Accumulation of phenolic compounds by in vitro cultures of Spanish Sage (Salvia lavandulifolia L.)

Spanish sage (S. lavandulifolia L.) grows wild in Spain and Southwest France and is used in perfumes and food flavouring [1]. Several reports indicate that extracts of this species have anticholinesterase, antioxidant and anti-inflammatory effects [2].

Our group has been studying the production of secondary metabolites by in vitro cultures of S. lavandulifolia, in order to evaluate its potential use for the production of phytomedicines. To our knowledge, this type of study, including the establishment of in vitro cultures, was not yet performed for this species. In this presentation the accumulation of phenolic compounds by in vitro cultures of S. lavandulifolia is reported. Aseptic shoot cultures were established from surface sterilized seeds and maintained by culturing nodal segments excised from the in vitro growing seedlings. Calli cultures were induced from leaves of these shoots. Suspension cultures were established by transference of calli to liquid medium. Dried biomass of in vitro shoots, calli and suspended cells was extracted with acetone and analyzed by HPLC-DAD. An extract from the aerial parts of in Nature growing S. lavandulifolia (in vivo plants) was also analyzed for comparison.

Total phenolics in the different samples, some of them unidentified, decreased according to the following order: in vivo plants > in vitro shoots > suspension cells > calli. Luteolin, apigenin, cirsimaritin and genkwanin were identified in both shoots and in vivo plants. Salvigenin, the main flavonoid from shoots, was not detected in Nature growing plants. This one, as well as other flavonoids, was not detected in calli neither in suspended cells. Phenolic diterpenes accumulated in higher amounts in Nature growing plants and in shoots and were not detected in suspended cells. The accumulation of phenolic acids, namely rosmarinic acid, on the contrary, was higher in suspended cells and shoots than in in vivo plants.

Acknowledgements: This work was sponsored by EU (FSE/FEDER) and Portuguese Republic Government (FCT) through the Grant SFRH/BD/18908/2004 and the Project SageBiotech (POCTI/AGR/62040/2004).

References: [1] Kintzios, SE (2000), Sage, The genus Salvia, Harwood academic publishers, Amsterdam, Netherlands. [2] Perry, NSL (2003) Biochem Pharmacol Behav 75: 651–659.