Optimization of Callus Induction and Cannabidiol (CBD) Production in Hemp (Cannabis sativa L.) through Culture Condition Regulation

Abstract

Abstract: Industrial-scale production of cannabidiol (CBD) from hemp (Cannabis sativa L.) faces challenges due to slow growth rates and low CBD yields. This study presents an optimized protocol for efficient callus induction in hemp as a fundamental step towards CBD production; the hemp was used as experimental material to study the callus growth and CBD content under different cultivation conditions. The experimental method was designed to optimize the induction condition and an HPLC method was developed to analyze the CBD content. When the culture conditions were 1/2 Murashige-Skoog medium, 1 mg/L 6-benzylaminopurine and 0.3 mg/L α-naphthaleneacetic acid, lighting for16 h and temperature of 25°C, the hemp callus induction rate reached a maximum of 97.22%, and the CBD content of hemp callus reached a maximum of 147.06 ± 2 μg/g. In this study, the effects of various factors, including hormone types and concentrations, culture medium compositions, light conditions, and temperature conditions, on the induction rate and CBD content of hemp callus were systematically investigated. By optimizing callus culture conditions, this work provides methodological insights into the initial induction of hemp callus. The results contribute to the growing body of research on hemp tissue culture and the biosynthesis of secondary metabolite.