激发子基因peaT1转化三生烟及其对TMV抗性的提高

Transformation of Elicitor-encoding Gene peaT1 in Tobacco (Nicotianatobacum cv.Samsun NN) and Enhancement of Resistance to TMV

通过根癌农杆菌(Agrobactrium tumefaciens)介导转化法,将含有激发子基因peaT1的植物表达载体pCAM-BIA2300-G4AS-peaT1转化三生烟,获得了转基因烟草植株。用PCR检测确认了阳性转化株,用Southern杂交、RT-PCR和Western杂交进一步证实了peaT1基因的整合、转录和表达。对T1代转基因阳性株进行TMV接种试验,结果显示,与非转基因对照相比,表达peaT1的烟草叶片枯斑数量减少,表明蛋白激发子基因peaT1的表达提高了转基因烟草对TMV的抗性。

Plant expression vector pCAMBIA2300-G4AS-peaT1 harboring elicitor-encoding gene peaT1 was constructed,then was transformed into tobacco（ Nicotiana tobacum cv. Samsun NN）by Agrobactrium tumefaciens method. The kanamyein-resistant regenerated plants were confirmed to be electropositive by PCR. Integration, transcription and expression of the peaT1 were further confirmed by Southern blotting,RT-PCR and Western blotting, respectively. Then, transgenic tobacco plants of T1 generation were inoculated with TMV, compared with TMV-infected wild-type SNN plants ,peaT1-expressed plants displayed reduced hypersensitive-response lesions ,the result idieated that the expression of peaT1 in tobacco plants improved the resistance to TMV.