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本研究利用花粉管通道法将携带有外源抗除草剂的bar基因导入红麻 ,分析了不同导入方法 (子房注射法、柱头滴加法 )和技术参数对受体植株当代结实率、T1代种子出苗率及转化率的影响。结果表明 :不同导入方法在受体当代结实率、T1代种子出苗率上存在较明显的差异。其中子房注射法结实率高于柱头滴加法 ,而出苗率则相反。外源DNA导入浓度和剂量对受体当代结实率、T1代种子出苗率则无显著影响 ;并确定除草剂PPT对红麻青皮 3号受体品种的幼苗期 ( 3— 4叶期 )的筛选浓度为 70 μg/ml,用该浓度PPT溶液筛选转化株 ,其T1代有 1 2 %的植株表现出对除草剂的抗性 ,其推断转化株的比率随外源DNA导入浓度及剂量的增大而提高 ,子房注射法较柱头滴加法其T1代有更高的推断转化株比率
In this study, pollen tube pathway was used to introduce the bar gene with external herbicide into kenaf. The effects of different methods of introduction (ovary injection, stigma dropping) and technical parameters on the seed setting rate, T1 generation Seed germination rate and conversion rate. The results showed that there were obvious differences in the seed setting rate and seedling emergence rate of T1 generation under different introduction methods. Among them, the seed setting rate of ovary injection was higher than that of stigma dropping, while the germination rate was the opposite. The concentration of exogenous DNA and the dosage did not affect the seed setting rate and the seedling emergence rate of T1 seedlings at the same time. The herbicide PPT was selected to screen the seedling stage (3-4 leaf stage) At a concentration of 70 μg / ml, the transformants were screened with this concentration of PPT solution, and 12% of the T1 plants showed resistance to the herbicide. The inferred transformant ratio increased with the concentration of the introduced DNA and dose Large and improved, ovary injection method than the stigma dropwise T1 generation have higher deduced transformant ratio