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Cucumber mosaic virus (CMV) and Alternariaspp. are among important and devastating plant pathogens with a wide host range. However, mixed infection of CMV and Alternariaspecies, is more common in greenhouses and fields. 2bis an important CMV gene that plays a critical role in CMV-host interactions and bZIP60is among the plant genes that affect by biotic stresses especially some plant viruses. In this research, the effect of silencing of 2b on bZIP60expression and resistance to A. atrawas evaluated. The results data showed induction of bZIP60in CMV infected plants by 18 folds while six-fold downregulation by A. atracompared to the healthy plants and also it showed a reduction in bZIP60expression to 1/2 fold in mixed infection plants in comparison with healthy plants. But transiently expression of 2b silencing gene construct in CMV infected plants reduced Alternaria necrosis leaf spot area and reduced the expression of bZIP60to 40 fold in mixed infected plants. However, the results of this research suggested the application of CMV-2b silencing construct for induction of resistance to both pathogens.