283 a Fig.1.a)Hairy roots cultured on hormone-free RC solid medium containing 3%sucrose.b)Hairy roots cultured in Determination of shikonin.Culture media,adsorbents and lyophilized hormone-free RC liquid medium and shikonin secretion. hairy roots were extracted with CHCl3.After drying over Na2S04 the c)Microscopic observation of hairy roots and shikonin extract was evaporated to dryness and the shikonin content was secretion.d)Shikonin adsorption on a XAD-2 column. determined by its UV absorption at 520 nm (Fujita et al.1983). light.Two-3 weeks after the inoculation,hairy roots (ca 5/ shoot)appeared at the infected sites.They were cut off and Results and discussion subcultured on MS solid medium containing carbenicillin. Within one week the hairy roots formed calli on the Transformation antibiotic medium.After the elimination of the bacteria the Axenic shoots of L.erythrorhizon were cultured for 3 weeks white calli were transferred to hormone-free MS solid at low light intensity (100 lux).The etiolated shoots were medium,where the formation of hairy roots with red spots infected directly with A.rhizogenes 15834 and kept at low in the older parts of the tissue was observed.However,283 Determination ofshikonin. Culture media, adsorbents and lyophilized hairy roots were extracted with CHCI3. After drying over Na2SO4 the extract was evaporated to dryness and the shikonin content was determined by its UV absorption at 520 nm (Fujita et al. 1983). Results and discussion Transformation Axenic shoots of L. erythrorhizon were cultured for 3 weeks at low light intensity (100 lux). The etiolated shoots were infected directly with A. rhizogenes 15834 and kept at low Fig. 1. a) Hairy roots cultured on hormone-free RC solid medium containing 3% sucrose, b) Hairy roots cultured in hormone-free RC liquid medium and shikonin secretion. c) Microscopic observation of hairy roots and shikonin secretion, d) Shikonin adsorption on a XAD-2 column. light. Two-3 weeks after the inoculation, hairy roots (ca 5 / shoot) appeared at the infected sites. They were cut off and subcultured on MS solid medium containing carbeniciUin. Within one week the hairy roots formed calli on the antibiotic medium. After the elimination of the bacteria the white calli were transferred to hormone-free MS solid medium, where the formation of hairy roots with red spots in the older parts of the tissue was observed. However