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可育的抗除草剂溴苯腈转基因小麦

SELF FERTILE TRANSGENIC WHEAT PLANTS EXPRESSING HERBICIDE BROMOXYNIL RESISTANCE

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【作者】 王小军刘玉乐田波HuangYong

【Author】 Wang Xiao Jun, 1Liu Yu le, 2Huang Yong and 1Tian Bo 1(Institute of Microbiology, Academia Sinica, Beijing 100080) 2(Department of Plant and Animal Science, University of Alaska, AK 99775, USA)

【机构】 中国科学院微生物研究所!北京100080DepartmentofPlantandAnimalScience!UniversityofAlaskaAK99775USA

【摘要】 报道了采用微粒轰击(Microprojectile bom bardm ent) 幼胚将除草剂抗性基因导入小麦(Triticumaestivum L.)的转化研究。实验共使用了13 个小麦品种, 从开花后14~18 d 的籽粒中剥取幼胚, 植物表达质粒含有CaMV 35S启动子控制的除草剂溴苯腈抗性基因bxn 以及筛选标记基因NTPⅡ。采用高压放电基因枪,用质粒DNA 包被的钨粒轰击预培养3 d 的幼胚。在含有卡那霉素类似物geneticin G418sulphate 的MS培养基上, 经过多步骤筛选和分化, 从800 多个幼胚中获得了16 株转化苗。除草剂抗性鉴定和Southern 杂交分析证明, 其中4 株为转基因植物,具有溴苯腈抗性, 并且自交可育。转化工作从分离幼胚到转化苗鉴定完毕, 最短时间为6 个月, 因此, 该方法是一项快速有效的基因导入技术

【Abstract】 Genetic manipulation of wheat (Triticum aestivum L.) by biotechnological approaches is currently limited by a lack of efficient and reliable transformation method. The authors report a reproducible protocol for rapid production of transgenic wheat via microprojectile bombardment. The experiment was carried out by using the immature embryo excised from caryopsis 14 to 18 days postanthesis and the plant expression plasmid carrying a CaMV 35S controlled bxn gene, for resistance to herbicide bromoxynil and a selectable marker gene NPTⅡ. After bombarding the precultured immature embryos isolated from 13 wheat varieties with plasmid DNA coated tungsten particle, these embryos were transferred on MS medium containing 10 mg/L geneticin G418 sulphate to select and regenerate transformants step by step. As a result, 16 transformed plants were obtained from a total of 849 bombarded embryos. The characterization of these plants by inoculation with herbicide bromoxynil and Southern analysis with bxn gene as a probe showed that 4 of the self fertile transformed plants contained the target gene and presented herbicide resistance. In several independent transformation experiments, the fastest one took only 6 months from embryo excision to characterization of regenerated plants. Therefore, this procedure is a rapid and efficient technique for delivering foreign DNA into wheat.

  • 【文献出处】 植物学报 ,Acta Botanica Sinica , 编辑部邮箱 ,1996年12期
  • 【分类号】Q943.2
  • 【被引频次】113
  • 【下载频次】146
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