基因枪法获得GNA转基因小麦植株的研究-中国农业科学.PDFVIP

　2001, 34( 1): 1- 4 Scientia Ag ricultura Sinica　　　 GNA 徐琼芳,李连城,陈　孝,马有志,叶兴国, 张增艳,徐惠君,辛志勇 (,,　100081) : 以小麦品种京 411作为基因 转化的靶材料,取其未成熟胚诱导愈伤组织,经过 10d 左右培养后,用含有 雪花莲凝集素( Galanthus nivalis agglutinin , GN A)和 bar基因的质粒 pBI121-2轰击 800 个胚性愈伤组织,在含有 4mg / Basta溶液的培养基上进行筛选 ,分化及生根培养,获得 67棵再生植株。 田间涂抹 Basta溶液( 50、75mg / )检 测和接麦蚜实验,提取转基因植株基因组 DNA,用扩增 GN A基因的引物经 PCR扩增 South ern杂交 ,结果表明利用 基因 转化已从 T 代获得了 8株含有编码 bar /GNA基因的转基因植株。 2 : GN A基因; bar基因; 基因 转化;转基因小麦 : 512. 1; 753　: 　: 0578-1752(2001) 01-0005-04 S Q A St udy on t he Obtaining of Transgenic Wheat s w it h GNA A lien Gene by Biolist ic Part icle - , - , , - , XU Qiong fang I iang cheng CHEN Xiao M A You zhi - , - , - , - YE Xing guo ZHAN G Zeng yan XU Hui jun XIN Zhi yong ( Key Laboratory of Agricultural Ministry f or Crop Genetics and Breeding , Institute of Crop Breeding and Cultivation, CAAS, Beij ing　100081) Abstract: The immature embryos of wheat Jing, 411 cv. , 12～ 14 days after pollination were taken, to be cultured on SD2 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 / Basta. According to the resistance tests both by inoculating wheat aphids onto the plants and applying Basta solution of 50mg / or 75mg / 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