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基因枪法获得GNA转基因小麦植株的研究
引用本文:徐琼芳,李连城,陈孝,马有志,叶兴国,张增艳,徐惠君,辛志勇.基因枪法获得GNA转基因小麦植株的研究[J].中国农业科学,2001,34(1):5-8.
作者姓名:徐琼芳  李连城  陈孝  马有志  叶兴国  张增艳  徐惠君  辛志勇
作者单位:中国农业科学院作物育种栽培研究所,
基金项目:国家转基因植物研究与产业化专项资助项目(J2000-D-015)
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。

关 键 词:GNA基因  bar基因  基因枪转化  转基因小麦
文章编号:0578-1752(2001)01-0005-04

Study on the Obtaining of Transgenic Wheats with GNA Alien Gene by Biolistic Particle
XU Qiong fang,LI Liang cheng,CHEN Xiao,MA You zhi,YE Xing guo,ZHANG Zeng yan,XU Hui jun,XIN Zhi yong.Study on the Obtaining of Transgenic Wheats with GNA Alien Gene by Biolistic Particle[J].Scientia Agricultura Sinica,2001,34(1):5-8.
Authors:XU Qiong fang  LI Liang cheng  CHEN Xiao  MA You zhi  YE Xing guo  ZHANG Zeng yan  XU Hui jun  XIN Zhi yong
Abstract: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.
Keywords:GNA gene  bar gene  Gene transferring of biolistic particle  Transgenic wheat
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