Effect of radiation on regeneration of Chinese narcissus and analysis of genetic variation with AFLP and RAPD markers

Abstract

The aim of our study was to establish an efficient in vitro propagation protocol for Chinese narcissus (Narcissus tazetta var. chinensis) to obtain variants of this species using γ-radiation treatment and evaluate the effectiveness of this system for variant induction using amplification fragment length polymorphism (AFLP) and randomly amplified polymorphic DNA (RAPD) analysis. Various doses (5–100 Gy) of gamma rays were applied to investigate the effect of radiation on adventitious bud formation from bulb-scales and the survival rate of plantlets. It was demonstrated that the regeneration of Chinese narcissus was very sensitive to gamma radiation even at low doses. The survival and multiplication rate significantly decreased with an increase of radiation dose. The optimal irradiation dose for survival and mutation induction was approximately 10 Gy. The genetic variations among the regenerants derived from irradiated explants were evaluated by DNA fingerprinting using RAPD and AFLP markers which detected a variation frequency of 8.33% and 15.48% respectively. The high frequency of mutants detected by molecular markers indicated that treatment of in vitro cultures with γ-rays may be an effective way to improve narcissus cultivars.