SSR-Marker Analysis—A Method for S. cerevisiae Strain Characterization and Its Application for Wineries
Abstract
:1. Introduction
2. Materials and Methods
2.1. Material
2.1.1. Commercially Available Saccharomyces Cerevisiae Strains
2.1.2. Wine Samples
2.1.3. Yeast Isolation and Preservation
2.2. Methods:
2.2.1. DNA Extraction
2.2.2. SSR-PCR
2.2.3. Fragment Length Determination by Agarose Gel Electrophoresis
2.2.4. Capillary Sequencing and Method Validation
2.2.5. Fermentation and Sensory Tests
3. Results
3.1. Primer Sets’ Composition and Method Validation
3.2. Yeast Differentiation
3.3. Fermentative Tests
3.4. Sensory Differences
4. Discussion
4.1. Method Validation and Primer Set Composition
4.2. Winery’s Own Yeast Differentiation
4.3. Fermentative Tests
4.4. Sensory Differences
5. Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Commercial Name | Abbreviation | Origin |
---|---|---|
Lalvin BM4*4 | Bm4 | Lallemand, Fredericia, Denmark |
Oenoferm Color | Oc | Erbslöh, Geisenheim, Germany |
Oenoferm Bouquet | Ob | Erbslöh, Geisenheim, Germany |
Oenoferm Klosterneuburg | Ok | Erbslöh, Geisenheim, Germany |
Oenoferm Freddo | Of | Erbslöh, Geisenheim, Germany |
Melody | Mel | Chr Hansen, Hoersholm, Denmark |
Siha Activhefe 8 | Sa8 | Eaton Technologies, Langenlonsheim, Germany |
1895c | 1895c | Swiss Wineyeast GmbH, Meilen, Switzerland |
Lalvin EC 1118 | Lec | Lallemand, Fredericia, Denmark |
Primer | Motif | ORF Coordinates | Sequences |
---|---|---|---|
ScAAT2 | TAA | YBL084c | FW:CAGTCTTATTGCCTTGAACGA RV:GTCTCCATCCTCCAAACAGCC |
ScAAt3 | TAA | YDR160w | FW:TGGGAGGAGGGAAATGGACAG RV:TTCAGTTACCCGCACAATCTA |
C5 | GT | VI-210250/210414 | FW:TGACACAATAGCAATGGCCTTCA RV:GCAAGCGACTAGAACAACAATCACA |
C3 | CAA | YGL139w | FW:CTTTTTATTTACGAGCGGGCCAT RV:AAATCTCATGCCTGTGAGGGGTAT |
C8 | TAA | YGL014w | FW:CAGGTCGTTCTAACGTTGGTAAAATG RV:GCTGTTGCTGTTGGTAGCATTACTGT |
C11 | GT | X-518870/519072 | FW:TTCCATCATAACCGTCTGGGATT RV:TGCCTTTTTCTTAGATGGGCTTTC |
YKR072c | GAC | YKR072c | FW:AGATACAGAAGATAAGAACGAAAA RV:TTATTGATGCTTATCTATTATACC |
SCYOR267c | TGT | YOR267c | FW:TACTAACGTCAACACTGCTGCCAA RV:GGATCTACTTGCAGTATACGGG |
YKL172w | GAA | YKL172w | FW:CAGGACGCTACCGAAGCTCAAAAG RV:ACTTTTGGCCAATTTCTCAAGAT |
ScAAT1 | TTA | XIII-86902/87140 | FW:AAGCGTAAGCAATGGTGTAGATACTT RV:CAAGCCTCTTCAAGCATGACCTTT |
C4 | TAA + TAG | XV-110701/110935 | FW:AGGAGAAAAATGCTGTTTATTCTGACC RV:TTTTCCTCCGGGACGTGAAATA |
C9 | TAA | YOR156c | FW:AAGGGTTCGTAAACATATAACTGGCA RV:TATAAGGGAAAAGAGCACGATGGC |
ScAAT5 | TAA | XVI-897051/8970210 | FW:AGCATAATTGGAGGCAGTAAAGCA RV:TCTCCGTCTTTTTTGTACTGCGTG |
C6 | CA | XVI-485898/485996 | FW:GTGGCATCATATCTGTCAATTTTATCAC RV:CAATCAAGCAAAAGATCGGCCT |
YPL009c | CTT | YPL009c | FW:AACCCATTGACCTCGTTACTATCGT RV:TTCGATGGCTCTGATAACTCCATTC |
Primer | Size Range by Agarose gel (bp) | Size Range by Capillary Sequencing (bp) | Primer Set |
---|---|---|---|
ScAAT2 | 310–390 | 376–385 | 1 |
ScAAt3 | 260 | 244–274 | |
C5 | 130–170 | / | |
SCYOR267c | 305–390 | 305–325 | |
C8 | 150 | 133–146 | |
C11 | 200–250 | / | |
YKL172w | 135 | 120–125 | 2 |
C4 | 240–320 | 248–260 | |
C9 | 100 | 90–100 | |
ScAAT5 | 160–180 | 156–162 | |
C6 | 100–120 | 100–102 | |
YPL009c | 250–290 | 276–311 |
Pattern | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 |
---|---|---|---|---|---|---|---|---|
A | 18 | 0 | 0 | 0 | 1 | 0 | 0 | 0 |
B | 0 | 1 | 0 | 0 | 0 | 0 | 0 | 1 |
C | 4 | 0 | 1 | 1 | 0 | 7 | 0 | 3 |
D | 1 | 0 | 1 | 0 | 0 | 0 | 0 | 0 |
E | 0 | 0 | 0 | 0 | 1 | 0 | 0 | 0 |
F | 0 | 1 | 0 | 0 | 0 | 1 | 0 | 0 |
G | 2 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
H | 1 | 0 | 0 | 0 | 1 | 1 | 0 | 0 |
I | 0 | 0 | 0 | 0 | 0 | 0 | 1 | 1 |
J | 1 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
Group | Fermentation Time (days) | Yeasts | Total Number of Yeast |
---|---|---|---|
Group 1 | 33 | Ymg1 | 1 |
Group 2 | 40 | Ymj1, Ymg4, Ymg7, Ymg9, Ymj4 | 5 |
Group 3 | 48 | Ymf1, Ymg5, Ymi3 | 3 |
Group 4 | 55 | Ymi1, Ymg2, Ymg8, Ymj2 | 4 |
Group 5 | 62 | Ymg3, Ymh4 | 2 |
Group 6 | 73 | Ymg6, Ymf3 | 2 |
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Rex, F.; Hirschler, A.; Scharfenberger-Schmeer, M. SSR-Marker Analysis—A Method for S. cerevisiae Strain Characterization and Its Application for Wineries. Fermentation 2020, 6, 101. https://doi.org/10.3390/fermentation6040101
Rex F, Hirschler A, Scharfenberger-Schmeer M. SSR-Marker Analysis—A Method for S. cerevisiae Strain Characterization and Its Application for Wineries. Fermentation. 2020; 6(4):101. https://doi.org/10.3390/fermentation6040101
Chicago/Turabian StyleRex, Friederike, Adeline Hirschler, and Maren Scharfenberger-Schmeer. 2020. "SSR-Marker Analysis—A Method for S. cerevisiae Strain Characterization and Its Application for Wineries" Fermentation 6, no. 4: 101. https://doi.org/10.3390/fermentation6040101