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Stimulation of the proline cycle and anthraquinone accumulation in Rubia tinctorum cell suspension cultures in the presence of glutamate and two proline analogs

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Abstract

Anthraquinone biosynthesis in Rubia tinctorum L. involves different metabolic routes. Chorismic acid, the end-product of the shikimate pathway, becomes the branch point between primary and secondary metabolism. It has been proposed that the proline cycle could be coupled with the pentose phosphate pathway (PPP), since the NADP+ generated by proline reduction from glutamate could act as a cofactor of the first enzymes of the PPP. This pathway generates erythrose-4-phosphate, the substrate of the shikimate pathway. The aim of the present work was to study the effect of the addition of glutamate and two proline analogs, azetidine-2-carboxylic acid and thiazolidine-4-carboxylic acid (T4C), on the PPP, the proline cycle, and anthraquinone production in R. tinctorum cell suspension cultures. The addition of 5 mM of glutamate enhanced both anthraquinone (up to 30%) and total phenolic content (12%), which correlated well with proline accumulation. Only the addition of 200 μM of T4C resulted in an increase in anthraquinone production, which was accompanied by a rise in the proline content. Neither the addition of glutamate nor proline analogs resulted in the induction of PPP, so this route was not a limiting factor as a carbon donor to the shikimate pathway.

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Acknowledgments

This work was supported by grants PICT 14-15112 and 14-33166, Agencia Nacional de Promoción Científica y Tecnológica (ANPCyT), Argentina, and by grant UBACyT B111, Universidad de Buenos Aires. J.R.T. and A.M.G. are researchers from Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Argentina. M.P. and C.V.Q. are fellows from CONICET.

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  1. Cátedra de Microbiología Industrial y Biotecnología, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Junín 956, 1113, Ciudad Autónoma de Buenos Aires, Argentina

    María Perassolo, Carla Verónica Quevedo, Ana María Giulietti & Julián Rodríguez Talou

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  1. María Perassolo
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  2. Carla Verónica Quevedo
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  3. Ana María Giulietti
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  4. Julián Rodríguez Talou
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Correspondence to Julián Rodríguez Talou.

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Perassolo, M., Quevedo, C.V., Giulietti, A.M. et al. Stimulation of the proline cycle and anthraquinone accumulation in Rubia tinctorum cell suspension cultures in the presence of glutamate and two proline analogs. Plant Cell Tiss Organ Cult 106, 153–159 (2011). https://doi.org/10.1007/s11240-010-9903-5

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