Abstract
To produce novel bioactive flavonoids, Gericudranin A and Gericudranin B, a cell culture system of Cudrania tricuspidata including callus induction and optimization of culture conditions was established. Friable calli were efficiently induced from the hypocotyl segments of seedlings on B5 medium supplemented with 1.0 mg/L NAA, 0.1 mg/L kinetin and 3% sucrose. Several factors were optimized for the Gericudranin production and the cell growth in suspension cultures. Low level of basal salt medium (1/8 MS), 1.0 mg/L IAA and 0.1 mg/L zeatin, and high level of sucrose (5%) were effective for the production of Gericudranins, whereas WPM with 1.0 mg/L NAA, 0.1 mg/L zeatin, and 5% sucrose were more effective for the cell growth. When cells were cultured on MS liquid medium supplemented with 1.0 mg/L IBA, about 2200 $\mu\textrm{g}$/g dry wt of Gericudranin A could be produced. The level might be about 10 times of the native inner bark. About 2350 $\mu\textrm{g}$/g dry wt of Gericudranin B was also produced on MS liquid medium with 5% sucrose, 1.0 mg/L NAA, 0.1 mg/L kinetin. The content was estimated about 3 times of the level of native inner bark.
꾸지뽕나무 (Cudrania tricuspidata)에 미량으로 함유되어 있는 신규 생리활성물질인 Gericudranin를 기내 배양세포에서 효율적으로 생산하는 시스템을 확립하기 위하여 캘러스 유도 및 여러 가지 배양조건 규명에 관한 실험을 수행하였다. 캘러스는 1.0 mg/L NAA, 0.1 mg/L kinetin, 3% sucrose가 함유된 B5 배지에 배양된 배축에서 효율적으로 유도되었다. Gericudranin 생산과 세포생장에 관여하는 여러 가지 요인을 조사한 결과, 세포생장을 위해서는 1.0mg/L NAA, 0.1 mg/L zeatin과 5% sucrose가 함유된 WPM배지가 좋았고, Gericudranin 생산을 위해서는 무기염 농도가 1/8로 희석된 MS배지, 1.0 mg/l NAA, 0.1 mg/l zeatin과 5% sucrose 처리구가가장 좋았다 Gericudranin A의 함량 (2200 $\mu\textrm{g}$/g dry wt)은 3% sucrose, 1.0 mg/L IBA가 함유된 1/2MS배지에서 모식물체의 수피의 함량보다 약 10배 높았으며, Gericudranin B의 함량 (2350 $\mu\textrm{g}$/g dry wt)은 sucrose 5%를 함유한 1/2 MS 배지에서 수피의 함량보다 약 3배 높았다.
