Construction and Application of Specific Fingerprint of Flue-cured Tobacco Variety Zhongyantexiang 301

Fingerprinting is an effective method for verifying the authenticity of plant varieties, providing crucial technical support for the expansion of cultivation and the industrial application of these varieties. In this study, single nucleotide polymorphism(SNP) variations were analyzed across 33 tobacco germplasm resources, including the new flue-cured tobacco variety Zhongyantexiang 301, taking advantage of the whole genome resequencing technology. This analysis identified a total of 268 specific SNP loci unique to Zhongyantexiang 301. After screening 62 tobacco germplasm resources from various market types—flue-cured, cigar, oriental, burley, and sun-cured tobacco—21 effective SNP markers were preliminarily developed. Field-fresh and primary flue-cured leaves of Zhongyantexiang 301, along with five other main cultivars from the Shandong growing area, were collected between 2022 and 2023 for variety authenticity identification using the 21 SNP markers. The results showed that 18 of these markers successfully detected the specific SNP loci of Zhongyantexiang 301 in all tobacco samples, demonstrating their effectiveness in authenticating the variety.