根癌农杆菌法转化康乃馨的条件探索

Exploration on the Genetic Transformation Conditions of Dianthus caryophyllus L.Mediated by Agrobacterium tumefaciens

[目的]为构建高效的康乃馨遗传转化体系奠定基础。[方法]以康乃馨叶片为受体材料, 通过抗G418 筛选, 探讨预培养时间、根癌农杆菌菌液浓度、共培养时间等因素对根癌农杆菌介导康乃馨遗传转化的影响。[结果]20mg/ ml G418 是康乃馨叶片转化的适宜浓度。随着预培养时间的增加, 康乃馨叶片的存活率降低, 预培养2 ～3 d 时抗性芽分化率较高。当根癌农杆菌OD600 为0.5 ～0.8 时, 抗性芽分化率较高, 适合康乃馨转化。共培养2 ～3 d 时, 康乃馨叶片生长状态好, 抗性芽分化率最高。[结论]预培养、共培养各2 ～3 d ,

[ Objective] Tne research aimed to lay the foundation for constructing the highly-efficient genetic transfomation system of Dianthus caryophyllus L. [Method] With the leaves of D. caryophyllus as receptor materials, the effects of such factors as preculture time, the eonen, of Agrobacterhan tumefaciens and coculture time on the genetic transfomation of D. caryophyllus mediated by A. tumefaciens were studied through G418 screening. [Result] 20 mgiml G418 was the suitable eonen, for the transfomation of D. caryophyllus leaves. With the increasing of precuhure time, the survival rote of D. caryophyllus leaves were reduced. When precuhure time was 2- 3 d, the differentiation rote of resistant buds were higher. When OD600 of A. tumefaciens was 0.5 -0.8, the differentiation rote of resistant buds were higher and favorable for the transformation of D, caryophyllus. When cocuhure time was 2- 3 d, the growth state of D. caryophyllus leaves were good and the differentiation rote of resistant buds were highest. [ Conclusion ] Both precultue and cocuhure for 2-3 d and the bacterial liquid conen. OD600 of 0.5-0.8 were the suitable conditions for the genetic transforrmtion of D. caryophyllus mediated by A. tumefaciens.