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为探讨利用基因工程技术进行小麦品质改良的可行性,用基因枪法对湖北省3个小麦品种鄂恩1号、鄂麦11号和鄂麦12号分别进行优质高分子量麦谷蛋白亚基(HMW-GS)基因1Ax1和1Dx5+1Dy10的转导。试验结果表明,经基因枪轰击的幼胚外植体的再生频率和转化频率明显比幼穗高;基因型间愈伤组织再生能力和转化频率的差异,直接影响着小麦转化的成功率;3个品种的幼胚发育时期与其再生频率均呈显著负相关(r=-0.93或-0.95);在可控环境条件下,花后12～14d的供试植株幼胚的转化频率最高(鄂恩1号、鄂麦11号和鄂麦12号分别为4.5%、2.9%和2%)。研究表明,鄂恩1号、鄂麦11号适龄期的幼胚试材,用等摩尔比的优势亚基基因1Dx5和1Dy10与选择标记基因以2:1摩尔比混合的沉淀物包裹金粉,经600～900psi氦气压轰击后,在含0.5mg·L-12,4-D的MS培养基中诱导愈伤、含0.1mg·L-12,4-D和5mg·L-1玉米素的R培养基中分化、3mg·L-1L-PPT选择压下继代分化筛选,可获得稳定可育的T0代转基因植株以及胚乳特异性表达的T1代转基因种子。
In order to explore the feasibility of improving the quality of wheat by using genetic engineering techniques, three high-quality glutenin subunits (HMW- GS) genes 1Ax1 and 1Dx5 + 1Dy10. The results showed that the frequency and frequency of regeneration of immature embryos bombarded by particle bombardment were significantly higher than those of young ears. The difference of regeneration ability and transformation frequency between genotypes had a direct impact on the success rate of wheat transformation. 3 There was a significant negative correlation (r = -0.93 or -0.95) between the developmental stages of immature embryos and the regeneration frequency of these embryos. The frequency of immature embryos was the highest at 12-14 days after flowering under controlled environmental conditions No. 1, Emei No. 11 and Emei No. 12 were 4.5%, 2.9% and 2% respectively). The results showed that the age-appropriate immature embryos of E’en 1 and E’mai 11 were coated with the precipitates mixed with 2: 1 molar ratio of 1Dx5 and 1Dy10, which are the dominant subunit genes of the same molar ratio, Callus was induced in MS medium containing 0.5mg · L-12,4-D after bombardment with helium at 600-900psi. R (0.1mg · L-12,4-D and 5mg · L-1 zeatin) Medium differentiation, 3mg · L-1L-PPT selection pressure subculture selection and screening can be stable and fertile T0 transgenic plants and endosperm-specific expression of T1 transgenic seed.