玉米皮固态发酵菌体蛋白饲料菌群qPCR检测方法的建立  

Establishment of a qPCR method for detection of microbial community in corn husk by solid-state fermentation

作　　者：刘再 燕亚平 崔金娜 刘伟 刘敏 孙晗 刘占英[1] LIU Zai;YAN Ya-ping;CUI Jin-na;LIU Wei;LIU Min;SUN Han;LIU Zhan-ying

机构地区：[1]内蒙古自治区发酵产业节能减排工程技术研究中心、生物发酵绿色制造内蒙古自治区工程研究中心、内蒙古工业大学化工学院,内蒙古呼和浩特010051

出　　处：《饲料研究》2023年第7期110-116,共7页Feed Research

基　　金：内蒙古自治区科技重大专项(项目编号:2019ZD021)。

摘　　要：试验旨在建立玉米皮固态发酵菌体蛋白饲料菌群qPCR检测方法。试验以玉米皮为发酵底物,枯草芽孢杆菌、酿酒酵母菌和产朊假丝酵母菌为发酵菌剂制备玉米皮菌体蛋白饲料,采用qPCR法定量测定固态发酵玉米皮菌体蛋白饲料中的菌群数量变化,并与平板计数法进行比较。结果显示,试验建立的枯草芽孢杆菌、酿酒酵母菌和产朊假丝酵母菌的标准曲线符合SYBR Green qPCR的检测要求,两种方式检测玉米皮固态发酵菌种数量变化趋势相同。将两种方法所得结果的数据进行线性拟合,得出两者间的线性关系y=1.301x-5.139、y=2.257x-8.431和y=2.941x-12.940,任取3批玉米皮菌体蛋白饲料进行验证试验,两者之间相似度达94%以上,表明本方法可以用于玉米皮固态发酵菌体蛋白饲料中菌群的定量检测。研究表明,试验建立的qPCR技术在玉米皮固态发酵菌种数量变化监测应用中,克服了平板计数法时间滞后和两种酵母菌不好区分等缺点。The study was to establish a fluorescence quantitative qPCR method for the detection of microbial community changes in corn husk solid fermentation microbial protein feed.In the study,corn husk was used as fermentation substrate,Bacillus subtilis,Saccharomyces cerevisiae and Candida prionogenes were used as fermentation agents to prepare corn husk mycelium protein feed.qPCR method was used to quantitatively determine the changes in the number of bacteria in solid fermentation corn husk mycelium protein feed,and compared with plate counting method.The results showed that the established standard curves of Bacillus subtilis,Saccharomyces cerevisiae and Candida prionogenes were in line with the requirements of SYBR Green qPCR,and the number of solid-state fermentation bacteria in corn husk detected by two methods showed the same trend.The linear relationship between the two methods was obtained by linear fitting,y=1.301x-5.139,y=2.257x-8.431 and y=2.941x-12.940.Three batches of corn husk mycorrhizal protein feed were selected for verification test,and the similarity between two was more than 94%,indicating that the method could be used for quantitative detection of the microflora in solid fermentation process of corn husk mycelium protein feed.The results show that the established qPCR technique overcomes the shortcomings of plate counting method,such as the time lag and the difficulty in distinguishing two kinds of yeast in the application of solid-state fermentation of corn husk.

关 键 词：固态发酵 菌群动态变化 qPCR 平板计数 

分 类 号：S816.17[农业科学—饲料科学]