpose: Podophyllum hexandrum Royle, a source of highly valued podophyllotoxin has been subjected to heavy collection from the wild. The ever-increasing demand of podophyllum is mainly due to two semi synthetic derivatives of podophyllotoxin that is etoposide and teniposide, which are used in the treatment of various types of cancer. The anti cancer lignan derivative podophyllotoxin in Podophyllum hexandrum is biosynthesized at very low quantities in intact plant, so the biotechnological production of podophyllotoxin has been considered essential. Method: The aseptically germinated embryos of Podophyllum hexandrum were developed on solid nutrient agar slab. For the growth of callus culture, Murashigae and Skoog media (MS media)) with various concentrations of BAP, NAA and GA3 adjusted to pH 5.8 was used. Podophyllotoxin content in the alcoholic extract of calli and plant root was analysed by HPLC and HPTLC and was also compared with cultivated Podophllum hexandrum root extracts. Result: A fully defined MS medium supplemented with Naphthalene acetic acid and 6benzylaminopurine (BAP) were effective for both initiation and sustained growth of callus tissue. The relative proportion of callus was markedly influenced by presence of plant growth regulators. The amount of Podophyllotoxin obtained from callus was 0.78 and 0.79 percent as characterized by HPLC and HPTLC respectively. Conclusion: The study revealed that callus culture may be a fruitful tool for the production of Podophyllotoxin resin, an anticancer entity.
Highlights
Podophyllum hexandrum Royle (Berberidaceae) known as the Indian podophyllum is a perennial herb, growing on the lower slopes of the Himalayas in scrub and forest from Afghanistan eastwards to central China[1,2,3,4]
In the present study, experiments were carried out on the tissue culture of Podophyllum hexandrum (Berberidaceae).Tissue culture study was started from aseptic seed germination, standardization of media, callus initiation and growth study followed by extraction, and estimation of Podophyllotoxin content in the callus cultures and from cultivated roots of the plant
HPLC Analysis The Qualitative tests for the identification of podophyllotoxin and the methods for their quantitative estimation were carried out by using HPLC
Summary
Podophyllum hexandrum Royle (Berberidaceae) known as the Indian podophyllum is a perennial herb, growing on the lower slopes of the Himalayas in scrub and forest from Afghanistan eastwards to central China[1,2,3,4]. Its insecticidal and phytotoxic activities are reported[7] These lignans are toxic for the treatment of neoplastic disease in humans. Podophyllotoxin is used as starting compound for the chemical synthesis of etoposide and teniposide; both being applied successfully as antitumor agents[8,9]. Their cytotoxic action is based on inhibition of topoisomerase II, while Podophyllotoxin acts as an inhibitor of the microtubule assembly. Podophyllotoxin content are prone to changes due to environment factors, type of fertilizers applied and stage of harvest These changes could be controlled by in-vitro culture of Podophyllum hexandrum for the synthesis of lignan Podophyllotoxin. This study demonstrates the potentiality of static culture in production of Podophyllotoxin
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