INDUCTION AND IDENTIFICATION OF AUTOTETRAPLOID FOR THE AUGMENTATION OF TANSHINONE IIA IN SALVIA MILTIORRHIZA BUNGE BY IN VITRO TREATMENT WITH COLCHICINE

Tóm tắt

Polyploidization technique has been successfully used for increasing the levels of quantitative and qualitative patterns of secondary metabolite production and exhibit enhanced vigour and superior performance in different medicinal plants. A protocol for the in vitro induction of Salvia miltiorrhiza Bunge tetraploids has been optimized to enhance the Tanshione IIA content, a major component of Salvia miltiorrhiza Bunge, inhibits platelet activation. In vitro leaves were used for treatment with different concentration of colchicine (0, 0.01, 0.02, 0.05, 0.1 w/v) along with treatment durations (7, 14, 21 and 28 days). The treated explants were then incubated on Murashige and Skoog (MS) medium having 0.5 mg/L N6-benzylaminopurine for shoot regeneration. The mutant types were isolated based on morphological characteristics and flow cytometry assays plantlets at in vitro and in vivo conditions. The tetraploids of S. miltiorrhiza were proficiently induced by the treatment of 0.05% colchicine for 14 days. The resulting tetraploid plants showed significantly enhanced agronomic traits, including the size of stomata and leaflet as well as root diameter, and fresh weight of root. In addition, an obvious reduction of length to width ratio was found in the 4x plants, including stomata frequency, leaflets, and roots. High-performance thin-layer chromatography showed a significant enhancement in the bioactive compound tanshinone IIA content of tetraploid plants (0.22% of dried sample) in comparison to diploid plants (0.14% of dried sample), signifying the prospective of this technique for the trade value improvement.