Tobacco producers have experienced problems with aerial rooting (negative gravitropism) in greenhouse grown seedlings. An experiment using eleven seed sources from different cultivars, different color of seed coating and several suppliers of coated seed, were grown in two soilless media. One soilless media resulted in significantly greater aerial rooting as compared to a standard soilless media. Cultivar NC72 produced fewer aerial roots as compared to NC71 and K346. Blue seed coating had less aerial rooting, while all sources of seed coating were similar in aerial rooting. Primed tobacco seed did not show a clear influence on aerial rooting. The difference in aerial rooting between the media is not understood at this time. This study illustrated that some seed sources also exhibited germination and growth problems when planted in some media. Past research has shown numerous factors including aeration, media, cultivar, seed priming, and light can influence aerial rooting.