Fuente: WIPO "tobacco agriculture" The invention discloses a group of molecular markers for breaking linkage drag of a TMV (Tobacco Mosaic Virus) resistance gene N upstream (5'-end) and application thereof. Serial numbers of the molecular markers for breaking the linkage drag of the TMV (Tobacco Mosaic Virus) resistance gene N upstream (5'-end) are TMn007, TMn025, TMn027, TMn085, TMn113, TMn131, TMn163, TMn183, TMn203, TMn223, TMn226, TMn233, TMn245, TMn249, TMn288, TMn302, TMn306 and TMn323-N. The group of molecular markers disclosed by the invention has the characteristics of stability, reliability, simplicity and high sensitivity and can be used for detecting and judging fragment breaking information of the linkage drag at different positions of the tobacco N gene upstream (5'-end), and a new TMV-resistant tobacco variety which contains an N gene and can fully break or partially break adverse linkage drag influences at different positions is cultivated.