花粉管法将外源除草剂基因导入红麻的有效方法及参数研究

Effective methods and parmeters of exotic genss introduced into kenaf for herbicide-resistant through pollen tube

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 。

Pollen tube pathway method was employed to introduce herbicide resistance bar gene into genome of Hibiscus cannabinus. Analysis was conducted on influences of different method for gene introductioo (micro-injecting ovary and dipping on stigma) and of different parameters,including DNA dosage and concentration, to setting rate of teceptive plants emergence rate of T 1 generation and transformation rate.The resultes showe that: Introduction methods showed different influence on setting rate of recepting plants and seeding emergence rate of T 1 generation,ovary injection method gave higher setting rate than stigma.But in seeding emergence, it was just in the oppositeside.Concentration and dosage of exogenic DNA showed in signi ficant influence on setting rate of recepting plants and seeding emergence rate of T 1 gemeration;PPT selective concentration for seeding of the tested variety was determined as 70μg/ml,which was adopted to scrape putative transgenic plants,12% of T 1 plants were resistant to PPT,and the putation transgenic plants rate was positived related to the concentration and dosage of introduced DNA. Ovary injection gave relatively higher transgenic rate than stigma dipping method.