Abstract: In order to determine the suitable seedling density on nursery trays for Enshi tobacco producing area for the Tobacco Plantlet Transplanting In Well Cellar (TPTWC) using floating seedling cultivation, we tested nursery trays with 153 holes, 200 holes, 288 holes and 500 holes for their effects on seedling growth and development, tobacco yield and quality. The results showed that nursery trays with 153 holes, 200 holes and 288 holes were all able to produce tobacco seedlings with good quality, healthy enough to meet the requirements of TPTWC. The 153 hole and 200 hole trays however, increased the cost of seedling productio. Seedling cultivation with the 500 hole tray had the lowest cost but the quality of tobacco seedlings was the worst. Overall, the 288 hole nursery tray had relatively low nursing cost, the best economic benefit, significantly less labor employment, reduced production cost, increased production efficiency, and more in line with the situation in Enshi. It is worth of popularizing and applying in tobacco production.