【摘要】 为了探索短柄草的遗传转化体系,以二倍体短柄草ABR 6为受体材料,通过对诱导培养基类型、潮霉素筛选浓度和根癌农杆菌侵染浓度等参数的优化,建立了农杆菌介导短柄草遗传转化体系。结果表明,来源于未成熟胚的胚性愈伤组织在LS培养基上诱导率最高,达76.27%,最佳Hpt筛选浓度为40 mg.L-1,最佳农杆菌侵染浓度为OD600=0.6,在此条件下ABR 6的转化效率可达5%;通过PCR检测12株抗性植株,发现7株能扩增出Hpt基因(845 bp)条带;通过荧光显微镜观察转基因植株叶片,发现绿色荧光蛋白的表达,进一步证实了转基因植株的可靠性。更多还原

【Abstract】 Using diploid Brachypodium distachyon line ABR 6 as transformation receptor,the genetic transformation system for model grass B.distachyon was established by agrobacterium-mediated method after optimizing the induction medium,Hpt concentration,Agrobacterium concentrations and other conditions.The results showed that the induction rate of embryogenic callus,derived from immature embryos of the line ABR 6 was 76.27% on the LS basal medium.The most suitable concentration of Hpt was 40 mg·L-1.The frequency of transformation was 5% when the optimum agrobacterium concentration was OD600=0.6.The PCR detection of the 12 hpt resistant plantlets indicated that 7 could amplify a 845 bp band of the target.The expression of green fluorescence protein in the leaves of transgenic plants under fluorescence microscope further confirmed the transgenic plants.更多还原