Enhanced rosmarinic acid production in cultured plants of two species of Mentha.

In the present investigation an attempt has been made to enhance rosmarinic acid level in plants, grown in vitro, of 2 species of Mentha in presence of 2 precursors in the nutrient media during culture. For in vitro culture establishment and shoot bud multiplication, MS basal media were used supplemented with different concentrations and combinations of different growth regulator like NAA (alpha-napthaleneacetic acid), BAP (6-benzylaminopurine). The medium containing NAA (0.25 mg/L) and BAP (2.5 mg/L) gave the highest potentiality of shoot formation (average 58.0 numbers of shoots) per explant for Mentha piperita L. and the medium containing BAP (2.0 mg/L) gave the highest potentiality of shoot (average 19.2 numbers of shoots) formation per explant for Mentha arvensis L. The complete plants were regenerated in above mentioned media after 8 weeks of subculture. For in vitro enhancement of rosmarinic acid production, the 2 precursors tyrosine (Tyr) and phenylalanine (Phe) were added in the nutrient media at different levels (0.5 mg/L to 15.0 mg/L). Tyrosine was found to be very effective for augmenting rosmarinic acid content in Mentha piperita L. It nearly increased the production up to 1.77 times. In case of Mentha arvensis L., phenylalanine significantly affected the production of rosmarinic acid and the production was nearly 2.03 times more than the control. No significant increase in biomass was observed after addition of these precursors indicating that the added amino acids acting as precursors for rosmarinic acid synthesis were readily utilized in producing rosmarinic acid without promoting growth. Total protein profile also revealed the presence of a specific band in polyacrylamide gel electrophoresis.