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Method for fast and quantitatively measuring microbial community composition in fermentation process of baijiu

A microbial community and fermentation process technology, applied in the field of rapid PSP-qPCR quantification, can solve the problems of high quality requirements for endonucleases, failure to achieve microbial community composition and high cost, and achieve improved wine production efficiency, low cost, significant effect

Active Publication Date: 2016-02-17
ANHUI RUISIWEIER TECH
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Problems solved by technology

But the cost is expensive, the purity of DNA and the quality of endonuclease are high, etc.
[0011] However, none of the above-mentioned techniques can quickly determine the composition and structure of the microbial community in the liquor fermentation process in about two hours on site.

Method used

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  • Method for fast and quantitatively measuring microbial community composition in fermentation process of baijiu
  • Method for fast and quantitatively measuring microbial community composition in fermentation process of baijiu
  • Method for fast and quantitatively measuring microbial community composition in fermentation process of baijiu

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Embodiment Construction

[0043] In this example, the method for rapidly determining the composition of the microbial community in the liquor fermentation process is to use phylum-specific primers in the PCR reaction system to specifically amplify the microbial flora of a specific phylum during the liquor fermentation process, and use qPCR The system performs quantitative amplification to obtain quantitative identification.

[0044] Prepare gate-specific primers as follows:

[0045] Step a, extracting the metagenomic group of the brewing microbial community sample as a PCR amplification template;

[0046] Step b, amplify the ribosomal DNA in the amplification template close to the full-length 16srDNA sequence, the amplification primers are 27F (5'-AGAGTTTGATCCTGGCTCAG-3') and 1492R (5'-GGTTACCTTGTTACGACTT-3'), to obtain amplification product;

[0047] Step c. Perform TA cloning, blue-white screening and white-white colony bidirectional sequencing sequentially after the amplification product is purifi...

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Abstract

The invention relates to a method for fast and quantitatively measuring the microbial community composition in the fermentation process of baijiu. The method is characterized in that phylum specific primers are used in a PCR system so that special amplification of microbial communities of a certain specific phylum can be performed in the fermentation process of baijiu, quantitative amplification is performed through a qPCR system, and quantitative recognition is achieved. According to the category of known microorganisms, primers of the bacteroidetes, proteobacteria, firmicutes, synergistetes and actinobacteria are respectively designed, and the microstructure change of microbial communities can be fast inspected in the baijiu fermentation process in the level of the phylum on the basis of quantitative amplification of the phylum specific primers.

Description

technical field [0001] The invention relates to a method for operating microbial communities in the field of biotechnology, and is a rapid PSP-qPCR quantitative method for determining the composition and structure of microbial communities in the liquor fermentation process based on phylum-specific primers (PSP). Background technique [0002] Natural complex microbial communities are generally composed of dozens, hundreds or even more different species of microorganisms, and the composition will change dynamically under different environmental conditions. Most of the bacterial species included in almost all natural microbial communities are poorly understood by humans, so it is still very difficult to quickly quantify the structural composition of natural microbial communities during liquor fermentation. Common techniques currently include: [0003] One is the traditional culture method; for example, most of the microorganisms in the pit mud belong to uncultivated bacteria, ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/06C12R1/01
CPCC12Q1/689C12Q1/6851
Inventor 何宏魁周庆伍李安军罗锡梅张会敏刘国英曹润洁汤知辉张治洲
Owner ANHUI RUISIWEIER TECH
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