J Integr Plant Biol. ›› 2008, Vol. 50 ›› Issue (8): 975-981.DOI: 10.1111/j.1744-7909.2008.00680.x

• Plant-environmental Interactions • Previous Articles     Next Articles

Establishment of Withania somnifera Hairy Root Cultures for the Production of Withanolide A

Hosakatte N. Murthy, Camelia Dijkstra, Paul Anthony, Daniel A. White, Mike R. Davey, J. Brian Power, Eun J. Hahn, Kee Y. Paek   

  • Received:2007-06-06 Accepted:2007-12-18

Abstract: Withania sominifera (Indian ginseng) was transformed by Agrobacterium rhizogenes. Explants from seedling roots, stems, hypocotyls, cotyledonary nodal segments, cotyledons and young leaves were inoculated with A. rhizogenes strain R1601. Hairy (transformed) roots were induced from cotyledons and leaf explants. The transgenic status of hairy roots was confirmed by polymerase chain reaction using nptII and rolB specific primers and, subsequently, by Southern analysis for the presence of nptII and rolB genes in the genomes of transformed roots. Four clones of hairy roots were established; these differed in their morphology. The doubling time of faster growing cultures was 8–14 d with a fivefold increase in biomass after 28 d compared with cultured, non-transformed seedling roots. MS-based liquid medium was superior for the growth of transformed roots compared with other culture media evaluated (SH, LS and N6), with MS-based medium supplemented with 40 g/L sucrose being optimal for biomass production. Cultured hairy roots synthesized withanolide A, a steroidal lactone of medicinal and therapeutic value. The concentration of withanolide A in transformed roots (157.4 μg/g dry weight) was 2.7-fold more than in non-transformed cultured roots (57.9 μg/g dry weight).

Key words: Agrobacterium rhizogenes, hairy (transformed) roots, Indian ginseng, transformation, withanolide A

Editorial Office, Journal of Integrative Plant Biology, Institute of Botany, CAS
No. 20 Nanxincun, Xiangshan, Beijing 100093, China
Tel: +86 10 6283 6133 Fax: +86 10 8259 2636 E-mail: jipb@ibcas.ac.cn
Copyright © 2022 by the Institute of Botany, the Chinese Academy of Sciences
Online ISSN: 1744-7909 Print ISSN: 1672-9072 CN: 11-5067/Q
备案号:京ICP备16067583号-22