Brassinosteroid substances in immature Cassia tora seeds

결명자의 brassinosteroid 활성물질

In order to explore the brassinosteroid-active component in Cassia tora, methanol extract of immature seeds was purified by sequences of solvent fractionation, silica gel adsorption chromatography, Sephadex LH-20 chromatography, charcoal adsorption chromatography and Bondesil chromatography. The activity of brassinosteroid was monitored by the rice inclination test and its presence could be confirmed in each purification step. The purified active components were separated by silica gel adsorption chromatography. Brassinosteroid substances in separated active fractions were identified as teasterone, castasterone, brassinolide by TLC and HPLC. Our work is probably the first report of endogenous brassinosteroid in Cassia tora. The content of brassinosteroid in Cassia tora as converted into brassinolide was $3.5{\sim}5.5\;ng/g$ fresh weight. The order of brassinosteroid contents was toward to be teasterone, castasterone, brassinolide.

결명자에 존재하는 brassinosteroid 활성물질을 탐색하기 위하여 미숙종자를 MeOH로 추출하고, 이 추출물을 rice inclination test를 지표로, 용매분획, silica gel adsorption chromatography, Sephadex LH-20 chromatography, charcoal adsorption chromatography, Bondesil chromatography 등의 수법으로 brassinosteroid 활성물질을 정제한 다음, silica gel 흡착 chromatography로 활성성분을 분리하고, 각 활성획분에 대해 TLC 및 $C_{18}$ column과 ODS column을 사용한 HPLC에 의해 활성본체를 구명한 결과, 결명자의 brassinosteroid 활성물질로 최초로 teasterone, castasterone, brassinolide 가 동정되었다. 결명자에 함유된 brassinosteroid의 함량은 brassinolide로 환산하여 생체중량g당 $3.5{\sim}5.5\;ng$ 정도였으며, 동정된 3종의 brassinosteroid는teasterone, castasterone, brassinolide순으로 존재하였다.