小麦组织培养和基因枪轰击影响因素探讨  

Study on influncing factors of tissue culture and biolistic bombardment in wheat

作　　者：刘伟华[1] 李文雄[2] 胡尚连[2] 徐香玲[1] 李集临[1] 

机构地区：[1]哈尔滨师范大学生物系,哈尔滨150080 [2]东北农业大学农学系,哈尔滨150030

出　　处：《西北植物学报》2002年第3期602-610,T012,共10页Acta Botanica Boreali-Occidentalia Sinica

摘　　要：本研究就基因枪法转化小麦过程中的组织培养和轰击参数等影响因素进行了探讨。结果表明 ,小麦幼胚是比幼穗或成熟胚更理想的转化受体。因供试小麦品种基因型不同 ,幼胚愈伤组织再生频率差异明显 ,辽 - 1 0为 7.84% ,91 B5 6 9为 1 3 .6 8% ,东农 7742为 5 4 .90 %。小麦本身对选择剂卡那霉素有较高的天然抗性 ,采用 G41 8对小麦幼胚愈伤组织进行筛选效果明显。G41 8的毒性作用有滞后特点 ,3个小麦品种对 G41 8的敏感性依次为辽 - 1 0 >91 B5 6 9>东农 7742。用 G41 8做选择剂筛选辽 - 1 0、91 B5 6 9和东农 7742抗性愈伤的适合浓度分别为2 5 mg/L、3 0 mg/L和 3 5 mg/L。此外 ,不同轰击参数影响金粉分布的范围和密度。轰击距离为 6 cm或 9cm时 ,内部金粉密度大而外围金粉密度小 ,差异极大。轰击距离为 1 2 cm时 ,内部和外围金粉密度差异小 。In the article,we studied the different factors influncing the tissue culture and biolistic bombardment in wheat transfer.Immature embryo used as acceptor in gene transfer is better than that of young spike because of quickly callus formation and good callus quality.The frequency of plant regeneration in calli from immature embryo of the three genotypes used in the study was quite different.It is 7.84% in Liao10,13.68% in 91B569,and 54.88% in NE7742.The study indicated that geneticin (G418) showed significant selection effect because of the natural resistance to kanamycin in wheat.The sensitivity of three genotypes to geneticin showed different.Liao-10 ranked the first,91B569 next,and NE7742 the last.The optimum concentration of G418 used in resistant calli selection was 25 mg/L,30 mg/L,and 35 mg/L for Liao-10,91B569,and NE7742,respectively.Besides,the effect of bombard factor on the distribution and density of gold particle was found.Under the condition both of 6 cm and 9 cm in distance of bombardment,the inside circle gained more gold particle than outside,and the different was markedly.However,well distribution of gold particle was found in 12 cm distance.

关 键 词：小麦 组织培养 基因枪轰击 影响因素 转基因技术 

分 类 号：S512.1[农业科学—作物学]