利用发根农杆菌转化短日照茄子。

Transformation of Solanum brevidens using Agrobacterium tumefaciens.

机构信息

Department of Horticulture, Iowa State University, 50011-1100, Ames, IA, USA.

出版信息

Plant Cell Rep. 1995 Dec;15(3-4):196-9. doi: 10.1007/BF00193719.

DOI:10.1007/BF00193719

Abstract

Leaf pieces of in vitro-cultured plantlets of the wild potato species Solanum brevidens Phil. were cocultivated with Agrobacterium tumefaciens that contained nptII and uidA genes on the disarmed plasmid pBI121. Independent transgenic shoots were regenerated from solidified and liquid medium that contained 50 mg l(-1) kanamycin. Two Agrobacterium strains were investigated for transformation efficiency. GV2260, which contained p35SGUSINT, resulted in a 11% transformation frequency, compared with 1% using LBA4404. Transformation rates were 7% in liquid culture and 3% on solidified medium. All kanamycinresistant, putatively transformed plantlets were confirmed positive by histochemical GUS assays. GUS activity in 22 independently transformed plants was quantified by fluorometric assay. Southern analysis of randomly selected transgenic plants showed that each transgenic plant contained at least one copy of the uidA gene.

摘要

离体培养的野生马铃薯种短柄薯叶片小块与携带 nptII 和 uidA 基因的无毒性质粒 pBI121 上的根癌农杆菌共培养。在含有 50mg/L 卡那霉素的固体和液体培养基中，独立的转化芽从固体和液体培养基中再生。研究了两种根癌农杆菌菌株的转化效率。含有 p35SGUSINT 的 GV2260 导致转化频率为 11%，而 LBA4404 为 1%。液体培养中的转化率为 7%，固体培养基中的转化率为 3%。所有卡那霉素抗性的、疑似转化的植株均通过组织化学 GUS 分析得到证实。通过荧光测定法对 22 个独立转化的植株中的 GUS 活性进行定量。随机选择的转基因植株的 Southern 分析表明，每个转基因植株至少含有一个 uidA 基因的拷贝。