Plant Cell, Tissue and Organ Culture

, Volume 93, Issue 1, pp 73–83 | Cite as

In vitro production of metabolism-enhancing phytoecdysteroids from Ajuga turkestanica

  • Diana M. Cheng
  • Gad G. Yousef
  • Mary H. Grace
  • Randy B. Rogers
  • J. Gorelick-Feldman
  • I. Raskin
  • Mary Ann Lila
Original Paper

Abstract

In order to develop a sustainable source of metabolism-enhancing phytoecdysteroids, cell suspension and hairy root cultures were established from shoot cultures of wild-harvested Ajuga turkestanica, a medicinal plant indigenous to Uzbekistan. Precursors of phytoecdysteroids (acetate, mevalonic acid cholesterol) or methyl jasmonate (an elicitor) were added to subculture media to increase phytoecdysteroid accumulation. In cell suspension cultures, 20-hydroxyecdysone (20E) content increased 3- or 2-fold with the addition of 125 or 250 μM methyl jasmonate, respectively, compared to unelicited cultures. Precursor addition, however, did not provoke phytoecdysteroid accumulation. In hairy root cultures, addition of sodium acetate, mevalonic acid, and methyl jasmonate, but not cholesterol, increased phytoecdysteroid content compared to unelicited cultures. Hairy root cultures treated with 150 mg l−1 sodium acetate, or 15 or 150 mg l−1 mevalonic acid, increased 20E content approximately 2-fold to 19.9, 20.4 or 21.7 μg mg−1, respectively, compared to control (10.5 μg mg−1). Older hairy root cultures, extracted after the seventh subculture cycle, also showed increases in 20E content (24.8 μg mg−1), turkesterone (0.9 μg mg−1) and cyasterone (8.1 μg mg−1) compared to control cultures maintained for a shorter duration of four subculture cycles. Doses of 10 or 20 μg ml−1 hairy root extract increased protein synthesis by 25.7% or 31.1%, respectively, in a C2C12 mouse skeletal cell line. These results suggest that sustainable production of metabolically active phytoecdysteroid can be achieved through hairy root culture systems.

Keywords

20-Hydroxyecdysone Cell suspension culture Cyasterone Hairy root Methyl jasmonate Mevalonic acid Turkesterone 

Abbreviations

20E

20-Hydroxyecdysone

CC

Column chromatography

CD3OD

Deuterated methanol

DAD

Diode array detector

DMEM

Dulbecco’s modified eagle’s media

DPM

Decays per minute

DW

Dry weight

ESI-MS

Electrospray ionization-mass spectroscopy

FW

Fresh weight

HPLC

High performance liquid chromatography

NMR

Nuclear magnetic resonance

TLC

Thin-layer chromatography

TMS

Tetramethylsilane

Supplementary material

11240_2008_9345_MOESM1_ESM.doc (306 kb)
(DOC 306 kb)

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Copyright information

© Springer Science+Business Media B.V. 2008

Authors and Affiliations

  • Diana M. Cheng
    • 1
  • Gad G. Yousef
    • 1
  • Mary H. Grace
    • 1
  • Randy B. Rogers
    • 1
  • J. Gorelick-Feldman
    • 2
  • I. Raskin
    • 2
  • Mary Ann Lila
    • 1
  1. 1.Department of Natural Resources and Environmental SciencesUniversity of IllinoisUrbanaUSA
  2. 2.Biotech Center, Cook CollegeRutgers UniversityNew BrunswickUSA

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