Abstract: 【Objective】 This experiment was carried out to establish an efficient anther culture system of mutlifoliate alfalfa, and get regenerate plantlet. It will also provide basic materials for the research of ploidy, genomics and molecular biology of alfalfa.【Method】 Firstly, Huaiyin alfalfa and a mutlifoliate alfalfa varieties named Grandeur were used as the parents to create a hybrid, and the anther of hybrid F1 was used as the explants. The ability of induction of callus after bud was studied by using the culture medium of NB added with 2,4-D (0.5 mg•L-1), NAA (0.3 mg•L-1) and KT (3.0 mg•L-1). The appropriate medium was screened for the differentiation of anther callus of alfalfa by orthogonal design L16 (45). In addition, the rooting ability from buds was studied by adding different concentrations of NAA into 1/2 B5 and 1/2 MS.【Result】The highest induction rate of alfalfa callus was in 15 to 28 days after budding, and the maximum value was 82.79%. The culture medium that was good for the differentiation of the callus of alfalfa anther was made up of BⅠwith CH (200 mg•L-1), 2-IP (0.5 mg•L-1), NAA (0.2 mg•L-1) and KT (1.0 mg•L-1), and the maximum regeneration rate was 54.05%. The best culture medium for adventitious bud was 1/2 B5 with NAA (0.3 mg•L-1), and the maximum rooting rate was 88.46%.【Conclusion】An efficient anther culture system of mutlifoliate alfalfa was established successfully, and the regenerated plants were obtained.

Key words: alfalfa , anther culture , medium , buds differentiation