Studies on Factors Affecting the Establishment of Gentiana davidii var. formosana (Hayata) T. N. Ho Cell Suspension Cultures

Abstract

Gentiana davidii var. formosana (Hayata) T. N. Ho (Gentianaceae), commonly known as long-dan in Chinese, is a perennial herb indigenous to Taiwan. It is distributed throughout the island, ranging from low to high elevations. The roots, which contain bitter-tasting secoiridoid glucosides, are used in traditional Chinese medicine. It is mainly used in the treatment of gastrointestinal tract diseases. Continuous collection of plant material from natural habitat has led to the depletion of Gentiana population. The purpose of this study was to establish the cell suspension cultures of Gentiana, which could be used for large-scale production of active principles such as gentiopicroside and swertiamarin. Callus was initiated by culturing stem explants of G. davidii var. formosana on Murashige and Skoog's (1962) basal medium supplemented with 0.2 mg/L 6-furfurylaminopurine (kinetin) and 1.0 mg/L a-naphthaleneacetic acid (NAA). Fast-growing suspension cell cultures were established by subculturing the callus in MS basal medium (pH 4.2-5.2) supplemented with 0.2 mg/L kinetin and 3% sucrose. The cultures were incubated on an orbital shaker (80-100 rev/min) at 25±1℃ and low light intensity (2.33 muE.m(-2).s(-1)).