根癌农杆菌(Agrobacterium tumefaciens)介导的马铃薯高效遗传转化体系筛选及优化

Screening and Optimization Agrobacterium tumefaciens-Mediated Potato Genetic Transformation System

马铃薯是重要的块茎类作物,根癌农杆菌(Agrobacterium tumefaciens)介导的遗传转化方法是马铃薯基因工程育种的重要技术,研究其遗传转化体系和转化效率有助于提高分子育种效率。针对利用根癌农杆菌介导的2种外植体遗传转化马铃薯普通栽培品种,通过筛选获得最佳转化体系和转化率。以4种马铃薯普通栽培品种(Favorita、Shepody、Atlantic、甘农薯2号)的茎段和试管薯薄片为受体材料,对根癌农杆菌介导的不同遗传转化体系的分化率和转化率进行分析。结果表明:Favorita、甘农薯2号和Atlantic茎段经S2(MS;6-BA 2.5mg·L^(-1),2,4-D0.6mg·L^(-1),Carb 400mg·L^(-1),Kan 50mg·L^(-1))和M2(MS;6-BA 2.5mg·L^(-1),IAA 0.25 mg·L^(-1),2,4-D0.25mg·L~(^(-1)),Carb 400mg·L^(-1),Kan 50mg·L^(-1))培养后获得的转化率显著高于其他培养基。Favorita茎段最大转化率显著高于试管薯薄片转化率;甘农薯2号和Shepody试管薯薄片转化率显著高于茎段转化率。茎段转化体系的最佳诱导愈伤培养基为S2,最佳分化培养为基为M2。Favorita适合采用茎段转化体系,甘农薯2号和Shepody适合采用试管薯薄片转化体系。不同品种的愈伤组织诱导培养基和分化培养基的差别主要是由基因型差异引起的,在提高遗传转化效率中应该针对品种进行转化体系的筛选。

Potato is an important tuber crop.Agrobacterium tumefaciens-mediated potato genetic transformation is an important gene engineering method to improve molecular breeding efficiency in potato.It is very important to get the best transformation system and transformation efficiency.Stem segments and microtuber discs from potato varieties of Gannongshu No.2,Favorita,Shepody and Atlantic were used as acceptor materials.The transformation system of Agrobacterium tumefaciens-mediated were analysis depend on differentiation rate and transformation efficiency.The transformation rate in S2（MS;6-BA2.5 mg·L^（-1）,2,4-D0.6 mg·L^（-1）,Carb 400 mg·L^（-1）,Kan 50 mg·L^（-1））,M2（MS;6-BA2.5 mg·L^（-1）,IAA0.25 mg·L^（-1）,2,4-D0.25 mg·L^（-1）,Carb 400 mg·L^（-1）,Kan 50 mg·L^（-1））medium of stem segments from Gannongshu No.2,Favorita andf Atlantic were significantly higher than other mediums.The stem segment of Favorita had higher transformation rate than microtuber discs of Favorita.The microtuber discs of Gannongshu No.2 and Shepody had higher transformation rate than stem segment of Gannongshu No.2 and Shepody,respectively.The results showed that the best callus-inducing rate potato stem segment was observed on medium S2.The optimum differentiation medium components of potato stem segment was M2.Favorita was suitable for stem segment transformation system,and Gannongshu No.2 and Shepody were suitable for microtuber discs transformation system.The different reasons between callus induction medium and differentiation medium were genotype difference in different varieties.Screening genetic transformation system should be carried out according to different varieties.