马铃薯高效再生及遗传转化体系的优化

Optimization of High Efficiency Regeneration and Genetic Transformation System in Potato

为优化马铃薯高效再生及遗传转化体系,研究鄂马铃薯3号茎段外植体在不同激素浓度配比下愈伤组织及分化情况,获得最佳愈伤组织及分化激素配比,再采用根癌农杆菌的遗传转化进行筛选,获得最佳转化体系,为马铃薯优质品种的基因工程育种提供依据。结果表明,不同激素浓度配比下愈伤组织及分化率差异较大,其中2.0 mg/L ZR+0.1 mg/L IAA激素配比下可高效再生出芽,出芽率高达100%,芽长势粗壮。通过构建植物表达载体pCambia1301,以GV3101为介导菌株,预培养2~3 d,共培养2~3 d,350 mg/L羧苄青霉素,50 mg/L卡那霉素筛选浓度转化效果较好,转化效率可达65%,其中转化苗在0.8 mg/L IBA+0.2 mg/L NAA激素配比的培养基中可形成再生植株,再生率达80%,苗茎粗壮,根系发达。

In order to optimize the efficient regeneration and genetic transformation system of potato,the effects by using stem segment of Hubei potato No.3 as explants were studied in different hormone concentration ratios,and the optimal callus and differentiation hormone ratio were obtained.Then,the genetic transformation of Agrobacterium tumefaciens was used for screening to obtain the best transformation system,which provided the basis for genetic engineering breeding of high quality potato varieties.The results showed that the callus and differentiation rate were great different under the ratio of different hormone concentration.The hormone ratio of 2.0 mg/L ZR and 0.1 mg/L IAA could regenerate the buds efficiently,the germination rate was as high as 100%,and the bud growth was strong.By constructing expression vector pCambia1301 in plant,GV3101-mediated strain,pre-cultured for 2-3 days,and co-cultured for 2-3 days,and 350 mg/L carbenicillin,and 50 mg/L kanamycin to screen the concentration was better.The transformation efficiency could reach 65%.The transformed seedlings could form regenerated plants in the medium of 0.8 mg/L IBA and 0.2 mg/L NAA hormone.The regeneration rate could reach 80%,the stems were thick and the roots were developed.