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Development of a Quantitative PCR for Detection of Lactobacillus plantarum Starters During Wine Malolactic Fermentation
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  • Development of a Quantitative PCR for Detection of Lactobacillus plantarum Starters During Wine Malolactic Fermentation
  • Development of a Quantitative PCR for Detection of Lactobacillus plantarum Starters During Wine Malolactic Fermentation
저자명
Cho. Gyu-Sung,KrauB. Sabrina,Huch. Melanie,Toit. Maret Du,Franz. Charles M.A.P.
간행물명
Journal of microbiology and biotechnology
권/호정보
2011년|21권 12호|pp.1280-1286 (7 pages)
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한국미생물생명공학회
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정기간행물|ENG|
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이 논문은 한국과학기술정보연구원과 논문 연계를 통해 무료로 제공되는 원문입니다.
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A quantitative, real-time PCR method was developed to enumerate Lactobacillus plantarum IWBT B 188 during the malolactic fermentation (MLF) in Grauburgunder wine. The qRT-PCR was strain-specific, as it was based on primers targeting a plasmid DNA sequence, or it was L. plantarum-specific, as it targeted a chromosomally located plantaricin gene sequence. Two 50 l wine fermentations were prepared. One was inoculated with 15 g/hl Saccharomyces cerevisiae, followed by L. plantarum IWBT B 188 at $3.6{ imes}10^6$ CFU/ml, whereas the other was not inoculated (control). Viable cell counts were performed for up to 25 days on MRS agar, and the same cells were enumerated by qRT-PCR with both the plasmid or chromosomally encoded gene primers. The L. plantarum strain survived under the harsh conditions in the wine fermentation at levels above $10^5$/ml for approx. 10 days, after which cell numbers decreased to levels of $10^3$ CFU/ml at day 25, and to below the detection limit after day 25. In the control, no lactic acid bacteria could be detected throughout the fermentation, with the exception of two sampling points where ca. $1{ imes}10^2$ CFU/ml was detected. The minimum detection level for quantitative PCR in this study was $1{ imes}10^2$ to $1{ imes}10^3$ CFU/ml. The qRT-PCR results determined generally overestimated the plate count results by about 1 log unit, probably as a result of the presence of DNA from dead cells. Overall, qRT-PCR appeared to be well suited for specifically enumerating Lactobacillus plantarum starter cultures in the MLF in wine.