玉米优良自交系31的基因枪转化.pdfVIP

摘要 利用PDS．1000／He型基因枪，将携带选择标记基因Bar、报告基因GUS、 自交系Z31的胚性愈伤组织，在含10mg／L草丁膦的选择培养基上筛选2个月 后，得到了304块抗性愈伤。将抗性愈伤转到分化培养基，分化出了85株再生 植株。PCR检测表明，26株携带GUS报告基因，10株携带选择标记基因 Southern验证，表明SOS3确实整合到了Z3l的基因组。RT—PCR检测表明，整 则检测到其中4株的叶片和根被染成了兰色，表明GUS在翻译水平的表达。但 RT．PCR未检测到目的基因SOS2和SOS3的表达。本文就研究中存在的诸如抗 性筛选中出现的逃逸现象、转化效率低以及转基因沉默等问题进行了讨论。 关键词：玉米；I型愈伤；遗传转化；基因枪 2 Abstract I calluseswereinducedfromimmatureofmaize Typeembryogenic embryos eliteinbredlineZ31．The containstheBARselective 1，which plasmidpCAMBIA330 marker GUS thesalt—resistantrelatedSOS2andSOS3 gene，thereportergene，and from deliveredintothecalluses thePDS1000／He genesArabidopsis，was byusing biolistic 2910callus werebombarded．Aftertwomonthsselection gun．Total pieces 1 were on ammonium，304 calluses 0mg／Lglufosinate putativelyglufosinate—resistant obtained．274resistantcallusesweretransferredontothe medium，and regeneration 85 were indicatedthe85 plantsregenerated．PCRscreeninganalysis that，of weredetectedGUS 0 BAR regeneratedplants，26plants positive，1plantspositive，5 SOS3 1 SOS2 conformed analysis plants positive，andplant positive．PCR—Southem thattheSOS3indeed intothe5 gene integrated regeneratedplants．RT-PCRanalysis the26 7