番茄Pti基因瞬时表达与互作检测

Transient expression of tomato Pti genes and their interaction detection

丁香假单胞菌所导致的细菌性斑点病是影响番茄产量和品质的重要病害,而番茄色氨酸苏氨酸激酶(Pto)是植物识别、防御这一病原菌的重要抗性蛋白。文章通过克隆番茄Pti(Pto interaction protein)基因Pti4、Pti5和Pti6,分别构建植物表达载体,利用农杆菌介导方法在烟草叶片中进行瞬时表达和western blotting检测,并进一步利用免疫共沉淀技术,在植物体系中检测了它们与番茄Pto蛋白的相互作用。实验结果表明,利用所构建Pti4、Pti5和Pti6基因植物表达载体,在烟草中获得了预期大小Pti4、Pti5和Pti6蛋白,并在植物体系中验证了它们与Pto蛋白的相互作用。该工作为深入研究Pti基因在番茄分子免疫途径的功能奠定了基础。

Bacterial speck caused by Pseudomonas syringae pv.tomato is an important disease affecting tomato yield and quality.Pto,a resistance protein,plays a critical role in recognizing and defending against this plant pathogen.In this paper,tomato Pti(Pto interaction protein)genes,Pti4,Pti5and Pti6,were cloned and constructed for plant expression vectors,respectively.Transient expression and western blotting of the genes were performed in tobacco leaves by agrobacterium-mediated method,and the interactions between Pti and Pto were detected by means of co-immunoprecipitation in plant system.The results indicated that expected bands of Pti4,Pti5and Pti6proteins were expressed and their interaction with Pto was confirmed in plants by using the constructed plant expression vectors of Pti4,Pti5and Pti6.This study will provide a foundation for further investigations on the function of Pti genes in tomato molecular immunity pathway.