Full Length Research Paper
Abstract
Artemisinin production from plant tissue cultures and induction of hairy roots in vitro have been considered to be a promising alternative, which offer a high degree of genetic stability, grow rapidly and produce the higher spectrum of secondary metabolites than wild type plants. Hairy root cultures developed from infection of different explants of in vitro germinated Artemisia annua L. plant with Agrobacterium rhizogenes LBA 9402 strain were selected on the basis of high artemisinin content and growth. Integration of the TL-DNA (rol gene) region of the pRi plasmid was confirmed by polymerase chain reaction (PCR) analysis of the gene located in this region. The effect of different environmental factors like temperature, pH, cultivation media and carbon source on growth and artemisinin production were studied in shake flask cultures. Detailed batch growth and production kinetics with sugar consumption profile were also established. Maximum volumetric productivity of 390 µg L-1 day-1 was obtained in hairy root cultures.
Key words: Agrobacterium, Artemisia annua L., artemisinin, hairy root cultures.
Abbreviation
ANOVA, Analysis of variance; CCD, central composite design; CTAB, cetyltrimethylammonium bromide; DCW, dry cell weight; DW, dry weight; FW, fresh weight; HPLC, High performance liquid chromatography; MS, Murashige and Skoog medium; PCR, polymerase chain
reaction; RSM, Response surface Methodology; YMB, Yeast mannitol broth.
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