不同窖龄窖泥细菌的系统发育多样性分析.PDF

※生物工程 食品科学 2013, Vol.34, No.11 177 不同窖龄窖泥细菌的系统发育多样性分析 1,2 2,* 2 2 王明跃 ，张文学 ，王海英，刘超兰 (1.阜阳职业技术学院，安徽 阜阳 236031；2.四川大学轻纺与食品学院，四川 成都 610065) 摘  要：提取20 、50 、150a窖龄窖泥样品总DNA ，构建窖泥细菌16S rDNA文库，经过16S rDNA扩增片段的限制 性酶切分析(ARDRA) 、DNA序列测定和系统发育关系分析，对不同窖龄窖泥细菌群落组成与多样性进行研究。结 果表明：398个细菌阳性克隆子分成37个可操作分类单元(OTUs) ，系统发育分析归为6个门，优势顺序为：厚壁菌 门(Firmicute)65.1%、拟杆菌门(Bacteroidetes)2 1.4%、绿弯菌门(Chloroflexi)4.8%、放线菌门(Actinobacteria)3.8%、 未分类细菌(Unclassified Bacteria)3.9%和变形菌门(Proteobacteria) 1.0%，窖泥中的大多数克隆子(73.9%)与未培养 细菌亲缘关系近。20a窖泥优势菌是Syntrophomonas 、Petrimonas 、Proteiniphilum ，50a和150a龄窖泥优势菌为 Natronoanaerobium和Unclassified Clostrida 。随着窖龄的增加，窖泥细菌多样性指数呈增加的趋势，从20～50a之间 变化幅度较大，50a和150a窖泥的菌群结构最为相似。本研究初步揭示了不同窖龄窖泥细菌系统发育多样性与窖泥 老熟老化的关系。 关键词：窖泥；16S rDNA序列；克隆文库；系统发育；多样性 Analysis of Bacterial Phylogenetic Diversity of Pit Muds with Different Cellar Ages 1,2 2,* 2 2 WANG Ming-yue ，ZHANG Wen-xue ，WANG Hai-ying ，LIU Chao-lan (1. Fuyang Vocational and Technical College, Fuyang 236031, China ；2. College of Light Industry, Textile and Food Engineering, Sichuan University, Chengdu 610065, China) Abstract ：Total DNAs were extracted from pit muds with different cellar ages, 20, 50 a and 150 a, respectively and were used for constructing 16S rDNA clone libraries. The bacterial composition and diversity from the pit muds were investigated by amplified rDNA restriction analysis (ARDRA), DNA sequencing and phylogenetic analysis. The results showed that 398 clones from the 16S rDNA library fell into 37 operational taxonomic units (OTUs) and 6 phyla including