新陆早33号高效遗传转化体系研究  

Study on Protocol of Efficiency Genetic Transformation in Xinluzao 33(Gossypium hirsutum L.)

作　　者：李鹏飞[1] 朱华国[1] 程文翰[1] 王凡龙[1] 张新宇[1] 孙杰[1] 

机构地区：[1]石河子大学农学院/新疆生产建设兵团绿洲生态农业重点实验室,新疆石河子832003

出　　处：《新疆农业科学》2013年第6期981-987,共7页Xinjiang Agricultural Sciences

基　　金：农业部转基因生物新品种培育重大专项(2011ZX08005);"十二五"科技支撑计划项目(2011BAD35B05);石河子大学动植物育种专项(gxjs2010-yz01)

摘　　要：【目的】以新疆北疆棉区主栽品种新陆早33号为材料,研究不同处理条件对转基因抗性胚性愈伤组织增殖、胚状体分化以及植株再生的影响,为新疆棉花遗传转化奠定基础。【方法】以新陆早33号下胚轴为外植体,利用农杆菌介导法转化抗草甘膦基因EPSPS,通过优化培养条件以提高抗性胚性愈伤组织获得完整再生植株的能力。【结果】在胚性愈伤组织增殖阶段,培养基中KNO_3浓度为3.8g/L,继代周期为11 d有助于胚性愈伤组织快速增殖和保持良好生长状态;将凝固剂Phytagel浓度提高到4.0g/L能明显促进胚状体分化;在子叶胚生根阶段以Phytagel为凝固剂同时加入1.0 g/L的活性炭有助于壮根、减少褐化和提高正常植株比例。【结论】通过对培养条件的优化,缩短了胚状体发生的周期,并获得了大量再生植株,建立了新陆早33号的高效遗传转化体系。[ Objective ] In treatments on resistant embryog cotton （Gossypium hirsutum L. ） order to carry out the transgenic protocol in cotton, the effect of different enic callus proliferation, somatic embryogenesis and regeneration of upland Xinluzao 33 was studied. [ Method] Hypocotyls of Xinluzao 33 were taken as explants, and glyphosate -resistant gene EPSPS was transformed by Agrobacterium -mediated transformation method. Somatic embryogenesis capacity and cotyledonary embryos culture methods were optimized . [ Result] During the period of embryogenic callus proliferation, when the concentration of the culture medium was 3.8 g/L KNO3, 11 days subculture cycle were suitable for fast proliferation and keeping better state, Phytagel in MSB medium supplemented with 4.0 g/L obviously promoted embryoid differentiation, and with 1.0 g/L activated carbon facilitated root development, reduced browning and improved the ratio of normal regenerated plant. [ Conclusion ] The cycle of regeneration via somatic embryogenesis was shortened and a large number of regenerated plants were obtained. The efficient genetic transformation protocol for Xinluzao 33 was constructed.

关 键 词：棉花 抗性胚性愈伤组织 体细胞胚胎发生 再生植株 

分 类 号：S562[农业科学—作物学] S503.53