Fuente: WIPO "tobacco agriculture" The invention discloses a tobacco genome gap filling method, device and equipment based on Poretols and Nextdenovo software and a medium, and the method comprises the following steps: S1, adopting the Poretols software under default parameter setting to trim and filter data to be processed to obtain processed data; s2, setting Nextdenovo software according to the core parameters shown in the table 1, and obtaining an initial assembly result; and S3, identifying an overlapping region of the genome gap of the initial assembly result to obtain filled tobacco genome data. By adopting the method to fill up the tobacco genome gaps, the genome quality is greatly improved, complete tobacco genome data without gaps or with few gaps is obtained, and the obtained complete tobacco genome data is beneficial to improving the accuracy of the obtained research result when being used for subsequent research.