AGROBACTERIUM RHIZOGENES MEDIATED HAIRY ROOT CULTURE AND GENETIC TRANSFORMATION OF AN ENDANGERED MEDICINAL PLANT OF DECALEPIS HAMILTONII WIGHT & ARN.

摘要

The total dry matter content of 2-hydroxy-4-methoxy benzaldehyde production by hairy root culture of Decalepis hamiltonii from in vitro internode and roots explants inoculated with Agrobacterium rhizogenes. The addition of acetosyringone containing basal Murashige and Skoog’s (MS) liquid medium fortified with α-naphthaleneacetic acid (NAA) and indole 3-butyric acid (IBA) are improved hairy root production. PCR analysis of Ri plasmid on transformed internode and root tissues showed DNA amplification at 780 bp amplified in internode, root derived hairy roots. The highest biomass of dry matter content (25.36 and 24.12 g) was observed on liquid MS medium containing 0.3 mg/L NAA, co-cultivated with A. rhizogenes on the internode and root explants. After that the internode and root explants were inoculated on MS medium supplemented with 0.3 mg/L IBA was improved biomass content of dry matter 23.56 and 22.91 g respectively. The dry matter content of biomass was extracted with 1 mg/ml methanol to measure the total content accumulation of 2-hydroxy-4-methoxy benzaldehyde in hairy roots cultures confirmed by using HPLC.