俄罗斯杂花苜蓿愈伤组织原生质体游离与培养条件的优化  

Optimization of Dissociation and Culture Conditions of Protoplasts of Callus of Russian Variegated Alfalfa

作　　者：张凌云[1] 师尚礼[1] 

机构地区：[1]甘肃农业大学草业学院/草业生态系统教育部重点实验室/中-美草地畜牧业可持续发展研究中心,甘肃兰州730070

出　　处：《中国草地学报》2013年第3期12-17,共6页Chinese Journal of Grassland

基　　金：国家牧草产业技术体系项目(CARS-35);农业部公益性行业(农业)科研专项"牧区优质高效饲草生产利用技术研究与示范"(201003023)

摘　　要：对俄罗斯杂花苜蓿(Medicago varia)原生质体游离和培养条件进行优化,建立其原生质体融合的体细胞杂交体系。以下胚轴诱导所得愈伤组织为材料进行原生质体游离,研究酶液组合、渗透压调节物甘露醇浓度、酶解时间、愈伤组织继代时间及预处理措施对原生质体游离效果的影响,以及激素浓度与培养密度对原生质体培养的影响。结果表明:最佳酶液组合为2%纤维素酶+0.5%果胶酶+0.3%离析酶,最佳渗透压为0.55mol/L,最佳酶解时间为12h,愈伤组织继代培养至14d的游离性最佳;固-液综合培养条件下最适激素组合为1.5mg/L 2,4-D+1.0mg/L 6-BA,最适原生质体培养密度为3×105个/mL。The objectives of this study were to optimize the dissociation and culture conditions of pro toplast of Medicago varia and establish the protoplast fusion for forming somatic hybridization system. The tested material was hypocotyl callus of Russian variegated alfalfa. And the influences of enzyme com- binations, osmotic regulation content, enzymolysis time and callus derived time for hydrolysis protoplast and the influences of studied. The results th sh e hormone concentration and the protoplasts densi owed that the optimum enzyme combinations were 0.5%Pectinase Y-23＋0.3% Macerozyme R-10, the optimum mannito optimum enzymolysis time was 12 hours and the optimum callus subcu ty for protoplasts culture were 2% cellulose onnzuka R-10＋ concentration was 0.55mol/L, the ur liquid comprehensive cultivation mode, the optimum hormone combination 6-BA and the optimum culture density was 3×10^5/mL. e time was 14 days; in the solid- was 1.5mg/L 2,4-D＋1.0mg/L

关 键 词：杂花苜蓿 愈伤组织 酶解条件 原生质体 

分 类 号：Q943.1[生物学—植物学]