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Characterization of Microbial Diversity and Community Structure in Fermentation Pit Mud of Different Ages for Production of Strong-Aroma Baijiu Cover

Characterization of Microbial Diversity and Community Structure in Fermentation Pit Mud of Different Ages for Production of Strong-Aroma Baijiu

Polish Journal of Microbiology
Volume 69 (2020): Issue 2 (June 2020)
By: XU-JIA WANG,  HONG-MEI ZHU,  ZHI-QIANG REN,  ZHI-GUO HUANG,  CHUN-HUI WEI and  JIE DENG  
Open Access
|May 2020

Figures & Tables

Fig. 1.

The production (A) and the FP (B) of strong-aroma Baijiu. A: The concentration of rice husks is about 20% of sorghum (dry weight); the main chemical composition of the fresh grain (sorghum and rice husks) are starch (48.1–50.4%), protein (5.3–8.3%), fat (3.2–3.7%), cellulose (8.3–10.2%), lignin (4.8–6.4%), pentosan (3.2–3.4%), tannin (0.1–0.5%), etc; the concentration of fresh grain is about 20% of mixed grain (dry weight); the concentration of Daqu is about 20% of sorghum; during fermentation, the moisture content of the grain is about 55–60%. B: The solid circle (●) represent sampling sites of FPM, the FP is surrounded by FPM.
The production (A) and the FP (B) of strong-aroma Baijiu. A: The concentration of rice husks is about 20% of sorghum (dry weight); the main chemical composition of the fresh grain (sorghum and rice husks) are starch (48.1–50.4%), protein (5.3–8.3%), fat (3.2–3.7%), cellulose (8.3–10.2%), lignin (4.8–6.4%), pentosan (3.2–3.4%), tannin (0.1–0.5%), etc; the concentration of fresh grain is about 20% of mixed grain (dry weight); the concentration of Daqu is about 20% of sorghum; during fermentation, the moisture content of the grain is about 55–60%. B: The solid circle (●) represent sampling sites of FPM, the FP is surrounded by FPM.

Fig. 2.

OTUs rarefaction curves at a 97% sequence similarity cutoff. OTUs from the amplicon libraries of bacteria (A), and archaea (B).
OTUs rarefaction curves at a 97% sequence similarity cutoff. OTUs from the amplicon libraries of bacteria (A), and archaea (B).

Fig. 3.

Phylum level prokaryotes taxonomic classification in the FPM samples as determined with the RDP classifier using an 85% confidence threshold.
Phylum level prokaryotes taxonomic classification in the FPM samples as determined with the RDP classifier using an 85% confidence threshold.

Fig. 4.

The relative FPM prokaryotic genera abundance, based on Roche 454 pyrosequencing. It includes the genera of bacteria (A) and genera of archaea (B). Variations in log2 scale-based genera abundance are shown by the scale bar, with “*” indicating the genera shared among all FPM samples that had the relative abundance >1%.
The relative FPM prokaryotic genera abundance, based on Roche 454 pyrosequencing. It includes the genera of bacteria (A) and genera of archaea (B). Variations in log2 scale-based genera abundance are shown by the scale bar, with “*” indicating the genera shared among all FPM samples that had the relative abundance >1%.

Fig. 5.

Concentration of phospholipid fatty acids (PLFAs) in different FPM samples. The concentration of each sample was determined by an internal standard (19:0). Error bars indicate standard deviations (n = 3).
Concentration of phospholipid fatty acids (PLFAs) in different FPM samples. The concentration of each sample was determined by an internal standard (19:0). Error bars indicate standard deviations (n = 3).

Fig. 6.

Concentrations of phospholipid fatty acids (PLFA) in different samples at the microbial groups level. Error bars show standard deviations (n = 3).
Concentrations of phospholipid fatty acids (PLFA) in different samples at the microbial groups level. Error bars show standard deviations (n = 3).

Fig. 7.

Prokaryotic community redundancy analysis (RDA) based upon Roche 454 pyrosequencing and chemical properties. The dominant community structure-related genera and the direction/magnitude of chemical properties are represented by arrows. It includes the genera of bacteria (A) and genera of archaea (B).
Prokaryotic community redundancy analysis (RDA) based upon Roche 454 pyrosequencing and chemical properties. The dominant community structure-related genera and the direction/magnitude of chemical properties are represented by arrows. It includes the genera of bacteria (A) and genera of archaea (B).

