| 摘要: |
| 【目的】优化小麦基因枪转化技术体系，为提高小麦基因枪转化效率提供参考。【方法】以小偃22为材料,采用基因枪法分析小麦幼胚愈伤组织诱导时间、微弹轰击金粉用量及筛选分化培养基类型对基因枪遗传转化频率的影响，并对转化植株进行了检测。【结果】在相同条件下，小麦幼胚经过9 d的愈伤组织诱导时再生植株率最高，达到4.24%；同一条件下，金粉用量为60 μg/枪的再生植株率最高，达到3.90%；使用1/2MS培养基添加NAA 1.0 mg/L和KT 1.0 mg/L进行转化细胞筛选与分化的效果最好，再生植株率为3.72%；对转化抗性再生植株的特异PCR检测表明，目的基因已整合到小麦染色体组中。【结论】小麦幼胚通过9 d的愈伤组织诱导，基因枪轰击金粉用量为60 μg/枪，使用筛选分化培养基1/2MS+KT 1.0 mg/L+NAA 1.0 mg/L+双丙氨膦2.0 mg/L+蔗糖30 g/L+Ag 5.0 g/L进行转化细胞的筛选与分化，可获得较高的抗性再生植株率。 |
| 关键词: 小麦幼胚 基因枪 愈伤组织 恢复培养 筛选分化 再生植株 |
| DOI： |
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| 基金项目:国家转基因生物新品种培育项目（2008ZX08002003，2009ZX08002008B）；陕西省13115重大科技专项（2009ZDKG-11）；陕西省科技攻关项目（2011K02-01） |
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| Study on influncing factors of biolistic bombardment in immature embryos of wheat |
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| Abstract: |
| 【Objective】The research was on the effect of several factors on regenerative plants rate,optimize wheat genetic transformation system and improve the efficiency of genetic transformation by gene gun.【Method】We analysed the effect of induction time of young embryo callus,dosage of gold dust and culture medium of screening and differentiation on regenerative plants rate by single factor experiment.【Result】When induction time of young embryo callus was 9 days,regenerative plants rate achieved 4.24%;when dosage of gold dust per gene gun was 60 μg,regenerative plants rate achieved 3.90%;and culture medium of screening and differentiation was 1/2MS+NAA 1.0 mg/L+KT 1.0 mg/L+Bialaphos 2.0 mg/L,regenerative plants rate achieved 3.72%.【Conclusion】The best optimum conditions of wheat genetic transformation system:Induction time of young embryo callus is 9 days,dosage of gold dust per gene gun 60 μg,and culture medium of screening and differentiation 1/2MS+NAA 1.0 mg/L+KT 1.0 mg/L+Bialaphos 2.0 mg/L+sucralose 30 g/L+Ag 5.0 g/L.PCR analysis preliminarily proved the object gene had been integrated into the wheat genome. |
| Key words: immature embryos of wheat gene gun callus recovery culture screening and differentiation regeneration plants |
