摘要
利用基因枪法将含有３个不同基因（ｈｐｔ，ｇｕｓ和ｇｎａ）的质粒ｐＷＲＧ１５１５和ｐＲＳＳＧＮＡ１共同转化粳稻品种鄂晚５号成熟胚诱导的愈伤组织．共再生出３５株独立转基因植株．ＰＣＲ／Ｓｏｕｔｈｅｒｎ印迹法分析发现，８３ ％的转基因植株含有所有３个外源基因．Ｗｅｓｔｅｒｎ印迹法分析发现 ７９％的含 ｇｎａ基因的转基因植株以不同水平表达ＧＮＡ。遗传分析证实外源基因在转基因植株后代中以孟德尔方式遗传．从其Ｒ１代亲本为孟德尔３：１方式遗传的Ｒ２代中，鉴定出２个含有所有３个外源基因的独立转基因植株纯系．这些纯系具有相似的外源基因表达量．褐飞虱喂养试验表明，这些纯系对褐飞虱具有显著的抑制作用．这些揭飞虱抗性提高的转基因纯系将应用于水稻抗虫育种中．实验证明，通过遗传转化和筛选可获得含在农业上有应用价值基因的转基因水稻纯系．
Mature seed-derived callus from japonica rice cv. Ewan 5 was co-transformed with pWRG1515 and pRSS- GNA1 containing three different genes (hpt, gus and gna) via particle bombardment. Thirty-five independent trans- genic rice plants were regenerated. Eighty-three percent of all transgenic plants contained all the three genes, revealed by PCR/ Southern blot analysis. Western blot analysis revealed that 79 % of gna -containing transgenic plants ex- pressed GNA at various levels. Genetic analysis confirmed Mendelian segregation of transgenes in progeny. From R2 generations with their R1 parent plants showing 3:1 Mendelian segregation ratios, two independent homozygous lines containing all the 3 transgenes were identified. The homozygous lines showed the uniform expression of all the trans- genes and had significant inhibition to brown planthopper by feeding test. These BPH-resistance enhanced homozygous lines will be incorporated into rice insect resistance breeding program. These results demonstrate that homozygous transgenic rice lines containing an agronomically useful gene can be obtained via genetic transformation and through ge- netic analysis-based selection.
出处
《复旦学报（自然科学版）》
CAS
CSCD
北大核心
2001年第5期477-482,共6页
Journal of Fudan University：Natural Science
基金
Supported by the Rockefeller Foundation, USA (Grant No. RF97001 #594)
Supported by China Na-tional High-Tech "863"program (
