| 摘要: |
| 以水稻基因组测序品种日本晴(Oryza sativa L.ssp.japonica)为材料，对根癌农杆茼介导的水稻转化系统进行了优化。结果表明，水稻种子经质量分数2.5％的次氯酸钠灭菌30 min后，愈伤组织诱导牢最高；所获得的胚性愈伤组织经过6～7 d的继代培养后转化率达到95％；感染液菌浓度OD600值为0.145～0.190时，转化效率高；将抗性愈伤组织进行7 d的暗培养可使分化效率提高到85％。应用本转化系统已经获得了大垃的转化体，GUS表达率达到30％，并且3个月即可获得转化植株。 |
| 关键词: 根癌农杆菌 水稻 遗传转化 |
| DOI： |
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| 基金项目:国家863计划项目(2001AA225051) |
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| Analysis of factors influencing highly efficient Agrobacterium-mediated rice transformation |
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| Abstract: |
| The rice transformation mediated by Agrobacterium tumefaciens was optimized by using Oryza sativa L.ssp.japonica cultivars,which is the matrerial of rice genome sequencing.The main results were as following:The frequency of callus induction is the highest when seeds sterilized in 2.5% sodium hypochlorite solution for 30 minutes;The transformation efficiency is the highest when embryogenic calli are subcultured for 6-7 days;the OD600=0.145-0.190 of bacteria concentration is the best for callus transformation;with the 7-day cultivation in dark,the differentiation efficiency of the resistant calli can be improved to 85%.A lot of transformants have been obtained by using the transformation system in three months.GUS gene expression frequency is up to 30%. |
| Key words: Agrobacterium tumefaciens Oryza sativa L. genetic transformation |
