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Molecular cloning of SoHPPR encoding a hydroxyphenylpyruvate reductase, and its expression in cell suspension cultures of Salvia officinalis

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Abstract

Rosmarinic acid (RA) is a naturally-derived compound present in medicinal plants, herbs and spices, and is well-known for its antioxidant, antibacterial and medicinal properties. It has been recently considered for use as a food preservative, nutraceutical, and for medicinal purposes. Undifferentiated cells (callus and cell suspensions) cultured in vitro are often used for the production of secondary metabolites, and for identifying biosynthetic pathways. In this work Salvia officinalis L. (common sage) cell suspension cultures were established, and a specific cell line was selected for the high antioxidant capacity of its methanolic extract, which was characterized by a high content of RA. Scavenger activity (DPPH test) and total RA content were evaluated during cell growth. Gene coding for Hydroxyphenylpyruvate reductase, the key-enzyme responsible for the RA metabolic biosynthesis, was cloned from common sage (SoHPPR). Its transcript expression level was monitored during cell suspension cultures, and showed a relationship with scavenger activity and RA yield. Our results suggest the potential use of this gene as a marker and target for the modulation of RA production in controlled conditions.

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Acknowledgments

This research was supported by the European project RD NUTRASNACK (EC FP6 contract No FOOD-CT-2005-023044), European Project INTERREG-ALCOTRA “AROMA” n°68,and by EU Project “PROFICIENCY” FP7- REGPOT-2009-1-245751.

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Authors and Affiliations

  1. Department of Agriculture, Food and Environment, University of Pisa, Via del Borghetto 80, 56124, Pisa, Italy

    S. Barberini, D. Raffi & La. Pistelli

  2. CRA-FSO Ornamental Plant Research Unit, CRA Consiglio per la Ricerca e la Sperimentazione in Agricoltura, Corso degli Inglesi 508, 18038, Sanremo, IM, Italy

    S. Barberini, M. Savona, D. Raffi & B. Ruffoni

  3. Department of Pharmacy, University of Pisa, Via Bonanno 33, 56126, Pisa, Italy

    M. Leonardi & Lu. Pistelli

  4. Institute of Soil Science and Plant Cultivation, State Research Institute, ul. Czartoryskich 8, 24-100, Pulawy, Poland

    A. Stochmal

  5. Institute of Plant Sciences and Genetics in Agriculture, The Hebrew University of Jerusalem, P.O. Box 12, 76100, Rehovot, Israel

    A. Vainstein

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  1. S. Barberini
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  2. M. Savona
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Corresponding author

Correspondence to B. Ruffoni.

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11240_2013_300_MOESM1_ESM.pdf

Supplementary Fig. 1 Multiple alignment of HPPR nucleotidic sequence in four Lamiaceae species performed in ClustalW: Perilla frutescens (GenBank accession number HM587131), Coleus blumei (GenBank accession number AJ507733), Salvia miltiorrhiza (GenBank accession number DQ266514) and Salvia officinalis (GenBank accession number JX566894). The regions in which primers were designed are underlined. (PDF 13 kb)

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Barberini, S., Savona, M., Raffi, D. et al. Molecular cloning of SoHPPR encoding a hydroxyphenylpyruvate reductase, and its expression in cell suspension cultures of Salvia officinalis . Plant Cell Tiss Organ Cult 114, 131–138 (2013). https://doi.org/10.1007/s11240-013-0300-8

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