农杆菌介导的紫花苜蓿遗传转化体系的建立与优化

Establishment and optimization of Agrobacterium-mediated transformation of alfalfa (Medicago sativa L.)

以'中苜一号'紫花苜蓿品种作为受体材料,建立了适用于农杆菌介导的转基因组培体系,并对GUS基因进行转化,经GUS组织化学染色,以获得抗性愈伤组织分析为目的,转化优化条件为:叶片预培养4～5 d,用农杆菌菌液(A600＝0.3～0.8)侵染20 min,然后在培养基上铺一层灭菌滤纸共培养7 d后清洗.

This paper established the regeneration system of somatic embryogenesis for the cultivar alfalfa “Zhongmu NO.1”genetic transformation. Manipulation system was optimized by Km-resistant calli assays and the histological detection of GUS activity, which GUS gene comed from plant expression vector pCAMBIA2301G. The condition of optimization was pre-culture for 4~5 days, infection with Agrobacterium tumefaciens (A600=0.3~0.8) for 20 min and washed after 7 days of culture.