Improvement of friable callus induction of Crocus sativus L. and establishment of a cell suspension culture system with high biomass

Purpose: This study aims to explore the potential of in vitro culture as a method for scaling up the production of saffron based medicinal compounds, the most expensive spice renowned. Emphasis is placed on the critical role of friable callus (FC) formation as a prerequisite for successful suspension culture. Research method: The research primarily investigates FC formation, focusing on the impact of varying strengths of Murashige and Skoog (MS) medium as well as combinations of NAA or 2,4-D and BA or Kin on compact callus. Subsequently, the study involves supplementing the MS medium with different concentrations of 2,4-D, kin, zeatin, glutamine, sucrose, and nitrogen to establish a cell suspension culture. Findings: The highest FC yield was achieved on a solid medium containing 2,4-D (1 mg l^-1 )+Kin (0.2 mg l^-1 ), resulting in a fresh weight (FW) of 0.413 g. Furthermore, MS combined with 2,4-D (1 mg l^-1 )+Kin (0.2 mg l^-1 )+glutamine (10 mg l^-1 ), as well as MS+2,4-D (0.5 mg l^-1 )+zeatin (0.3 mg l^-1 )+glutamine (10 mg l^-1 ), demonstrated the highest FW under suspension conditions. The study also identified that 30 g l^-1 sucrose and 30 µM were optimal for inducing maximum FW. Research limitations: Cell biomass is influenced by several factors that should to be optimized. Originality/Value: This research concludes that a cell suspension system holds promise for rapidly generating sufficient cell biomass to produce valuable secondary metabolites within a limited timeframe and space. Notably, the system successfully increased biomass from 0.2 to 1.2 g, underscoring its potential for efficient saffron-based product development.