基因枪法获得Phs-r转基因小麦植株的研究

实验以普通小麦豫麦18-64品种为供试材料,采用基因枪法对报告基因(Bar基因)和目的基因(PHS-r基因)两个重组质粒进行了共转化,获得了再生植株,并对转化植株进行了检测。结果表明:以预培养15d的幼胚愈伤组织作为受体材料其转化频率明显高于当天剥取和预培养3～4d的幼胚;在基因枪轰击前6h和轰击后18h,用0.5moL/L的甘露醇进行渗透处理,其转化频率会有明显的提高;实验通过基因枪转化与再生培养,共获得了12株再生植株,PPT检测且均表现出抗性,其中6株经PCR检测呈阳性,表明PHS-r基因已整合到小麦基因组中并正常表达。

Study on Obtaining Transgenic Wheats with Phs-r Gene by Particle Bombardment

In this paper,the tested materials was YuMai 18-64 cultivar.The two plasmids with PHS-r gene and Bar gene were co-transformed by particle bombardment.The regenerated plantlets were obtained and the transformed plants were analyzed.The results showed that the immature embryos pre-cultured for fifteen days had a higher transformation frequency than that of pre-cultured three to four days and immature embryos.0.5M mannitol treatment from 6h before to 18h after bombardment could increase the transformation frequency.Twelve regenerated plantlets were obtained by optimum plant regeneration culture and PPT screening,which all were positive in Bialaphos resistance and six plants were positive individuals by the PCR assay.