16SrDNA克隆文库法分析拜城油鸡盲肠细菌多样性研究  

Study on Baicheng Soy Sauce Chicken Caecun Bacterial Diversity through 16SrDNA Clone Library Analysis

作　　者：陈荣[1] 刘利林[1] 格明古丽·木哈台[1] 王永[1] 

机构地区：[1]塔里木大学动物科学学院,新疆生产建设兵团塔里木畜牧科技重点实验室,新疆阿拉尔843300

出　　处：《家畜生态学报》2017年第2期18-21,共4页Journal of Domestic Animal Ecology

基　　金：兵团塔里木畜牧科技重点实验室开放课题(HS201205);国家自然科学基金(31160521);塔里木大学校长基学校长基金(TDZKGG201504)

摘　　要：为了研究拜城油鸡盲肠细菌物种多样性,本研究采用基于16SrDNA基因序列系统发育分析的免培养方法进行细菌物种多样性分析,以盲肠内容物样品总DNA为模板,利用细菌通用引物扩增16SrDNA基因,构建16SrDNA基因克隆文库,并对文库中的插入序列进行RFLP分析。结果表明:随机挑选的252个克隆为阳性克隆,阳性率为65.7%,经过RFLP筛选出95个不同图谱的重组克隆并测序,比对分析后发现:拜城油鸡盲肠细菌分属于厚壁菌门(Firmicutes)(65.26%)、拟杆菌门(Bacteroidetes)27.37%)和变形菌门(Proteobacteria)(4.21%)。在纲水平有梭菌纲(Clostridia)、拟杆菌纲(Bacteroidia)和ε-变形菌纲(Epsilonproteobacteria)相对含量最多,分别占总菌的64.21%、26.32%和4.21。在科水平上以瘤胃菌科(Ruminococcaceae)(21.05%)、毛螺菌科(Lachnospiraceae)(20%)和理研菌科(Rikenellaceae)(12.63%)相对含量最多,为优势科。在属水平上仅有31.58%的序列可以鉴定到属,Lachnoclostridium的相对含量最多,占总菌的12.63%,其次为另枝菌属(Alistipes)和瘤胃梭菌属(Ruminiclostridium)。拜城油鸡盲肠内容物中的细菌组成具有丰富的多样性,并有新细菌分类单位的潜在资源,值得进一步进行开发研究。In order to study Baicheng chicken cecal bacterial species diversity, 16S rDNA gene sequence for phylogenetic analysis with free culture method for analysis of bacterial species diversity was conducted in the present research. With total DNA of cecum sample as template and universal primer of 16SrDNA gene for amplification,a 16s rDNA clone library was established,and the insertion sequence were analyzed with RFLP. The results showed that 252 randomly selected clones were positive, with a rate of 65. 7%. RFLP screened 95 different profiles of recombinant clone and sequenced to find that,three phyla,4 classes, 9 families and 8 genera were identified via taxonomic summary. The dominant phyla among them were Firmicutes （65.26%） ,Bacteroidetes （27. 37%） and Proteobacteria （4. 21%）. The most abundant classes were Clostridia,Bacteroidia and Epsilonproteobacteria, accounting for 64. 21 % , 26. 32% and 4. 21, respectively. Ruminococcaceae （21. 05%） , Lachnospiraceae （20%） and Rikenellaceae （12. 63%） were the pre-dominant families. Only 31. 58% of the sequences at the genus level can be identified, the most abuadant genera were Lachnoclostridium （12. 63%） , followed by Alistipes and Ruminiclostridium. Baicheng Soy Sauce Chicken had a rich diversity ef bacterial composition in cecal ccntents and new potential bacterial fax-on, ready for further research and development.

关 键 词：拜城油鸡 盲肠 细菌 16SrDNA 多样性 

分 类 号：S811.5[农业科学—畜牧学]