农杆菌介导大豆萌动种子遗传转化的影响因素研究

目的]提高农杆菌介导大豆遗传转化效率。方法]以萌发1 d的半片大豆种子为目标组织,通过GUS瞬时表达检测,研究乙酰丁香酮、菌液的浓度及侵染时间对转化效率的影响。结果]在共培养基中添加200μmol/L的乙酰丁香酮时,GUS瞬时表达效率最高。当农杆菌生长到对数生长期(D600 nm=0.5)时,转化效率最高,平均每个外植体上出现9.4个蓝色斑点。农杆菌侵染时间不宜过长,以30~60 min较好。结论]乙酰丁香酮浓度、农杆菌菌液浓度和侵染时间均影响农杆菌介导大豆遗传转化效率。

Study on the Factors Affecting the Genetic Transformation Efficiency of Soybean Germinating Seeds Mediated by Agrobacterium tumefacines

Objective] The aim of the research was to enhance the genetic transformation efficiency of soybean mediated by Agrobacterium tumefacines.Method] With half piece of soybean seeds germinated for 1 d as target tissue,the effects of acetosyringone and A.tumefacines liquid concn.and infection time on the transformation efficiency were studied through GUS transient expression detection.Result] When 200 μmol/L acetosyringone was added to the co-culture medium,GUS transient expression efficiency was highest.When A. tumefacines grew up to logarithmic growth stage(D600 nm of 0.5),the transformation efficiency was highest and 9.4 blue spots averagely appeared in each explant.Too long infection time was not suitable,with 30～60 min being better.Conclusion] The genetic transformation efficiency of soybean mediated by A.tumefacines was affected by acetosyringone concn.,the liquid concn.of A.tumefacines and the infection time