Wine Yeast Population Dynamics During Inoculated and Spontaneous Fermentations in Three British Columbia Wineries

Transcription

1 Wine Yeast Population Dynamics During Inoculated and Spontaneous Fermentations in Three British Columbia Wineries MSc Candidate: Jessica Lange Supervisor: Dr. Daniel Durall July 7 th, 22

2 Please note: Darryl Brooker s presentation titled S. cerevisiae Population Dynamics: A Practical Application is included at the end of this PowerPoint

3 CONTACT Jessica Lange, BSc (Microbiology) UBCO MSc Candidate e.mail: phone: Supervisor e.mail:

4 Research Objective To determine the population dynamics of wine yeast species and strains in inoculated and spontaneous fermentations of three Okanagan wineries

5 Wineries of Study Quails Gate Cedar Creek Road3 Four tanks sampled in each winery (2 vintage): 3 Inoculated fermentation tanks Spontaneous fermentation tank Vitis vinifera L. var Pinot noir

6 Fermentation Stages Sampled CS ER M F 2-25 ºBrix 5-2 ºBrix -5 ºBrix < 5 ºBrix

7 DNA Extraction and Amplification Target Region Non-Saccharomyces isolates: Amplification of the ITS and the D/D2 regions of rdna Primers: ITS/ITS4 and NL/NL4 DNA Sequencing Species level discrimination S. cerevisiae isolates: Amplification of six loci specific to S. cerevisiae genome Primers: C4, C5, C, YPL, SCAATc, SCY9c (Legras et al. 25) DNA Fingerprinting Strain level discrimination

8 What is Microsatellite Fingerprinting? After amplifying the six different loci of the S. cerevisiae genome (DNA), it looks like this: C5 C SCA C4 SCY YPL The peaks differ between strains by shifting left or right The red, green, black, and blue peak pattern is the fingerprint Fingerprints like this were made for all the commercial strains reported utilized within the wineries of study I compared the fingerprints of the yeast I obtained from the tanks to the commercial fingerprints I constructed achieved S. cerevisiae strain identity Fingerprints are very strain specific

9 Winery Information Number of different strains used in the wineries: Cedar Creek: 32 commercial S. cerevisiae strains Quails Gate: 3 commercial S. cerevisiae strains Road3: 7 commercial S. cerevisiae strains Age of the wineries: Cedar Creek: 983 (29 years) Quails Gate: 989 (23 years) Road3: 998 (4 years)

10 Quails Gate Results Tank (Lalvin RC22) Tank 2 (Lalvin RC22) Species/StrainFrequency of Occurrence.75.5 Tank 3 (Lalvin RC22).75.5 Spontaneous H. uvarum Lalvin RC22 Lalvin ICV-D254 Lalvin A. Plus Lalvin CY379 RS P. Cuvee QG-UN QG-UN2 QG-UN3 QG-UN4 QG-UN5 Species/strainFrequency of Occurrence

11 Cedar Creek Results Species/StrainFrequencyof Occurrence.75.5 Tank (L2TD 73/735) Occurrence.75.5 Tank 2 (Enoferm AMH) H. uvarum S. uvarum Tolurospora Metschnikowia Wickerhamomyces Lalvin ICV-D254 Lalvin RC22 Enoferm AMH Zymaflore VL Zymaflore FX Species/StrainFrequencyof Occurrence.75.5 Tank 3 (Lalvin RC22).75.5 Spontaneous Zymaflore X5 Lallemand EC8 Lallemand R256 Lallemand D47 CC-UN

12 Road3 Results Species/Strain Frequency of Occurrence Species/Strain Frequency of Occurrence Tank (Lalvin RC22) Tank 3 (Lalvin RC22) Tank 2 (Lalvin RC22) Spontaneous H. uvarum S. uvarum Tolurospora Metschnikowia Wickerhamomyces Pichia S. bayanus Lalvin RC22 Fermol Chardonnay R3-UN R3-UN2 R3-UN3 R3-UN4 R3-UN5 R3-UN6 R3-UN7 R3-UN8 R3-UN9 R3-UN R3-UN R3-UN2 R3-UN3 R3-UN4 R3-UN5

13 Future Studies Our lab is currently observing the interactions and metabolomics of Lalvin RC22 and ICV-D254 through co-inoculations at varying ratios and its effects on Pinot noir sensorial attributes

