【摘要】 通过对以马铃薯 (Solanum tuberosum L .)块茎为受体的农杆菌转化系统进行优化 ,筛选出使三个品种 (鲁引 1号、克新 3号、优金 )的受体均可得到高频分化的培养基 :MS+2 .0 mg/L BA+3.0mg/L ZT+0 .5mg/L NAA+0 .5mg/L GA.对几种农杆菌转化条件的比较研究发现 ,菌体用 Ms+0 .5mg/L GA液体重悬并稀释 4倍 ,与受体于 2 8℃ ,10 0 r/min振荡侵染 2 0～ 30 min,共培养时在分化培养基上加 30 μ mol/L AS(乙酰丁香酮 ) ,16 h弱光照共培养 3d,可使马铃薯品种鲁引 1号的转化效率较常规方法提高 5～ 7倍 .将禽流感病毒 (H5N1)基因与 35S启动子及 nos终止子构建表达载体 p130 1H5N1,直接法转入农杆菌 EHA10 5,用此菌株转化马铃薯得到再生植株 .基因组总 DNA经 PCR和 Southern杂交证明目的基因已整合到马铃薯基因组中更多还原

【Abstract】 To optimize the genetic transformation conditions of potato( Solanum tuberosum L. ),several conditions of Agro bacterium mediated gene transfer were determined.The medium(MS+2.0 mg/L BA+3.0 mg/L ZT+0.5mg/L NAA+0.5mg/L GA)is high efficient for discs regenerating of three cultivars(Luying No.1、Kexing No.3 and Youjing). Agro bacterium resuspended and diluted by solution MS+0.5 mg/L GA, invaded potato discs swirring at 100 r/min 28 ℃ for 20~30 minites.Following cocultivation, the explants were transfered to the medium as same as the above medium but with the addition of 30 μ mol/L AS and under 16 hr weak light for 3 days.Compared with routine method,the transformation effciency increased by 5~7 times.The expression vector p1301H5N1,which contained CaMV35S promoter and gene H5N1 and Tnos,was directly transfered into Agrobacterium strain EHA105.After incubating the discs of tuber,28 individuls of regenerated resistant plants were obtained.The results of PCR and Southern blotting of potato genomic DNA demonstrated that the target gene was integrated into the genome of potato.更多还原