Abstract: To clarify the main species and infection types of tobacco viral diseases in Anshun tobacco growing areas of Guizhou Province, 306 diseased samples with symptoms such as mosaic, necrotic veins, abnormal shapes, lightning spots, and yellow spots were collected from Anshun tobacco growing areas in 2023. RT-PCR was performed using specific primers for 5 major viral diseases of tobacco mosaic virus (TMV), cucumber mosaic virus (CMV), potato Y virus (PVY), tobacco vein banding mosaic virus (TVBMV) and chilli veinal mottle virus (ChiVMV), and the detection rates of the viruses were calculated and the mixed infection types were analyzed. The results showed that the total detection rate of the viral diseases was 89.87%, and among all the diseases, TMV (81.70%) ranked the highest, followed by ChiVMV (67.97%), and CMV ranked the lowest (16.34%). The main type of infection was mixed infection with a detection rate of 68.30%. Among them, ChiVMV+TMV was the main type with a detection rate of 22.88%, followed by ChiVMV+TMV+TVBMV, ChiVMV+TMV+PVY, and ChiVMV+TMV+CMV with detection rates of 11.11%, 10.46%, and 9.15%, respectively. TMV and ChiVMV were the dominant viral species of the tobacco viral diseases from the area, mixed infection occurred frequently and ChiVMV+TMV was the main mixed type.