Plant Biotechnology

Electronic Journal of Biotechnology ISSN: 0717-3458 Vol. 8 No. 3, Issue of December 15, 2005
© 2005 by Pontificia Universidad Católica de Valparaíso -- Chile Received May 5, 2005 / Accepted June 23, 2005
DOI: 10.2225/vol8-issue3-fulltext-6
RESEARCH ARTICLE

Effects of elicitor and copper sulfate on grindelic acid production in submerged cultures of Grindelia pulchella

Xenia E. Hernandez
Area de Química Orgánica (INTEQUI-CONICET)
Facultad de Química, Bioquímica y Farmacia
Universidad Nacional de San Luis
Chacabuco y Pedernera, 5700
San Luis, Argentina
Tel: 54 2652439909
Fax: 54 2652426711

Alejandro A. Orden
Area de Química Orgánica (INTEQUI-CONICET)
Facultad de Química, Bioquímica y Farmacia
Universidad Nacional de San Luis
Chacabuco y Pedernera, 5700
San Luis, Argentina
Tel: 54 2652439909
Fax: 54 2652426711
E-mail:aaorden@unsl.edu.ar

Oscar S. Giordano
Area de Química Orgánica (INTEQUI-CONICET)
Facultad de Química, Bioquímica y Farmacia
Universidad Nacional de San Luis
Chacabuco y Pedernera, 5700
San Luis, Argentina
Tel: 54 2652439909
Fax: 54 2652426711
E-mail:ogiord@unsl.edu.ar

Marcela Kurina*
Area de Química Orgánica (INTEQUI-CONICET)
Facultad de Química, Bioquímica y Farmacia
Universidad Nacional de San Luis
Chacabuco y Pedernera, 5700
San Luis, Argentina
Tel: 54 2652439909
Fax: 54 2652426711
E-mail: mkurina@unsl.edu.ar

*Corresponding author

Financial support: UNSL: PROIPRO 0201; CONICET: PID 2429; FONCyT; ANPCyT: PICT2002 Nº 06-10710.

Keywords: Cu2+, dimethylsulfoxide permeabilization, grindelane diterpenes, jasmonic acid.

Abbreviations: 

BA: N6-benzylaminopurine
CDCl3: chloroform-d
2,4-D: 2,4-dichlorophenoxyacetic acid
DMSO: dimethylsulfoxide
DW: dry weight
Et2O: Ethyl ether
GC: gas chromatography
GC-MS: gas chromatography-mass espectrometry
IBA: indole-3-butyric acid
JA: jasmonic acid
K: kinetin
MS: Murashige and Skoog (1962) medium containing 3% (w/v) sucrose
NAA: 1-naphthalene acetic acid
NMR: nuclear magnetic resonance
TLC: thin layer chromatography

 
Abstract
Full Text

Grindelia pulchella callus and cell suspension cultures were established from seedling leaves. When several phytoregulator supplementations were assayed in solid Murashige and Skoog medium containing 3% (w/v) of sucrose (MS medium), combinations of indole-3-butyric acid (IBA) and N6-benzylaminopurine (BA) resulted the most appropriate conditions to generate fast growing friable calli with detectable levels of grindelic acid. Moreover, the same basal media supplemented with 20.0 µM IBA/4.4 µM BA was found to be optimal for cell growth in submerged cultures (µmax = 0.26 days-1) while the addition of 20.0 µM IBA/18.0 µM BA resulted in a relative higher metabolite production (4.55 mg/gDW) when the inocula was 5% (v/v). Furthermore, three different stress factors and combinations of them were used to elicit cell suspensions. These experiments demonstrated that the combination of CuSO4 and dimethylsulfoxide (DMSO) increase the grindelic acid production to 2.63 mg/gDW in the elicited essay versus 0.756 mg/gDW in the control, at expense of cell growth. In contrast, the addition of jasmonic acid (JA) alone and combined with DMSO neither affected cell growth nor grindelic acid accumulation.

 
Supported by UNESCO / MIRCEN network