根癌农杆菌介导的抗除草剂转基因甘薯植株的获得

以甘薯品种栗子香的胚性悬浮细胞为受体材料, 用根癌农杆菌介导法，获得了表达bar基因的抗除草剂转基因甘薯植株。农杆菌菌株LBA4404携带含有bar基因的双元载体pCAMBIA3300。共计450个胚性细胞团用于遗传转化。在添加2 mg/L 2,4－D、100 mg/L Carb和0.5 mg/L PPT 的固体MS培养基上选择培养8周后，得到了19个PPT抗性愈伤组织。将这19个抗性愈伤组织转移到添加1 mg/L ABA、100 mg/L Carb和0.5 mg/L PPT 的固体MS 培养基上，其中的10个抗性愈伤组织共再生出103株拟转基因植株。PCR分析表明，其中的69株为转基因植株。Southern blot分析表明，bar基因稳定整合到转基因植株的基因组中。除草剂喷洒试验结果表明，转基因植株具有高度除草剂抗性。

Generation of Transgenic Herbicide-Resistant Sweetpotato Plants by Agrobacterium tumefaciens-mediated Transformation

Production of transgenic sweetpotato plants expressing the bar gene for herbicide resistance was achieved using Agrobacterium tumefaciens-mediated transformation and sweetpotato cv. Lizixiang embryogenic suspension cultures. Total 450 embryogenic cell aggregates were cocultivated with the A. tumefaciens strain LBA4404 harboring a binary vector pCAMBIA3300 with the bar gene. Eight weeks after selection on MS medium supplemented with2.0 mg/L 2,4-D, 100 mg/L Carb and 0.5 mg/L PPT, 19 PPT-resistant embryogenic calluses were produced. After transferred to MS medium supplemented with 1.0 mg/L ABA, 100 mg/L Carb and 0.5 mg/L PPT, 10 of them formed 103 putatively transgenic plants. PCR analysis indicated that 69 plants were transgenic. Stable integration of the bar gene into the genome of transgenic plants was confirmed by Southern blot analysis. Transgenic plants exhibited functional expression of the bar gene by in vivo assay for herbicide resistance.