运用PCR-DGGE分析比较瘤胃中不同饲料固相粘附微生物区系  

Comparison of Microbial Communities Adherent to Three Kinds of Feeds in Dairy Cows by PCR-DGGE

作　　者：冯薇[1,2] 王加启[1] 刘开朗[1] 卜登攀[1] 李旦[1] 赵圣国[1] 杨开伦[2] 

机构地区：[1]中国农业科学院北京畜牧兽医研究所动物营养学国家重点实验室,北京100193 [2]新疆农业大学动物科学学院,乌鲁木齐830052

出　　处：《畜牧兽医学报》2010年第12期1556-1562,共7页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：动物营养学国家重点实验室自主研究课题(2004DA125184(团)0801)

摘　　要：通过研究奶牛瘤胃中苜蓿、青贮玉米和羊草3种饲料固相粘附微生物菌群结构,分析比较附着于不同粗饲料微生物区系的差异。选择3头健康且体质量相近的装有永久性瘤胃瘘管的中国荷斯坦奶牛,将苜蓿、青贮玉米和羊草分别装入尼龙袋,在瘤胃中孵育24h后取出,PBS洗脱获得固相粘附微生物,提取总DNA,采用变性梯度凝胶电泳技术(DGGE)分析群落结构,选取清晰条带测序后构建系统发育树。不同样品的DGGE图谱条带的数目和条带颜色深浅有一定差异;苜蓿与青贮玉米和羊草间相比相似性较低,青贮玉米与羊草间的相似性较高;3种饲料及瘤胃内容物固相粘附微生物的Simpson多样性指数差异显著(P<0.05),Shannon-Wiener多样性指数差异不显著(P>0.05);3种饲料固相粘附微生物条带近源种分别归属Butyrivibrio sp.、Fibrobacter sp.、Prevotella sp.、Succiniclasticum sp.、Pseudobutyrivibrio sp.5个属。3种饲料固相粘附微生物的种群构成存在差异。In order to get the information of microbes involved in coarse degradation,microbial communities adherent to alfalfa hay,corn silage and Chinese wild hay in the rumen of Holstein dairy cows were analyzed.Three healthy Chinese Holstein dairy cows with permanent rumen cannula and similar weight were selected,three kinds of feeds were put in nylon bags,respectively,then incubated in the rumen for 24 h,the microbial communities adherent to feeds were eluted with PBS method and compared by DGGE.Some different and the same bands were selected and sequenced.Phylogenetic tree was built by Mega 4.1.DGGE profiles showed the number and intensity of bands from different samples were different;The identity between alfalfa hay and corn silage,Chinese wild hay was low,but the identity between corn silage and Chinese wild hay was high;Simpson index of different samples were different significantly（P0.05）,Shannon-Wiener index was not different significantly（P0.05）;The phylogenetic tree derived from 16S rDNA sequence was found that the closest relatives belonged to Butyrivibrio sp.,Fibrobacter sp.,Prevotella sp.,Succiniclasticum sp.,Pseudobutyrivibrio sp..The results showed that the microbial communities adherent to three feeds were different.

关 键 词：固相粘附微生物 DGGE 瘤胃 

分 类 号：S816.3[农业科学—饲料科学]