In Vitro Tetraploidization Towards Enhancement Of Charantin Biosynthesis In Momordica Charantia (L.)

Abstract

A practical and reliable method for in vitro tetraploidization of Bitter Gourd [Momordica charantia (L.)] has been established to enhance the production of charantin. Shoot tip from in vitro-grown culture (2n = 22) were exposed to the anti-mitotic chemical, i.e., colchicine, at various concentrations (0, 0.01, 0.05, 0.1, 0.2 and 0.3% along with 2% DMSO) for 12, 24, 36, and 48 h. The treated explants were then incubated and proliferated on Murashige and Skoog (MS) medium fortified with 1.5 mg L-1 benzyladenine and 0.5 mg L-1 naphthalene acetic acid, followed by root induction in 1.0 mg L-1 indole-3 acetic acid enriched 1/2MS medium. Treatment of shoot tips with 0.1% colchicine for 24 h supported the highest tetraploid induction efficiency (33.56 ±0.22%). Morphological, stomatal, and cytological characteristics along with the secondary metabolite content of the in vitro tetraploids were compared to that of diploids. The recovered tetraploid plants possessed superior plant height, stem diameter, leaf size, and increased length and width of stomata but decreased stomatal frequency. The tetraploid plants demonstrated twice the chromosome number (2n = 4x = 44) in respect to diploids as confirmed through cytology, spectrophotometry and flow cytometry. Highperformance liquid chromatography showed 1.09 times enhancement of charantin content in tetraploid plants than that of diploid plants, signifying the prospective of this technique for the trade value improvement.