ORIGINAL PAPER Fungal elicitor-mediated enhancement in phenylpropanoid and naphtodianthrone contents of Hypericum perforatum L. cell cultures Sonja Gadzovska Simic 1,2 • Oliver Tusevski 2 • Ste ´phane Maury 1 • Christophe Hano 3 • Alain Delaunay 1 • Brigitte Chabbert 4 • Fre ´de ´ric Lamblin 3 • Eric Laine ´ 3 • Claude Joseph 1 • Daniel Hage `ge 3 Received: 24 December 2014 / Accepted: 28 March 2015 / Published online: 4 April 2015 Ó Springer Science+Business Media Dordrecht 2015 Abstract Hypericum perforatum cell suspensions were evaluated for their growth, phenylpropanoid and naphto- dianthrone productions, and antioxidant activity after treatments with fungal elicitors Fusarium oxysporum, Phoma exigua and Botrytis cinerea. Elicited cells displayed a reduced biomass production, a rapid stimulation of sec- ondary metabolites production and a modification of cell redox state compared to control. Cells responded strongly towards the applied elicitors through the enhanced pro- duction of naphtodianthrones. Hypericin and pseudohy- pericin production was significantly increased (up to fourfold) in the early growth phase and remained stable all along the post-elicitation period. Significant increase in contents of total phenolics, total flavonoids and total an- thocyanins was observed during the entire period of culti- vation, while total flavanols were enhanced at the end of post-elicitation. The enzymatic activities of phenylalanine ammonia lyase and chalcone isomerase were remarkably elevated in elicited cells confirming a strong activation of phenylpropanoid/flavonoid pathways. The fingerprint pro- file of Fourier transform infrared spectroscopy spectra from the cell walls showed a little variation in lignin accumu- lation between elicited and control samples. With regards to the antioxidant state, an early up-regulation of per- oxidase activity was observed in elicited cells, whereas non-enzymatic properties and catalase activity were en- hanced at the end of post-elicitation. These findings suggest the involvement of an efficient antioxidant defense system in the adaptive response of cells to fungal elicitation. Al- together, these results indicated that H. perforatum elicited cells represent a promising experimental system for scale- up production of naphtodianthrones for medicinal uses. Keywords Antioxidant activity Á Cell suspensions Á Fungal elicitors Á Hypericum perforatum L. Á Naphtodianthrones Á Phenylpropanoids Abbreviations CAT Catalase CHI Chalcone isomerase FTIR Fourier transform infrared spectroscopy HYP Hypericin JA Jasmonic acid NEAOP Non-enzymatic antioxidant properties PAL Phenylalanine ammonia lyase PHYP Pseudohypericin POD Peroxidase ROS Reactive oxygen species SA Salicylic acid TA Total anthocyanins TFL Total flavanols TF Total flavonoids TP Total phenolics & Sonja Gadzovska Simic sonjag@pmf.ukim.mk 1 Laboratoire de Biologie des Ligneux et des Grandes Cultures (LBLGC) INRA USC1328, ITP SBCV, UPRES EA 1207, University of Orle ´ans, rue de Chartres, BP6759, 45067 Orle ´ans Cedex 2, France 2 Laboratory for Plant Cell and Tissue Culture, Faculty of Natural Sciences and Mathematics, Institute of Biology, University ‘‘Ss. Cyril and Methodius’’, Archimedova str. 3, 1000 Skopje, Macedonia 3 Laboratory of Wood and Crop Biology, Antenne Scientifique Universitaire de Chartres (ASUC), Universite ´ d’Orle ´ans, 21 rue de Loigny la Bataille, 28000 Chartres, France 4 INRA Parois Ve ´ge ´tales et Mate ´riaux Fibreux UMR FARE, 2 esplanade Roland Garros, BP224, 51686 Reims, France 123 Plant Cell Tiss Organ Cult (2015) 122:213–226 DOI 10.1007/s11240-015-0762-y