14 Cell density during fermentation (CFUs/mL) Fermentation duration (days)

15 Acknowledgements Funding: NSERC & Quail s Gate Estate Winery Collaborative Research Development (CRD) Grant Winery Establishments & Pinot noir samples provided by: Grant Stanley of Quail s Gate Darryl Brooker of Cedar Creek Michael Bartier & Bailey Williamson of Road3 (present during the 2 year) Lallemand, Lalvin, AEB Fermol, Anchor, Laffort companies Commercial S. cerevisiae donations Supervisor: Dr. Daniel Durall Assistance in the lab: Hanna Pawluck & Liz Halverson UBCO FADSS technician: Sheri Maxwell

16 S. CEREVISIAE POPULATION DYNAMICS A PRACTICAL APPLICATION DARRYL BROOKER

17 PROCESSING Each block was hand harvested into 7lb bins and delivered to the winery within 2hrs of picking Both blocks were destemmed with minimal crushing (no rollers) 4ppm SO2 was added to each tank Cooling applied to each tank + dry ice and temperature was lowered to <8oC Tanks were warmed to approx. 5oC for ferment and yeast was added (where indicated) at 25ppm following manufacturers rehydration protocol All tanks were pumped over during cold soak and hand plunged during fermentation/post ferment maceration

18 SPONTANEOUS TANK (PNCC) - Population Dynamics This was a spontaneous fermentation, S. uvarum was dominant in the cold soak D254, which was obviously not added, was the dominant yeast in the EARLY stage occurring 6 days after destemming D254 continued to dominate the ferment through mid and end, with three other yeasts present (X5, Rhone 256 and RC22) Even though this tank was a spontaneous fermentation, it appears that common wine yeasts used in the winery were responsible for the majority of alcoholic fermentation This tank fermented to dryness (3.22g/L) with a relatively low Volatile Acidity (.44g/L) on pressing off

19 TANK (PNCC2) - Population Dynamics This was a double inoculant fermentation T. delbrueckii (a known vineyard yeast) followed by S. cerevisiae Very similar yeast population during cold soak as batch Much greater yeast diversity during the early, mid and end Great to see that T. delbrueckii was present during the early, given it was added just before this sample was taken T. delbrueckii was not present during the mid and end stage, which supports the notion that it is not an alcohol tolerant yeast I found it interesting that three non-saccharomyces yeast were present during early ferment and as expected all yeast were S. cerevisiae by mid and end ferment This tank fermented to dryness (3.46g/L) with a slightly higher Volatile Acidity (.48g/L vs.44g/l) than batch, however it did take longer (2 days vs 6 days for )

20 TANK 3 (PNCC4) - Population Dynamics This tank was inoculated with RC22 Very similar yeast population during cold soak as previous batches This tank fermented warmer than batch and 2 (27oC) and the fastest to ferment to dryness (5 days) RC22 remained relatively dominant in the early, mid and end stages, even in the presence of D254 Presence of white yeast in this ferment, being X5 and D47 which were both used in a nearby white wine cellar This tank fermented to dryness (2.4g/L) with the lowest Volatile Acidity (.38g/L)

21 TANK 2 (PNCC42) - Population Dynamics This tank was inoculated with AMH, known as a weak fermenter Very similar yeast population during cold soak as previous batches AMH was detected at a low rate in the early stage of ferment and was not present in the mid and end stages This tank was dominated by RC22 in the early stage and D254 in the latter stages of ferment it appears that AMH had little effect on ferment This tank had the highest ferment temperature of 28oC This tank fermented to dryness (2.39g/L) with a relatively low Volatile Acidity (.4g/L)

22 My take home points The known vineyard yeast H. uvarum is relatively resistant to SO2 and dominated all cold soaks after the addition of 4ppm SO2 S. cerevisiae was only present in small amounts during the cold soak likely due to the high dose of SO2 Strong yeast such as D254 and RC22 present in the winery dominated most of the Pinot Noir ferments by mid/end stage Yeast dispersion by air was obvious as white yeasts used in a different part of the cellar were present in all ferments

23 My take home points continued The spontaneous fermentation showed the lowest diversity in yeast population The two stage inoculation using T. delbrueckii showed the greatest diversity in yeast population Weaker yeasts (AMH) may struggle to survive in the presence of strong fermenting yeast, even if the strong fermenting yeast are not the inoculum Yeast competition was obvious in all tanks and diversity/population may be able to be manipulated through SO2 additions, cold soak duration, ferment temperature and inoculant choice