Introduction
Abiotic elicitors affect both, biomass and secondary metabolite production in cell suspension cultures. In this work we have studied CuSO4 effect on the accumulation of grindelic acid in Grindelia pulchella cell suspension cultures.
Material and Methods
Cell suspension cultures
MS media supplemented with indolbutiric acid and bencilamine purine was employed. Samples of 20 ml were taken, after filtration biomass dry weight was evaluated.
Grindelic acid production evaluation
Liquid media was acidified to pH 5.00 with HCl 10% and submitted to liquid-liquid extraction procedure with Et2O (x3). Cells were extracted by reflux in MeOH 4h (x3). The methanolic extract was dried, recovered with distilled acidified water (pH 5.00) and extracted with Et2O (x3). Samples were methylated with CH3N2 and evaluated by GC. Each assay was repeated three times.
Elicitation with CuSO4
CuSO4 (final concentration 1 y 2 mM sterilized by filtration) was added at day 7.
Results and Discussion
Cu SO4 addition in both concentrations inhibited the biomass production. This effect may be attributed to the fact that heavy metals (Cu or Cd) cause an increment in the catabolic activity or suppress the lipid biosynthesis.
Grindelic acid accumulation, in cultures elicited with CuSO4 (1mM), was completely inhibited. On the other hand the addition of CuSO4 (2mM) induced a grindelic acid accumulation dismissing at the early times of the cell cycle but increased the production in the stationary phase.
Figure 1.
CuSO4 effect on biomass production [Biomass (g dry weigth/ml)].
Figure 2.
CuSO4 effect on grindelic acid accumulation [Production (mg/ g cell)].
References and Notes
- García, M.; Hernández, X.E.; Sosa, M.; Tonn, C.E. Jornadas de la Sociedad de Biología de Córdoba; 1999; 28 de junio al 3 de julio; Córdoba, Argentina. [Google Scholar]
- Hernández, X.E.; Kurina Sanz, M.B.; Giordano, O.S. Biotechnol. Lett. 1997, 19(12), 1223.
- Hernández, X.E.; Kurina Sanz, M.B.; Giordano, O.S. Congreso Nacional de Biotecnología: Talca. Chile, Setiembre 1998.
- Ovariti, O; Boussama, N.; Zarrouk, M.; Cherif, A.; Ghorbal, M.H. Phytochemistry 1997, 45(7), 1343.
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