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分析韩国黑山羊瘤胃液和瘤胃颗粒中的产甲烷古菌群落。

Analysis of methanogenic archaeal communities of rumen fluid and rumen particles from Korean black goats.

机构信息

Department of Animal Science and Technology, College of Bio industry Science, Sunchon National University.

出版信息

Anim Sci J. 2011 Oct;82(5):663-72. doi: 10.1111/j.1740-0929.2011.00890.x. Epub 2011 Jun 15.

Abstract

Molecular diversity of methanogens in the rumen of Korean black goats was investigated with 16S rRNA gene clone libraries using methanogen-specific primers. The libraries were composed of rumen fluid-associated methanogens (FAM) and rumen particle-associated methanogens (PAM) from rumen-fistulated Korean black goats. Among the 141 clones of the FAM library, the sequences were mostly related to two phyla, the Methanobacteriaceae family (77.3%) and the Thermoplasmatales family (22.7%); and among the 68 clones of the PAM library, sequences were also mainly clustered in the two phyla, the Thermoplasmatales family (63.24%) and the Methanobacteriaceae family (35.29%). Most of the sequenced clones in the two libraries were closely related to uncultured methanogenic archaeon. Quantitative real-time PCR revealed that PAM (8.97 log 10) had significantly higher (P < 0.01) density of methanogens by the methanogenic 16S rRNA gene copies than FAM (7.57 log 10). The two clone libraries also showed difference in Shannon index (FAM library 1.70 and PAM library 1.59) and Chao 1 estimator (FAM library 18 and PAM library 17 operational taxonomic units). Apparent differences found in the microbial community from the two 16S rRNA gene libraries could be a result of such factors as the chemical and physical nature of the target material surface, types or component of diets, the interaction between the methanogens and other microbes, and age of the experimental goats.

摘要

采用甲烷菌特异性引物,通过 16S rRNA 基因克隆文库的方法,研究了韩国黑山羊瘤胃中甲烷菌的分子多样性。文库由瘤胃液相关甲烷菌(FAM)和瘤胃颗粒相关甲烷菌(PAM)组成,来源于瘤胃切开的韩国黑山羊。在 FAM 文库的 141 个克隆中,序列主要与甲烷杆菌科(77.3%)和热原体目(22.7%)两个门有关;在 PAM 文库的 68 个克隆中,序列也主要聚类在两个门,即热原体目(63.24%)和甲烷杆菌科(35.29%)。两个文库中的大多数测序克隆与未培养的产甲烷古菌密切相关。定量实时 PCR 显示,PAM(8.97 log 10)的产甲烷 16S rRNA 基因拷贝数的甲烷菌密度显著高于 FAM(7.57 log 10)(P < 0.01)。两个克隆文库在 Shannon 指数(FAM 文库为 1.70,PAM 文库为 1.59)和 Chao1 估计值(FAM 文库为 18,PAM 文库为 17 个操作分类单位)上也存在差异。从两个 16S rRNA 基因文库中发现的微生物群落的明显差异可能是由于目标材料表面的化学和物理性质、饮食的类型或成分、甲烷菌与其他微生物的相互作用以及实验山羊的年龄等因素造成的。

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