表达bar基因的抗除草剂转基因甘薯的获得  

Development of Transgenic Sweetpotato Expressing bar Gene for Herbicide Resistance

作　　者：臧宁[1] 翟红[1] 王玉萍[1] 于波[1] 何绍贞[1] 刘庆昌[1] 

机构地区：[1]中国农业大学植物遗传育种系,农业部作物基因组学与遗传改良重点开放实验室,北京市作物遗传改良重点实验室,教育部作物杂种优势研究与利用重点实验室,北京100094

出　　处：《分子植物育种》2007年第4期475-479,共5页Molecular Plant Breeding

基　　金："十一五"国家科技支撑计划(2006BAD01A06);国家杰出青年科学基金(30225028)资助。

摘　　要：用农杆菌介导法将bar基因导入甘薯主栽品种徐薯18的胚性悬浮细胞中,获得了抗除草剂的转基因植株。农杆菌菌株EHA105携带的双元载体pCAMBIA3300上含有bar基因。来自于徐薯18胚性悬浮细胞的直径为0.7～1.3mm的280个胚性细胞团用于遗传转化。共培养3d后,首先在含有2mg/L2,4-D、100mg/LCarb的液体MS培养基中培养1周,然后将胚性细胞团转移到添加2mg/L2,4-D、100mg/LCarb和0.3mg/LPPT的固体MS培养基上进行选择培养。选择8周后,将获得的37个PPT抗性愈伤组织转移到添加1mg/LABA、100mg/LCarb和0.3mg/LPPT的固体MS培养基上,其中的34个愈伤组织诱导得到体细胞胚并发芽形成小植株,共获得了164株拟转基因植株。PCR分析表明,其中的123株为转基因植株。Southernblot和Northernblot分析表明,bar基因稳定整合到转基因植株的基因组中并正确表达。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。Transgenic sweetpotato plants expressing the bar gene for herbicide resistance was achieved by using Agrobacterium tumefaciens-mediated transformation and cv. Xushu18 embryogenic suspension cultures. Cell aggregates 0.7-1.3 mm in size from embryogenic suspension cultures were cocultivated with the Agrobacterium. tumefaciens strain EHA105 harboring a binary vector pCAMBIA3300 with the bar gene. Three days after cocultivation, the inoculated 280 cell aggregates were cultured in MS medium with 2.0 mg/L 2,4-D and 100 mg/L Carb for one week, and then were cultured on MS medium supplemented with 2.0 mg/L 2,4-D, 100 mg/L Carb and 0.3 mg/L PPT for the selection culture. Eight weeks after selection, 37 PPT-resistant embryogenic calluses were produced. After transferred to MS medium supplemented with 1.0 mg/L ABA, 100 mg/L Carb and 0.3 mg/L PPT, 34 calluses formed 164 putatively transgenic plantlets via somatic embryogenesis. PCR analysis indicated that 123 of the 164 plantlets were transgenic plants. Stable integration of the bar gene into the genome of transgenic plants was confirmed by Southern blot and Northern blot analyses. Transgenic plants exhibited functional expression of the bar gene by in vivo assay for herbicide resistance.

关 键 词：甘薯(Ipomoea batatas(L.)Lam.) 农杆菌介导转化 胚性悬浮细胞 除草剂 转基因植株 

分 类 号：S531[农业科学—作物学]