CAJ | 학술논문

采用荧光定量PCR技术检测圈养羚牛粪样中3种纤维分解菌：白色瘤胃球菌、黄色瘤胃球菌、产琥珀酸丝状杆菌的数量。提取羚牛粪便中微生物总DNA，分别以3种纤维分解菌特异性引物进行模板片段扩增，10倍系列稀释，检测Ct值，建立标准曲线。提取各待测样品DNA，进行荧光定量PCR反应测定Ct值，并与标准曲线比较得出定量结果。结果显示，所构建的各标准曲线均具有很高的相关性（R2＞0.99），说明成功构建了标准曲线。1号牛粪样中白色瘤胃球菌、黄色瘤胃球菌、产琥珀酸丝状杆菌的数量分别为3.09×107、2.24×106、5.13×107拷贝数/g，2号牛粪样中三种菌数量分别为4.57×106、2.24×106、2.88×107拷贝数/g，3号牛粪样中三种菌数量分别为5.12×106、1.86×106、8.91×107拷贝数/g。结果表明，3头羚牛之间的纤维分解菌的数量存在差异，但均以产琥珀酸丝状杆菌的数量最高，为主要的纤维分解菌。
채용형광정량PCR기술검측권양령우분양중3충섬유분해균：백색류위구균、황색류위구균、산호박산사상간균적수량。제취령우분편중미생물총DNA，분별이3충섬유분해균특이성인물진행모판편단확증，10배계렬희석，검측Ct치，건립표준곡선。제취각대측양품DNA，진행형광정량PCR반응측정Ct치，병여표준곡선비교득출정량결과。결과현시，소구건적각표준곡선균구유흔고적상관성（R2＞0.99），설명성공구건료표준곡선。1호우분양중백색류위구균、황색류위구균、산호박산사상간균적수량분별위3.09×107、2.24×106、5.13×107고패수/g，2호우분양중삼충균수량분별위4.57×106、2.24×106、2.88×107고패수/g，3호우분양중삼충균수량분별위5.12×106、1.86×106、8.91×107고패수/g。결과표명，3두령우지간적섬유분해균적수량존재차이，단균이산호박산사상간균적수량최고，위주요적섬유분해균。
Detected three kinds of cellulolytic bacteria in Budorcas taxicolor’s fecal by quantitative PCR (q-PCR), namely Ruminococcus albus,Ruminococcus flavefaciens,Fibrobacter succinogenes.Extracted total DNA of the bacteria from the Budorcas taxicolor’s fecal samples,the specific primers of the three cellulolytic bacterias for PCR amplification,serial dilution of 10-fold for the PCR products,and detected the Ct by q-PCR,and established the standard curve of each sample. Extracted the DNA of each sample,which would be taken in the reactions to check out the Ct,the results were obtained by comparing them with the standard curve.The results showed that:all the standard curve had a high correlation (R2>0.99). The amounts of Ruminococcus albus,Ruminococcus flavefaciens,Fibrobacter succinogenes in sample Ⅰ were 3.09 × 107,2.24×106,5.13×107 copies/g,the amounts in sampleⅡwere 4.57×106,2.24×106,2.88×107 copies/g,and the amounts in sampleⅢwere 5.12×106,1.86×106,8.91×107 copies/g.The cellulolytic bacterias amounts of three samples were different,but the number of Fibrobacter succinogenes was the highest of all samples,and it was the major cellulolytic bacteria.