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中国油料作物学报 ›› 2014, Vol. 36 ›› Issue (5): 586-.doi: 10.7505/j.issn.1007-9084.2014.05.005

• 遣传育种 • 上一篇    下一篇

抗病基因KR3在大豆中的转化研究

王浩铭(1987-),男,广东揭阳人,硕士研究生,主要从事大豆转基因工作,E-mail: wanghaoming1987@gmail.com;§贺红利具有同等贡献   

  1.  
    1.浙江大学农业与生物技术学院,浙江 杭州,310058; 2.东北师范大学分子表观遗传学教育部重点实验室,吉林 长春,130024
  • 出版日期:2014-10-28 发布日期:2015-01-12
  • 通讯作者: 唐桂香(1966-),E-mail: tanggx@zju.edu.cn
  • 基金资助:

    国家转基因生物新品种培育科技重大专项(2013ZX08004-004-008);浙江省重大科技专项(2012C12902);国家自然科学基金(31071443);浙江省科技厅公益技术(2012C32001)

KR3 disease resistance gene transformation in soybean

  1. 1.Institute of Crop Science, College of Agriculture and Biotechology, Zhejiang University, Hangzhou Zhejiang 310058, China; 2. Molecular Epigenetics of the Ministry of Education (MOE), Northeast Normal University, Changchun Jilin 130024, China
  • Online:2014-10-28 Published:2015-01-12

摘要:

采用农杆菌介导子叶节转化法将抗性基因KR3导入大豆中,共转化386个外植体,获得转基因生根苗15株,经草丁膦涂抹、Bar试纸条和PCR鉴定,最终获得12株KR3转基因阳性苗,农杆菌介导转化效率3.11%。RT-PCR结果显示转基因植株KR3基因表达量显著高于对照(非转基因植株);花叶病毒摩擦接种表明,转基因植株对花叶病毒能产生一定抗性,而非转基因植株则明显感病;摩擦接种三周后ELISA检测进一步表明在所检测的12株T1KR3转基因植株中未检测出SMV,而非转基因大豆植株SMV检出率为75%。研究结果表明,提高KR3基因的表达可以提高大豆的抗病毒性。

关键词: 大豆, 农杆菌介导转化, KR3基因, 花叶病毒病

Abstract:

 To improve soybean mosaic virus (SMV) resistance, KR3 gene was transformed using Agrobacterium tumefaciens mediated transformation method. Result showed that 12 transgenic soybean plants from 386 transformed explants were identified by coating leaves with glufosinate-ammonium, detecting with Bar protein quick dip stick and PCR analysis. The final soybean transformation efficiency was 3.11%. RT-PCR results showed that the KR3 expression in transgenic plants was higher than that of non-transgenic plants. 7 days after mechanical SMV inoculation, transgenic plants showed higher resistance. By DAS-ELISA, SMV detection rate was 75% in non-transgenic plants, and was zero among 12 positive transgenic T1 plants. The results suggested that KR3 expression in soybean improved the resistance to SMV.

Key words: Glycine max L., Agrobacterium tumefaciens mediated transformation;KR3 gene, Soybean mosaic virus