Somaye Amini - Department of Horticultural Science, Gorgan University of Agricultural Sciences and Natural Resources, Gorgan, Iran
Seyed Mahdi Ziaratnia - Department of Food Biotechnology, Research Institute of Food Science and Technology (RIFST), Mashhad, Iran
Ghadir Rajabzadeh - Department of Food Nanotechnology, Research Institute of Food Science and Technology (RIFST), Mashhad, Iran

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 ۲,۴-D and BA or Kin on compact callus. Subsequently, the study involves supplementing the MS medium with different concentrations of ۲,۴-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 ۲,۴-D (۱ mg l-۱)+Kin (۰.۲ mg l-۱), resulting in a fresh weight (FW) of ۰.۴۱۳ g. Furthermore, MS combined with ۲,۴-D (۱ mg l-۱)+Kin (۰.۲ mg l-۱)+glutamine (۱۰ mg l-۱), as well as MS+۲,۴-D (۰.۵ mg l-۱)+zeatin (۰.۳ mg l-۱)+glutamine (۱۰ mg l-۱), demonstrated the highest FW under suspension conditions. The study also identified that ۳۰ g l-۱ sucrose and ۳۰ 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 ۰.۲ to ۱.۲ g, underscoring its potential for efficient saffron-based product development.

Callogenesis, Glutamine, MS medium, Saffron, Solid medium