Co-Dominant InDel Marker for Identifying Burley Tobacco

To scientifically and efficiently identify and detect Burley tobacco materials contained in the raw materials of tobacco involved in the case, a specific molecular marker was developed in this study. We designed 2,718 pairs of TMb series InDel primers by bioinformatics analysis of large amounts of genomic data. K326 (P1), TN90 (P2), first-generation hybrids of the two parents (F1), the green leaf vein pool (B1) and the white leaf vein pool (B2), 5 genome materials were used to screen the Burly tobacco specific markers by bulked segregation analysis. One hundred tobacco materials were used to validate the specificity of screened markers. The results indicated that a co-dominant marker TMb0192 closely linked to the white tobacco leaf vein gene was obtained by bulked segregation analysis. The primer pair TMb0192 could effectively amplify a target band of 169 bp in 50 tobacco materials, while a target band of 181 bp in 50 non-Burly tobacco materials, pointing to the primer pair being specific to Burly tobacco. The marker TMb0192 could quickly and effectively identify and detect between Burly tobacco and non- Burly tobacco materials and improved scientificity and accuracy of the identification and detection of tobacco-making raw materials, which provide molecular evidence at the genome level for the tobacco industry's monopoly on anti-counterfeiting and anti-smuggling work.