Establishment of regeneration system of callus pathway for Iris sanguinea Donn ex Horn

Abstract

Iris is a perennial flowering plant, usually cultivated through seeds or bulbs. However, due to the limitations of traditional reproduction, the establishment of a callus regeneration system is particularly important, and callus induction and plant regeneration are the keys to transgenic technology. The callus regeneration system was studied using the I. sanguinea flower stem as explants. The experiment investigated the impact of different disinfectant solution concentrations, disinfection exposure times, and hormone concentrations on the explant growth status at various culture stages. The research analyzed the impact of the experimental variables on explant regeneration to determine the optimum callus regeneration system for I. sanguinea. In this research, it was determined that when I. sanguinea flower stems, that were used for explants, were treated with 75% alcohol for 30 s and 2% NaClO for 8 min for disinfection, there was a 54.48% callus induction rate. The callus induction medium was Murashige and Skoog medium (MS) + 6-benzylaminopurine (6-BA) 1.0 mg/L + 1-naphthylacetic acid (NAA) 0.2 mg/L + 2,4-dichlorophenoxyacetic acid (2,4-D) 2.0 mg/L for 40 d. The optimum medium for callus multiplication was MS + 6-BA 0.5 mg/L + 2,4-D 0.5 mg/L, while the optimum medium for callus adventitious bud induction was MS + 6-BA 1.0 mg/L + NAA 0.2 mg/L + kinetin (KT) 0.5 mg/L, 60 d. The adventitious bud differentiation rate from flower stem calli was 46.03%.