苜蓿愈伤组织原生质体游离与培养  

Dissociation and Culture of Callus Protoplasts of Medicago sativa L.

作　　者：王娟[1] 李玉珠[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：《草地学报》2010年第2期258-262,共5页Acta Agrestia Sinica

基　　金：国家科技支撑计划资助(2007BAD52B06);国家科技支撑计划资助(2006BAD01A19-6);国家牧草产业技术体系专项(chye031231)资助

摘　　要：对阿尔冈金苜蓿(Medicago sativa L. cv. Algonguin)原生质体游离和培养的最佳条件进行研究,建立其细胞融合原生质体杂交体系。以下胚轴诱导的愈伤组织为材料,研究了愈伤组织继代培养时间、酶液组合、酶解时间及酶液渗透压对原生质体游离的影响,并探讨了愈伤组织原生质体在KM8P培养基上的分裂情况。结果表明:继代12 d的愈伤组织游离性最好,最佳酶液组合为2%纤维素酶+0.5%果胶+1%半纤维素酶、最佳酶解时间为12h、最佳酶液渗透压为0.55 mol/L;最适宜培养苜蓿愈伤组织原生质体的培养基为KM8P+2.0 mg/L2,4-D+1.0mg/LNAA。The objectives of this study were to optimize the isolation and culture conditions for protoplast of Medicago sativa L.cv.Algonguin and establish the cell fusion and protoplast hybridization system.Using hypocotyls callus as tested materials,the influences of callus derived time,enzyme combination,and enzymolysis time on protoplast dissociation were studied and the division of callus protoplasts on KM8P basic medium was discussed.The results show that the callus subcultured 12 days was the optimum free material and the optimum condition for high yield of viable protoplast was as follows： enzyme combination containing 2% cellulase Onozuka R-10＋0.5% pectinase＋1.0% hemicellulase,mannitol concentration at 0.55 mol/L,and enzymolysis time of 12 h.The optimal medium for callus protoplast culture was KM8P＋2.0 mg/L 2,4-D＋1.0 mg/L NAA.

关 键 词：苜蓿 愈伤组织 原生质体 酶解条件 

分 类 号：S541[农业科学—作物学] Q943.1[生物学—植物学]