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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 ,均匀度好
In this study, the factors influencing tissue culture and bombardment parameters during the transformation of wheat by particle gun were discussed. The results showed that wheat immature embryos are more ideal transformants than spikelets or mature embryos. The frequency of regeneration of immature embryos was significantly different due to different genotypes of wheat cultivars. The average frequency of calli regeneration was 7.84% in Liaoning-Liaoning Province, 13.68% in 91 B5 6 9 and 54.9% in Dongnong 7742. The wheat itself has higher natural resistance to the selective agent kanamycin, and the effect of selecting the callus of wheat immature embryo using G41 8 is obvious. The toxicity of G41 8 lagged. The sensitivity of three wheat cultivars to G41 8 was Liao - 1 0> 91 B5 6 9> Dong Nong 7742. The selection of G41 8 as a selection agent for the selection of Liao - 10,91 B5 6 9 and Dongnong 7742 resistant callus were 25 mg / L, 30 mg / L and 35 mg / L, respectively. In addition, different bombardment parameters affect the range and density of the powder distribution. When the bombardment distance is 6 cm or 9 cm, the density of the internal gold powder is large and the density of the peripheral gold powder is small, with great differences. When the bombardment distance is 1 2 cm, the density difference between the inside and the outside is small and the uniformity is good