摘要:采用常规分离培养方法从霉变烟叶和正常烟叶中分离细菌,扩增16S rDNA片段,用限制性内切酶MaeⅠ和HaeⅢ对PCR产物进行限制性酶切长度多态性分析,根据酶切图谱进行并菌后,对不同PCR产物进行测序,确定分离出菌株的种属关系,构建系统发育树.结果表明,霉变烟叶中细菌数量(7.0333×108CFU/g)大约是正常烟叶(7.1193×106CFU/g)的100倍.从霉变烟叶中获得11个属15个种,其中特有种9个(Staphylococcus sciuri, Staphylococcus equorum subsp.equorum,Oceanobacillus oncorhynchi subsp.Incaldanensis, Pseudomonas alcaligenes,Pseudomonas geniculata,Corynebacterium casei,Tetrathiobacter kashmirensi,Brevundimonas diminuta, Erwinia sp.);正常烟叶中获得10个属15个种,特有种9个(Bacillus aerophilus,Bacillus licheniformi,Bacillus altitudini,Bacillus clausii, Enterobacter pulveris,Stenotrophomonas maltophilia,Cellulosimicrobium cellulans, Aquabacterium parvum,Sphingopyxis aejonensis),两种烟叶中的共有种6个(Bacillus pumilus,Bacillus subtili,Staphylococcu aprophyticus, Acinetobacter johnsonii, Cronobacter sakazakii, Novosphingobium panipatense).