基于RAPD技术对窖泥梭菌近似菌株的分类研究

Using Random Amplification of Polymorphic DNA to Classify Clostridium Strains Isolated from Pit Mud

对实验室已经进行16S rDNA鉴定的来源于浓香型白酒窖泥的近似梭菌进行随机扩增多态性DNA标记(random amplified polymorphic DNA,RAPD)分型,19株梭菌基于16S rDNA序列分析可以分为3类,分别与拜氏梭菌(Clostridium beijerinckii)、丁酸梭菌(Clostridium butyricum)和广西梭菌(Clostridium guangxiense)近似。结果表明,通过筛选获得4个具有良好的分型效果的随机引物,基于其RAPD扩增图谱的0/1矩阵分析可将这19株梭菌分成10类。其中与拜氏梭菌类菌群近似的11株菌株分为6类,包括2-1类、4-30类、C2-2类、G3-1类、G3-5类和JL3-1类;将与丁酸梭菌类菌群近似的6株菌分为2类,包括3-3类和JL3-4类;将广西梭菌类菌群近似的2株菌分为2类包括B2-1类、F1-1类。

Randomly amplified polymorphic DNA(RAPD)was used to distinguish between similar strains of Clostridium isolated from the pit mud of strong-flavor Baijiu fermentation cellars,based on 16S rDNA sequences.Nineteen similar Clostridium strains were divided into the following three categories based on the analysis:those similar to Clostridium beijerinckii,C.butyricum,or C.guangxiense.Four random primers with good distinguishing capabilities were identified by screening experiments.Based on the 0/1 matrix analysis of RAPD amplification electrophoretograms,the 19 Clostridium strains were grouped into 10 categories.Eleven of these strains were similar to C.beijerinckii and were divided further into the following six categories:2-1,4-30,C2-2,G3-1,G3-5,and JL3-1.Six strains were similar to C.butyricum and divided into two categories,3-3 and JL3-4,while two strains similar to C.guangxiense were divided into the categories B2-1 and F1-1.