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金刚砂辅助农杆菌进行玉米基因转化研究
Transgenic corn research by Emery-assisted Agrobacterium-mediated Transformation
投稿时间:2019-01-01  修订日期:2019-07-06
中文关键词:农杆菌  玉米  金刚砂  EPSPS基因
英文关键词:agrobacterium  corn  emery  EPSPS gene
基金项目:广东大学生科技创新培育专项资金“金刚砂辅助农杆菌进行玉米基因转化创新方法研究”( pdjh2017b0257)。
作者单位E-mail
储坤 仲恺农业工程学院生命科学院 liangxuelian2005@sina.com 
聂小熊 仲恺农业工程学院生命科学院  
冯华鲒 仲恺农业工程学院生命科学院  
韩瑰丽 仲恺农业工程学院生命科学院  
梁雪莲 仲恺农业工程学院生命科学院 liangxuelian2005@sina.com 
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中文摘要:
      近10年来,农杆菌介导的遗传转化具有稳定、高效等优点,被广泛应用到育种和分子生物学研究中,发展迅猛。但农杆菌对玉米的转化作用有限,运用传统的农杆菌转化方法处理玉米种子时,存在操作复杂、转化率低下等缺点。生物学家发现,利用超声波处理萌动玉米种子可以提高农杆菌介导的转化率,取得了一定的研究成果。本研究对前人超声波处理方式进行了改进,在超声波处理时,加入金刚砂以辅助EPSPS基因转导到玉米基因中。800粒处理种子播种出苗93株,玉米苗经大田glyphosate筛选有27株具有抗性,继续进行PCR电泳检测显示23株阳性,初步证明EPSPS基因已经转化到玉米植株中,转化率为29%。本研究初步证明,金刚砂辅助处理玉米基因转化具有可行性,利于生产实际操作。
英文摘要:
      Over the past decade, agrobacterium mediated genetic transformation has been widely used in breeding and molecular biology research because of its stability and efficiency. However, when using traditional agrobacterium mediated transformation in corn, the transformation of agrobacterium tumefaciens is limited, and it has many disadvantages, such as complex operation, low conversion rate, and so on. Biologists have found that the use of ultrasonic-assisted agrobacterium-mediated transformation in germinating corn seed can improve the agrobacterium mediated transformation rate. In this study, the ultrasonication-assisted Agrobacterium treatment method was modified with the addition of emery to increase the treatment intensity to the apical meristem cells of germinating seeds. The emerged 93 plants from 800 control seeds were sprayed three times with glyphosate solution, and 27 of them survived, their leaves were taken with a PCR electrophoresis test and 23 shown positive, which suggested that epsps gene was introduced into the maize cultivar, and the transformation rate was 29%. Finally, which has been shown that the method has the advantages of circumventing the tedious tissue culture procedures and being easy to apply.
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