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Podophyllotoxin is isolated mainly from the rhizomes of Podophyllumplants, and serves as the main precursor for synthesis of anticancer drugs, such as VP-16 and VM-26. VP-16 and VM-26 are used for curing lung cancer, testicular cancer, neuroblastoma, hepatoma and other tumors. However, these plants are all near-extinction species due to over-collection and their own biological characteristics. The chemical synthesis of podophyllotoxin is so complicated that its price is unbelievably high. This paper discusses the current status of the biosynthetic pathway of podophyllotoxin and that of the podophyllotoxin production using several biotechnological approaches such as plant organ cultures, plant cell cultures with both flasks and bioreactors, hairy root cultures, bioconversions and metabolic regulations.