以泸州老窖1、50、100和400年窖泥为研究对象,采用变性梯度凝胶电泳(DGGE)研究浓香型白酒窖泥古菌的群落结构及其多样性。DGGE图谱显示,同窖池窖泥多样性指数(H)和均匀度指数(EH)均以窖底泥较高。不同窖池同部位窖泥的H值差异较大,窖壁泥和窖底泥H值分别为2.54~2.86和2.76~3.23;EH值差异较小,且随窖龄延长而上升,分别为0.985~0.991和0.991~0.995。此外,同窖池窖泥的群落结构相似性系数(S C)水平较高,为0.36~0.74;其次是窖底泥,为0.42~0.63;窖壁泥为0.36~0.52。测序结果显示,与DGGE条带序列亲缘最近的种属均为产甲烷菌,归于甲烷短杆菌属(Methanobrevibacter)、甲烷杆菌属(Methanobacterium)、甲烷鬃菌属(Methanosaeta)和甲烷囊菌属(Methanoculleus)。其中,甲烷短杆菌属(Methanobrevibacter)在不同窖泥中占优势,优势度为7.23%~39.64%。实验结果表明,浓香型白酒窖泥古菌群落结构及其多样性因窖龄和空间分布的不同而存在差异,其中以同窖池古菌种类较相似,窖底泥群落结构较稳定;窖泥中蕴藏着产甲烷菌资源,具有进一步开发研究的价值。

In this study,the pit mud of Luzhou-flavour liquor which had been used for 1,50,100 and 400 years were taken from a factory affiliated to the Luzhou Laojiao Group. Eight mixed samples were sampled from the wall and bottom layers of the pit mud in the cellars. PCR-DGGE was employed to analyse Archaea community structure and diversity in the samples. DGGE analysis showed that both of the diversity index( H) and the evenness index( EH) of the bottom-layer sample was higher than those of the wall-layer samples from the same cellar. However,in different cellar,the discrepancy of H values between samples taken from the same position was significant,and the variation of H values of the wall and the bottom layers were between 2. 54 to 2. 86 and 2. 76 to 3. 23,respectively. In addition,with the increase of pit age,the EH values gradually increased from 0. 985 to 0. 991 in the wall-layer sample,and from 0. 991 to 0. 995 in the bottom-layer sample. Moreover,those samples taken from the same cellar exhibited a higher level of the similarity coefficient( S C),ranging form 0. 36 to 0. 74,and the S C values of samples from the wall and the bottom layers were between 0. 42 to 0. 63 and 0. 36 to 0. 52,respectively.