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Establishment of Withania somnifera Hairy Root Cultures for the Production of Withanolide A
Author(s) -
Murthy Hosakatte N.,
Dijkstra Camelia,
Anthony Paul,
White Daniel A.,
Davey Mike R.,
Power J. Brian,
Hahn Eun J.,
Paek Kee Y.
Publication year - 2008
Publication title -
journal of integrative plant biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.734
H-Index - 83
eISSN - 1744-7909
pISSN - 1672-9072
DOI - 10.1111/j.1744-7909.2008.00680.x
Subject(s) - withanolide , withania somnifera , hypocotyl , agrobacterium , biology , seedling , explant culture , hairy root culture , inoculation , botany , murashige and skoog medium , dry weight , horticulture , transformation (genetics) , in vitro , gene , biochemistry , medicine , alternative medicine , pathology
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 npt II and ro lB specific primers and, subsequently, by Southern analysis for the presence of npt II and rol B 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).