Uncultivated bacterial diversity and phylogenetic analysis in flue-cured tobacco
There are lots of uncultivated bacteria in flue-cured tobacco, the species and roles of these bacteria in aging process of tobacco are unknown. Bacterial diversity of the flue-cured tobacco K326 (C3F) was studied using the culture-independent method and 16S rDNA phylogenetic analysis in this study. Total DNA of bacteria in the flue-cured tobacco was isolated using SDS-CTAB method. 16S rDNA was amplified and RFLP analysis was carried out for identification and phylogenetic analysis of bacteria in flue-cured tobacco K326. Results showed that Pseudomonas sp. and Bacillus sp. were the primary groups in tobacco K326. Some species, such as Alcaligenes sp. and Phyllobacterium sp., which were not found in pure culture, were identified by culture-independent method. These results provided a basis for studying the roles of uncultivated bacteria in aging process and gave new ideas and methods for systematically analyzing the microorganisms' diversities in the flue-cured tobacco.