The future field-based production of transgenic tobacco for traditional or new applications will rely on robust containment measures based on sound risk assessment. Information concerning the persistence of tobacco seeds in the environment is an essential contributor to that risk analysis. Using three different commercial tobacco cultivars we examined the viability of seed which had been buried at four different soil depths in the field, sampling annually over a 6-year period. Our findings indicate that tobacco seed can persist in the natural environment over this time frame, and suggest that monitoring for volunteer plants should extend beyond 1 year following the production of transgenic tobacco outdoors.