穿刺胚加金刚砂超声波辅助农杆菌转化花生
The Transgenic Plants of Peanut Were Obtained by Puncture and Emery Grit Assisted Agrobacterium Mediated Transformation

DOI: 10.12677/HJAS.2019.912172, PP. 1204-1212

Keywords: 花生，穿刺胚，金刚砂，超声波，农杆菌，EPSPS
Peanut, Puncturing Embryo, Emery Grit, Ultrasonic, Agrobacterium, EPSPS

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Abstract:

论文尝试了一种穿刺胚加金刚砂超声波辅助农杆菌进行植物萌发种子基因转化的方法，其目的是直接以植物萌发种子作为农杆菌侵染受体，无需进行组织培养和植株再生过程。本实验，首先用清水浸泡花生种子12小时后，采用穿刺胚方法加金刚砂超声波处理萌动的种子，再将处理种子与农杆菌菌液(含AS)共培养24小时，然后播种。经过几周的喷洒草甘膦除草剂药液，田间筛选出具有良好抗性的植株，收获具有抗性植株的种子(T1代)。T1代继续培养，幼苗叶片提取DNA，进行PCR分子检测，通过PCR检测电泳图中条带证明获得了弱阳性转化植株。本论文最后建立了适合于花生的遗传转化系统，探讨了穿刺和超声波在花生转基因中的影响。
The paper relates to an ultrasonic assisted Agrobacterium mediated gene transformation method for germination seed, which aims to directly take the germination seed as the Agrobacterium infection receptor without the need for tissue culture and plant regeneration process. In this experiment, the seeds soaked in water for 12 hours, then the embryoes were punctured and ultrasonic treated with emery grit, and the treated seeds were cultured with Agrobacterium liquid (containing AS) for 24 hours, finally were sowed. In the process of seedling, glyphosate was sprayed for several times. Some plants has good resistant in field screening. The resistant plant seeds (T1) were cultured. But PCR gel electrophoresis strips of transformed plants were not very clear, proved weakly positive. Finally, a genetic transformation system suitable for peanut with emery grit was established, and the effect of puncturing and sonication in peanut was studied. And some plants with field resistance to glyphosate were obtained.

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