Liquid shoot culture of Salvia officinalis L. for micropropagation and production of antioxidant compounds; effect of triacontanol

Izabela Grzegorczyk,Halina Wysokińska

doi:10.5586/asbp.2008.013

Abstract

Liquid shoot culture of Salvia officinalis L. in MS medium containing IAA (0.1 mg l-1) and BAP (0.45 mg l-1) was developed and evaluated in relation to shoot multiplication and antioxidant compound (carnosic acid, carnosol and rosmarinic acid) accumulation. In the liquid medium, on average, 3 new shoots per explant (shoot tip) were obtained within 3 weeks. The shoots produced 8.2±0.02 mg of diterpenoids and 31.2±0.29 mg of rosmarinic acid per gram of dry weight. Shoot proliferation and diterpenoid content increased when triacontanol (5, 10 or 20 pg l-1) was added to the liquid medium. In optimum conditions (at 20 pg l-1 TRIA) almost 7 shoots were formed per explant after 3 weeks. An increase in diterpenoid production (expressed as the sum of carnosol and carnosic acid) ranged from 30% to 50% and dependended on triacontanol concentration tested. The level of diterpenoids in triacontanol-treated shoots was similar to the content of compounds in commercial herbal product (dried leaves of S. officinalis) (10-12 mg g-1 dry wt). Triacontanol did not increase rosmarinic acid production, but the content of the phenolic as compound in shoots grown in liquid culture (31 mg g-1 dry wt) was even 24 times higher compared to samples of dried leaves of S. officinalis plants. We also demonstrated that the highest amounts of CA, Car and RA were accumulated in young, top parts of sage shoots. This observation could be useful for improving the selection of material for the extraction of natural antioxidants from S. officinalis.

Highlights

Salvia officinalis L. is an aromatic perennial herb native to Southern Europe and Asia Minor
We have previously described micropropagation of S. officinalis using shoot tips grown on agar-solidified medium and showed that multiple shoots and in vitro regenerated plants were able to synthesize carnosol (Car), carnosic acid (CA) and rosmarinic acid (RA) (Grzegorczyk et al 2005 and 2006)
Shoot tips (0.5 cm) of multiple shoot culture of S. officinalis on MS (Murashige and Skoog 1962) agar (0.7%) medium supplemented with IAA (0.1 mg l-1) and BAP (0.45 mg l-1) were inoculated into liquid medium of the same composition, but excluding agar

Summary

Introduction

Salvia officinalis L. (sage) is an aromatic perennial herb native to Southern Europe and Asia Minor. We have previously described micropropagation of S. officinalis using shoot tips grown on agar-solidified medium and showed that multiple shoots and in vitro regenerated plants were able to synthesize carnosol (Car), carnosic acid (CA) and rosmarinic acid (RA) (Grzegorczyk et al 2005 and 2006).

Results

Conclusion