根癌农杆菌介导的紫花苜蓿遗传转化体系的优化

Optimization of Agrobacterium Tumefaciens-Mediated Genetic Transformation System of Alfalfa

为进一步提高根癌农杆菌介导的紫花苜蓿遗传转化效率,本试验以紫花苜蓿(Medicago sativa L.)品种‘中苜1号’为试验材料,通过筛选高效再生的植株,以筛选出的植株叶片为外植体,并通过调节培养基激素配比,在农杆菌侵染过程中添加谷氨酰胺、进行冷处理等措施优化苜蓿由农杆菌介导的遗传转化体系。试验结果表明:转化过程中以嫩叶作为外植体,菌液和重悬液中添加150μmol·L^(-1)乙酰丁香酮,侵染过程中使用300μmol·L^(-1)的谷氨酰胺将瞬时转化效率从32%提高至92%,转化效率提高到31%。研究紫花苜蓿的遗传转化体系可为紫花苜蓿高效遗传改良及基因功能研究提供了技术支持。

In order to improve Agrobacterium tumefaciens-mediated genetic transformation efficiency of alfalfa(Medicago sativa L.),we used alfalfa variety‘Zhongmu No.1’as plant material.The genetic transformation system of alfalfa mediated by agrobacterium was optimized by selecting the highly regeneration plant,using young leaves as explants,adjusting the hormone ratio in the medium,adding glutamine,and giving a cold treatment during agrobacterium infection.The results showed that the transient transformation efficiency of‘Zhongmu No.1’increased from 32%to 92%and the stable transformation efficiency increased to 31%by adding 150μmol·L^(-1) acetosyringone to the bacterial suspensions solution,and using 300μmol·L^(-1) glutamine during infection.The genetic transformation system of alfalfa reported in this paper provides technical support for efficient genetic improvement and gene functional study of alfalfa.