Diversity shifts of rumen bacteria induced by dietary forages in dairy cows and quantification of the changed bacteria using a new primer design strategy

作     者：JIN Di ZHAO Sheng-guo ZHANG Yang-dong SUN Peng BU Deng-pan Yves Beckers WANG Jia-qi 

作者机构：State Key Laboratory of Animal NutritionInstitute of Animal ScienceChinese Academy of Agricultural Sciences Animal Science UnitGembloux Agro-Bio TechUniversity of Liège 

基　　金：supported by the National Natural Science Foundation of China(31261140365) the National Basic Research Program of China(2011CB100804) the Agricultural Science and Technology Innovation Program of Chinese Academy of Agricultural Sciences(ASTIP-IAS12) 

出 版 物：《Journal of Integrative Agriculture》 (农业科学学报（英文版）)

年 卷 期：2016年第15卷第11期

页      码：2596-2603页

摘      要：The partial 16 S r RNA gene sequences（100 to 500 bp） were widely used to reveal rumen bacterial composition influenced by diets, while quantification of the changed uncultured bacteria was inconvenient due to difficult designing of specific primers based on short sequences. This study evaluated the effect of forage resources on rumen bacterial diversity and developed new strategy for primer design based on short sequences to quantify the changed uncultured bacteria. Denaturing gradient gel electrophoresis（DGGE） analysis and subsequent band sequencing were used to reveal the distinct rumen bacteria composition in cows fed with two forage sources（single corn stover vs. mixed forages including alfalfa hay and corn silage）. The bacterial diversity in the rumen of dairy cows fed with corn stover was lower than that with mixed forages（P0.05）. The bacterium named R-UB affiliating to uncultured Succinivibrionaceae was identified, and it was abundant in the rumen of cows fed with mixed forages compared to corn stover. The full length 16 S r RNA gene sequences with identity of 97% to the R-UB 16 S r RNA gene sequence were obtained from Gen Bank and used to design specific primers to quantify uncultured bacterium R-UB. All sequences of amplicon from the new primers were of 100% identity to R-UB sequences indicating the high specificity of new primers. Quantitative PCR confirmed that abundance of R-UB in the rumen of cows fed with corn stover was lower than those fed with mixed forages（P0.01）. New strategy for designing primers based on partial 16 S r RNA genes to quantify targeted uncultured bacteria was successfully developed. The rumen bacteria descending significantly in the cows fed corn stover compared to those fed mixed forages was identified as uncultured R-UB from Succinivibrionaceae.

主 题 词：rumen bacterial diversity forage source primers qPCR 

学科分类：0905[农学-林学类] 09[农学] 

核心收录：

D　O　I：10.1016/S2095-3119(16)61346-1

馆 藏 号：203208220...