基因枪介导的冬小麦临汾5064遗传转化体系构建  

Construction of A Genetic Transformation System Mediated by Biolistic Particle for Winter Wheat Linfen 5064

作　　者：乔麟轶 李锐[1] 郝宇琼 乔玲[2] 李欣[1] 张晓军 畅志坚 郑兴卫 QIAO Linyi;LI Rui;HAO Yuqiong;QIAO Ling;LI Xin;ZHANG Xiaojun;CHANG Zhijian;ZHENG Xingwei(College of Agriculture,Shanxi Agricultural University/Shanxi Key Laboratory of Crop Genetics and Molecular Improvement,Taiyuan 030031,China;Institute of Wheat Research,Shanxi Agriculture University,Linfen 041000,China)

机构地区：[1]山西农业大学农学院/作物遗传与分子改良山西省重点实验室,山西太原030031 [2]山西农业大学小麦研究所,山西临汾041000

出　　处：《山西农业科学》2023年第12期1347-1352,共6页Journal of Shanxi Agricultural Sciences

基　　金：山西省科技重大专项计划“揭榜挂帅”项目(202201140601025);山西省重点研发计划项目(202102140601001)。

摘　　要：以中国小麦优质骨干亲本临汾5064幼胚为试验材料,对诱导、分化、生根培养基中的激素浓度和基因枪轰击压力等关键参数进行对比分析,旨在总结出临汾5064的最适遗传转化体系,为山西小麦育种提供技术平台。结果表明,临汾5064幼胚在3种激素浓度的诱导培养基中均可形成愈伤组织。其中,在含2.0 mg/L 2,4-D的诱导培养基中,参试幼胚具有最高的出愈率(83.0%)。临汾5064愈伤组织在4种激素浓度的分化培养基中均有再生芽产生,当分化培养基包含0.2 mg/L 2,4-D和0.2 mg/L 6-BA时,愈伤组织具有最高的分化率(70.0%)。临汾5064分化愈伤组织在4种激素浓度的生根培养基中均有根生成,当生根培养基包含0.1 mg/L 2,4-D且不添加6-BA时,分化愈伤组织具有最多的生根率(77.5%),并且形成较多数目的根。利用上述优化激素浓度的培养基对临汾5064幼胚开展基因枪介导的遗传转化,在轰击步骤设置3个压力处理组分别导入测试质粒。结果显示,在900 psi轰击压力下获得192株再生植株,再生植株获得率为28.3%,其中有4株成功扩增出目标条带,具有最高的遗传转化率(0.50%)。In order to summarize the optimal genetic transformation system for Linfen 5064 and provide a technical platform for Shanxi wheat breeding,in this study,the immature embryo of Linfen 5064,a high-quality backbone parent of Chinese wheat,was used as the test material,the key parameters including hormone concentrations of induction,differentiation as well as rooting culture media and bombardment pressure was compared and analyzed.The results showed that Linfen 5064 immature embryos could form callus in all the three concentrations of hormone in inducing culture medium.Among them,in the induction medium containing 2.0 mg/L 2,4-D,the tested immature embryos showed the highest callus induction rate of 83.0%.Linfen 5064 callus produced regenerated buds in four differentiation culture mediums with diverse hormone concentrations.When the differentiation medium contained 0.2 mg/L 2,4-D and 0.2 mg/L 6-BA,the callus had the highest differentiation rate of 70.0%.The differentiated callus of Linfen 5064 exhibited root formation in four hormone concentrations of rooting medium.When the rooting medium contained 0.1 mg/L 2,4-D without 6-BA,the differentiated callus had the highest rooting rate of 77.5%and formed a majority of roots.Using the optimized hormone concentration culture medium mentioned above,genetic transformation mediated by biolistic particle was carried out on the immature embryo of Linfen 5064.Three pressure treatment groups were set up in the bombardment step to introduce the test plasmids separately.The results showed that 192 regenerated plants were obtained under 900 psi bombardment pressure,with a regeneration rate of 28.3%.Among them,four plants carrying target bands were successfully amplified,with the highest genetic transformation rate of 0.50%.

关 键 词：山西小麦 临汾5064 基因枪 遗传转化 

分 类 号：S512.11[农业科学—作物学]