Determination of the fungal community of pit mud in fermentation cellars for Chinese strong-flavor liquor, using DGGE and Illumina MiSeq sequencing

Maoke Liu; Yuming Tang; Ke Zhao; Ying Liu; Xiaojiao Guo; Daoqun Ren; Wanchun Yao; Xinhui Tian; Yunfu Gu; Bin Yi; Xiaoping Zhang

doi:10.1016/j.foodres.2016.11.037

Determination of the fungal community of pit mud in fermentation cellars for Chinese strong-flavor liquor, using DGGE and Illumina MiSeq sequencing

Food Res Int. 2017 Jan:91:80-87. doi: 10.1016/j.foodres.2016.11.037. Epub 2016 Dec 1.

Authors

Maoke Liu¹, Yuming Tang², Ke Zhao³, Ying Liu², Xiaojiao Guo², Daoqun Ren², Wanchun Yao², Xinhui Tian², Yunfu Gu³, Bin Yi⁴, Xiaoping Zhang⁵

Affiliations

¹ College of Resource, Sichuan Agricultural University, Wenjiang 611130, People's Republic of China; Institute of Rice and Sorghum Sciences, Sichuan Academy of Agricultural Sciences, Deyang 618000, People's Republic of China.
² Institute of Rice and Sorghum Sciences, Sichuan Academy of Agricultural Sciences, Deyang 618000, People's Republic of China.
³ College of Resource, Sichuan Agricultural University, Wenjiang 611130, People's Republic of China.
⁴ Center of National Engineering Research of Solid-State Brewing, Luzhou Laojiao Company Limited, Luzhou 646100, People's Republic of China.
⁵ College of Resource, Sichuan Agricultural University, Wenjiang 611130, People's Republic of China. Electronic address: zhangxiaopingscau@126.com.

PMID: 28290330
DOI: 10.1016/j.foodres.2016.11.037

Abstract

Chinese strong-flavor liquor (CSFL) is fermented in cellars lined with pit mud (PM). This PM, specific fermented clay, contains microbes that play important roles in CSFL production. However, there is limited information about fungal community structure and cellar-age-related changes in PM. In this study, PM samples were removed from the cellars used for 5 and 100years and characterized using denaturing gradient gel electrophoresis (DGGE) and Illumina MiSeq sequencing. Both methods revealed there were no significant differences in fungal species diversity (Shannon index, Chao1, and observed species) between the 5- and 100-year PM samples (p>0.05), but the communities were more stable in the 100-year PM samples (UPGMA). Illumina MiSeq sequencing allowed identification of 111 fungal genera belonging to 4 phyla (Ascomycota, Zygomycota, Basidiomycota, and Chytridiomycota) in the PM samples, with the predominant phylum being Ascomycota. The results also indicated that the compositions of dominant genera in the PM samples were significantly changed during long-term CSFL fermentation. There were relatively more Rhizopus, Phoma, and Trichosporon in the 5-year PM samples, and Aspergillus and Candida were most highly represented in the 100-year PM samples (p<0.05). Of these, Candida increased its relative abundance significantly in the 100-year samples (p<0.05). Overall, the results provide novel insights into the fungal community associated with CSFL production, and may suggest why fermentation in a cellar with PM that has been in usage for a longer time allows better quality CSFL production.

Keywords: DGGE; ITS region; Illumina MiSeq sequencing; Pit mud; Strong-flavor liquor.

Publication types

Comparative Study
Research Support, Non-U.S. Gov't

MeSH terms

Alcoholic Beverages / microbiology*
Biodiversity
Bioreactors / microbiology*
China
Clay / microbiology*
DNA, Ribosomal Spacer / genetics*
Denaturing Gradient Gel Electrophoresis*
Fermentation*
Fungi / classification
Fungi / genetics*
Fungi / metabolism
High-Throughput Nucleotide Sequencing*
Microbiota / genetics*
Phylogeny
Ribotyping / methods*
Time Factors

Substances

DNA, Ribosomal Spacer
Clay