Highly efficient plant regeneration via somatic embryogenesis from cell suspension cultures of Boesenbergia rotunda

Boesenbergia rotunda is a perennial ginger species rich in flavonoids, flavones, and cyclohexenyl chalcone derivatives. Several of these secondary metabolites have shown promising antiviral and anticancer activities, and thus, it is important to optimize methods for robust production of clonal materials. In this study, cell suspensions were established and their growth capacities were evaluated in liquid media supplemented with varying growth regulator compositions. The highest settled cell volume of 6.1?±?0.3 ml with a specific growth rate of 0.0892?±?0.0035 was achieved by maintaining cells in Murashige and Skoog liquid media supplemented with 1.0 mg L^?1 of 2,4-dichlorophenoxyacetic acid and 0.5 mg L^?1 6-benzyladenine, representing a 12-fold increase in cell volume during the culture period. A somatic embryogenesis rate of 1,433.33?±?387.84 somatic embryos per milliliter of settled cells was achieved with an inoculation cell density of 50 μl settled cell volume and on growth regulator-free agar plates. Around half (53.5?±?7.9%) of the somatic embryos germinated into complete plantlets on media supplemented with 3 mg L^?1 6-benzyladenine and 1 mg L^?1 α-naphthaleneacetic acid. The plantlets were successfully transferred to soil and grown in the greenhouse. Phytochemical profiling via high-performance liquid chromatography analysis revealed that regenerated plantlets retained the capacity to produce and accumulate bioactive compounds. Hence, this protocol will be helpful for metabolic engineering and functional studies of genes and enzymes involved in the biosynthetic pathway of valuable compounds in B. rotunda.