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[期刊论文]

Development of Reverse Transcription Quantitative Real-Time PCR (RT-qPCR) Assays for Monitoring Saccharomycopsis fibuligera, Rhizopus oryzae, and Monascus purpureus During the Traditional Brewing of Hong Qu Glutinous Rice Wine

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author:

Lv, Xu-Cong (Lv, Xu-Cong.) [1] (Scholars:吕旭聪) | Jia, Rui-Bo (Jia, Rui-Bo.) [2] | Chen, Jing-Hao (Chen, Jing-Hao.) [3] | Unfold

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EI Scopus SCIE

Abstract:

pies/mu L for S. fibuligera and 10(2) gene copies/mu L for R. oryzae and M. purp In this study, reverse transcription quantitative real-time PCR (RT-qPCR) assays using species-specific primers were developed for the quantification of the three major fungal species (Saccharomycopsis fibuligera, Rhizopus oryzae, and Monascus purpureus) during the traditional brewing of Hong Qu glutinous rice wine. Species-specific primers (Sfi-F/Sfi-R, Ro-F/Ro-R, and Mp-F/Mp-R) targeting the ITS-5.8S rRNA gene and the beta-tubulin gene sequences were designed and their specificity was evaluated by RT-qPCR with the total RNA from fungal species closely related to the traditional brewing of Hong Qu glutinous rice wine. Results of RT-qPCR using species-specific primers showed 100 % inclusivity (positive signal in the presence of target complementary DNA (cDNA)) and 100 % exclusivity (negative signals in the presence of non-target template). The specificity of each primer set was also tested when the target cDNA was mixed with non-target cDNA, and results showed that there was no significant difference (P > 0.05) in threshold cycle (Ct) values obtained from pure target cDNA and target cDNA mixed with non-target cDNA. The detection limits of the established qPCR assays were determined to be 10(1) gene coureus, respectively. Good correlations were obtained for the tested species between 10(2) and 10(8) gene copies/mu L by qPCR analysis. Finally, the established RT-qPCR assays were applied to quantify viable S. fibuligera, R. oryzae, and M. purpureus during the traditional brewing of Hong Qu glutinous rice wine. This study standardized the RT-qPCR assays for dominant fungi associated with the traditional brewing of Hong Qu glutinous rice wine, and it also proved the usefulness of RT-qPCR assays for the quantification of live cells in multi-species samples. Results would show great value in scientifically understanding of the brewing mechanism of Hong Qu glutinous rice wine.

Keyword:

Dominant microorganisms Hong Qu glutinous rice wine Reverse transcription quantitative real-time PCR (RT-qPCR) Species-specific primers Traditional brewing

Community:

  • [ 1 ] [Lv, Xu-Cong]Fujian Agr & Forestry Univ, Natl Engn Res Ctr, JUNCAO Technol, Fuzhou 350002, Fujian, Peoples R China
  • [ 2 ] [Jia, Rui-Bo]Fujian Agr & Forestry Univ, Natl Engn Res Ctr, JUNCAO Technol, Fuzhou 350002, Fujian, Peoples R China
  • [ 3 ] [Chen, Jing-Hao]Fujian Agr & Forestry Univ, Natl Engn Res Ctr, JUNCAO Technol, Fuzhou 350002, Fujian, Peoples R China
  • [ 4 ] [Zhou, Wen-Bin]Fujian Agr & Forestry Univ, Natl Engn Res Ctr, JUNCAO Technol, Fuzhou 350002, Fujian, Peoples R China
  • [ 5 ] [Li, Yan]Fujian Agr & Forestry Univ, Natl Engn Res Ctr, JUNCAO Technol, Fuzhou 350002, Fujian, Peoples R China
  • [ 6 ] [Liu, Bin]Fujian Agr & Forestry Univ, Natl Engn Res Ctr, JUNCAO Technol, Fuzhou 350002, Fujian, Peoples R China
  • [ 7 ] [Lv, Xu-Cong]Fujian Agr & Forestry Univ, Coll Food Sci, Fuzhou 350002, Fujian, Peoples R China
  • [ 8 ] [Jia, Rui-Bo]Fujian Agr & Forestry Univ, Coll Food Sci, Fuzhou 350002, Fujian, Peoples R China
  • [ 9 ] [Chen, Jing-Hao]Fujian Agr & Forestry Univ, Coll Food Sci, Fuzhou 350002, Fujian, Peoples R China
  • [ 10 ] [Zhou, Wen-Bin]Fujian Agr & Forestry Univ, Coll Food Sci, Fuzhou 350002, Fujian, Peoples R China
  • [ 11 ] [Li, Yan]Fujian Agr & Forestry Univ, Coll Food Sci, Fuzhou 350002, Fujian, Peoples R China
  • [ 12 ] [Liu, Bin]Fujian Agr & Forestry Univ, Coll Food Sci, Fuzhou 350002, Fujian, Peoples R China
  • [ 13 ] [Chen, Shao-Jun]Fujian Agr & Forestry Univ, Coll Food Sci, Fuzhou 350002, Fujian, Peoples R China
  • [ 14 ] [Tian, Yu-Ting]Fujian Agr & Forestry Univ, Coll Food Sci, Fuzhou 350002, Fujian, Peoples R China
  • [ 15 ] [Lv, Xu-Cong]Fuzhou Univ, Inst Food Sci & Technol, Coll Biol Sci & Technol, Xue Yuan Rd, Fuzhou 350002, Fujian, Peoples R China
  • [ 16 ] [Xu, Bing-Xin]Fuzhou Univ, Inst Food Sci & Technol, Coll Biol Sci & Technol, Xue Yuan Rd, Fuzhou 350002, Fujian, Peoples R China
  • [ 17 ] [Liang, Yi-Ting]Fuzhou Univ, Inst Food Sci & Technol, Coll Biol Sci & Technol, Xue Yuan Rd, Fuzhou 350002, Fujian, Peoples R China
  • [ 18 ] [Rao, Ping-Fan]Fuzhou Univ, Inst Food Sci & Technol, Coll Biol Sci & Technol, Xue Yuan Rd, Fuzhou 350002, Fujian, Peoples R China
  • [ 19 ] [Ni, Li]Fuzhou Univ, Inst Food Sci & Technol, Coll Biol Sci & Technol, Xue Yuan Rd, Fuzhou 350002, Fujian, Peoples R China

Reprint 's Address:

  • 吕旭聪 倪莉

    [Lv, Xu-Cong]Fujian Agr & Forestry Univ, Natl Engn Res Ctr, JUNCAO Technol, Fuzhou 350002, Fujian, Peoples R China;;[Lv, Xu-Cong]Fujian Agr & Forestry Univ, Coll Food Sci, Fuzhou 350002, Fujian, Peoples R China;;[Lv, Xu-Cong]Fuzhou Univ, Inst Food Sci & Technol, Coll Biol Sci & Technol, Xue Yuan Rd, Fuzhou 350002, Fujian, Peoples R China;;[Ni, Li]Fuzhou Univ, Inst Food Sci & Technol, Coll Biol Sci & Technol, Xue Yuan Rd, Fuzhou 350002, Fujian, Peoples R China

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Source :

FOOD ANALYTICAL METHODS

ISSN: 1936-9751

Year: 2017

Issue: 1

Volume: 10

Page: 161-171

2 . 2 4 5

JCR@2017

2 . 6 0 0

JCR@2023

ESI Discipline: AGRICULTURAL SCIENCES;

ESI HC Threshold:157

JCR Journal Grade:2

CAS Journal Grade:3

Cited Count:

WoS CC Cited Count: 4

SCOPUS Cited Count: 4

30 Days PV: 1

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