根癌农杆菌介导的抗除草剂转基因甘薯植株的获得  

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

作　　者：臧宁[1] 张美彦[1] 翟红[1] 李雪琴[1] 王玉萍[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：《农业生物技术学报》2008年第1期103-107,共5页Journal of Agricultural Biotechnology

基　　金：“十一五”国家科技支撑计划(No.2006BAD01A06);农业部行业计划(No.nyhyzx07-012);国家杰出青年科学基金(No.30225028)

摘　　要：以甘薯(Ipomoea batatas(L.)Lam.)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。根癌农杆菌(Agrobacterium tumefaciens)菌株LBA4404携带有bar基因的双元载体pCAM-BIA3300,共计450个胚性细胞团用于遗传转化。在添加2mg/L2,4-D、100mg/L Carb和0.5mg/L PPT(phosphinothricin)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1mg/L ABA、100mg/LCarb和0.5mg/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基因稳定地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 sweetpotato plants expressing bar gene for herbicide resistance was achieved using Agrobacterium tumefaciens-mediated transformation and sweetpotato （Ipomoea batatas （L.） Lam.） cv. Lizixiang embryogenic suspension cultures. Total 450 embryogenic cell aggregates were cocultivated with A. tumefaciens strain LBA4404 harboring a binary vector pCAMBIA3300 with the bar gene. Eight weeks after selection on MS medium supplemented with 2.0 mg/L 2,4-D, 100 mg/L Carb and 0.5 mg/L PPT （phosphinothricin）, 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 putative 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. Experiment result showed that transgenic plants exhibited functional expression of the bar gene by in vivo assay for herbicide resistance.

关 键 词：甘薯 农杆菌介导转化 胚性悬浮细胞 除草剂 转基因植株 

分 类 号：S188[农业科学—农业基础科学]