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. 2020;69(1):1-11.
doi: 10.33073/pjm-2020-004.

Assessing the Microbial Communities in Four Different Daqus by Using PCR-DGGE, PLFA, and Biolog Analyses

Yuxi Ling  1 , Wenying Li  2 , Tong Tong  1 , Zuming Li  1 , Qian Li  1 , Zhihui Bai  3 , Guijun Wang  4 , Jiahao Chen  4 , Yuguang Wang  4
Affiliations
Free PMC article

Assessing the Microbial Communities in Four Different Daqus by Using PCR-DGGE, PLFA, and Biolog Analyses

Yuxi Ling et al. Pol J Microbiol. 2020.
Free PMC article

Abstract

Daqu made from raw wheat, barley or pea is used as an inoculum for the fermentation of Chinese Baijiu. In this study, the microbial communities of four different types of Daqus (sauce-flavor Wuling Daqu, sauce and strong-flavor Baisha Daqu, strong-flavor Deshan Daqu, and light-flavor Niulanshan Daqu) were analyzed by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE), phospholipid fatty acid (PLFA) analysis, and Biolog EcoPlates analysis (Biolog). Clear differences were seen between the microbial communities of the four Daqus. PCR-DGGE showed differences in the number and brightness of bands between the Daqus, indicating the presence of unique bacterial species in Deshan Daqu, Wuling Daqu, and Niulanshan Daqu. Lactobacillus sanfranciscensis, Bacillus thermoamylovorans, and some unclassified bacteria were unique to Wuling Daqu, Deshan Daqu, and Niulanshan Daqu, respectively. Moreover, some bacterial species were observed in all four Daqus. A total of 26 PLFAs between C12 to C20 were detected from the four Daqus by PLFA analysis. Wuling Daqu had the highest total and fungal biomasses, Baisha Daqu had the highest bacterial biomass, and Niulanshan Daqu had the highest ratio of fungal biomass to bacterial biomass. The Biolog results indicated differences in the carbon source use and mode of the four Daqus, and also demonstrated that each Daqu had varying abilities to utilize different types of carbon sources. The cluster analysis of the three methods showed that the microbial communities of the four Daqus were different. This study also demonstrates the applicability of the three analytical methods in the evaluating of the microbial communities of Daqus.

Daqu made from raw wheat, barley or pea is used as an inoculum for the fermentation of Chinese Baijiu. In this study, the microbial communities of four different types of Daqus (sauce-flavor Wuling Daqu, sauce and strong-flavor Baisha Daqu, strong-flavor Deshan Daqu, and light-flavor Niulanshan Daqu) were analyzed by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE), phospholipid fatty acid (PLFA) analysis, and Biolog EcoPlates analysis (Biolog). Clear differences were seen between the microbial communities of the four Daqus. PCR-DGGE showed differences in the number and brightness of bands between the Daqus, indicating the presence of unique bacterial species in Deshan Daqu, Wuling Daqu, and Niulanshan Daqu. Lactobacillus sanfranciscensis, Bacillus thermoamylovorans, and some unclassified bacteria were unique to Wuling Daqu, Deshan Daqu, and Niulanshan Daqu, respectively. Moreover, some bacterial species were observed in all four Daqus. A total of 26 PLFAs between C12 to C20 were detected from the four Daqus by PLFA analysis. Wuling Daqu had the highest total and fungal biomasses, Baisha Daqu had the highest bacterial biomass, and Niulanshan Daqu had the highest ratio of fungal biomass to bacterial biomass. The Biolog results indicated differences in the carbon source use and mode of the four Daqus, and also demonstrated that each Daqu had varying abilities to utilize different types of carbon sources. The cluster analysis of the three methods showed that the microbial communities of the four Daqus were different. This study also demonstrates the applicability of the three analytical methods in the evaluating of the microbial communities of Daqus.

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Conflict of interest statement

Conflict of interest

The authors do not report any financial or personal connections with other persons or organizations, which might negatively affect the contents of this publication and/or claim authorship rights to this publication.

Figures

Fig. 1.
Fig. 1.
PCR-DGGE profiles (A) and clustering analysis (B) of the bacterial communities in four typical Daqu samples. (a-m represent the specific bands excised; 1–4 represent Wuling Daqu, Baisha Daqu, Deshan Daqu, and Niulanshan Daqu, respectively.
Fig. 2.
Fig. 2.
Total biomass, bacterial biomass, fungi biomass (A) and the ratio of fungi biomass to bacteria biomass (B) of Daqu samples.
Fig. 3.
Fig. 3.
Principal component analysis (PCA) showing variations in the PLFA pattern in different types of Daqu (A); clustering analysis (B) of the four Daqus on PLFAs content.
Fig. 4.
Fig. 4.
The AWCD of five types of carbon sources in four Daqus communities, including all carbon sources (A), monosaccharides and their derivatives (B), disaccharides and polysaccharides (C), amino acid substrate and its derivatives (D), fatty acids and lipids (E), and metabolites and secondary metabolites (F).
Fig. 5.
Fig. 5.
Principal component analysis (A) and clustering analysis (B) based carbon source utilization patterns of microbial communities.
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