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Are polyamines directly involved in silymarin production in the milk thistle [Silybum marianum (L.) Gaernt]?

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Abstract

The possible role of polyamines (PAs) in the regulation of silymarin (Sm) production in milk thistle [Silybum marianum (L.) Gaernt] cell suspension cultures was studied in a young cell culture line (H2 line) and in a synchronized cell line (>3 years; H1 line). The effect of two exogenous PAs, putrescine (Put) and spermidine (Spd), and a number of metabolic inhibitors (L-canavanine, DL-α-difluoromethylornithine, methylglyoxal-bis-guanylhydrazone, cyclohexylamine) on the production of Sm during the growth cycle were analyzed. The results suggest that PAs are not directly involved in the Sm synthesis pathway. In our cell culture system, Sm production and PA contents were determined by the age of the suspension culture cells: with increasing age, the suspension culture cells showed a decreasing capacity to reach the stationary phase during prolonged subculture that was associated with a decreased production of Sm, a steady increase in PA content, and a constant Put/Spd ratio. The synchronization of dividing cells from the S. marianum H1 line did not modify this behaviour. In young cell suspensions, maximum Sm production occurred in the stationary phase, concurrent with the cellular PA contents reaching their minimum value. At the start of the stationary phase, the high percentage of cells in the growth phases (G0/G1) and a transient increase in the Put/Spd ratio were accompanied by maximum Sm production and a blockade of cell division.

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Abbreviations

CHA:

Cyclohexylamine

DFMA:

DL-α-Difluoromethylarginine

DFMO:

DL-α-Difluoromethylornithine

DHP:

1-7 Diaminoheptane

LC:

L-Canavanine

MGBG:

Methylglyoxal-bis-guanylhydrazone

PAs:

Polyamines

PCA:

Perchloric acid

PH:

Fraction pellet-hydrolysed

Put:

Putrescine

S:

Fraction soluble

SH:

Fraction soluble-hydrolysed

Sm:

Silymarin

Spd:

Spermidine

Spm:

Spermine

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Acknowledgements

This work was supported by the Spanish grant of the Ministerio de Ciencia y Tecnología (MCYT) Plan Nacional del I + D+I and FEDER funds. (Ref: BF12000-1362). We are very grateful to Patrick M. Woster, PhD (Wayne State University, USA) for his kind gift of the DFMO product.

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  1. Departamento de Fisiología Vegetal, Universidad de Salamanca, Plaza de los Doctores de la Reina s/n, 37007, Salamanca, Spain

    Margarita Cacho, Alexis Torres Domínguez & Juana-A. Elena-Rosselló

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  1. Margarita Cacho
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  2. Alexis Torres Domínguez
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  3. Juana-A. Elena-Rosselló
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Correspondence to Margarita Cacho.

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Cacho, M., Torres Domínguez, A. & Elena-Rosselló, JA. Are polyamines directly involved in silymarin production in the milk thistle [Silybum marianum (L.) Gaernt]?. Plant Cell Tiss Organ Cult 103, 361–368 (2010). https://doi.org/10.1007/s11240-010-9789-2

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