Callus Production and Analysis of Isoflavonoids and Some Terpenes from Bituminaria bituminosa

Abstract

Background: Bituminaria bituminosa is a long-day plant. The plant has a high pastoral value and palatability. It is grazed by cattle, goats and sheep. Leaves and stems contain four furanocoumarins’ psoralen, angelicin, xanthotoxin and bergapten. The seed is covered by the fruit. Germination was not observed in beaked fruits. A cut with a scalpel can increase the germination rate. According to the literature there are not enough studies about the culture media and convenient conditions. Methods: In our study, we tried six medium for callus production with different hormone concentrations including 2,4-D, BAP and kinetin. In this study, a protocol for callus induction and analysis of isoflavonoids and terpenes of Bituminaria bituminosa has been developed. Result: Maximum callus induction frequency was observed on Murashige and Skoog (MS) medium supplemented with 0.75 mg/L 2,4-D + 2.5 mg/L kinetin in young primary leaves. 0.75 mg/L 2,4-D+2.5 mg/L kinetin medium is more successful in the production ofses of flavonoids that can be produced in vitro and is more successful in producing phenols. In terpene production, it is seen that 1.25 mg/L 2,4-D+3.5 mg/L BAP, which is the best callus producing medium containing BAP, is the most successful medium among all tested mediums.