| 刘 丹, 蔡高磊, 张 立,王建锋, 康振生,韩德俊.农杆菌介导短柄草遗传转化体系的建立[J].麦类作物学报,2011,31(4):598 |
| 农杆菌介导短柄草遗传转化体系的建立 |
| Establishment of Agrobacterium mediated Transformation System in Brachypodium distachyon Line ABR 6 |
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| DOI：10.7606/j.issn.1009-1041.2011.04.004 |
| 中文关键词: 短柄草 农杆菌转化 Hpt基因 |
| 英文关键词:Brachypodium distachyon Agrobacterium transformation Hpt gene |
| 基金项目:国家自然科学基金项目（30871603）；高等学校学科创新引智计划资助项目(B07049)。 |
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| 中文摘要: |
| 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。 |
| 英文摘要: |
| 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. |
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