摘要
为了解天然放牧和舍饲条件下内蒙古绒山羊瘤胃中甲烷菌的种群组成。选取甲烷菌mcrA基因的特异性引物序列,利用PCR技术对从瘤胃液中抽提的细菌总DNA进行扩增,并建立了甲烷菌特异性的mcrA基因文库。用限制性内切酶Taq I对该文库特异性片段进行了限制性酶切片断长度多态性分析(Restriction fragment lengthpolymor-phism,RFLP)。放牧和舍饲内蒙古绒山羊瘤胃中甲烷菌的mcrA基因片段被分成6个不同的RFLP类型,并由此建立了DNA指纹图谱;系统发育树分析表明,其属于甲烷杆菌目和甲烷微菌目,且绒山羊瘤胃中的甲烷菌大部分属于甲烷短杆菌。放牧绒山羊瘤胃内甲烷菌种类比舍饲绒山羊更加丰富,6种甲烷菌所占比例均较高,而舍饲绒山羊只有两种优势菌比例较高,其他几种所占比例极低。
In order to analyze the methanogen species composition in the rumen from grazing and barn feeding cashmere goat respectively. A pair of mcrA gene specific primers was designed,and total microbial genomic DNA of rumen liquid was extracted by the direct method of PCR-amplification template. Then a mcrA gene specific DNA clone library was subsequently constructed. And RFLP analysis of mcrA gene was conducted using enzymes（Taq I） to digest mcrA gene from the library. The randomly selected cloned fragments were finally clustered into 6 different phylotypes, and contruct the DNA fingerprint patterns from it. Through the phylogenetic trees analyzing,Methanogen in cashmere goat rumen belong to methanobacteriales and methanomicrobiales,and the preponderant methanogen was methanobrevibacter. The methanogen in grazing cashmere goat rumen was more abundant than the one in barnbreeding. The grazing cashmere goat had a higher proportion of six kinds of methanogen than the barn feeding cashmere goat,the latter only have two kinds of methanogen which were higher proportion,the others were low proportion.
出处
《华北农学报》
CSCD
北大核心
2010年第5期228-233,共6页
Acta Agriculturae Boreali-Sinica
基金
现代农业产业技术体系建设专项资金资助(nycytx-39)
