‘新牧4号’紫花苜蓿原生质体游离和培养因素的研究  

Protoplasts Isolation and Culture Factors of Medicago sativa L.‘Xinmu No.4'

作　　者：陈爱萍[1] 刘慧敏[1] 李瑶[1] 冯维卫 

机构地区：[1]新疆草地资源与生态实验室/新疆农业大学草业与环境科学学院,乌鲁木齐830052

出　　处：《中国农学通报》2017年第1期137-141,共5页Chinese Agricultural Science Bulletin

基　　金：草业科学国家重点学科项目"提高苜蓿原生质体细胞分裂频率的研究"(XJCYQ-2012-02)

摘　　要：主要探讨影响苜蓿原生质体游离和培养的因素,为原生质体培养体系的建立提供依据。以‘新牧4号’紫花苜蓿的子叶、下胚轴和愈伤组织为材料,研究了酶液的p H值,酶解时间,酶液组合和不同培养方法对原生质体游离和培养效果的影响。结果表明:当酶液p H 5.8时,有活力的原生质体的产量最高,同时细胞碎片少;子叶、下胚轴和愈伤组织适宜的酶解时间均为10 h;酶液组合为2%纤维素酶+0.5%果胶酶+0.3%或0.5%离析酶,游离出的有活力的原生质体产量较高;采用液体浅层培养和固液双层培养,均观察到原生质体发生分裂,但固液双层培养法更有利于‘新牧4号’紫花苜蓿原生质体的分裂和培养。The factor of alfalfa protoplasts isolation and culture were discussed, which could provide a basis forthe establishment of protoplast culture system. Using cotyledons, hypocotyls and hypocotyl callus of Medicago sativa L.‘Xinmu No.4＇as tested materials, the effects of p H value of enzyme, enzymolysis time, enzymecombination, different culture methods on protoplasts isolation and culture were studied. The results showedthat when p H value of enzyme was 5.8, the yield of viable protoplast was the highest, at the same time, celldebris was fewer. The optimum enzymolysis time of cotyledons, hypocotyls and hypocotyl callus was 10 h. Theenzyme concentration was 2.0% cellulase ＋ 0.5% pectinase ＋ 0.3%（or 0.5%） macerozyme, the yield of viableprotoplast was relatively high. Using liquid medium and solid-liquid double layer medium, protoplast divisionwas observed, but solid-liquid double layer culture was more favorable for protoplast division and culture of Medicago sativa L.‘Xinmu No.4＇.

关 键 词：'新牧4号’紫花苜蓿 P H值 酶解时间 酶液组合 培养方法 

分 类 号：S541.9[农业科学—作物学]