Cardillo Alejandra Beatriz, Rodriguez Talou Julián, Giulietti Ana María
Cátedra de Biotecnología-Instituto Nanobiotec (UBA/Conicet), Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Junin 954 Ciudad, Autonoma de Buenos Aires, Buenos Aires, Argentina.
Methods Mol Biol. 2016;1391:173-86. doi: 10.1007/978-1-4939-3332-7_12.
Brugmansia candida (syn. Datura candida) is a South American native plant that produces tropane alkaloids. Hyoscyamine, 6β-hydroxyhyoscyamine (anisodamine), and scopolamine are the most important ones due to their anticholinergic activity. These bioactive compounds have been historically and widely applied in medicine and their demand is continuous. Their chemical synthesis is costly and complex, and thereby, these alkaloids are industrially produced from natural producer plants. The production of these secondary metabolites by plant in vitro cultures such as hairy roots presents certain advantages over the natural source and chemical synthesis. It is well known that hairy roots produced by Agrobacterium rhizogenes infection are fast-growing cultures, genetically stable and able to grow in hormone-free media. Additionally, recent progress achieved in the scaling up of hairy root cultures makes this technology an attractive tool for industrial processes. This chapter is focused on the methods for the induction and establishment of B. candida hairy roots. In addition, the scaling up of hairy root cultures in bioreactors and tropane alkaloid analysis is discussed.
白花曼陀罗(同义词:白曼陀罗)是一种原产于南美洲的植物,可产生托烷生物碱。由于其抗胆碱活性,莨菪碱、6β-羟基莨菪碱(山莨菪碱)和东莨菪碱是其中最重要的生物碱。这些生物活性化合物在历史上已被广泛应用于医学领域,并且其需求持续存在。它们的化学合成成本高昂且复杂,因此,这些生物碱是通过天然生产植物进行工业化生产的。通过植物体外培养(如毛状根)生产这些次生代谢产物相对于天然来源和化学合成具有一定优势。众所周知,由发根农杆菌感染产生的毛状根生长迅速、遗传稳定且能够在无激素培养基中生长。此外,毛状根培养放大技术的最新进展使该技术成为工业生产过程的一个有吸引力的工具。本章重点介绍白花曼陀罗毛状根的诱导和建立方法。此外,还讨论了生物反应器中毛状根培养的放大以及托烷生物碱分析。