Abstract: Fungi levels in four tobacco cultivars in Guizhou province were tested on tobacco seedlings that incubated on sterile filter paper. The fungi were isolated and identified. Results showed that fungi levels in K326, Guiyan 4, Jiucaiping 2, and Nanjiang 3 were 15%, 77.78%, 21.67% and 61.39%, respectively. The fungi in those four cultivars included Alternaria sp., Phomopsis sp., Acremonium sp., Zopfiella sp., Aspergillus sp., Fusarium sp., Penicillium sp., Gibberella sp., Cladosporium sp., Bipolaris sp., and Stagonosporopsis cucurbitacearum. Forty isolates were isolated all together, in between about 22 different fungi. However, Alternaria sp., Cladosporium sp., Fusarium sp. and Gibberella sp. were the dominant fungi. Alternaria sp. were found on all four cultivars; Fusarium sp. were found on Guiyan 4, Jiucaiping 2 and Nanjiang 3, and was dominant on cultivar Nanjing 3. Cladosporium sp. was only found on cultivar Guiyan 4. The number of fungi and number of species were the highest in cultivar Guiyan 4 and Nanjiang 3. Those findings provide important information for tobacco seed production and process.