腐败干酪中微生物种类的分离与鉴定
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摘要
本文选取腐败干酪,用传统的微生物培养法培养、分离和鉴定,同时通过18S rDNA高通量测序法,根据稀释曲线、OTU聚类以及样品的多样性分析来反映腐败干酪中的微生物菌群。通过传统的微生物培养法得到3种菌株,其中两种为青霉菌属,一种为酵母菌属。通过高通量测序得出腐败干酪中微生物菌群主要有青霉菌属、德巴利(氏)酵母属、耐碱酵母属、丝孢酵母属、木拉克酵母属、双足囊菌属以及未分类的真菌等。
In this paper, spoilage cheese was selected and cultured, isolated and identified by traditional microbial culture method. Meanwhile,the microbial flora in spoilage cheese was reflected according to dilution curve, OTU clustering and diversity analysis of samples through 18 S high-throughput sequencing of rDNA. Three strains were obtained by traditional microbial culture, two of which were Penicillium and one was yeast. The results of high-throughput sequencing indicated that the microbial flora in spoilage cheese mainly included Penicillium, De-baryomyces, Alkali resistant yeast, Trichosporon, Mrakia, Dipodascus and unclassified fungi, etc.
In this paper, spoilage cheese was selected and cultured, isolated and identified by traditional microbial culture method. Meanwhile,the microbial flora in spoilage cheese was reflected according to dilution curve, OTU clustering and diversity analysis of samples through 18 S high-throughput sequencing of rDNA. Three strains were obtained by traditional microbial culture, two of which were Penicillium and one was yeast. The results of high-throughput sequencing indicated that the microbial flora in spoilage cheese mainly included Penicillium, De-baryomyces, Alkali resistant yeast, Trichosporon, Mrakia, Dipodascus and unclassified fungi, etc.
引文
[1]李红娟,抗真菌乳酸菌的筛选及特性研究[D],中国农业科学院2011.
[2] FILTENBORG O, FRISVAD J C, THRANE U.Moulds in food spoilage[J]. International Journal of Food Microbiology,1996, 33(9):85-102.
[3] LOUREIRO V, QUEROL A. The prevalence and control of spoilage yeasts in foods and beverages[J]. Trends in Food Scienceand Technology,1999, 10, 356-365.
[4]倪奎奎.全株水稻青贮饲料中微生物菌群以及发酵品质分析[D].郑州大学, 2016.(下转第36页)
[5] BJORNSDOTTIR-BUTLER K, JONES J L, BENNER R, et al.Development of a real-time PCR assay with an internal amplification control for detection of Gram-negative histamine producing bacteria in fish[J]. Food Microbiology, 2011, 28(3):356-363.
[6] JIANG Y, GAO F, XU X L, et al. Changes in the composition of the bacterial flora on tray-packaged pork during chilled storage analyzed by PCR-DGGE and real-time PCR[J]. Journal of Food Science,2011, 76(1):27-33.
[7]许爱清.茯砖茶的真菌菌群特性及其整肠功能研究[D].湖南农业大学,2011.
[8] LIU W T,MARSH T L,CHENG H,et al.Character-ization of microbial diversity by determing terminalrestriction fragment length polymorphisms of genes en-coding 165 rRNA[J].App; Environ Microbiol,1997,63(11):4516-4522.
[9]任南琪,赵阳国,高崇洋. TRFLP在微生物群落结构与动态分析中的应用[J].哈尔滨工业大学报, 2007, 39(4):552-556.
[10] CARSTEN RUSS,MICHAEL G ROSS,CARNCIRO et al.Pacific biosciences sequencing technology for genotyping and variation discovery in human data[J].BMC Genomics,2012,13:375-381.
[11] POLKA J, REBECCHI A,PISACANE V, et al. Bacterial diversity in typical Italian salami at different ripening stages as revealed by highthroughput sequencing of 16S r RNA amplicons[J]. Food Microbiology, 2015, 48(6):342-356.
[12]李院,魏新元,王静,等.抑制青霉菌乳酸菌的分离鉴定及抑菌物质分析[J].食品科学,2015,(21):150-155.
[2] FILTENBORG O, FRISVAD J C, THRANE U.Moulds in food spoilage[J]. International Journal of Food Microbiology,1996, 33(9):85-102.
[3] LOUREIRO V, QUEROL A. The prevalence and control of spoilage yeasts in foods and beverages[J]. Trends in Food Scienceand Technology,1999, 10, 356-365.
[4]倪奎奎.全株水稻青贮饲料中微生物菌群以及发酵品质分析[D].郑州大学, 2016.(下转第36页)
[5] BJORNSDOTTIR-BUTLER K, JONES J L, BENNER R, et al.Development of a real-time PCR assay with an internal amplification control for detection of Gram-negative histamine producing bacteria in fish[J]. Food Microbiology, 2011, 28(3):356-363.
[6] JIANG Y, GAO F, XU X L, et al. Changes in the composition of the bacterial flora on tray-packaged pork during chilled storage analyzed by PCR-DGGE and real-time PCR[J]. Journal of Food Science,2011, 76(1):27-33.
[7]许爱清.茯砖茶的真菌菌群特性及其整肠功能研究[D].湖南农业大学,2011.
[8] LIU W T,MARSH T L,CHENG H,et al.Character-ization of microbial diversity by determing terminalrestriction fragment length polymorphisms of genes en-coding 165 rRNA[J].App; Environ Microbiol,1997,63(11):4516-4522.
[9]任南琪,赵阳国,高崇洋. TRFLP在微生物群落结构与动态分析中的应用[J].哈尔滨工业大学报, 2007, 39(4):552-556.
[10] CARSTEN RUSS,MICHAEL G ROSS,CARNCIRO et al.Pacific biosciences sequencing technology for genotyping and variation discovery in human data[J].BMC Genomics,2012,13:375-381.
[11] POLKA J, REBECCHI A,PISACANE V, et al. Bacterial diversity in typical Italian salami at different ripening stages as revealed by highthroughput sequencing of 16S r RNA amplicons[J]. Food Microbiology, 2015, 48(6):342-356.
[12]李院,魏新元,王静,等.抑制青霉菌乳酸菌的分离鉴定及抑菌物质分析[J].食品科学,2015,(21):150-155.