农杆菌介导CBF基因转化木纳格葡萄的分析

Analysis of Genetic Transformation of Munage Grape with CBF Gene Mediated by Agrobacterium

为了建立木纳格葡萄稳定、可行的遗传转化体系,本研究以木纳格葡萄的叶片和叶柄为受体材料,分析卡那霉素(Kan)、头孢霉素(Cef)浓度以及筛选方式对木纳格葡萄遗传转化效率的影响,且通过农杆菌介导法转化CBF基因,并对获得的抗性植株进行PCR检测。结果表明,以叶片、叶柄为外植体,Cef的最适浓度为300 mg/L,Kan的临界浓度为5 mg/L,延迟筛选法更适合遗传转化,可减少抑制作用。最终共获得12株抗性植株,经过PCR检测发现5株为阳性转基因植株,初步证明CBF基因已经导入到木纳格葡萄中,遗传转化体系建立成功。该遗传转化体系的建立,为葡萄的基因改良提供了技术支持。

In order to establish a stable and feasible genetic transformation system of Munage grape,in this study,we used the leaves and petioles of Munage grape as materials,and analyzed the effect of genetic transformation efficiency on the Munage grape by kanamycin concentration,cephalosporin concentration and selection methods,then CBF gene were transformated by Agrobacterium tumefaciens,and the resistant plants were detected by PCR.The results showed that when we used the leaves and petioles as explants,the optimal concentration of cephalosporin was 300 mg/L,the critical concentration of Kanamycin was 5 mg/L,and the methods of delayed screening was more suitable for genetic,which could reduce inhibition.Finally,we obtained 12 resistant plants,and 5 positive transgenic plants were detected by PCR,which means CBF gene has been introduced into Munage grape,and the genetic transformation system has been established successfully,which provides technical support for grape genetic improvement in the future.