PRODUCTION OF PHYTOECDYSTEROIDS BY HAIRY ROOT CULTURES OF AJUGA-REPTANS VAR ATROPURPUREA

Over 20 'hairy root' clones of Ajuga reptans var. atropurpurea (Labiatae) were obtained from transformation with Agrobacterium rhizogenes MAFF 03-01724. Growth opine synthesis and production of phytoecdysteroids in the clones have been examined. Mikimopine, a specific opine synthesized in the tissue transformed with strain MAFF 03-01724, was detected in most of the clones, and four phytoecdysteroids, 20-hydroxyecdysone, norcyasterone B, cyasterone and isocyasterone, were detected in all of the clones. It was found that the production of phytoecdysteroids was closely related to the growth of hairy roots, by monitoring the changes in the culture. One of the rapidly growing and well branching hairy root lines, Ar-4, was selected and used for fermenter experiments. On an air-lift type culture of Ar-4 for 45 days, the weight of the root tissue was increased by 230 times and the content of 20-hydroxyecdysone reached 0.12% on a dry weight basis. This content is 4 times higher than those of the original roots.