[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 ofploidy,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 culturemedium of NB added with 2,4-D (0.5 mg.L1),NAA (0.3 mg·L-1) and KT (3.0 mg.L1).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 ofB I with CH (200 mg·L1),2-IP (0.5 mg·L1),NAA (0.2 mg·L1) and KT (1.0 mg·L1),and the maximum regeneration rate was 54.05％.The best culture medium for adventitious bud was 1/2 B5 with NAA (0.3 mg·L1),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.%[目的]建立多叶苜蓿高效花药培养体系,获得再生植株,为苜蓿倍性、基因组学及分子生物学的研究提供基础材料.[方法]以淮阴苜蓿与多叶苜蓿品种飞马(Grandeur)杂交F1的花药为外植体,使用NB+2,4-D(0.5 mg·L-1) +NAA(0.3 mg·L-1) +6-BA(0.5 mg·L-1)+KT(3.0 mg·L-1)研究苜蓿现蕾后花药愈伤组织的诱导能力；利用正交设计L16 (45)筛选适宜苜蓿花药愈伤组织分化的培养基；并用1/2B5和1/2MS添加不同浓度的NAA研究适宜的生根培养基.[结果]苜蓿现蕾后15-28 d愈伤组织诱导率达到最高,最高为82.79％.适宜苜蓿花药愈伤组织分化的培养基组合为B I+ CH (200 mg·L-1) +2-IP (0.5 mg·L-1)+NAA (0.2 mg·L-1)+KT (1.0 mg·L-1),分化率可达54.05％.不定芽在1/2 B5+NAA(0.3 mg·L-1)培养基上生根效果最好,生根率可达88.46％.[结论]成功建立了苜蓿高效花药培养体系,并获得再生植株.
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