水稻花粉管通道法导入含Xa21总基因的研究简报

• 作 者: 宁波市农科所基因转化课题组
• 出 处: 宁波农业科技
• 发表日期: 1996年01期
• 关键词: 花粉管通道法 Xa21 总基因 抗白叶枯病 转化处理 DNA 基因克隆 分子杂交 基因片段 外源

采用花粉管通道法.将含Xa21基因(广谱抗白叶枯病基因)的水稻(oryza saztiva)品种1188的总DNA及PBI121质粒DNA导入丙1067、宁67、繁7、宁波2号、293、浙733和舟903等七个品种(系),收获转基因的当代水稻种子并进行大田选育。将外源DNA含PBI121的转化处理后代种子发牙,在14天苗龄时分别提取水稻幼苗总DNA而后经多种限制性内切酶酶切,并以gus基因片段为探针进行Southern blot杂交分析,证实了外源gus基因已整合到水稻丙1067等7个品种(系)中。另外,用六针刺孔法和剪叶法白叶枯病接种试验,D2代转基因水稻种子的氨基酸测定及考种结果表明外源抗白叶枯病Xa21基因已整合到丙1067等7个品种(系)中。

The total DNA and PBI121 plasmid DNA of oryza saztiva cultivar 1188 containing Xa21 gene (broad-spectrum bacterial leaf blight resistance gene) was introduced into C10677, Ning67, Zhong7, Ningbo2, 293 , Zhejiang 733 and Zhou 903 seven varieties (lines), the harvest of genetically modified contemporary rice seeds and field selection. The exogenous DNA containing PBI121 transformation treatment offspring tooth hair, respectively, at 14 days of seedling age were extracted total DNA of rice seedlings and then digested by a variety of restriction endonucleases and gus gene fragment as a probe for Southern blot hybridization Analysis confirmed that the exogenous gus gene has been integrated into rice C107 7 varieties (lines). In addition, with the six-needle puncture and leaf-cut bacterial blight inoculation test, the amino acid determination of D2 transgenic rice seeds and test results showed that Xa21 gene of resistant to bacterial blight was integrated into 7 varieties (Department) in.