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采用单分子实时测序技术结合碘化丙啶(propidium monoazide,PMA)研究传统发酵乳制品中的细菌微生物组。

Investigating the bacterial microbiota of traditional fermented dairy products using propidium monoazide with single-molecule real-time sequencing.

机构信息

Key Laboratory of Dairy Biotechnology and Engineering, Ministry of Education, Key Laboratory of Dairy Products Processing, Ministry of Agriculture, Inner Mongolia Agricultural University, Huhhot 010018, P. R. China.

Key Laboratory of Dairy Biotechnology and Engineering, Ministry of Education, Key Laboratory of Dairy Products Processing, Ministry of Agriculture, Inner Mongolia Agricultural University, Huhhot 010018, P. R. China.

出版信息

J Dairy Sci. 2019 May;102(5):3912-3923. doi: 10.3168/jds.2018-15756. Epub 2019 Mar 7.


DOI:10.3168/jds.2018-15756
PMID:30852020
Abstract

Traditional fermented dairy foods have been the major components of the Mongolian diet for millennia. In this study, we used propidium monoazide (PMA; binds to DNA of nonviable cells so that only viable cells are enumerated) and single-molecule real-time sequencing (SMRT) technology to investigate the total and viable bacterial compositions of 19 traditional fermented dairy foods, including koumiss from Inner Mongolia (KIM), koumiss from Mongolia (KM), and fermented cow milk from Mongolia (CM); sample groups treated with PMA were designated PKIM, PKM, and PCM. Full-length 16S rRNA sequencing identified 195 bacterial species in 121 genera and 13 phyla in PMA-treated and untreated samples. The PMA-treated and untreated samples differed significantly in their bacterial community composition and α-diversity values. The predominant species in KM, KIM, and CM were Lactobacillus helveticus, Streptococcus parauberis, and Lactobacillus delbrueckii, whereas the predominant species in PKM, PKIM, and PCM were Enterobacter xiangfangensis, Lactobacillus helveticus, and E. xiangfangensis, respectively. Weighted and unweighted principal coordinate analyses showed a clear clustering pattern with good separation and only minor overlapping. In addition, a pure culture method was performed to obtain lactic acid bacteria resources in dairy samples according to the results of SMRT sequencing. A total of 102 LAB strains were identified and Lb. helveticus (68.63%) was the most abundant, in agreement with SMRT sequencing results. Our results revealed that the bacterial communities of traditional dairy foods are complex and vary by type of fermented dairy product. The PMA treatment induced significant changes in bacterial community structure.

摘要

传统发酵乳制品是蒙古族数千年来的主要饮食组成部分。在本研究中,我们使用吖啶橙单染(PMA;结合非存活细胞的 DNA,因此只计数存活细胞)和单分子实时测序(SMRT)技术,研究了 19 种传统发酵乳制品的总菌和活菌组成,包括内蒙古马奶酒(KIM)、蒙古马奶酒(KM)和蒙古发酵牛奶(CM);用 PMA 处理的样品组分别命名为 PKIM、PKM 和 PCM。全长 16S rRNA 测序在 PMA 处理和未处理的样品中鉴定出 121 个属和 13 个门中的 195 个细菌种。PMA 处理和未处理的样品在细菌群落组成和α多样性值方面存在显著差异。KM、KIM 和 CM 中的主要种为瑞士乳杆菌、副猪链球菌和德氏乳杆菌,而 PKM、PKIM 和 PCM 中的主要种为襄汾肠杆菌、瑞士乳杆菌和襄汾肠杆菌。加权和非加权主坐标分析显示出清晰的聚类模式,分离良好,仅有微小重叠。此外,根据 SMRT 测序结果,采用纯培养方法从乳制品样品中获得乳酸菌资源。共鉴定出 102 株乳酸菌,其中乳杆菌属(68.63%)最为丰富,与 SMRT 测序结果一致。我们的研究结果表明,传统乳制品的细菌群落是复杂的,且因发酵乳制品的类型而异。PMA 处理会引起细菌群落结构的显著变化。

相似文献

[1]
Investigating the bacterial microbiota of traditional fermented dairy products using propidium monoazide with single-molecule real-time sequencing.

J Dairy Sci. 2019-3-7

[2]
Diversity of lactic acid bacteria associated with traditional fermented dairy products in Mongolia.

J Dairy Sci. 2011-7

[3]
Molecular identification and quantification of lactic acid bacteria in traditional fermented dairy foods of Russia.

J Dairy Sci. 2015-8

[4]
Microbial diversity analysis of jiaoke from Xilingol, Inner Mongolia.

J Dairy Sci. 2020-5-7

[5]
Bacterial microbiota of Kazakhstan cheese revealed by single molecule real time (SMRT) sequencing and its comparison with Belgian, Kalmykian and Italian artisanal cheeses.

BMC Microbiol. 2017-1-9

[6]
Profiling of the viable bacterial and fungal microbiota in fermented feeds using single-molecule real-time sequencing.

J Anim Sci. 2020-2-1

[7]
Investigating bacterial population structure and dynamics in traditional koumiss from Inner Mongolia using single molecule real-time sequencing.

J Dairy Sci. 2016-10

[8]
Comparison of Bacterial Microbiota in Raw Mare's Milk and Koumiss Using PacBio Single Molecule Real-Time Sequencing Technology.

Front Microbiol. 2020-10-27

[9]
Population structure of Lactobacillus helveticus isolates from naturally fermented dairy products based on multilocus sequence typing.

J Dairy Sci. 2015-5

[10]
High-throughput sequencing for the detection of the bacterial and fungal diversity in Mongolian naturally fermented cow's milk in Russia.

BMC Microbiol. 2015-2-22

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