应用16S rRNA基因序列分析青海放牧牦牛瘤胃细菌多样性  

Analysis of Qinghai grazing Yak rumen bacteria diversity using 16S rRNA gene sequence

作　　者：崔占鸿[1] 曹连宾 郝力壮[1] 孙璐[1] 刘书杰[1] 

机构地区：[1]青海大学畜牧兽医科学院青海省高原放牧家畜营养与生态国家重点实验室培育基地青海省高原放牧家畜动物营养与饲料科学重点实验室,青海西宁810016

出　　处：《青海大学学报（自然科学版）》2015年第6期1-7,29,共8页Journal of Qinghai University(Natural Science)

基　　金：国家973计划前期研究专项项目课题(2012CB722906);青海省科技支撑计划项目(2014-NS-112)

摘　　要：以青海省海北藏族自治州刚察县全放牧牦牛瘤胃内容物为样本,使用细菌通用引物进行瘤胃细菌16S rRNA基因序列的扩增,以16S rRNA序列分析技术分析牦牛瘤胃细菌多样性,并构建16S rRNA基因克隆文库。结果表明：获得114个非嵌合体序列属于93个OTUs（相似性≥97%作为一个OTUs）,其中64个OTUs与已知16S rRNA序列相似性≥97%,占总代表序列的68.82%;有27个OTUs与已知16S rRNA序列相似性处于90%-96%,占总代表序列的29.03%;有2个OTUs与已知16S rRNA序列相似性〈90%,占总代表序列的2.15%。这93个OTUs序列已向Genebank提交,序列号为（KP455685-KP455713,KP765451-KP765514）。将93个OTUs代表序列与已知序列构建系统发育树的分析表明,青海放牧牦牛瘤胃细菌主要分为LGCGPB（the low G＋C Gram positive bacteria）和CFB（the Cytophaga-Flexibacter-Bacteroides group）两大类;而瘤胃内纤维降解菌主要属于LGCGPB这一类。同时在瘤胃内容物中还检测到了14株已培养瘤胃细菌,其中6株属于纤维降解菌,说明了全放牧条件下牦牛瘤胃内纤维降解菌的优势地位。Taking the rumen contents of all- year grazing yak from Gangcha County of Haibei Tibetan Autonomous Prefecture,Qinghai,as materials,the yak rumen bacteria diversity,rumen advantage bacterium group were analyzed by using bacteria universal primers for amplification of rumen bacterial 16 S rRNA sequence and analysis of phylogenetic tree. The results show that the obtained114 non- chimeric sequences belong to 93 OTUs（ similarity as a OTUs of 97% or higher）,of which the similarity of 64 OTUs and the known 16 S rRNA sequence are 97% or higher,accounting for 68. 82% of the general representative sequence. The similarity of 27 OTUs and the known 16 S rRNA sequence are from 90% to 96%,accounting for 29. 03% of the general representative se-quence. The similarity of 2 OTUs and the known 16 S rRNA sequence are lower than 90%,accounting for 2. 15% of the general representative sequences. The sequence of 29 OTUs（ the similarity with known 16 s rRNA sequence were 96% or less） has been submitted to Genebank,accession numbers（ KP455685- KP455713）. Using 93 OTUs representative with known sequences to construct phylogenetic tree,the rumen bacteria are divided into LGCGPB（ the low G ＋ C gram positive bacteria） and CFB（ the Cytophaga- Flexibacter- Bacteroides group） of the two big kinds,and the bacteria for degrading fibers mainly belong to LGCGPB. At the same time,this study also detected14 strains trained bacteria in the rumen contents,and six strains of them belong to fiber degradation bacteria,which showed that fiber degrading bacteria are the dominant position in the yak rumen under the grazing conditions.

关 键 词：放牧牦牛 16S RRNA基因序列 瘤胃 细菌多样性 青海 

分 类 号：S823.85[农业科学—畜牧学]