根癌农杆菌介导油菜CBF1基因转化拟南芥

Transformation of CBF1 Gene from Rape into Arabidopsis thaliana Mediated by Agrobacterium tumefaciens

[目的]为油菜CBF1基因应用于作物抗逆遗传改良提供理论依据。[方法]采用根癌农杆菌介导法将油菜CBF1基因转入拟南芥,筛选抗性转基因拟南芥植株,进行PCR检测和GUS染色。[结果]与不抗潮霉素的野生型拟南芥幼苗相比,对潮霉素具有抗性的转基因拟南芥幼苗的叶片比较大,根特别长。PCR检测表明38株抗性拟南芥植株中有29株能扩增出与目的基因大小一致的条带,阳性率为76.3%,而野生型拟南芥植株没有扩增出相应条带。阳性PCR扩增产物的测序结果表明油菜CBF1基因已经转入到拟南芥。GUS染色表明有22株转基因拟南芥植株呈现阳性。[结论]油菜CBF1基因可以整合到拟南芥基因组并能稳定表达。

[Objective] The purpose was to supply theoretical basis for applying CBF1 gene from rape on the genetic improvement of crop resistance. [Method] CBF1 gene from rape was transformed into Arabidopsis thaliana by Agrobacterium tumefaciens-mediated method to screen its transgenic plants with resistance and conduct PCR detection and GUS staining. [Result] Compared with wild A. thaliana seedlings without resistance to hygromycin, the leaf blades of its transgenic seedlings with resistance to hygromycin were bigger and their roots were especially long. PCR detection showed that among 38 A. thaliona plants with resistance, 29 plants could amplify the bands with sizes according with aim gene and their positive rate was 76.3%, but the wild A. thaliana plants didn＇t amplify corresponding bands. The sequencing results on the amplified products of positive PCR showed that CBF1 gene from rape had been transformed into A. thaliana. GUS staining showed that 22 transgenic A. thaliana plants showed positive. [Conclusion] CBF1 gene from rape could be integrated into A. thaliana genome and stably expressed.