The invention discloses identification and an application of a tomato SolyWRKY54 transcription factor in regulating and controlling tomato yellow leaf curl virus. According to the invention, a WRKY transcription factor gene SolyWRKY54, which responds to the tomato yellow leaf curl virus, is identified from tomatoes; protein coded by the gene belongs to III sub-family in a WRKY transcription factorfamily. A semiquantitative PCR result shows that a susceptible variety, namely 'Jinpeng No.1', is higher than a disease-resistant variety, namely 'Zheza 301', in virus accumulation. Based upon fluorescent quantitative PCR analysis, it is indicated that the expression amount of the SolyWRKY54 gene is reduced in the disease-resistant tomato variety, namely 'Zheza 301'. Through a virus-induced genesilencing experiment, it further provides a negative regulation function of the SolyWRKY54 gene in a process of responding to the tomato yellow leaf curl virus. The SolyWRKY54 transcription factor gene, which is identified by the invention, is beneficial to analyzing and researching a resistance mechanism of tomatoes responding to the tomato yellow leaf curl virus; and the SolyWRKY41 transcriptionfactor is applicable to tobacco disease-resistance breeding.