The Amaranthaceae family comprises of many species, which are used in nutrition and in traditional folk medicine for the treatment of several diseases such as infections, inflammation and fever. Gomphrena, Pfaffia and Alternanthera species are used in the extraction of natural pigments such as betaxanthin and betalains for application as food colorants and antioxidants. Pfaffia paniculata (Brazilian ginseng or Suma) has been indicated as a tonic, as well as having aphrodisiac, analgesic, and antidiabetic properties and may act against cancer. Gomphreneae is the major tribe of Amaranthaceae and previous chemical analyses have demonstrated the occurrence of anthraquinones, aurone, betacyanins, betaxanthins, betalains, chromoalkaloids, ecdysteroids, flavonoids, protoalkaloids, saponins, steroids and triterpenes. Biotechnological investigation with Amaranthaceae plants from the Gomphreneae tribe, demonstrated their potential for bioprospection of bioactive natural compounds such as flavonoids, steroids, terpenoides and saponins. Plant cell cultures, nowadays, are an important strategy for bioprospection of natural products. The in vitro large-scale production of bioactive compounds or extracts used as phytotherapics, pharmaceutical products, food additives and cosmetics should be encouraged because of their scientific, economical or ecological importance. Therefore, the present chapter reviews the literature data of the bioactive chemical constituents and biotechnological production of secondary metabolites in Amaranthaceae plants (Gomphreneae tribe), species that have many pharmacological properties and other applications. © 2012 Bentham Science Publishers. 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