Production Stability of Active Polysaccharides of Dendrobium huoshanense using Long-Term Cultures of Protocorm-Like Bodies

Abstract

In this study, the production stability of active polysaccharides in protocorm-like bodies (PLBs) induced from the seedling segments of Dendrobium huoshanense C. Z. Tang et S. J. Cheng was investigated during long-term subculture. Subcultures were conducted once every 30 days. With an average inoculum of 39 g/L fresh PLBs, the increase in biomass ranged from 95.7 g/L to 103.9 g/L in fresh weight and 3.2 g/L to 3.4 g/L in dry weight during eighteen continuous subcultures while polysaccharide content in PLBs was from 0.8 mg/g Fw (mg polysaccharide per gram PLBs in fresh weight) to 1.0 mg/g Fw. In addition, polysaccharides from all cultures showed a similar potential of stimulating interferon γ (IFN-γ) release in the supernatant of splenocytes and tumor necrosis factor alpha (TNF-α) release in the supernatant of peritoneal macrophages. To elucidate the genetic basis of polysaccharide production stability in long-term subculture of PLBs, the genetic fingerprints by RAPD were further analyzed using plantlets from PLB development. Results showed that there is no evidence of genetic variation both within the plantlets from the different subcultures of PLBs and between long-term subcultures and the donor plants.

Abbreviations

ANOVA:one-way analysis of variance

HPLC:high performance liquid chromatography

IFN-γ:interferon γ

KT:kinetin

MS:Murashige and Skoog (1962) medium

NAA:α-naphthaleneacetic acid

PLBs:protocorm-like bodies

RAPD:random amplified polymorphic DNA

TFA:trifluoroacetic acid

TNF-α:tumor necrosis factor alpha

Key words

Dendrobium huoshanense - Orchidaceae - PLBs - polysaccharide - RAPD

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