Organogenic and Embryogenic Callus Proliferation Method and Squalene Enrichment in Alocasia cuprea Callus with Amino Acid Elicitors

Abstract

This method is designed for the initial and continued proliferation of Alocasia cuprea culture through organogenesis and embryogenesis pathways using a combination of basic media with five types of amino acids and resulting in an increase in the bioactive compound squalene. The method includes stages: initial proliferation using organogenic and embryogenic callus explants, and continued proliferation using basic media MS + 2 mg / l TDZ + 1 mg / l BAP (TB9) and MS + 2 mg / l TDZ + 1 mg / l 2,4-D (IA9) combined with the addition of amino acids (L-arginine, L-asparagine, L-cysteine, L-glutamine, and L-proline) with a concentration of 100 mg / L for three subcultures with a duration of 6 weeks per subculture. The initial proliferation results showed that the highest organogenic callus initiation was obtained from organogenic callus explants on TB9 media, while the highest embryogenic callus was obtained from embryogenic callus cultured on IA9 media.Further proliferation showed that TB9 medium with L-arginine produced organogenic callus with the highest productivity. TB9 medium was dominated by white and pink organogenic callus and shoots with greater weight compared to the yellow embryogenic callus produced in IA9 medium with the addition of amino acids. Further proliferation produced callus with higher squalene content than the control without amino acids. This callus proliferation method has the potential to provide mass seed production and increase squalene yield.