Fuente: WIPO "tobacco agriculture" The invention discloses an editing method for seamless insertion or replacement of long gene segments in tobacco. Preparing tobacco protoplast from the tobacco leaves; carrying out gene editing on the tobacco protoplast by utilizing CRISPR/Cas9 and donor DNA, and carrying out insertion and replacement of an exogenous fragment; and regenerating the tobacco protoplast subjected to gene editing into a plant. According to the method disclosed by the invention, the resistance gene N'in the tobacco is modified by finding out a core segment, responsible for TMV resistance, of the N 'alta gene from wild tobacco, and replacing a corresponding region of the tobacco N' gene, so that a cultivated tobacco material with TMV resistance character is obtained.