基因枪法获得GNA转基因小麦植株的研究  

Study on the Obtaining of Transgenic Wheats with GNA Alien Gene by Biolistic Particle

作　　者：徐琼芳[1] 李连城[1] 陈孝[1] 马有志[1] 叶兴国[1] 张增艳[1] 徐惠君[1] 辛志勇[1] 

机构地区：[1]中国农业科学院作物育种栽培研究所

出　　处：《中国农业科学》2001年第1期5-8,T002,共5页Scientia Agricultura Sinica

基　　金：国家转基因植物研究与产业化专项资助项目!( J2 0 0 0 -D-0 15 )

摘　　要：以小麦品种京 411作为基因枪转化的靶材料 ,取其未成熟胚诱导愈伤组织 ,经过 10 d左右培养后 ,用含有雪花莲凝集素 (Galanthus nivalis agglutinin ,GNA)和 bar基因的质粒 p BI12 1- 2轰击 80 0个胚性愈伤组织 ,在含有4mg/ L Basta溶液的培养基上进行筛选 ,分化及生根培养 ,获得 6 7棵再生植株。田间涂抹 Basta溶液 (5 0、75 m g/ L)检测和接麦蚜实验 ,提取转基因植株基因组 DNA,用扩增 GNA基因的引物经 PCR扩增 Southern杂交 ,结果表明利用基因枪转化已从 T2 代获得了 8株含有编码 bar/ GNA基因的转基因植株。The immature embryos of wheat, Jing 411 cv., 12~14 days after pollination, were taken to be cultured on SD 2 medium for callus induction. As the receptor of target genes of galanthus nivalis agglutinin and bar, 800 wheat calli were bombardmented by biolistic particle coated with the DNA of plasmid pBI121 2 after 10 days culture. 67 plantlets were finally regenerated from the bombardment calli on the selection medium containing 4mg/L Basta. According to the resistance tests both by inoculating wheat aphids onto the plants and applying Basta solution of 50mg/L or 75mg/L on the wheat leaves in field, and the molecular analysis such as of PCR and Southern of the genomic DNA extracting from the plants, 8 plants were identified to be the transgenic ones containing the target genes.

关 键 词：GNA基因 BAR基因 基因枪转化 转基因小麦 

分 类 号：S512.1[农业科学—作物学] [生物学—分子生物学] Q753