The preference of sequencing results based on different variable regions V2-V3 and V6-V8 a of the 16S rRNA gene_

Variable regionFPM age (yr)TotalBacteriaArchaea
readsOTUreadsOTUreadsOTU
V2-V3    510977 ± 684126 ± 121267 ± 10451 ± 99710 ± 56275 ± 7
  3010562 ± 513100 ± 111009 ± 8447 ± 89553 ± 41053 ± 5
10011172 ± 75175 ± 6938 ± 5327 ± 510233 ± 72048 ± 4
V6-V8    518893 ± 886594 ± 1717826 ± 844583 ± 131067 ± 8911 ± 3
  3017409 ± 395428 ± 1516543 ± 354419 ± 11866 ± 68  9 ± 2
10016894 ± 806319 ± 2415537 ± 758312 ± 201357 ± 133  7 ± 2

Prokaryotic diversity indices based on a cutoff of 97% similarity of the 16S rRNA gene sequences a _

FPM age (yr)BacteriaArchaea
Chao1Observed OTUShannonChao1Observed OTUShannon
    5634.0 ± 84.4583 ± 132.49 ± 0.5697.3 ± 12.275 ± 71.374 ± 0.049
  30501.6 ± 64.6419 ± 112.16 ± 0.3178.5 ± 6.353 ± 51.159 ± 0.068
100435.8 ± 62.1312 ± 271.93 ± 0.3474.2 ± 4.748 ± 41.068 ± 0.087

Basic physiocochemical properties of fermentation pit mud_

VariableValues for fermentation pit age (yr) a
530100
Moisture(%)35.15 ± 6.4239.62 ± 1.1144.97 ± 2.83
Humic matter (%)10.54 ± 1.5913.82 ± 0.0910.90 ± 0.52
Total acidity(%)  1.54 ± 0.09  1.81 ± 0.05  1.13 ± 0.15
Acetic acid(mg/g)  3.98 ± 0.36  3.44 ± 0.46  4.21 ± 0.67
Lactic acid(mg/g)94.22 ± 1.3028.81 ± 7.0521.33 ± 3.15
Butyric acid(mg/g)  2.87 ± 0.34  2.36 ± 0.26  2.79 ± 0.38
Caproic acid(mg/g)15.15 ± 5.1229.62 ± 4.1945.97 ± 5.13
Total nitrogen(%)  0.94 ± 0.09  1.41 ± 0.08  2.16 ± 0.11
NH4 +(mg/g)14.12 ± 1.7721.02 ± 2.0325.32 ± 1.66
Available phosphorus(mg/g)  4.91 ± 0.52  9.15 ± 0.78  9.15 ± 0.41
Available calcium(mg/g)  9.51 ± 1.02  8.18 ± 1.91  7.41 ± 0.52

PLFA markers used for taxonomic groups_

Taxonomic groupPLFA groupSpecific PLFA markersReference
BacteriaMultiple groupsSum of i15:0, a15:0, i16:0, i17:0, a17:0, cy17:0, cy19:0, 16:1ω7, 18:1ω7, and 17:1ω9(Frostegård and Bååth 1996)
Gram-positive bacteriaBranched PLFAsSum of i15:0, a15:0, i16:0, i17:0, and a17:0(O’Leary 1988)
Gram-negative bacteriaCyclopropyl and mono PLFAsSum of cy17:0, 16:1ω7, 18:1ω7, and 17:1ω9(Moore-Kucera and Dick 2008)
Actinomycetes10Me-PLFAsSum of 10Me16:0, 10Me17:0, and 10Me18:0(Moore-Kucera and Dick 2008)
AnaerobesCyclopropylSum of cy17:0, cy19:0ω7, cy19:0ω9(Li et al. 2017)
DOI: https://doi.org/10.33073/pjm-2020-018 | Journal eISSN: 2544-4646 | Journal ISSN: 1733-1331
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Language: English
Page range: 151 - 164
Submitted on: Jan 6, 2020
Accepted on: Mar 29, 2020
Published on: May 12, 2020
Published by: Polish Society of Microbiologists
In partnership with: Paradigm Publishing Services
Publication frequency: 4 issues per year
Keywords:
strong-aroma Baijiu,
fermentation pit mud,
microbial community,
Roche 454 pyrosequencing,
PLFAs
Related subjects:
Life sciences,
Microbiology and virology

© 2020 XU-JIA WANG, HONG-MEI ZHU, ZHI-QIANG REN, ZHI-GUO HUANG, CHUN-HUI WEI, JIE DENG, published by Polish Society of Microbiologists
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License.

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