Anatabine is a minor alkaloid found in tobacco and constitutes about 3 % of the total alkaloids. It is a precursor for N-nitrosoanatabine (NAT), one of four tobacco-specific nitrosamines (TSNAs) found in cured tobacco leaf. Depending on the tobacco type, agronomic practices, and environmental conditions, NAT content in the lamina ranges from 35-50 % of total TSNAs. Therefore, reduction or elimination of anatabine content in tobacco could significantly reduce total TSNA content in tobacco products. Since regulation of anatabine synthesis is not completely understood, identifying molecular markers associated with the anatabine trait will be useful in identifying and/or developing tobacco varieties that are low in anatabine and NAT.

A novel group of mutant lines has been developed and characterized that exhibit 80-95 % reduction of anatabine while maintaining normal levels of other tobacco alkaloids. Microsatellite markers closely linked to low anatabine were identified and used to genotype F₂ populations, revealing that the mutation is recessive. The low-anatabine trait was mapped to linkage group six, 0.3 centimorgan (cM) away from PT60878 on the high-density tobacco map. The results from this study will be useful in isolating the locus that regulates anatabine synthesis through map-based cloning, as well as identifying candidate gene assays and will help in elucidating the late stages of the alkaloid biosynthetic pathway.